Search results for: respiratory infection

Authors: Rebin Mzhda Mohammed, Hoshmand Ali Hama Agha

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Excess skin, often consequential to substantial weight loss or the aging process, introduces physical discomfort, obstructs daily activities, and undermines an individual's self-esteem. As these challenges become increasingly prevalent, the need to explore viable solutions grows in significance. Extra skin removal surgery, colloquially known as body contouring surgery, has emerged as a compelling intervention to ameliorate the physical and psychological burdens of excess skin. This study undertakes a comprehensive review to illuminate the intricacies of extra skin removal surgery, encompassing its diverse procedures, associated risks, benefits, and psychological implications on patients. The methodological approach adopted involves a systematic and exhaustive review of pertinent scholarly literature sourced from reputable databases, including PubMed, Google Scholar, and specialized cosmetic surgery journals. Articles are meticulously curated based on their relevance, credibility, and recency. Subsequently, data from these sources are synthesized and categorized, facilitating a comprehensive understanding of the subject matter. Qualitative analysis serves to unravel the nuanced psychological effects, while quantitative data, where available, are harnessed to underpin the study's conclusions. In terms of major findings, the research underscores the manifold advantages of extra skin removal surgery. Patients experience a notable improvement in physical comfort, amplified mobility, enhanced self-confidence, and a newfound ability to don clothing comfortably. Nonetheless, the benefits are juxtaposed with potential risks, encompassing infection, scarring, hematoma, delayed healing, and the challenge of achieving symmetry. A salient discovery is the profound psychological impact of the surgery, as patients consistently report elevated body image satisfaction, heightened self-esteem, and a substantial enhancement in overall quality of life. In summation, this research accentuates the pivotal role of extra skin removal surgery in ameliorating the intricate interplay of physical and psychological difficulties posed by excess skin. By elucidating the diverse procedures, associated risks, and psychological outcomes, the study contributes to a comprehensive and informed comprehension of the surgery's multifaceted effects. Therefore, individuals contemplating this transformative surgical option are equipped with comprehensive insights, ultimately fostering informed decision-making, guided by the expertise of medical professionals.

Keywords: extra skin removal surgery, body contouring, abdominoplasty, brachioplasty, thigh lift, body lift, benefits, risks, psychological effects

Procedia PDF Downloads 54

Authors: A. F. C. Nassar, D. K. Tessler, L. Okuda, C. Del Fava, D. P. Chiebao, A. H. C. N. Romaldini, A. P. Alvim, M. J. Sanchez-Vazquez, M. S. Rosa, J. C. Pompei, R. Harakava, M. C. S. Araujo, G. H. F. Marques, E. M. Pituco

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Glanders is a zoonotic disease of Equidae caused by the bacterium Burkholderia mallei presented in acute or chronic clinical forms with inflammatory nodules in the respiratory tract, lymphangitis and caseous lymph nodes. There is not a treatment with veterinary drugs to this life-threatening disease; thus, its occurrence must be notified to official animal health services and any infected animal must be eliminated. This study aims to detect B. mallei from horses euthanized in outbreaks of glanders in Brazil, providing a better understanding of the bacterial characteristics and determine a proper protocol for isolation. The work was carried out with the collaboration of the Ministry of Agriculture and the Sao Paulo State Animal Health Department, while its procedures were approved by the Committee of Ethics in Animal Experimentation from the Instituto Biologico (CETEA n°156/2017). To the present time, 16 horses from farms with outbreaks of glanders detected by complement fixation test (CFT) serology method were analyzed. During the necropsy, samples of possibly affected organs (lymph nodes, lungs, heart, liver, spleen, kidneys and trachea) were collected for bacterial isolation, molecular tests and pathology. Isolation was performed using two enriched mediums, a potato infusion agar with 5% sheep blood, 4% glycerol and antibiotics (penicilin100U/ mL), and another with the same ingredients except the antibiotic. A PCR protocol was modified for this study using primers design to identify a region of the Flip gen of B. mallei. Thru isolation, 12.5% (2/16) animals were confirmed positive using only the enriched medium with antibiotic and confirmed by PCR: from mediastinal and submandibular lymph nodes and lungs in one animal and from mediastinal lymph node in the other. The detection of the bacterium using PCR showed positivity of 100% (16/16) horses from 144 samples of organs. Pathology macroscopic lesions observed were catarrhal nasal discharge, fetlock ulcers, emaciation, lymphangitis in limbs, suppurative lymphangitis, lymph node enlargement, star shaped liver, and spleen scars, adherence of the renal capsule, pulmonary hemorrhage, and miliary nodules. Microscopic lesions were suppurative bronchopneumonia with microabscesses and Langhans giant cells in lungs; lymph nodes with abscesses and intense lymphoid reaction; hemosiderosis and abscesses in spleen. Positive samples on PCR will be sequenced later and analyzed comparing with previous records in the literature. A throughout description of the recent acute cases of glanders occurring in Brazil and characterization of the bacterium related will contribute to advances in the knowledge of the pathogenicity, clinical symptoms, and epidemiology of this zoonotic disease. Acknowledgment: This project is sponsored by FAPESP.

