Search results for: Ajay Ivan
Commenced in January 2007
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Edition: International
Paper Count: 241

Search results for: Ajay Ivan

31 Refractory Cardiac Arrest: Do We Go beyond, Do We Increase the Organ Donation Pool or Both?

Authors: Ortega Ivan, De La Plaza Edurne

Abstract:

Background: Spain and other European countries have implemented Uncontrolled Donation after Cardiac Death (uDCD) programs. After 15 years of experience in Spain, many things have changed. Recent evidence and technical breakthroughs achieved in resuscitation are relevant for uDCD programs and raise some ethical concerns related to these protocols. Aim: To rethink current uDCD programs in the light of recent evidence on available therapeutic procedures applicable to victims of out-of-hospital cardiac arrest (OHCA). To address the following question: What is the current standard of treatment owed to victims of OHCA before including them in an uDCD protocol? Materials and Methods: Review of the scientific and ethical literature related to both uDCD programs and innovative resuscitation techniques. Results: 1) The standard of treatment received and the chances of survival of victims of OHCA depend on whether they are classified as Non-Heart Beating Patients (NHBP) or Non-Heart-Beating-Donors (NHBD). 2) Recent studies suggest that NHBPs are likely to survive, with good quality of life, if one or more of the following interventions are performed while ongoing CPR -guided by suspected or known cause of OHCA- is maintained: a) direct access to a Cath Lab-H24 or/and to extra-corporeal life support (ECLS); b) transfer in induced hypothermia from the Emergency Medical Service (EMS) to the ICU; c) thrombolysis treatment; d) mobile extra-corporeal membrane oxygenation (mini ECMO) instituted as a bridge to ICU ECLS devices. 3) Victims of OHCA who cannot benefit from any of these therapies should be considered as NHBDs. Conclusion: Current uDCD protocols do not take into account recent improvements in resuscitation and need to be adapted. Operational criteria to distinguish NHBDs from NHBP should seek a balance between the technical imperative (to do whatever is possible), considerations about expected survival with quality of life, and distributive justice (costs/benefits). Uncontrolled DCD protocols can be performed in a way that does not hamper the legitimate interests of patients, potential organ donors, their families, the organ recipients, and the health professionals involved in these processes. Families of NHBDs’ should receive information which conforms to the ethical principles of respect of autonomy and transparency.

Keywords: uncontrolled donation after cardiac death resuscitation, refractory cardiac arrest, out of hospital cardiac, arrest ethics

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30 Building on Previous Microvalving Approaches for Highly Reliable Actuation in Centrifugal Microfluidic Platforms

Authors: Ivan Maguire, Ciprian Briciu, Alan Barrett, Dara Kervick, Jens Ducrèe, Fiona Regan

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With the ever-increasing myriad of applications of which microfluidic devices are capable, reliable fluidic actuation development has remained fundamental to the success of these microfluidic platforms. There are a number of approaches which can be taken in order to integrate liquid actuation on microfluidic platforms, which can usually be split into two primary categories; active microvalves and passive microvalves. Active microvalves are microfluidic valves which require a physical parameter change by external, or separate interaction, for actuation to occur. Passive microvalves are microfluidic valves which don’t require external interaction for actuation due to the valve’s natural physical parameters, which can be overcome through sample interaction. The purpose of this paper is to illustrate how further improvements to past microvalve solutions can largely enhance systematic reliability and performance, with both novel active and passive microvalves demonstrated. Covered within this scope will be two alternative and novel microvalve solutions for centrifugal microfluidic platforms; a revamped pneumatic-dissolvable film active microvalve (PAM) strategy and a spray-on Sol-Gel based hydrophobic passive microvalve (HPM) approach. Both the PAM and the HPM mechanisms were demonstrated on a centrifugal microfluidic platform consisting of alternating layers of 1.5 mm poly(methyl methacrylate) (PMMA) (for reagent storage) sheets and ~150 μm pressure sensitive adhesive (PSA) (for microchannel fabrication) sheets. The PAM approach differs from previous SOLUBON™ dissolvable film methods by introducing a more reliable and predictable liquid delivery mechanism to microvalve site, thus significantly reducing premature activation. This approach has also shown excellent synchronicity when performed in a multiplexed form. The HPM method utilises a new spray-on and low curing temperature (70°C) sol-gel material. The resultant double layer coating comprises a PMMA adherent sol-gel as the bottom layer and an ultra hydrophobic silica nano-particles (SNPs) film as the top layer. The optimal coating was integrated to microfluidic channels with varying cross-sectional area for assessing microvalve burst frequencies consistency. It is hoped that these microvalving solutions, which can be easily added to centrifugal microfluidic platforms, will significantly improve automation reliability.

Keywords: centrifugal microfluidics, hydrophobic microvalves, lab-on-a-disc, pneumatic microvalves

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29 Development of Instructional Material Using Scientific Approach to Make the Nature of Science (NOS) and Critical Thinking Explicit on Chemical Bonding and Intermolecular Forces Topics

Authors: Ivan Ashif Ardhana, Intan Mahanani

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Chemistry education tends to change from triplet representation among macroscopic, microscopic, and symbolic to tetrahedron shape. This change set the aspect of human element on the top of learning. Meaning that students are expected to solve the problems involving the ethic, morality, and humanity through the class. Ability to solve the problems connecting either theories or applications is called scientific literacy which have been implemented in curriculum 2013 implicitly. Scientific literacy has an aspect of nature science and critical thinking. Both can be integrated to learning using scientific approach and scientific inquiry. Unfortunately, students’ ability of scientific literacy in Indonesia is far from expectation. A survey from PISA had proven it. Scientific literacy of Indonesian students is always at bottom five position from 2002 till 2012. Improving a scientific literacy needs many efforts against them. Developing an instructional material based on scientific approach is one kind of that efforts. Instructional material contains both aspect of nature of science and critical thinking which is instructed explicitly to improve the students’ understanding about science. Developing goal is to produce a prototype and an instructional material using scientific approach whose chapter is chemical bonding and intermolecular forces for high school students grade ten. As usual, the material is subjected to get either quantitative mark or suggestion through validation process using validation sheet instrument. Development model is adapted from 4D model containing four steps. They are define, design, develop, and disseminate. Nevertheless, development of instructional material had only done until third step. The final step wasn’t done because of time, cost, and energy limitations. Developed instructional material had been validated by four validators. They are coming from chemistry lecture and high school’s teacher which two at each. The result of this development research shown the average of quantitative mark of students’ book is 92.75% with very proper in criteria. Given at same validation process, teacher’s guiding book got the average mark by 96.98%, similar criteria with students’ book. Qualitative mark including both comments and suggestions resulted from validation process were used as consideration for the revision. The result concluded us how the instructional materials using scientific approach to explicit nature of science and critical thinking on the topic of chemical bonding and intermolecular forces are very proper if they are used at learning activity.

Keywords: critical thinking, instructional material, nature of science, scientific literacy

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28 Development of Microsatellite Markers for Dalmatian Pyrethrum Using Next-Generation Sequencing

Authors: Ante Turudic, Filip Varga, Zlatko Liber, Jernej Jakse, Zlatko Satovic, Ivan Radosavljevic, Martina Grdisa

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Microsatellites (SSRs) are highly informative repetitive sequences of 2-6 base pairs, which are the most used molecular markers in assessing the genetic diversity of plant species. Dalmatian pyrethrum (Tanacetum cinerariifolium /Trevir./ Sch. Bip) is an outcrossing diploid (2n = 18) endemic to the eastern Adriatic coast and source of the natural insecticide pyrethrin. Due to the high repetitiveness and large size of the genome (haploid genome size of 9,58 pg), previous attempts to develop microsatellite markers using the standard methods were unsuccessful. A next-generation sequencing (NGS) approach was applied on genomic DNA extracted from fresh leaves of Dalmatian pyrethrum. The sequencing was conducted using NovaSeq6000 Illumina sequencer, after which almost 400 million high-quality paired-end reads were obtained, with a read length of 150 base pairs. Short reads were assembled by combining two approaches; (1) de-novo assembly and (2) joining of overlapped pair-end reads. In total, 6.909.675 contigs were obtained, with the contig average length of 249 base pairs. Of the resulting contigs, 31.380 contained one or multiple microsatellite sequences, in total 35.556 microsatellite loci were identified. Out of detected microsatellites, dinucleotide repeats were the most frequent, accounting for more than half of all microsatellites identifies (21,212; 59.7%), followed by trinucleotide repeats (9,204; 25.9%). Tetra-, penta- and hexanucleotides had similar frequency of 1,822 (5.1%), 1,472 (4.1%), and 1,846 (5.2%), respectively. Contigs containing microsatellites were further filtered by SSR pattern type, transposon occurrences, assembly characteristics, GC content, and the number of occurrences against the draft genome of T. cinerariifolium published previously. After the selection process, 50 microsatellite loci were used for primer design. Designed primers were tested on samples from five distinct populations, and 25 of them showed a high degree of polymorphism. The selected loci were then genotyped on 20 samples belonging to one population resulting in 17 microsatellite markers. Availability of codominant SSR markers will significantly improve the knowledge on population genetic diversity and structure as well as complex genetics and biochemistry of this species. Acknowledgment: This work has been fully supported by the Croatian Science Foundation under the project ‘Genetic background of Dalmatian pyrethrum (Tanacetum cinerariifolium /Trevir/ Sch. Bip.) insecticidal potential’ - (PyrDiv) (IP-06-2016-9034).