Keywords: equines, bacterial isolation, zoonosis, PCR, pathology

Procedia PDF Downloads 119

Authors: Tim J. Walsh, Lindsay E. Young, John A. Schneider

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Background: The disproportionate rate of HIV infection among Black men who have sex with men (BMSM) in the United States suggest the importance of Preexposure Prophylaxis (PrEP) interventions for this population. As such, there is an urgent need for innovative outreach strategies that extend beyond the traditional patient-provider relationship to reach at-risk populations. Training members of the BMSM community as peer change agents (PCAs) is one such strategy. An important piece of this training is goal-setting. Goal-setting not only encourages PCAs to define the parameters of the intervention according to their lived experience, it also helps them plan courses of action. Therefore, the aims of this mixed methods study are: (1) Characterize the goals that BMSM set at the end of their PrEP training and (2) Assess the relationship between goal types and PCA engagement. Methods: Between March 2016 and July 2016, preliminary data were collected from 68 BMSM, ages 18-33, in Chicago as part of an ongoing PrEP intervention. Once enrolled, PCAs participate in a half-day training in which they learn about PrEP, practice initiating conversations about PrEP, and identify strategies for supporting at-risk peers through the PrEP adoption process. Training culminates with a goal-setting exercise, whereby participants establish a goal related to their role as a PCA. Goals were coded for features that either emerged from the data itself or existed in extant goal-setting literature. The main outcomes were (1) number of PrEP conversations PCAs self-report during booster conversations two weeks following the intervention and (2) number of peers PCAs recruit into the study that completed the PrEP workshop. Results: PCA goals (N=68) were characterized in terms of four features: Specificity, target population, personalization, and purpose defined. To date, PCAs report a collective 52 PrEP conversations. 56, 25, and 6% of PrEP conversations occurred with friends, family, and sexual partners, respectively. PCAs with specific goals had more PrEP conversations with at-risk peers compared to those with vague goals (58% vs. 42%); PCAs with personalized goals had more PrEP conversations compared to those with de-personalized goals (60% vs. 53%); and PCAs with goals that defined a purpose had more PrEP conversations compared to those who did not define a purpose (75% vs. 52%). 100% of PCAs with goals that defined a purpose recruited peers into the study compared to 45 percent of PCAs with goals that did not define a purpose. Conclusion: Our preliminary analysis demonstrates that BMSM are motivated to set and work toward a diverse set of goals to support peers in PrEP adoption. PCAs with goals involving a clearly defined purpose had more PrEP conversations and greater peer recruitment than those with goals lacking a defined purpose. This may indicate that PCAs who define their purpose at the outset of their participation will be more engaged in the study than those who do not. Goal-setting may be considered as a component of future HIV prevention interventions to advance intervention goals and as an indicator of PCAs understanding of the intervention.

Keywords: HIV prevention, MSM, peer change agent, preexposure prophylaxis

Procedia PDF Downloads 181

Authors: Gokulan Vethanayakam, Daniel Aston

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Introduction: The ReSPECT process supports healthcare professionals when making patient-centered decisions in the event of an emergency. It has been widely adopted by the NHS in England and allows patients to express thoughts and wishes about treatments and outcomes that they consider acceptable. It includes (but is not limited to) cardiopulmonary resuscitation decisions. ReSPECT conversations should ideally occur prior to ICU admission and should be documented in the eight sections of the nationally-standardised ReSPECT form. This audit evaluated the use of ReSPECT on a busy cardiothoracic ICU in an NHS Trust where established policies advocating its use exist. Methods: This audit was a retrospective review of ReSPECT forms for a sample of high-risk patients admitted to ICU at the Royal Papworth Hospital between January 2021 and March 2022. Patients all received one of the following interventions: Veno-Venous Extra-Corporeal Membrane Oxygenation (VV-ECMO) for severe respiratory failure (retrieved via the national ECMO service); cardiac or pulmonary transplantation-related surgical procedures (including organ transplants and Ventricular Assist Device (VAD) implantation); or elective non-transplant cardiac surgery. The quality of documentation on ReSPECT forms was evaluated using national standards and a graded ranking tool devised by the authors which was used to assess narrative aspects of the forms. Quality was ranked as A (excellent) to D (poor). Results: Of 230 patients (74 VV-ECMO, 104 transplant, 52 elective non-transplant surgery), 43 (18.7%) had a ReSPECT form and only one (0.43%) patient had a ReSPECT form completed prior to ICU admission. Of the 43 forms completed, 38 (88.4%) were completed due to the commencement of End of Life (EoL) care. No non-transplant surgical patients included in the audit had a ReSPECT form. There was documentation of balance of care (section 4a), CPR status (section 4c), capacity assessment (section 5), and patient involvement in completing the form (section 6a) on all 43 forms. Of the 34 patients assessed as lacking capacity to make decisions, only 22 (64.7%) had reasons documented. Other sections were variably completed; 29 (67.4%) forms had relevant background information included to a good standard (section 2a). Clinical guidance for the patient (section 4b) was given in 25 (58.1%), of which 11 stated the rationale that underpinned it. Seven forms (16.3%) contained information in an inappropriate section. In a comparison of ReSPECT forms completed ahead of an EoL trigger with those completed when EoL care began, there was a higher number of entries in section 3 (considering patient’s values/fears) that were assessed at grades A-B in the former group (p = 0.014), suggesting higher quality. Similarly, forms from the transplant group contained higher quality information in section 3 than those from the VV-ECMO group (p = 0.0005). Conclusions: Utilisation of the ReSPECT process in high-risk patients is yet to be well-adopted in this trust. Teams who meet patients before hospital admission for transplant or high-risk surgery should be encouraged to engage with the ReSPECT process at this point in the patient's journey. VV-ECMO retrieval teams should consider ReSPECT conversations with patients’ relatives at the time of retrieval.

Keywords: audit, critical care, end of life, ICU, ReSPECT, resuscitation

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Authors: Obasanjo Afolabi Bolarinwa