Keywords: genome assembly, NGS, SSR, Tanacetum cinerariifolium

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27 Analysis of the Outcome of the Treatment of Osteoradionecrosis in Patients after Radiotherapy for Head and Neck Cancer

Authors: Petr Daniel Kovarik, Matt Kennedy, James Adams, Ajay Wilson, Andy Burns, Charles Kelly, Malcolm Jackson, Rahul Patil, Shahid Iqbal

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Introduction: Osteoradionecrosis (ORN) is a recognised toxicity of radiotherapy (RT) for head and neck cancer (HNC). Existing literature lacks any generally accepted definition and staging system for this toxicity. Objective: The objective is to analyse the outcome of the surgical and nonsurgical treatments of ORN. Material and Method: Data on 2303 patients treated for HNC with radical or adjuvant RT or RT-chemotherapy from January 2010 - December 2021 were retrospectively analysed. Median follow-up to the whole group of patients was 37 months (range 0–148 months). Results: ORN developed in 185 patients (8.1%). The location of ORN was as follows; mandible=170, maxilla=10, and extra oral cavity=5. Multiple ORNs developed in 7 patients. 5 patients with extra oral cavity ORN were excluded from treatment analysis as the management is different. In 180 patients with oral cavity ORN, median follow-up was 59 months (range 5–148 months). ORN healed in 106 patients, treatment failed in 74 patients (improving=10, stable=43, and deteriorating=21). Median healing time was 14 months (range 3-86 months). Notani staging is available in 158 patients with jaw ORN with no previous surgery to the mandible (Notani class I=56, Notani class II=27, and Notani class III=76). 28 ORN (mandible=27, maxilla=1; Notani class I=23, Notani II=3, Notani III=1) healed spontaneously with a median healing time 7 months (range 3–46 months). In 20 patients, ORN developed after dental extraction, in 1 patient in the neomandible after radical surgery as a part of the primary treatment. In 7 patients, ORN developed and spontaneously healed in irradiated bone with no previous surgical/dental intervention. Radical resection of the ORN (segmentectomy, hemi-mandibulectomy with fibula flap) was performed in 43 patients (all mandible; Notani II=1, Notani III=39, Notani class was not established in 3 patients as ORN developed in the neomandible). 27 patients healed (63%); 15 patients failed (improving=2, stable=5, deteriorating=8). The median time from resection to healing was 6 months (range 2–30 months). 109 patients (mandible=100, maxilla=9; Notani I=3, Notani II=23, Notani III=35, Notani class was not established in 9 patients as ORN developed in the maxilla/neomandible) were treated conservatively using a combination of debridement, antibiotics and Pentoclo. 50 patients healed (46%) with a median healing time 14 months (range 3–70 months), 59 patients are recorded with persistent ORN (improving=8, stable=38, deteriorating=13). Out of 109 patients treated conservatively, 13 patients were treated with Pentoclo only (all mandible; Notani I=6, Notani II=3, Notani III=3, 1 patient with neomandible). In total, 8 patients healed (61.5%), treatment failed in 5 patients (stable=4, deteriorating=1). Median healing time was 14 months (range 4–24 months). Extra orally (n=5), 3 cases of ORN were in the auditory canal and 2 in mastoid. ORN healed in one patient (auditory canal after 32 months. Treatment failed in 4 patients (improving=3, stable=1). Conclusion: The outcome of the treatment of ORN remains in general, poor. Every effort should therefore be made to minimise the risk of development of this devastating toxicity.

Keywords: head and neck cancer, radiotherapy, osteoradionecrosis, treatment outcome

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26 Study of the Impact of Synthesis Method and Chemical Composition on Photocatalytic Properties of Cobalt Ferrite Catalysts

Authors: Katerina Zaharieva, Vicente Rives, Martin Tsvetkov, Raquel Trujillano, Boris Kunev, Ivan Mitov, Maria Milanova, Zara Cherkezova-Zheleva

Abstract:

The nanostructured cobalt ferrite-type materials Sample A - Co0.25Fe2.75O4, Sample B - Co0.5Fe2.5O4, and Sample C - CoFe2O4 were prepared by co-precipitation in our previous investigations. The co-precipitated Sample B and Sample C were mechanochemically activated in order to produce Sample D - Co0.5Fe2.5O4 and Sample E- CoFe2O4. The PXRD, Moessbauer and FTIR spectroscopies, specific surface area determination by the BET method, thermal analysis, element chemical analysis and temperature-programmed reduction were used to investigate the prepared nano-sized samples. The changes of the Malachite green dye concentration during reaction of the photocatalytic decolorization using nanostructured cobalt ferrite-type catalysts with different chemical composition are included. The photocatalytic results show that the increase in the degree of incorporation of cobalt ions in the magnetite host structure for co-precipitated cobalt ferrite-type samples results in an increase of the photocatalytic activity: Sample A (4 х10-3 min-1) < Sample B (5 х10-3 min-1) < Sample C (7 х10-3 min-1). Mechanochemically activated photocatalysts showed a higher activity than the co-precipitated ferrite materials: Sample D (16 х10-3 min-1) > Sample E (14 х10-3 min-1) > Sample C (7 х10-3 min-1) > Sample B (5 х10-3 min-1) > Sample A (4 х10-3 min-1). On decreasing the degree of substitution of iron ions by cobalt ones a higher sorption ability of the dye after the dark period for the co-precipitated cobalt ferrite materials was observed: Sample C (72 %) < Sample B (78 %) < Sample A (80 %). Mechanochemically treated ferrite catalysts and co-precipitated Sample B possess similar sorption capacities, Sample D (78 %) ~ Sample E (78 %) ~ Sample B (78 %). The prepared nano-sized cobalt ferrite-type materials demonstrate good photocatalytic and sorption properties. Mechanochemically activated Sample D - Co0.5Fe2.5O4 (16х10-3 min-1) and Sample E-CoFe2O4 (14х10-3 min-1) possess higher photocatalytic activity than that of the most common used UV-light catalyst Degussa P25 (12х10-3 min-1). The dependence of the photo-catalytic activity and sorption properties on the preparation method and different degree of substitution of iron ions by cobalt ions in synthesized cobalt ferrite samples is established. The mechanochemical activation leads to formation of nano-structured cobalt ferrite-type catalysts (Sample D and Sample E) with higher rate constants than those of the ferrite materials (Sample A, Sample B, and Sample C) prepared by the co-precipitation procedure. The increase in the degree of substitution of iron ions by cobalt ones leads to improved photocatalytic properties and lower sorption capacities of the co-precipitated ferrite samples. The good sorption properties between 72 and 80% of the prepared ferrite-type materials show that they could be used as potential cheap absorbents for purification of polluted waters.

Keywords: nanodimensional cobalt ferrites, photocatalyst, synthesis, mechanochemical activation

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25 Pediatric Drug Resistance Tuberculosis Pattern, Side Effect Profile and Treatment Outcome: North India Experience

Authors: Sarika Gupta, Harshika Khanna, Ajay K Verma, Surya Kant

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Background: Drug-resistant tuberculosis (DR-TB) is a growing health challenge to global TB control efforts. Pediatric DR-TB is one of the neglected infectious diseases. In our previously published report, we have notified an increased prevalence of DR-TB in the pediatric population at a tertiary health care centre in North India which was estimated as 17.4%, 15.1%, 18.4%, and 20.3% in (%) in the year 2018, 2019, 2020, and 2021. Limited evidence exists about a pattern of drug resistance, side effect profile and programmatic outcomes of Paediatric DR-TB treatment. Therefore, this study was done to find out the pattern of resistance, side effect profile and treatment outcome. Methodology: This was a prospective cohort study conducted at the nodal drug-resistant tuberculosis centre of a tertiary care hospital in North India from January 2021 to December 2022. Subjects included children aged between 0-18 years of age with a diagnosis of DR-TB, on the basis of GeneXpert (rifampicin [RIF] resistance detected), line probe assay and drug sensitivity testing (DST) of M. tuberculosis (MTB) grown on a culture of body fluids. Children were classified as monoresistant TB, polyresistant TB (resistance to more than 1 first-line anti-TB drug, other than both INH and RIF), MDR-TB, pre-XDR-TB and XDR-TB, as per the WHO classification. All the patients were prescribed DR TB treatment as per the standard guidelines, either shorter oral DR-TB regimen or a longer all-oral MDR/XDR-TB regimen (age below five years needed modification). All the patients were followed up for side effects of treatment once per month. The patient outcomes were categorized as good outcomes if they had completed treatment and cured or were improving during the course of treatment, while bad outcomes included death or not improving during the course of treatment. Results: Of the 50 pediatric patients included in the study, 34 were females (66.7%) and 16 were male (31.4%). Around 33 patients (64.7%) were suffering from pulmonary TB, while 17 (33.3%) were suffering from extrapulmonary TB. The proportions of monoresistant TB, polyresistant TB, MDR-TB, pre-XDR-TB and XDR-TB were 2.0%, 0%, 50.0%, 30.0% and 18.0%, respectively. Good outcome was reported in 40 patients (80.0%). The 10 bad outcomes were 7 deaths (14%) and 3 (6.0%) children who were not improving. Adverse events (single or multiple) were reported in all the patients, most of which were mild in nature. The most common adverse events were metallic taste 16(31.4%), rash and allergic reaction 15(29.4%), nausea and vomiting 13(26.0%), arthralgia 11 (21.6%) and alopecia 11 (21.6%). Serious adverse event of QTc prolongation was reported in 4 cases (7.8%), but neither arrhythmias nor symptomatic cardiac side effects occurred. Vestibular toxicity was reported in 2(3.9%), and psychotic symptoms in 4(7.8%). Hepatotoxicity, hypothyroidism, peripheral neuropathy, gynaecomastia, and amenorrhea were reported in 2 (4.0%), 4 (7.8%), 2 (3.9%), 1(2.0%), and 2 (3.9%) respectively. None of the drugs needed to be withdrawn due to uncontrolled adverse events. Conclusion: Paediatric DR TB treatment achieved favorable outcomes in a large proportion of children. DR TB treatment regimen drugs were overall well tolerated in this cohort.

Keywords: pediatric, drug-resistant, tuberculosis, adverse events, treatment

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24 Reproductive Governmentality in Mexico: Production, Control and Regulation of Contraceptive Practices in a Public Hospital

Authors: Ivan Orozco

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Introduction: Forced contraception constitutes part of an effort to control the life and reproductive capacity of women through public health institutions. This phenomenon has affected many Mexican women historically and still persists nowadays. The notion of reproductive governmentality refers to the mechanisms through which different historical configurations of social actors (state institutions, churches, donor agents, NGOs, etc.) use legislative controls, economic incentives, moral mandates, direct coercion, and ethical incitements, to produce, monitor and control reproductive behaviors and practices. This research focuses on the use of these mechanisms by the Mexican State to control women's contraceptive practices in a public hospital. Method: An Institutional Ethnography was carried out, with the objective of knowing women's experiences from their own perspective, as they occur in their daily lives, but at the same time, discovering the structural elements that shape the discourses that promote women's contraception, even against their will. The fieldwork consisted in an observation of the dynamics between different participants within a public hospital and the conduction of interviews with the medical and nursing staff in charge of family planning services, as well as women attending the family planning office. Results: Public health institutions in Mexico are state tools to control and regulate reproduction. There are several strategies that are used for this purpose, for example, health personnel provide insufficient or misleading information to ensure that women agree to use contraceptives; health institutions provide economic incentives to the members of the health staff who reach certain goals in terms of contraceptive placement; young women are forced to go to the family planning service, regardless of the reason they went to the clinic; health campaigns are carried out, consisting of the application of contraceptives outside the health facilities, directly in the communities of people who visit the hospital less frequently. All these mechanisms seek for women to use contraceptives, from the women’s perspective; however, the reception of these discourses is ambiguous. While, for some women, the strategies become coercive mechanisms to use contraceptives against their will, for others, they represent an opportunity to take control over their reproductive lives. Conclusion: Since 1974, the Mexican government has implemented campaigns for the promotion of family planning methods as a means to control population growth. Although it is established in several legislations that the counselling must be carried out with a gender and human rights perspective, always respecting the autonomy of people, these research testify that health personnel uses different strategies to force some women to use contraceptive methods, thereby violating their reproductive rights.