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This study assesses the levels of knowledge of sexually transmitted infections among unmarried youths in Nigeria; examines the pattern of high risk sex among unmarried youths in Nigeria; investigate the socio-demographic factors (age, place of residence, religion, level of education, wealth index and employment status) affecting the practice of high-risk sexual behaviour and ascertain the relationships between knowledge of sexually transmitted infections and practice of high risk sex. The goal of the study is to identify the factors associated with the practice of high risk sex among youth. These were with a view to identifying critical actions needed to reduce high risk sexual behaviour among youths. The study employed secondary data. The data for the study were extracted from the 2013 Nigeria Demographic and Health Survey (NDHS). The 2013 NDHS collected information from 38,948 Women ages 15-49 years and 17,359 men ages 15-49. A total of 7,744 female and 6,027 male respondents were utilized in the study. In order to adjust for the effect of oversampling of the population, the weighting factor provided by Measure DHS was applied. The data were analysed using frequency distribution and logistic regression. The results show that both male (92.2%) and female (93.6%) have accurate knowledge of sexually transmitted infections. The study also revealed that prevalence of high risk sexual behavior is high among Nigerian youths; this is evident as 77.7% (female) and 78.4% (male) are engaging in high risk sexual behavior. The bivariate analysis shows that age of respondent (χ2=294.2; p < 0.05), religion (χ2=136.64; p < 0.05), wealth index (χ2=17.38; p < 0.05), level of education (χ2=34.73; p < 0.05) and employment status (χ2=94.54; p < 0.05) were individual factors significantly associated with high risk sexual behaviour among male while age of respondent (χ2=327.07; p < 0.05), place of residence (χ2=6.71; p < 0.05), religion (χ2=81.04; p < 0.05), wealth index (χ2=7.41; p < 0.05), level of education (χ2=18.12; p < 0.05) and employment status (χ2=51.02; p < 0.05) were individual factors significantly associated with high risk sexual behaviour among female. Furthermore, the study shows that there is a relationship between knowledge of sexually transmitted infections and high risk sex among male (χ2=38.32; p < 0.05) and female (χ2=18.37; p < 0.05). At multivariate level, the study revealed that individual characteristics such as age, religion, place of residence, wealth index, levels of education and employment status were statistically significantly related with high risk sexual behaviour among male and female (p < 0.05). Lastly, the study shows that knowledge of sexually transmitted infection was significantly related to high risk sexual behaviour among youths (p < 0.05). The study concludes that there is a high level of knowledge of sexually transmitted infections among unmarried youths in Nigeria. The practice of high risk sex is high among unmarried youths but higher among male youths. The prevalence of high risk sexual activity is higher for males when they are at disadvantage and higher for females when they are at advantage. Socio-demographic factors like age of respondents, religion, wealth index, place of residence, employment status and highest level of education are factors influencing high risk sexual behaviour among youths.

Keywords: high risk sex, wealth index, sexual behaviour, knowledge

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Authors: Aakriti Chetan Shah, Elle Sein

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Background/purpose: Group B Streptococcus(GBS) is the leading cause of severe early onset infection in newborns, with the incidence of Early Onset Group B Streptococcus (EOGBS) in the UK and Ireland rising from 0.48 to 0.57 per 1000 births from 2000 to 2015. A WHO study conducted in 2017, has shown that 38.5% of cases can result in stillbirth and infant deaths. This is an important problem to consider as 20% of women worldwide have GBS colonisation and can suffer from these detrimental effects. Current Royal College of Obstetricians and Midwives (RCOG) guidelines do not recommend bacteriological screening for pregnant women due to its low sensitivity in antenatal screening correlating with the neonate having GBS but advise a patient information leaflet be given to pregnant women. However, a Healthcare Safety Investigation Branch (HSIB) 2019 learning report found that only 50% of trusts and health boards reported giving GBS information leaflets to all pregnant mothers. Therefore, this audit aimed to assess current practices of information dissemination about GBS at Chelsea & Westminster (C&W) Hospital. Methodology: A quantitative cross-sectional study was carried out using a questionnaire based on the RCOG GBS guidelines and the HSIB Learning report. The study was conducted in antenatal clinics at Chelsea & Westminster Hospital, from 29th January 2021 to 14th February 2021, with twenty-two practicing obstetricians and midwives participating in the survey. The main outcome measure was the proportion of obstetricians and midwives who disseminate information about GBS to pregnant women, and the reasons behind why they do or do not. Results: 22 obstetricians and midwives responded with 18 complete responses. Of which 12 were obstetricians and 6 were midwives. Only 17% of clinical staff routinely inform all pregnant women about GBS, and do so at varying timeframes of the pregnancy, with an equal split in the first, second and third trimester. The primary reason for not informing women about GBS was influenced by three key factors: Deemed relevant only for patients at high risk of GBS, lack of time in clinic appointments and no routine NHS screening available. Interestingly 58% of staff in the antenatal clinic believe it is necessary to inform all women about GBS and its importance. Conclusion: It is vital for obstetricians and midwives to inform all pregnant women about GBS due to the high prevalence of incidental carriers in the population, and the harmful effects it can cause for neonates. Even though most clinicians believe it is important to inform all pregnant women about GBS, most do not. To ensure that RCOG and HSIB recommendations are followed, we recommend that women should be given this information at 28 weeks gestation in the antenatal clinic. Proposed implementations include an information leaflet to be incorporated into the Mum and Baby app, an informative video and end-to-end digital clinic documentation to include this information sharing prompt.