Keywords: feminist research, forced contraception, institutional ethnography, reproductive. governmentality

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23 Survey of Prevalence of Noise Induced Hearing Loss in Hawkers and Shopkeepers in Noisy Areas of Mumbai City

Authors: Hitesh Kshayap, Shantanu Arya, Ajay Basod, Sachin Sakhuja

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This study was undertaken to measure the overall noise levels in different locations/zones and to estimate the prevalence of Noise induced hearing loss in Hawkers & Shopkeepers in Mumbai, India. The Hearing Test developed by American Academy Of Otolaryngology, translated from English to Hindi, and validated is used as a screening tool for hearing sensitivity was employed. The tool is having 14 items. Each item is scored on a scale 0, 1, 2 and 3. The score 6 and above indicated some difficulty or definite difficulty in hearing in daily activities and low score indicated lesser difficulty or normal hearing. The subjects who scored 6 or above or having tinnitus were made to undergo hearing evaluation by Pure tone audiometer. Further, the environmental noise levels were measured from Morning to Evening at road side at different Location/Hawking zones in Mumbai city using SLM9 Agronic 8928B & K type Digital Sound Level Meter) in dB (A). The maximum noise level of 100.0 dB (A) was recorded during evening hours from Chattrapati Shivaji Terminal to Colaba with overall noise level of 79.0 dB (A). However, the minimum noise level in this area was 72.6 dB (A) at any given point of time. Further, 54.6 dB (A) was recorded as minimum noise level during 8-9 am at Sion Circle. Further, commencement of flyovers with 2-tier traffic, sky walks, increasing number of vehicular traffic at road, high rise buildings and other commercial & urbanization activities in the Mumbai city most probably have resulted in increasing the overall environmental noise levels. Trees which acted as noise absorbers have been cut owing to rapid construction. The study involved 100 participants in the age range of 18 to 40 years of age, with the mean age of 29 years (S.D. =6.49). 46 participants having tinnitus or have obtained the score of 6 were made to undergo Pure Tone Audiometry and it was found that the prevalence rate of hearing loss in hawkers & shopkeepers is 19% (10% Hawkers and 9 % Shopkeepers). The results found indicates that 29 (42.6%) out of 64 Hawkers and 17 (47.2%) out of 36 Shopkeepers who underwent PTA had no significant difference in percentage of Noise Induced Hearing loss. The study results also reveal that participants who exhibited tinnitus 19 (41.30%) out of 46 were having mild to moderate sensorineural hearing loss between 3000Hz to 6000Hz. The Pure tone Audiogram pattern revealed Hearing loss at 4000 Hz and 6000 Hz while hearing at adjacent frequencies were nearly normal. 7 hawkers and 8 shopkeepers had mild notch while 3 hawkers and 1 shopkeeper had a moderate degree of notch. It is thus inferred that tinnitus is a strong indicator for presence of hearing loss and 4/6 KHz notch is a strong marker for road/traffic/ environmental noise as an occupational hazard for hawkers and shopkeepers. Mass awareness about these occupational hazards, regular hearing check up, early intervention along with sustainable development juxtaposed with social and urban forestry can help in this regard.

Keywords: NIHL, noise, sound level meter, tinnitus

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22 Effect of Antimony on Microorganisms in Aerobic and Anaerobic Environments

Authors: Barrera C. Monserrat, Sierra-Alvarez Reyes, Pat-Espadas Aurora, Moreno Andrade Ivan

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Antimony is a toxic and carcinogenic metalloid considered a pollutant of priority interest by the United States Environmental Protection Agency. It is present in the environment in two oxidation states: antimonite (Sb (III)) and antimony (Sb (V)). Sb (III) is toxic to several aquatic organisms, but the potential inhibitory effect of Sb species for microorganisms has not been extensively evaluated. The fate and possible toxic impact of antimony on aerobic and anaerobic wastewater treatment systems are unknown. For this reason, the objective of this study was to evaluate the microbial toxicity of Sb (V) and Sb (III) in aerobic and anaerobic environments. Sb(V) and Sb(III) were used as potassium hexahydroxoantimonate (V) and potassium antimony tartrate, respectively (Sigma-Aldrich). The toxic effect of both Sb species in anaerobic environments was evaluated on methanogenic activity and the inhibition of hydrogen production of microorganisms from a wastewater treatment bioreactor. For the methanogenic activity, batch experiments were carried out in 160 mL serological bottles; each bottle contained basal mineral medium (100 mL), inoculum (1.5 g of VSS/L), acetate (2.56 g/L) as substrate, and variable concentrations of Sb (V) or Sb (III). Duplicate bioassays were incubated at 30 ± 2°C on an orbital shaker (105 rpm) in the dark. Methane production was monitored by gas chromatography. The hydrogen production inhibition tests were carried out in glass bottles with a working volume of 0.36 L. Glucose (50 g/L) was used as a substrate, pretreated inoculum (5 g VSS/L), mineral medium and varying concentrations of the two species of antimony. The bottles were kept under stirring and at a temperature of 35°C in an AMPTSII device that recorded hydrogen production. The toxicity of Sb on aerobic microorganisms (from a wastewater activated sludge treatment plant) was tested with a Microtox standardized toxicity test and respirometry. Results showed that Sb (III) is more toxic than Sb (V) for methanogenic microorganisms. Sb (V) caused a 50% decrease in methanogenic activity at 250 mg/L. In contrast, exposure to Sb (III) resulted in a 50% inhibition at a concentration of only 11 mg/L, and an almost complete inhibition (95%) at 25 mg/L. For hydrogen-producing microorganisms, Sb (III) and Sb (V) inhibited 50% of this production with 12.6 mg/L and 87.7 mg/L, respectively. The results for aerobic environments showed that 500 mg/L of Sb (V) do not inhibit the Allivibrio fischeri (Microtox) activity or specific oxygen uptake rate of activated sludge. In the case of Sb (III), this caused a loss of 50% of the respiration of the microorganisms at concentrations below 40 mg/L. The results obtained indicate that the toxicity of the antimony will depend on the speciation of this metalloid and that Sb (III) has a significantly higher inhibitory potential compared to Sb (V). It was shown that anaerobic microorganisms can reduce Sb (V) to Sb (III). Acknowledgments: This work was funded in part by grants from the UA-CONACYT Binational Consortium for the Regional Scientific Development and Innovation (CAZMEX), the National Institute of Health (NIH ES- 04940), and PAPIIT-DGAPA-UNAM (IN105220).

Keywords: aerobic inhibition, antimony reduction, hydrogen inhibition, methanogenic toxicity

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21 Species Profiling of Scarab Beetles with the Help of Light Trap in Western Himalayan Region of Uttarakhand

Authors: Ajay Kumar Pandey

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White grub (Coleoptera: Scarabaeidae), locally known as Kurmula, Pagra, Chinchu, is a major destructive pest in western Himalayan region of Uttarakhand state of India. Various crops like cereals (up land paddy, wheat, and barley), vegetables (capsicum, cabbage, tomato, cauliflower, carrot etc) and some pulse (like pigeon pea, green gram, black gram) are grown with limited availability of primary resources. Among the various limitations in successful cultivation of these crops, white grub has been proved a major constraint in for all crops grown in hilly area. The losses incurred due to white grubs are huge in case of commercial crops like sugarcane, groundnut, potato, maize and upland rice. Moreover, it has been proved major constraint in potato production in mid and higher hills of India. Adults emerge in May-June following the onset of monsoon and thereafter defoliate the apple, apricot, plum, and walnut during night while 2nd and 3rd instar grubs feed on live roots of cultivated as well as non cultivated crops from August to January. Survey was conducted in hilly (Pauri and Tehri) as well as plain area (Haridwar district) of Uttarakhand state. Collection of beetle was done from various locations from August to September of five consecutive years with the help of light trap and directly from host plant. The grub was also collected by excavating one square meter area from different locations and reared in laboratory to find out adult. During the collection, the diseased or dead cadaver were also collected and brought in the laboratory and identified the causal organisms. Total 25 species of white grub was identified out of which Holotrichia longipennis, Anomala dimidiata, Holotrichia lineatopennis, Maladera insanabilis, Brahmina sp. make complex problem in different area of Uttarakhand where they cause severe damage to various crops. During the survey, it was observed that white grubs beetles have variation in preference of host plant, even in choice of fruit and leaves of host plant. It was observed that, a white grub species, which identified as Lepidiota mansueta Burmeister., was causing severe havoc to sugarcane crop grown in major sugarcane growing belt of Haridwar district. The study also revealed that Bacillus cereus, Beauveria bassiana, Metarhizium anisopliae, Steinernema, Heterorhabditis are major disease causing agents in immature stage of white grub under rain-fed condition of Uttarakhand which caused 15.55 to 21.63 percent natural mortality of grubs with an average of 18.91 percent. However, among the microorganisms, B. cereus found to be significantly more efficient (7.03 percent mortality) then the entomopathogenic fungi (3.80 percent mortality) and nematodes (3.20 percent mortality).

Keywords: Lepidiota, profiling, Uttarakhand, whitegrub

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20 Evolution of Antimicrobial Resistance in Shigella since the Turn of 21st Century, India

Authors: Neelam Taneja, Abhishek Mewara, Ajay Kumar

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Multidrug resistant shigellae have emerged as a therapeutic challenge in India. At our 2000 bed tertiary care referral centre in Chandigarh, North India, which caters to a large population of 7 neighboring states, antibiotic resistance in Shigella is being constantly monitored. Shigellae are isolated from 3 to 5% of all stool samples. In 1990 nalidixic acid was the drug of choice as 82%, and 63% of shigellae were resistant to ampicillin and cotrimoxazole respectively. Nalidixic acid resistance emerged in 1992 and rapidly increased from 6% during 1994-98 to 86% by the turn of 21st century. In the 1990s, the WHO recommended ciprofloxacin as the drug of choice for empiric treatment of shigellosis in view of the existing high level resistance to agents like chloramphenicol, ampicillin, cotrimoxazole and nalidixic acid. First resistance to ciprofloxacin in S. flexneri at our centre appeared in 2000 and rapidly rose to 46% in 2007 (MIC>4mg/L). In between we had an outbreak of ciprofloxacin resistant S.dysenteriae serotype 1 in 2003. Therapeutic failures with ciprofloxacin occurred with both ciprofloxacin-resistant S. dysenteriae and ciprofloxacin-resistant S. flexneri. The severity of illness was more with ciprofloxacin-resistant strains. Till 2000, elsewhere in the world ciprofloxacin resistance in S. flexneri was sporadic and uncommon, though resistance to co-trimoxazole and ampicillin was common and in some areas resistance to nalidixic acid had also emerged. Fluoroquinolones due to extensive use and misuse for many other illnesses in our region are thus no longer the preferred group of drugs for managing shigellosis in India. WHO presently recommends ceftriaxone and azithromycin as alternative drugs to fluoroquinolone-resistant shigellae, however, overreliance on this group of drugs also seems to soon become questionable considering the emerging cephalosporin-resistant shigellae. We found 15.1% of S. flexneri isolates collected over a period of 9 years (2000-2009) resistant to at least one of the third-generation cephalosporins (ceftriaxone/cefotaxime). The first isolate showing ceftriaxone resistance was obtained in 2001, and we have observed an increase in number of isolates resistant to third generation cephalosporins in S. flexneri 2005 onwards. This situation has now become a therapeutic challenge in our region. The MIC values for Shigella isolates revealed a worrisome rise for ceftriaxone (MIC90:12 mg/L) and cefepime (MIC90:8 mg/L). MIC values for S. dysenteriae remained below 1 mg/L for ceftriaxone, however for cefepime, the MIC90 has raised to 4 mg/L. These infections caused by ceftriaxone-resistant S. flexneri isolates were successfully treated by azithromycin at our center. Most worrisome development in the present has been the emergence of DSA(Decreased susceptibility to azithromycin) which surfaced in 2001 and has increased from 4.3% till 2011 to 34% thereafter. We suspect plasmid-mediated resistance as we detected qnrS1-positive Shigella for the first time from the Indian subcontinent in 2 strains from 2010, indicating a relatively new appearance of this PMQR determinant among Shigella in India. This calls for a continuous and strong surveillance of antibiotic resistance across the country. The prevention of shigellosis by developing cost-effective vaccines is desirable as it will substantially reduce the morbidity associated with diarrhoea in the country

Keywords: Shigella, antimicrobial, resistance, India

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19 Concealing Breast Cancer Status: A Qualitative Study in India

Authors: Shradha Parsekar, Suma Nair, Ajay Bailey, Binu V. S.

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Background: Concealing of cancer-related information is seen in many low-and-middle-income countries and may be associated with multiple factors. Comparatively, there is lack of information about, how breast cancers diagnosed women disclose cancer-related information to their social contacts and vice versa. To get more insights on the participant’s experience, opinions, expectations, and attitudes, a qualitative study is a suitable approach. Therefore, this study involving in-depth interviews was planned to lessen this gap. Methods: Interviews were conducted separately among breast cancer patients and their caregivers with semi-structured qualitative interview guide. Purposive and convenient sampling was being used to recruit patients and caregivers, respectively. Ethical clearance and permission from the tertiary hospital were obtained and participants were selected from the Udupi district, Karnataka, India. After obtaining a list of breast cancer diagnosed cases, participants were contacted in person and their willingness to take part in the study was taken. About 39 caregivers and 35 patients belonging to different breast cancer stages were recruited. Interviews were recorded with prior permission. Data was managed by Atlas.ti 8 software. The recordings were transcribed, translated and coded in two cycles. Most of the patients belonged to stage II and III cancer. Codes were grouped together into to whom breast cancer status was concealed to and underneath reason for the same. Main findings: followings are the codes and code families which emerged from the data. 1) Concealing the breast cancer status from social contacts other than close family members (such as extended family, neighbor and friends). Participants perceived the reasons as, a) to avoid questions which people probe (which doesn’t have answers), b) to avoid people paying courtesy visit (to inquire about the health as it is Indian culture to visit the sick person) making it inconvenient for patient and caregivers have to offer something and talk to them, c) to avoid people getting shocked (react as if cancer is different from other diseases) or getting emotional/sad, or getting fear of death d) to avoid getting negative suggestion or talking anything in front of patient as it may affect patient negatively, e) to avoid getting stigmatized, f) to avoid getting obstacle in child’s marriage. 2) Participant concealed the breast cancer status of young children as they perceived that it may a) affect studies, b) affect emotionally, c) children may get scared. 3) Concealing the breast cancer status from patients as the caregivers perceived that they have fear of a) worsening patient’s health, b) patient getting tensed, c) patient getting shocked, and d) patient getting scared. However, some participants stressed important in disclosing the cancer status to social contact/patient to make the people aware of the disease. Conclusion: The news of breast cancer spreads like electricity in the wire, therefore, patient or family avoid it for many reasons. Although, globally, due to physicians’ ethical obligations, there is an inclination towards more disclosure of cancer diagnosis and status of prognosis to the patient. However, it is an ongoing argument whether patient/social contacts should know the status especially in a country like India.

Keywords: breast cancer, concealing cancer status, India, qualitative study

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18 Magnetron Sputtered Thin-Film Catalysts with Low Noble Metal Content for Proton Exchange Membrane Water Electrolysis

Authors: Peter Kus, Anna Ostroverkh, Yurii Yakovlev, Yevheniia Lobko, Roman Fiala, Ivan Khalakhan, Vladimir Matolin

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Hydrogen economy is a concept of low-emission society which harvests most of its energy from renewable sources (e.g., wind and solar) and in case of overproduction, electrochemically turns the excess amount into hydrogen, which serves as an energy carrier. Proton exchange membrane water electrolyzers (PEMWE) are the backbone of this concept. By fast-response electricity to hydrogen conversion, the PEMWEs will not only stabilize the electrical grid but also provide high-purity hydrogen for variety of fuel cell powered devices, ranging from consumer electronics to vehicles. Wider commercialization of PEMWE technology is however hindered by high prices of noble metals which are necessary for catalyzing the redox reactions within the cell. Namely, platinum for hydrogen evolution reaction (HER), running on cathode, and iridium for oxygen evolution reaction (OER) on anode. Possible way of how to lower the loading of Pt and Ir is by using conductive high-surface nanostructures as catalyst supports in conjunction with thin-film catalyst deposition. The presented study discusses unconventional technique of membrane electron assembly (MEA) preparation. Noble metal catalysts (Pt and Ir) were magnetron sputtered in very low loadings onto the surface of porous sublayers (located on gas diffusion layer or directly on membrane), forming so to say localized three-phase boundary. Ultrasonically sprayed corrosion resistant TiC-based sublayer was used as a support material on anode, whereas magnetron sputtered nanostructured etched nitrogenated carbon (CNx) served the same role on cathode. By using this configuration, we were able to significantly decrease the amount of noble metals (to thickness of just tens of nanometers), while keeping the performance comparable to that of average state-of-the-art catalysts. Complex characterization of prepared supported catalysts includes in-cell performance and durability tests, electrochemical impedance spectroscopy (EIS) as well as scanning electron microscopy (SEM) imaging and X-ray photoelectron spectroscopy (XPS) analysis. Our research proves that magnetron sputtering is a suitable method for thin-film deposition of electrocatalysts. Tested set-up of thin-film supported anode and cathode catalysts with combined loading of just 120 ug.cm⁻² yields remarkable values of specific current. Described approach of thin-film low-loading catalyst deposition might be relevant when noble metal reduction is the topmost priority.

Keywords: hydrogen economy, low-loading catalyst, magnetron sputtering, proton exchange membrane water electrolyzer

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17 Ethnobotanical Study of Traditional Medicinal Plants Used by Indigenous Tribal People of Kodagu District, Central Western Ghats, Karnataka, India

Authors: Anush Patric, M. Jadeyegowda, M. N. Ramesh, M. Ravikumar, C. R. Ajay

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Kodagu district which is situated in Central Western Ghats regions falls in one of the hottest of hot spots of biodiversity which is recognised by UNESCO. The district has one of the highest densities of community managed sacred forests in the world with rich floral and faunal diversity. It is a habitat for more than ten different types of Ethnic Indigenous tribal groups commonly called ‘Girijanas’ (Soligas, Yarvas, Jenukuruba, Bettakuruba etc.), who are having the rich knowledge of medicinal value of the plants that are commonly available in the forest. The tribal men of this region are the treasure house of the traditional plant knowledge and health care practices. An ethnobotanical survey was undertaken in tribal areas of the district to collect information about some of the indigenous medicinal plant knowledge of tribal people by semi-structured interviews, ranking exercises and field observations on their native habitat in order to evaluate the potential medicinal uses of local plants. The study revealed that, the ethnobotanical information of 83 plant species belonging to 45 families, of the total 83 species documented, most plants used in the treatment were trees (11 species), shrubs (41 species), herbs (22 species) and rarely climbers (9 species) which are used in the treatment of Hyperacidity, Respiratory disorders, Snake bite Abortifacient, Anthelmintic, Paralysis, Antiseptic, Fever, Chest pain, Stomachic, Jaundice, Piles, Asthma, Malaria, Renal disorders, Malaria and many other diseases. Maximum of 6 plant species each of Acanthaceae, Apiaceae and were used for drug preparation, followed by Asclepiadaceae, Liliaceae, Fabaceae, Verbenaceae, Caesalpinaceae, Bombaceae, Papilonaceae, Solanaceae, Rubiaceae, Myrtaceae, Amaranthaceae, Asteraceae, Ascelepidaceae, Cucurbitaceae, Apocyanaceae, and Solanaceae etc. In our present study, only medicinal plants and their local medicinal uses are recorded and presented. Information was obtained by local informants having the knowledge about medicinal plants. About 23 local tribes were interviewed. For each plant, necessary information like botanical name, family of plant species, local name and uses are given. Recent trend shows a decline in the number of traditional herbal healers in the tribal areas since the younger generation is not interested to continue this tradition. Hence, there is an urgent need to record and preserve all information on plants used by different ethnic/tribal communities for various purposes before it reaches to verge of extinction. In addition, several wild medicinal plants are declining in numbers due to deforestation and forest fires. There is need for phytochemical analysis and conservation measures to be taken for conserving medicinal plant species which is far better than allopathic medicines and these do not cause any side effects as they are the natural disease healers. So, conservation strategies have to be practiced in all levels and sectors by creating awareness about the value of such medicinal plants, and it is necessary to save the disappearing plants to strengthen the document and to conserve them for future generation.

Keywords: diseases, ethnic groups, folk medicine, Kodagu, medicinal plants

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16 Regional Dynamics of Innovation and Entrepreneurship in the Optics and Photonics Industry

Authors: Mustafa İlhan Akbaş, Özlem Garibay, Ivan Garibay

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The economic entities in innovation ecosystems form various industry clusters, in which they compete and cooperate to survive and grow. Within a successful and stable industry cluster, the entities acquire different roles that complement each other in the system. The universities and research centers have been accepted to have a critical role in these systems for the creation and development of innovations. However, the real effect of research institutions on regional economic growth is difficult to assess. In this paper, we present our approach for the identification of the impact of research activities on the regional entrepreneurship for a specific high-tech industry: optics and photonics. The optics and photonics has been defined as an enabling industry, which combines the high-tech photonics technology with the developing optics industry. The recent literature suggests that the growth of optics and photonics firms depends on three important factors: the embedded regional specializations in the labor market, the research and development infrastructure, and a dynamic small firm network capable of absorbing new technologies, products and processes. Therefore, the role of each factor and the dynamics among them must be understood to identify the requirements of the entrepreneurship activities in optics and photonics industry. There are three main contributions of our approach. The recent studies show that the innovation in optics and photonics industry is mostly located around metropolitan areas. There are also studies mentioning the importance of research center locations and universities in the regional development of optics and photonics industry. These studies are mostly limited with the number of patents received within a short period of time or some limited survey results. Therefore the first contribution of our approach is conducting a comprehensive analysis for the state and recent history of the photonics and optics research in the US. For this purpose, both the research centers specialized in optics and photonics and the related research groups in various departments of institutions (e.g. Electrical Engineering, Materials Science) are identified and a geographical study of their locations is presented. The second contribution of the paper is the analysis of regional entrepreneurship activities in optics and photonics in recent years. We use the membership data of the International Society for Optics and Photonics (SPIE) and the regional photonics clusters to identify the optics and photonics companies in the US. Then the profiles and activities of these companies are gathered by extracting and integrating the related data from the National Establishment Time Series (NETS) database, ES-202 database and the data sets from the regional photonics clusters. The number of start-ups, their employee numbers and sales are some examples of the extracted data for the industry. Our third contribution is the utilization of collected data to investigate the impact of research institutions on the regional optics and photonics industry growth and entrepreneurship. In this analysis, the regional and periodical conditions of the overall market are taken into consideration while discovering and quantifying the statistical correlations.