Keywords: group B Streptococcus, early onset sepsis, Antenatal care, Neonatal morbidity, GBS

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Authors: Syed Asif Hassan, Tabrej Khan

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Tuberculosis (TB) caused by bacillus Mycobacterium tuberculosis (Mtb) continues to take a disturbing toll on human life and healthcare facility worldwide. The global burden of TB remains enormous. The alarming rise of multi-drug resistant strains of Mycobacterium tuberculosis calls for an increase in research efforts towards the development of new target specific therapeutics against diverse strains of M. tuberculosis. Therefore, the discovery of new molecular scaffolds targeting new drug sites should be a priority for a workable plan for fighting resistance in Mycobacterium tuberculosis (Mtb). Mtb non-acylated lipoprotein LprG (Rv1411c) has a Toll-like receptor 2 (TLR2) agonist actions that depend on its association with triacylated glycolipids binding specifically with the hydrophobic pocket of Mtb LprG lipoprotein. The detection of a glycolipid carrier function has important implications for the role of LprG in Mycobacterial physiology and virulence. Therefore, considering the pivotal role of glycolipids in mycobacterial physiology and host-pathogen interactions, designing competitive antagonist (chemotherapeutics) ligands that competitively bind to glycolipid binding domain in LprG lipoprotein, will lead to inhibition of tuberculosis infection in humans. In this study, a unified approach involving ligand-based virtual screening protocol USRCAT (Ultra Shape Recognition) software and molecular docking studies using Auto Dock Vina 1.1.2 using the X-ray crystal structure of Mtb LprG protein was implemented. The docking results were further confirmed by DSX (DrugScore eXtented), a robust program to evaluate the binding energy of ligands bound to the Ligand binding domain of the Mtb LprG lipoprotein. The ligand, which has the higher hypothetical affinity, also has greater negative value. Based on the USRCAT, Lipinski’s values and molecular docking results, [(2R)-2,3-di(hexadecanoyl oxy)propyl][(2S,3S,5S,6R)-3,4,5-trihydroxy-2,6-bis[[(2R,3S,4S,5R,6S)-3,4,5-trihydroxy-6 (hydroxymethyl)tetrahydropyran-2-yl]oxy]cyclohexyl] phosphate (XPX) was confirmed as a promising drug-like lead compound (antagonist) binding specifically to the hydrophobic domain of LprG protein with affinity greater than that of PIM2 (agonist of LprG protein) with a free binding energy of -9.98e+006 Kcal/mol and binding affinity of -132 Kcal/mol, respectively. A further, in vitro assay of this compound is required to establish its potency in inhibiting molecular evasion mechanism of MTB within the infected host macrophages. These results will certainly be helpful in future anti-TB drug discovery efforts against Multidrug-Resistance Tuberculosis (MDR-TB).

Keywords: antagonist, agonist, binding affinity, chemotherapeutics, drug-like, multi drug resistance tuberculosis (MDR-TB), RV1411c protein, toll-like receptor (TLR2)

Procedia PDF Downloads 256

Authors: Shilpee Jain, Sachin Latiyan, Kaushik Suneet

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Unlike traditional robots, medical microbots are not only smaller in size, but they also possess various unique properties, for example, biocompatibility, stability in the biological fluids, navigation opposite to the bloodstream, wireless control over locomotion, etc. The idea behind their usage in the medical field was to build a minimally invasive method for addressing the post-operative complications, including longer recovery time, infection eruption and pain. Herein, the present study demonstrates the fabrication of dual nature magneto-conducting Fe3O4 magnetic nanoparticles (MNPs) and SU8 derived carbon-based Janus microbots for the efficient intracellular delivery of biomolecules. The low aspect ratio with feature size 2-5 μm microbots were fabricated by using a photolithography technique. These microbots were pyrolyzed at 900°C, which converts SU8 into amorphous carbon. The pyrolyzed microbots have dual properties, i.e., the half part is magneto-conducting and another half is only conducting for sufficing the therapeutic payloads efficiently with the application of external electric/magnetic field stimulations. For the efficient intracellular delivery of the microbots, the size and aspect ratio plays a significant role. However, on a smaller scale, the proper control over movement is difficult to achieve. The dual nature of Janus microbots allowed to control its maneuverability in the complex fluids using external electric as well as the magnetic field. Interestingly, Janus microbots move faster with the application of an external electric field (44 µm/s) as compared to the magnetic field (18 µm/s) application. Furthermore, these Janus microbots exhibit auto-fluorescence behavior that will help to track their pathway during navigation. Typically, the use of MNPs in the microdevices enhances the tendency to agglomerate. However, the incorporation of Fe₃O₄ MNPs in the pyrolyzed carbon reduces the chances of agglomeration of the microbots. The biocompatibility of the medical microbots, which is the essential property of any biosystems, was determined in vitro using HeLa cells. The microbots were found to compatible with HeLa cells. Additionally, the intracellular uptake of microbots was higher in the presence of an external electric field as compared to without electric field stimulation. In summary, the cytocompatible Janus microbots were fabricated successfully. They are stable in the biological fluids, wireless controllable navigation with the help of a few Guess external magnetic fields, their movement can be tracked because of autofluorescence behavior, they are less susceptible to agglomeration and higher cellular uptake could be achieved with the application of the external electric field. Thus, these carriers could offer a versatile platform to suffice the therapeutic payloads under wireless actuation.

Keywords: amorphous carbon, electric/magnetic stimulations, Janus microbots, magnetic nanoparticles, minimally invasive procedures

Procedia PDF Downloads 107

Authors: Mumuni Sumaila, Viness Pillay, Yahya E. Choonara, Pradeep Kumar, Pierre P. Kondiah

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Tuberculosis (TB) remains a serious challenge to public health globally, despite every effort put together to curb the disease. Current TB therapeutics available have proven to be inefficient due to a multitude of drawbacks that range from serious adverse effects/drug toxicity to inconsistent bioavailability, which ultimately contributes to the emergence of drug-resistant TB. An effective ‘cargo’ system designed to cleverly deliver therapeutic doses of anti-TB drugs to infection sites and in a sustained-release manner may provide a better therapeutic choice towards winning the war against TB. In the current study, we investigated mannose-functionalized lipopolysaccharide hybrid nanoparticles for safety and efficacy towards macrophage-targeted simultaneous delivery of the two first-line anti-TB drugs, rifampicin (RF) and isoniazid (IS). RF-IS-loaded lipopolysaccharide hybrid nanoparticles were fabricated using the solvent injection technique (SIT), incorporating soy lecithin (SL) and low molecular weight chitosan (CS) as the lipid and polysaccharide components, respectively. Surface-functionalized nanoparticles were obtained through the reaction of the aldehyde group of mannose with free amine functionality present at the surface of the nanoparticles. The functionalized nanocarriers were spherical with average particle size and surface charge of 107.83 nm and +21.77 mV, respectively, and entrapment efficiencies (EE) were 53.52% and 69.80% for RF and IS, respectively. FTIR spectrum revealed high-intensity bands between 1663 cm⁻¹ and 1408 cm⁻¹ wavenumbers (absent in non-functionalized nanoparticles), which could be attributed to the C=N stretching vibration produced by the formation of Schiff’s base (–N=CH–) during the mannosylation reaction. In vitro release studies showed a sustained-release profile for RF and IS, with less than half of the total payload released over a 48-hour period. The nanocarriers were biocompatible and safe, with more than 80% cell viability achieved when incubated with RAW 264.7 cells at concentrations 30 to 500 μg/mL over a 24-hour period. Cellular uptake studies (after a 24-hour incubation period with the murine macrophage cells, RAW 264.7) revealed a 13- and a 9-fold increase in intracellular accumulation of RF and IS, respectively, when compared with the unformulated RF+IS solution. A 6- and a 3-fold increase in intracellular accumulation of RF and IS, respectively, were observed when compared with the non-functionalized nanoparticles. Furthermore, fluorescent microscopy images showed nanoparticle internalization and accumulation within the RAW 264.7 cells, which was more significant in the mannose-functionalized system compared to the non-functionalized nanoparticles. The overall results suggested that the fabricated mannose-functionalized lipopolysaccharide nanoparticles are a safe and promising platform for macrophage-targeted delivery of anti-TB therapeutics. However, in vivo pharmacokinetic/pharmacodynamics studies are required to further substantiate the therapeutic efficacy of the nanosystem.