Keywords: entrepreneurship, industrial clusters, optics, photonics, emerging industries, research centers

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15 Using Convolutional Neural Networks to Distinguish Different Sign Language Alphanumerics

Authors: Stephen L. Green, Alexander N. Gorban, Ivan Y. Tyukin

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Within the past decade, using Convolutional Neural Networks (CNN)’s to create Deep Learning systems capable of translating Sign Language into text has been a breakthrough in breaking the communication barrier for deaf-mute people. Conventional research on this subject has been concerned with training the network to recognize the fingerspelling gestures of a given language and produce their corresponding alphanumerics. One of the problems with the current developing technology is that images are scarce, with little variations in the gestures being presented to the recognition program, often skewed towards single skin tones and hand sizes that makes a percentage of the population’s fingerspelling harder to detect. Along with this, current gesture detection programs are only trained on one finger spelling language despite there being one hundred and forty-two known variants so far. All of this presents a limitation for traditional exploitation for the state of current technologies such as CNN’s, due to their large number of required parameters. This work aims to present a technology that aims to resolve this issue by combining a pretrained legacy AI system for a generic object recognition task with a corrector method to uptrain the legacy network. This is a computationally efficient procedure that does not require large volumes of data even when covering a broad range of sign languages such as American Sign Language, British Sign Language and Chinese Sign Language (Pinyin). Implementing recent results on method concentration, namely the stochastic separation theorem, an AI system is supposed as an operate mapping an input present in the set of images u ∈ U to an output that exists in a set of predicted class labels q ∈ Q of the alphanumeric that q represents and the language it comes from. These inputs and outputs, along with the interval variables z ∈ Z represent the system’s current state which implies a mapping that assigns an element x ∈ ℝⁿ to the triple (u, z, q). As all xi are i.i.d vectors drawn from a product mean distribution, over a period of time the AI generates a large set of measurements xi called S that are grouped into two categories: the correct predictions M and the incorrect predictions Y. Once the network has made its predictions, a corrector can then be applied through centering S and Y by subtracting their means. The data is then regularized by applying the Kaiser rule to the resulting eigenmatrix and then whitened before being split into pairwise, positively correlated clusters. Each of these clusters produces a unique hyperplane and if any element x falls outside the region bounded by these lines then it is reported as an error. As a result of this methodology, a self-correcting recognition process is created that can identify fingerspelling from a variety of sign language and successfully identify the corresponding alphanumeric and what language the gesture originates from which no other neural network has been able to replicate.

Keywords: convolutional neural networks, deep learning, shallow correctors, sign language

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14 Digital Technology Relevance in Archival and Digitising Practices in the Republic of South Africa

Authors: Tashinga Matindike

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By means of definition, digital artworks encompass an array of artistic productions that are expressed in a technological form as an essential part of a creative process. Examples include illustrations, photos, videos, sculptures, and installations. Within the context of the visual arts, the process of repatriation involves the return of once-appropriated goods. Archiving denotes the preservation of a commodity for storage purposes in order to nurture its continuity. The aforementioned definitions form the foundation of the academic framework and premise of the argument, which is outlined in this paper. This paper aims to define, discuss and decipher the complexities involved in digitising artworks, whilst explaining the benefits of the process, particularly within the South African context, which is rich in tangible and intangible traditional cultural material, objects, and performances. With the internet having been introduced to the African Continent in the early 1990s, this new form of technology, in its own right, initiated a high degree of efficiency, which also resulted in the progressive transformation of computer-generated visual output. Subsequently, this caused a revolutionary influence on the manner in which technological software was developed and uterlised in art-making. Digital technology and the digitisation of creative processes then opened up new avenues of collating and recording information. One of the first visual artists to make use of digital technology software in his creative productions was United States-based artist John Whitney. His inventive work contributed greatly to the onset and development of digital animation. Comparable by technique and originality, South African contemporary visual artists who make digital artworks, both locally and internationally, include David Goldblatt, Katherine Bull, Fritha Langerman, David Masoga, Zinhle Sethebe, Alicia Mcfadzean, Ivan Van Der Walt, Siobhan Twomey, and Fhatuwani Mukheli. In conclusion, the main objective of this paper is to address the following questions: In which ways has the South African art community of visual artists made use of and benefited from technology, in its digital form, as a means to further advance creativity? What are the positive changes that have resulted in art production in South Africa since the onset and use of digital technological software? How has digitisation changed the manner in which we record, interpret, and archive both written and visual information? What is the role of South African art institutions in the development of digital technology and its use in the field of visual art. What role does digitisation play in the process of the repatriation of artworks and artefacts. The methodology in terms of the research process of this paper takes on a multifacted form, inclusive of data analysis of information attained by means of qualitative and quantitative approaches.

Keywords: digital art, digitisation, technology, archiving, transformation and repatriation

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13 Species Profiling of White Grub Beetles and Evaluation of Pre and Post Sown Application of Insecticides against White Grub Infesting Soybean

Authors: Ajay Kumar Pandey, Mayank Kumar

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White grub (Coleoptera: Scarabaeidae) is a major destructive pest in western Himalayan region of Uttarakhand. Beetles feed on apple, apricot, plum, walnut etc. during night while, second and third instar grubs feed on live roots of cultivated as well as non-cultivated crops. Collection and identification of scarab beetles through light trap was carried out at Crop Research Centre, Govind Ballab Pant University Pantnagar, Udham Singh Nagar (Uttarakhand) during 2018. Field trials were also conducted in 2018 to evaluate pre and post sown application of different insecticides against the white grub infesting soybean. The insecticides like Carbofuran 3 Granule (G) (750 g a.i./ha), Clothianidin 50 Water Dispersal Granule (WG) (120 g a.i./ha), Fipronil 0.3 G (50 g a.i./ha), Thiamethoxam 25 WG (80 g a.i./ha), Imidacloprid 70 WG (300 g a.i./ha), Chlorantraniliprole 0.4% G(100 g a.i./ha) and mixture of Fipronil 40% and Imidacloprid 40% WG (300 g a.i./ha) were applied at the time of sowing in pre sown experiment while same dosage of insecticides were applied in standing soybean crop during (first fortnight of July). Commutative plant mortality data were recorded after 20, 40, 60 days intervals and compared with untreated control. Total 23 species of white grub beetles recorded on the light trap and Holotrichia serrata Fabricious (Coleoptera: Melolonthinae) was found to be predominant species by recording 20.6% relative abundance out of the total light trap catch (i.e. 1316 beetles) followed by Phyllognathus sp. (14.6% relative abundance). H. rosettae and Heteronychus lioderus occupied third and fourth rank with 11.85% and 9.65% relative abundance, respectively. The emergence of beetles of predominant species started from 15th March, 2018. In April, average light trap catch was 382 white grub beetles, however, peak emergence of most of the white grub species was observed from June to July, 2018 i.e. 336 beetles in June followed by 303 beetles in the July. On the basis of the emergence pattern of white grub beetles, it may be concluded that the Peak Emergence Period (PEP) for the beetles of H. serrata was second fortnight of April for the total period of 15 days. In May, June and July relatively low population of H. serrata was observed. A decreasing trend in light trap catch was observed and went on till September during the study. No single beetle of H. serrata was observed on light trap from September onwards. The cumulative plant mortality data in both the experiments revealed that all the insecticidal treatments were significantly superior in protection-wise (6.49-16.82% cumulative plant mortality) over untreated control where highest plant mortality was 17.28 to 39.65% during study. The mixture of Fipronil 40% and Imidacloprid 40% WG applied at the rate of 300 g a.i. per ha proved to be most effective having lowest plant mortality i.e. 9.29 and 10.94% in pre and post sown crop, followed by Clothianidin 50 WG (120 g a.i. per ha) where the plant mortality was 10.57 and 11.93% in pre and post sown treatments, respectively. Both treatments were found significantly at par among each other. Production-wise, all the insecticidal treatments were found statistically superior (15.00-24.66 q per ha grain yields) over untreated control where the grain yield was 8.25 & 9.13 q per ha. Treatment Fipronil 40% + Imidacloprid 40% WG applied at the rate of 300 g a.i. per ha proved to be most effective and significantly superior over Imidacloprid 70WG applied at the rate of 300 g a.i. per ha.

Keywords: bio efficacy, insecticide, soybean, white grub

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12 Mobile Genetic Elements in Trematode Himasthla Elongata Clonal Polymorphism

Authors: Anna Solovyeva, Ivan Levakin, Nickolai Galaktionov, Olga Podgornaya

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Animals that reproduce asexually were thought to have the same genotypes within generations for a long time. However, some refuting examples were found, and mobile genetic elements (MGEs) or transposons are considered to be the most probable source of genetic instability. Dispersed nature and the ability to change their genomic localization enables MGEs to be efficient mutators. Hence the study of MGEs genomic impact requires an appropriate object which comprehends both representative amounts of various MGEs and options to evaluate the genomic influence of MGEs. Animals that reproduce asexually seem to be a decent model to study MGEs impact in genomic variability. We found a small marine trematode Himasthla elongata (Himasthlidae) to be a good model for such investigation as it has a small genome size, diverse MGEs and parthenogenetic stages in the lifecycle. In the current work, clonal diversity of cercaria was traced with an AFLP (Amplified fragment length polymorphism) method, diverse zones from electrophoretic patterns were cloned, and the nature of the fragments explored. Polymorphic patterns of individual cercariae AFLP-based fingerprints are enriched with retrotransposons of different families. The bulk of those sequences are represented by open reading frames of non-Long Terminal Repeats containing elements(non-LTR) yet Long-Terminal Repeats containing elements (LTR), to a lesser extent in variable figments of AFLP array. The CR1 elements expose both in polymorphic and conservative patterns are remarkably more frequent than the other non-LTR retrotransposons. This data was confirmed with shotgun sequencing-based on Illumina HiSeq 2500 platform. Individual cercaria of the same clone (i.e., originated from a single miracidium and inhabiting one host) has a various distribution of MGE families detected in sequenced AFLP patterns. The most numerous are CR1 and RTE-Bov retrotransposons, typical for trematode genomes. Also, we identified LTR-retrotransposons of Pao and Gypsy families among DNA transposons of CMC-EnSpm, Tc1/Mariner, MuLE-MuDR and Merlin families. We detected many of them in H. elongata transcriptome. Such uneven MGEs distribution in AFLP sequences’ sets reflects the different patterns of transposons spreading in cercarial genomes as transposons affect the genome in many ways (ectopic recombination, gene structure interruption, epigenetic silencing). It is considered that they play a key role in the origins of trematode clonal polymorphism. The authors greatly appreciate the help received at the Kartesh White Sea Biological Station of the Russian Academy of Sciences Zoological Institute. This work is funded with RSF 19-74-20102 and RFBR 17-04-02161 grants and the research program of the Zoological Institute of the Russian Academy of Sciences (project number AAAA-A19-119020690109-2).

Keywords: AFLP, clonal polymorphism, Himasthla elongata, mobile genetic elements, NGS

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11 Pre-conditioning and Hot Water Sanitization of Reverse Osmosis Membrane for Medical Water Production

Authors: Supriyo Das, Elbir Jove, Ajay Singh, Sophie Corbet, Noel Carr, Martin Deetz

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Water is a critical commodity in the healthcare and medical field. The utility of medical-grade water spans from washing surgical equipment, drug preparation to the key element of life-saving therapy such as hydrotherapy and hemodialysis for patients. A properly treated medical water reduces the bioburden load and mitigates the risk of infection, ensuring patient safety. However, any compromised condition during the production of medical-grade water can create a favorable environment for microbial growth putting patient safety at high risk. Therefore, proper upstream treatment of the medical water is essential before its application in healthcare, pharma and medical space. Reverse Osmosis (RO) is one of the most preferred treatments within healthcare industries and is recommended by all International Pharmacopeias to achieve the quality level demanded by global regulatory bodies. The RO process can remove up to 99.5% of constituents from feed water sources, eliminating bacteria, proteins and particles sizes of 100 Dalton and above. The combination of RO with other downstream water treatment technologies such as Electrodeionization and Ultrafiltration meet the quality requirements of various pharmacopeia monographs to produce highly purified water or water for injection for medical use. In the reverse osmosis process, the water from a liquid with a high concentration of dissolved solids is forced to flow through an especially engineered semi-permeable membrane to the low concentration side, resulting in high-quality grade water. However, these specially engineered RO membranes need to be sanitized either chemically or at high temperatures at regular intervals to keep the bio-burden at the minimum required level. In this paper, we talk about Dupont´s FilmTec Heat Sanitizable Reverse Osmosis membrane (HSRO) for the production of medical-grade water. An HSRO element must be pre-conditioned prior to initial use by exposure to hot water (80°C-85°C) for its stable performance and to meet the manufacturer’s specifications. Without pre-conditioning, the membrane will show variations in feed pressure operations and salt rejection. The paper will discuss the critical variables of pre-conditioning steps that can affect the overall performance of the HSRO membrane and demonstrate the data to support the need for pre-conditioning of HSRO elements. Our preliminary data suggests that there can be up to 35 % reduction in flow due to initial heat treatment, which also positively affects the increase in salt rejection. The paper will go into detail about the fundamental understanding of the performance change of HSRO after the pre-conditioning step and its effect on the quality of medical water produced. The paper will also discuss another critical point, “regular hot water sanitization” of these HSRO membranes. Regular hot water sanitization (at 80°C-85°C) is necessary to keep the membrane bioburden free; however, it can negatively impact the performance of the membrane over time. We will demonstrate several data points on hot water sanitization using FilmTec HSRO elements and challenge its robustness to produce quality medical water. The last part of this paper will discuss the construction details of the FilmTec HSRO membrane and features that make it suitable to pre-condition and sanitize at high temperatures.

Keywords: heat sanitizable reverse osmosis, HSRO, medical water, hemodialysis water, water for Injection, pre-conditioning, heat sanitization

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10 Relationship between Illegal Wildlife Trade and Community Conservation: A Case Study of the Chepang Community in Nepal

Authors: Vasundhara H. Krishnani, Ajay Saini, Dibesh Karmacharya, Salit Kark

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Illegal Wildlife Trade is one of the most pressing global conservation challenges. Unregulated wildlife trade can threaten biodiversity, contribute to habitat loss, limit sustainable development efforts, and expedite species declines and extinctions. In low-income and middle-income countries, such as Nepal and other countries in Asia and Africa, many of the people engaged in the early stages of illegal wildlife trade, which includes the hunting and transportation of wildlife, belong to Indigenous tribes and local communities.These countries primarily rely on punitive measures to prevent and suppress Illegal Wildlife Trade. For example, in Nepal, people involved in wildlife crimes can often be sentenced to incarceration and a hefty fine and serve up to 15 years in prison. Despite these harsh punitive measures, illegal wildlife trade remains a significant conservation challenge in many countries. The aim of this study was to examine factors affecting the participation of Indigenous communities in Illegal Wildlife Trade while recording the experiences of members of the Indigenous Chepang community, some of whom were imprisoned for their alleged involvement in rhino poaching. Chepangs, belonging to traditionally a hunter-gatherer community, are often considered an isolated and marginalized Indigenous community, some of whom live around the Chitwan National Park in Nepal. Established in 1973, Chitwan National Park is situated in the Chitwan Valley of Nepal and was one of the first regions that was declared as a protected area in Nepal, aiming to protect the one-horned rhinoceros as a flagship species. Conducted over a period of three years, this study used semi-structured interviews and focus group discussions to collect data from Illegal Wildlife Trade offenders, family members of offenders, community Elders, NGO personnel, community forest representatives, Chepang community representatives, and Government school teachers from the region surrounding Chitwan National Park. The study also examined the social, cultural, health, and financial impacts that the imprisonment of offenders had on the families of the community members, especially women and children. The results suggest that involvement of the members of the Chepang community living around Chitwan National Park in the poaching of the one-horned rhinoceros (Rhinoceros unicornis) can be attributed to a range of factors, some of which include: lack of livelihood opportunities, lack of awareness regarding wildlife rules and regulations and poverty.This work emphasises the need for raising awareness and building programs to enhance alternative livelihood training and empower indigenous and marginalised communities that provide sustainable alternatives. Furthermore, the issue needs to be addressed as a community solution which includes all community members. We suggest this multi-pronged approach can benefit wildlife conservation by reducing illegal poaching and wildlife trade, as well as community conservation in regions with similar challenges. By actively involving and empowering local communities, the communities become key stakeholders in the conservation process. This involvement contributes to protecting wildlife and natural ecosystems while simultaneously providing sustainable livelihood options for local communities.

Keywords: alternative livelihoods, chepang community, illegal wildlife trade, low-and middle-income countries, nepal, one-horned rhinoceros

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9 Insufficiency of Cardioprotection at Adaptation to Chronic Hypoxia and at Remote Postconditioning in Young and Aged Rats with Metabolic Syndrome, the Role of Metabolic Disorders or Opioid Signaling

Authors: Natalia V. Naryzhnaya, Alexandr V. Mukhomedzyanov, Ivan A. Derkachev, Boris K. Kurbatov, Leonid N. Maslov

Abstract:

Background: Techniques of adaptation to hypoxia and remote postconditioning (RPost) have great prospects for use in the clinic. However, recent studies have shown low efficacy of remote postconditioning in patients with AMI. We hypothesize that the reasons for this inefficiency may be metabolic disorders, which are very common, especially in patients with cardiovascular disease, and age of patients. The purpose of the study was to reveal the effectiveness of adaptation to chronic hypoxia and RPost. To determine the possible relationship between the decrease in the effectiveness of projective impacts and disorders of carbohydrate and lipid metabolism. Design: The study was carried out on Wistar rats 60 day old. MetS was induced by high-carbohydrate, high-fat diet (HСHFD). Modeling MS led to the formation of obesity, hypertension, impaired lipid and carbohydrate metabolism, hyperleptinemia, and moderate stress. Groups with adaptation to chronic hypoxia were subjected to hypoxia for 21 days at 12% O2 and 0.3% CO2 after complete of HСHFD. All animals were subjected to 45 min coronary occlusion and 120 min reperfusion. Groups with RPost, immediately after the end of ischemia, tourniquets were applied to the hind limbs in the area of the hip joint (3 times in the mode of 5 min ischemia, 5 min reperfusion). Results: RPost led to a twofold reduction of infarct size in rats with intact metabolism (р < 0.0001), while in rats with MetS, a decrease in infarct size during RPost was 25 % (p = 0.00003). A direct correlation was found between of infarct size during RPost and the serum leptin level of rats with MetC (r = 0.85). The presented data suggested that a decrease in the efficiency of remote postconditioning in rats with diet-induced metabolic syndrome depends on serum leptin. Chronic hypoxia resulted in a 38% reduced in infarct size in metabolically intact rats. The decrease of cardioprotection was observed in rats with chronic hypoxia and MetS. Infarct size showed a direct correlation with impaired glucose tolerance (AUC, glucose tolerance test, r = 0.034) and serum triglyceride levels (r = 0.39). Our study showed the dependence of cardioprotection in rats with metabolic syndrome during chronic hypoxia and DPost on opioids in the blood serum and myocardium, protein kinase C and NO synthase activity. Conclusion: The results obtained showed that the infarct-limiting efficiency of adaptation to hypoxia and remote postconditioning is reduced or completely absent in animals with metabolic syndrome. The increase in the infarction, in this case, directly depends on the disturbances in carbohydrate. lipid metabolism and opioids signaling. Funding: Investigation of effectiveness of chronic hypoxia at the metabolic syndrome was carried out within the support of Russian Science Foundation Grant 22-15-00048. Studies of the mechanisms of arterial hypertension in induced metabolic syndrome were carried out within the framework of the state assignment (122020300042-4). The work was performed using the Center for Collective Use "Medical Genomics".

Keywords: chronic hypoxia, opioids, remote postconditioning, metabolic syndrome

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8 Language Skills in the Emergent Literacy of Spanish-Speaking Children with Autism Spectrum Disorders

Authors: Adriana Salgado, Sandra Castaneda, Ivan Perez

Abstract:

Learning to read and write is a complex process involving several cognitive skills, contextual, and cultural environments. The basis of this development is linguistic skills, such as the ability to name and understand vocabulary, retell a story, phonological awareness, letter knowledge, among others. In children with autism spectrum disorder (ASD), one of the main concerns is related to language disorders. Nevertheless, most of the children with ASD are able to decode written information but have difficulties in reading comprehension. The research of these processes in the Spanish-speaking population is limited. However, the increasing prevalence of this diagnosis (1 in 115 children) in Mexico has implications at different levels. Educational research is an important area of interest in ASD children, such as emergent literacy. Reading and writing expand the possibilities of academic, cultural, and social information access. Taking this information into account, the objective of this research was to identify the relationship between language skills, alphabet knowledge, phonological awareness, and early reading and writing in ASD Spanish-speaking children. The method used for this research was based on tasks that were selected, adapted and in some cases designed to measure initial reading and writing, as well as language skills (naming, receptive vocabulary, and narrative skills), phonological awareness (similar phonological word pairs, beginning sound awareness and spelling) and letter knowledge, in a sample of 45 children (38 boys and 7 girls) with prior diagnosis of ASD. Descriptive analyses, as well as bivariate correlations, cluster analysis, and canonical correspondence, were obtained for the data results. Results showed that variability was large; however, it was possible to characterize the sample in low, medium, and high score groups regarding children performance. The low score group (46.7% of the sample), had a null or deficient performance in language skills and phonological awareness, some could identify up to five letters of the alphabet, showed no early reading skills but they could scribble. The middle score group was characterized by a highly variable performance in different tasks, with better language skills in receptive and naming vocabulary, some narrative, letter knowledge, and phonological awareness (beginning sound awareness) skills. The high score group, (24.4% of the sample) had the best performance in language skills in relation to the sample data, as well as in the rest of the measured skills. Finally, scores were canonically correlated between naming, receptive vocabulary, narrative, phonological awareness, letter knowledge and initial learning of reading and writing skills for the high score group and letter knowledge, naming and receptive vocabulary for the lower score group, which is consistent with previous research in typical and ASD children. In conclusion, the obtained data is consistent with previous studies. Despite large variability, it was possible to identify performance profiles and relations based on linguistic, phonological awareness, and letter knowledge skills. These skills were predictor variables of the initial development of reading and writing. The above has implications for a future program and strategies development that may benefit the acquisition of reading and writing in ASD children.

Keywords: autism, autism spectrum disorders, early literacy, emergent literacy

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7 The Role of a Biphasic Implant Based on a Bioactive Silk Fibroin for Osteochondral Tissue Regeneration

Authors: Lizeth Fuentes-Mera, Vanessa Perez-Silos, Nidia K. Moncada-Saucedo, Alejandro Garcia-Ruiz, Alberto Camacho, Jorge Lara-Arias, Ivan Marino-Martinez, Victor Romero-Diaz, Adolfo Soto-Dominguez, Humberto Rodriguez-Rocha, Hang Lin, Victor Pena-Martinez

Abstract:

Biphasic scaffolds in cartilage tissue engineering have been designed to influence not only the recapitulation of the osteochondral architecture but also to take advantage of the healing ability of bone to promote the implant integration with the surrounding tissue and then bone restoration and cartilage regeneration. This study reports the development and characterization of a biphasic scaffold based on the assembly of a cartilage phase constituted by fibroin biofunctionalized with bovine cartilage matrix; cellularized with differentiated pre-chondrocytes from adipose tissue stem cells (autologous) and well attached to a bone phase (bone bovine decellularized) to mimic the structure of the nature of native tissue and to promote the cartilage regeneration in a model of joint damage in pigs. Biphasic scaffolds were assembled by fibroin crystallization with methanol. The histological and ultrastructural architectures were evaluated by optical and scanning electron microscopy respectively. Mechanical tests were conducted to evaluate Young's modulus of the implant. For the biological evaluation, pre-chondrocytes were loaded onto the scaffolds and cellular adhesion, proliferation, and gene expression analysis of cartilage extracellular matrix components was performed. The scaffolds that were cellularized and matured for 10 days were implanted into critical 3 mm in diameter and 9-mm in depth osteochondral defects in a porcine model (n=4). Three treatments were applied per knee: Group 1: monophasic cellular scaffold (MS) (single chondral phase), group 2: biphasic scaffold, cellularized only in the chondral phase (BS1), group 3: BS cellularized in both bone and chondral phases (BS2). Simultaneously, a control without treatment was evaluated. After 4 weeks of surgery, integration and regeneration tissues were analyzed by x-rays, histology and immunohistochemistry evaluation. The mechanical assessment showed that the acellular biphasic composites exhibited Young's modulus of 805.01 kPa similar to native cartilage (400-800 kPa). In vitro biological studies revealed the chondroinductive ability of the biphasic implant, evidenced by an increase in sulfated glycosaminoglycan (GAGs) and type II collagen, both secreted by the chondrocytes cultured on the scaffold during 28 days. No evidence of adverse or inflammatory reactions was observed in the in vivo trial; however, In group 1, the defects were not reconstructed. In group 2 and 3 a good integration of the implant with the surrounding tissue was observed. Defects in group 2 were fulfilled by hyaline cartilage and normal bone. Group 3 defects showed fibrous repair tissue. In conclusion; our findings demonstrated the efficacy of biphasic and bioactive scaffold based on silk fibroin, which entwined chondroinductive features and biomechanical capability with appropriate integration with the surrounding tissue, representing a promising alternative for osteochondral tissue-engineering applications.

Keywords: biphasic scaffold, extracellular cartilage matrix, silk fibroin, osteochondral tissue engineering

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6 Gas-Phase Noncovalent Functionalization of Pristine Single-Walled Carbon Nanotubes with 3D Metal(II) Phthalocyanines

Authors: Vladimir A. Basiuk, Laura J. Flores-Sanchez, Victor Meza-Laguna, Jose O. Flores-Flores, Lauro Bucio-Galindo, Elena V. Basiuk

Abstract:

Noncovalent nanohybrid materials combining carbon nanotubes (CNTs) with phthalocyanines (Pcs) is a subject of increasing research effort, with a particular emphasis on the design of new heterogeneous catalysts, efficient organic photovoltaic cells, lithium batteries, gas sensors, field effect transistors, among other possible applications. The possibility of using unsubstituted Pcs for CNT functionalization is very attractive due to their very moderate cost and easy commercial availability. However, unfortunately, the deposition of unsubstituted Pcs onto nanotube sidewalls through the traditional liquid-phase protocols turns to be very problematic due to extremely poor solubility of Pcs. On the other hand, unsubstituted free-base H₂Pc phthalocyanine ligand, as well as many of its transition metal complexes, exhibit very high thermal stability and considerable volatility under reduced pressure, which opens the possibility for their physical vapor deposition onto solid surfaces, including nanotube sidewalls. In the present work, we show the possibility of simple, fast and efficient noncovalent functionalization of single-walled carbon nanotubes (SWNTs) with a series of 3d metal(II) phthalocyanines Me(II)Pc, where Me= Co, Ni, Cu, and Zn. The functionalization can be performed in a temperature range of 400-500 °C under moderate vacuum and requires about 2-3 h only. The functionalized materials obtained were characterized by means of Fourier-transform infrared (FTIR), Raman, UV-visible and energy-dispersive X-ray spectroscopy (EDS), scanning and transmission electron microscopy (SEM and TEM, respectively) and thermogravimetric analysis (TGA). TGA suggested that Me(II)Pc weight content is 30%, 17% and 35% for NiPc, CuPc, and ZnPc, respectively (CoPc exhibited anomalous thermal decomposition behavior). The above values are consistent with those estimated from EDS spectra, namely, of 24-39%, 27-36% and 27-44% for CoPc, CuPc, and ZnPc, respectively. A strong increase in intensity of D band in the Raman spectra of SWNT‒Me(II)Pc hybrids, as compared to that of pristine nanotubes, implies very strong interactions between Pc molecules and SWNT sidewalls. Very high absolute values of binding energies of 32.46-37.12 kcal/mol and the highest occupied and lowest unoccupied molecular orbital (HOMO and LUMO, respectively) distribution patterns, calculated with density functional theory by using Perdew-Burke-Ernzerhof general gradient approximation correlation functional in combination with the Grimme’s empirical dispersion correction (PBE-D) and the double numerical basis set (DNP), also suggested that the interactions between Me(II) phthalocyanines and nanotube sidewalls are very strong. The authors thank the National Autonomous University of Mexico (grant DGAPA-IN200516) and the National Council of Science and Technology of Mexico (CONACYT, grant 250655) for financial support. The authors are also grateful to Dr. Natalia Alzate-Carvajal (CCADET of UNAM), Eréndira Martínez (IF of UNAM) and Iván Puente-Lee (Faculty of Chemistry of UNAM) for technical assistance with FTIR, TGA measurements, and TEM imaging, respectively.

Keywords: carbon nanotubes, functionalization, gas-phase, metal(II) phthalocyanines

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5 The Use of Non-Parametric Bootstrap in Computing of Microbial Risk Assessment from Lettuce Consumption Irrigated with Contaminated Water by Sanitary Sewage in Infulene Valley

Authors: Mario Tauzene Afonso Matangue, Ivan Andres Sanchez Ortiz

Abstract:

The Metropolitan area of Maputo (Mozambique Capital City) is located in semi-arid zone (800 mm annual rainfall) with 1101170 million inhabitants. On the west side, there are the flatlands of Infulene where the Mulauze River flows towards to the Indian Ocean, receiving at this site, the storm water contaminated with sanitary sewage from Maputo, transported through a concrete open channel. In Infulene, local communities grow salads crops such as tomato, onion, garlic, lettuce, and cabbage, which are then commercialized and consumed in several markets in Maputo City. Lettuce is the most daily consumed salad crop in different meals, generally in fast-foods, breakfasts, lunches, and dinners. However, the risk of infection by several pathogens due to the consumption of lettuce, using the Quantitative Microbial Risk Assessment (QMRA) tools, is still unknown since there are few studies or publications concerning to this matter in Mozambique. This work is aimed at determining the annual risk arising from the consumption of lettuce grown in Infulene valley, in Maputo, using QMRA tools. The exposure model was constructed upon the volume of contaminated water remaining in the lettuce leaves, the empirical relations between the number of pathogens and the indicator of microorganisms (E. coli), the consumption of lettuce (g) and reduction of pathogens (days). The reference pathogens were Vibrio cholerae, Cryptosporidium, norovirus, and Ascaris. The water quality samples (E. coli) were collected in the storm water channel from January 2016 to December 2018, comprising 65 samples, and the urban lettuce consumption data were collected through inquiry in Maputo Metropolis covering 350 persons. A non-parametric bootstrap was performed involving 10,000 iterations over the collected dataset, namely, water quality (E. coli) and lettuce consumption. The dose-response models were: Exponential for Cryptosporidium, Kummer Confluent hypergeomtric function (1F1) for Vibrio and Ascaris Gaussian hypergeometric function (2F1-(a,b;c;z) for norovirus. The annual infection risk estimates were performed using R 3.6.0 (CoreTeam) software by Monte Carlo (Latin hypercubes), a sampling technique involving 10,000 iterations. The annual infection risks values expressed by Median and the 95th percentile, per person per year (pppy) arising from the consumption of lettuce are as follows: Vibrio cholerae (1.00, 1.00), Cryptosporidium (3.91x10⁻³, 9.72x 10⁻³), nororvirus (5.22x10⁻¹, 9.99x10⁻¹) and Ascaris (2.59x10⁻¹, 9.65x10⁻¹). Thus, the consumption of the lettuce would result in greater risks than the tolerable levels ( < 10⁻³ pppy or 10⁻⁶ DALY) for all pathogens, and the Vibrio cholerae is the most virulent pathogens, according to the hit-single models followed by the Ascaris lumbricoides and norovirus. The sensitivity analysis carried out in this work pointed out that in the whole QMRA, the most important input variable was the reduction of pathogens (Spearman rank value was 0.69) between harvest and consumption followed by water quality (Spearman rank value was 0.69). The decision-makers (Mozambique Government) must strengthen the prevention measures related to pathogens reduction in lettuce (i.e., washing) and engage in wastewater treatment engineering.

Keywords: annual infections risk, lettuce, non-parametric bootstrapping, quantitative microbial risk assessment tools

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4 Hybrid Materials on the Basis of Magnetite and Magnetite-Gold Nanoparticles for Biomedical Application

Authors: Mariia V. Efremova, Iana O. Tcareva, Anastasia D. Blokhina, Ivan S. Grebennikov, Anastasia S. Garanina, Maxim A. Abakumov, Yury I. Golovin, Alexander G. Savchenko, Alexander G. Majouga, Natalya L. Klyachko

Abstract:

During last decades magnetite nanoparticles (NPs) attract a deep interest of scientists due to their potential application in therapy and diagnostics. However, magnetite nanoparticles are toxic and non-stable in physiological conditions. To solve these problems, we decided to create two types of hybrid systems based on magnetite and gold which is inert and biocompatible: gold as a shell material (first type) and gold as separate NPs interfacially bond to magnetite NPs (second type). The synthesis of the first type hybrid nanoparticles was carried out as follows: Magnetite nanoparticles with an average diameter of 9±2 nm were obtained by co-precipitation of iron (II, III) chlorides then they were covered with gold shell by iterative reduction of hydrogen tetrachloroaurate with hydroxylamine hydrochloride. According to the TEM, ICP MS and EDX data, final nanoparticles had an average diameter of 31±4 nm and contained iron even after hydrochloric acid treatment. However, iron signals (K-line, 7,1 keV) were not localized so we can’t speak about one single magnetic core. Described nanoparticles covered with mercapto-PEG acid were non-toxic for human prostate cancer PC-3/ LNCaP cell lines (more than 90% survived cells as compared to control) and had high R2-relaxivity rates (>190 mМ-1s-1) that exceed the transverse relaxation rate of commercial MRI-contrasting agents. These nanoparticles were also used for chymotrypsin enzyme immobilization. The effect of alternating magnetic field on catalytic properties of chymotrypsin immobilized on magnetite nanoparticles, notably the slowdown of catalyzed reaction at the level of 35-40 % was found. The synthesis of the second type hybrid nanoparticles also involved two steps. Firstly, spherical gold nanoparticles with an average diameter of 9±2 nm were synthesized by the reduction of hydrogen tetrachloroaurate with oleylamine; secondly, they were used as seeds during magnetite synthesis by thermal decomposition of iron pentacarbonyl in octadecene. As a result, so-called dumbbell-like structures were obtained where magnetite (cubes with 25±6 nm diagonal) and gold nanoparticles were connected together pairwise. By HRTEM method (first time for this type of structure) an epitaxial growth of magnetite nanoparticles on gold surface with co-orientation of (111) planes was discovered. These nanoparticles were transferred into water by means of block-copolymer Pluronic F127 then loaded with anti-cancer drug doxorubicin and also PSMA-vector specific for LNCaP cell line. Obtained nanoparticles were found to have moderate toxicity for human prostate cancer cells and got into the intracellular space after 45 minutes of incubation (according to fluorescence microscopy data). These materials are also perspective from MRI point of view (R2-relaxivity rates >70 mМ-1s-1). Thereby, in this work magnetite-gold hybrid nanoparticles, which have a strong potential for biomedical application, particularly in targeted drug delivery and magnetic resonance imaging, were synthesized and characterized. That paves the way to the development of special medicine types – theranostics. The authors knowledge financial support from Ministry of Education and Science of the Russian Federation (14.607.21.0132, RFMEFI60715X0132). This work was also supported by Grant of Ministry of Education and Science of the Russian Federation К1-2014-022, Grant of Russian Scientific Foundation 14-13-00731 and MSU development program 5.13.

Keywords: drug delivery, magnetite-gold, MRI contrast agents, nanoparticles, toxicity

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3 Development of Anti-Fouling Surface Features Bioinspired by the Patterned Micro-Textures of the Scophthalmus rhombus (Brill)

Authors: Ivan Maguire, Alan Barrett, Alex Forte, Sandra Kwiatkowska, Rohit Mishra, Jens Ducrèe, Fiona Regan

Abstract:

Biofouling is defined as the gradual accumulation of Biomimetics refers to the use and imitation of principles copied from nature. Biomimetics has found interest across many commercial disciplines. Among many biological objects and their functions, aquatic animals deserve a special attention due to their antimicrobial capabilities resulting from chemical composition, surface topography or other behavioural defences, which can be used as an inspiration for antifouling technology. Marine biofouling has detrimental effects on seagoing vessels, both commercial and leisure, as well as on oceanographic sensors, offshore drilling rigs, and aquaculture installations. Sensor optics, membranes, housings and platforms can become fouled leading to problems with sensor performance and data integrity. While many anti-fouling solutions are currently being investigated as a cost-cutting measure, biofouling settlement may also be prevented by creating a surface that does not satisfy the settlement conditions. Brill (Scophthalmus rhombus) is a small flatfish occurring in marine waters of Mediterranean as well as Norway and Iceland. It inhabits sandy and muddy coastal waters from 5 to 80 meters. Its skin colour changes depending on environment, but generally is brownish with light and dark freckles, with creamy underside. Brill is oval in shape and its flesh is white. The aim of this study is to translate the unique micro-topography of the brill scale, to design marine inspired biomimetic surface coating and test it against a typical fouling organism. Following extensive study of scale topography of the brill fish (Scophthalmus rhombus) and the settlement behaviour of the diatom species Psammodictyon sp. via SEM, two state-of-the-art antifouling surface solutions were designed and investigated; A brill fish scale bioinspired surface pattern platform (BFD), and generic and uniformly-arrayed, circular micropillar platform (MPD), with offsets based on diatom species settlement behaviour. The BFD approach consists of different ~5 μm by ~90 μm Brill-replica patterns, grown to a 5 μm height, in a linear array pattern. The MPD approach utilises hexagonal-packed cylindrical pillars 10.6 μm in diameter, grown to a height of 5 μm, with vertical offset of 15 μm and horizontal offset of 26.6 μm. Photolithography was employed for microstructure growth, with a polydimethylsiloxane (PDMS) chip-based used as a testbed for diatom adhesion on both platforms. Settlement and adhesion tests were performed using this PDMS microfluidic chip through subjugation to centrifugal force via an in-house developed ‘spin-stand’ which features a motor, in combination with a high-resolution camera, for real-time observing diatom release from PDMS material. Diatom adhesion strength can therefore be determined based on the centrifugal force generated at varying rotational speeds. It is hoped that both the replica and bio-inspired solutions will give comparable anti-fouling results to these synthetic surfaces, whilst also assisting in determining whether anti-fouling solutions should predominantly be investigating either fully bioreplica-based, or a bioinspired, synthetically-based design.

Keywords: anti-fouling applications, bio-inspired microstructures, centrifugal microfluidics, surface modification

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2 Design of DNA Origami Structures Using LAMP Products as a Combined System for the Detection of Extended Spectrum B-Lactamases

Authors: Kalaumari Mayoral-Peña, Ana I. Montejano-Montelongo, Josué Reyes-Muñoz, Gonzalo A. Ortiz-Mancilla, Mayrin Rodríguez-Cruz, Víctor Hernández-Villalobos, Jesús A. Guzmán-López, Santiago García-Jacobo, Iván Licona-Vázquez, Grisel Fierros-Romero, Rosario Flores-Vallejo

Abstract:

The group B-lactamic antibiotics include some of the most frequently used small drug molecules against bacterial infections. Nevertheless, an alarming decrease in their efficacy has been reported due to the emergence of antibiotic-resistant bacteria. Infections caused by bacteria expressing extended Spectrum B-lactamases (ESBLs) are difficult to treat and account for higher morbidity and mortality rates, delayed recovery, and high economic burden. According to the Global Report on Antimicrobial Resistance Surveillance, it is estimated that mortality due to resistant bacteria will ascend to 10 million cases per year worldwide. These facts highlight the importance of developing low-cost and readily accessible detection methods of drug-resistant ESBLs bacteria to prevent their spread and promote accurate and fast diagnosis. Bacterial detection is commonly done using molecular diagnostic techniques, where PCR stands out for its high performance. However, this technique requires specialized equipment not available everywhere, is time-consuming, and has a high cost. Loop-Mediated Isothermal Amplification (LAMP) is an alternative technique that works at a constant temperature, significantly decreasing the equipment cost. It yields double-stranded DNA of several lengths with repetitions of the target DNA sequence as a product. Although positive and negative results from LAMP can be discriminated by colorimetry, fluorescence, and turbidity, there is still a large room for improvement in the point-of-care implementation. DNA origami is a technique that allows the formation of 3D nanometric structures by folding a large single-stranded DNA (scaffold) into a determined shape with the help of short DNA sequences (staples), which hybridize with the scaffold. This research aimed to generate DNA origami structures using LAMP products as scaffolds to improve the sensitivity to detect ESBLs in point-of-care diagnosis. For this study, the coding sequence of the CTM-X-15 ESBL of E. coli was used to generate the LAMP products. The set of LAMP primers were designed using PrimerExplorerV5. As a result, a target sequence of 200 nucleotides from CTM-X-15 ESBL was obtained. Afterward, eight different DNA origami structures were designed using the target sequence in the SDCadnano and analyzed with CanDo to evaluate the stability of the 3D structures. The designs were constructed minimizing the total number of staples to reduce costs and complexity for point-of-care applications. After analyzing the DNA origami designs, two structures were selected. The first one was a zig-zag flat structure, while the second one was a wall-like shape. Given the sequence repetitions in the scaffold sequence, both were able to be assembled with only 6 different staples each one, ranging between 18 to 80 nucleotides. Simulations of both structures were performed using scaffolds of different sizes yielding stable structures in all the cases. The generation of the LAMP products were tested by colorimetry and electrophoresis. The formation of the DNA structures was analyzed using electrophoresis and colorimetry. The modeling of novel detection methods through bioinformatics tools allows reliable control and prediction of results. To our knowledge, this is the first study that uses LAMP products and DNA-origami in combination to delect ESBL-producing bacterial strains, which represent a promising methodology for diagnosis in the point-of-care.

Keywords: beta-lactamases, antibiotic resistance, DNA origami, isothermal amplification, LAMP technique, molecular diagnosis

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