Keywords: anti-tuberculosis therapeutics, hybrid nanosystem, lipopolysaccharide nanoparticles, macrophage-targeted delivery

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Authors: Heiko Jessen, Laura Tanus, Slobodan Ruzicic

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Objectives: The efficacy of Stribild®, an integrase strand transfer inhibitor (INSTI) -based STR, has been evaluated in randomized clinical trials and it has demonstrated durable capability in terms of achieving sustained suppression of HIV-1 RNA-levels. However, differences in monitoring frequency, existing selection bias and profile of patients enrolled in the trials, may all result in divergent efficacy of this regimen in routine clinical settings. The aim of this study was to assess the virologic outcomes, safety and tolerability profile of Stribild® in a routine clinical setting. Methods: This was a retrospective monocentric analysis on HIV-1-infected patients, who started with or were switched to Stribild®. Virological failure (VF) was defined as confirmed HIV-RNA>50 copies/ml. The minimum time of follow-up was 24 weeks. The percentage of patients remaining free of therapeutic failure was estimated using the time-to-loss-of-virologic-response (TLOVR) algorithm, by intent-to-treat analysis. Results: We analyzed the data of 197 patients (56 ART-naïve and 141 treatment-experienced patients), who fulfilled the inclusion criteria. Majority (95.9%) of patients were male. The median time of HIV-infection at baseline was 2 months in treatment-naïve and 70 months in treatment-experienced patients. Median time [IQR] under ART in treatment-experienced patients was 37 months. Among the treatment-experienced patients 27.0% had already been treated with a regimen consisting of two NRTIs and one INSTI, whereas 18.4% of them experienced a VF. The median time [IQR] of virological suppression prior to therapy with Stribild® in the treatment-experienced patients was 10 months [0-27]. At the end of follow-up (median 33 months), 87.3% (95% CI, 83.5-91.2) of treatment-naïve and 80.3% (95% CI, 75.8-84.8) of treatment-experienced patients remained free of therapeutic failure. Considering only treatment-experienced patients with baseline VL<50 copies/ml, 83.0% (95% CI, 78.5-87.5) remained free of therapeutic failure. A total of 17 patients stopped treatment with Stribild®, 5.4% (3/56) of them were treatment-naïve and 9.9% (14/141) were treatment-experienced patients. The Stribild® therapy was discontinued in 2 (1.0%) because of VF, loss to follow-up in 4 (2.0%), and drug-drug interactions in 2 (1.0%) patients. Adverse events were in 7 (3.6%) patients the reason to switch from therapy with Stribild® and further 2 (1.0%) patients decided personally to switch. The most frequently observed adverse events were gastrointestinal side effects (20.0%), headache (8%), rash events (7%) and dizziness (6%). In two patients we observed an emergence of novel resistances in integrase-gene. The N155H evolved in one patient and resulted in VF. In another patient S119R evolved either during or shortly upon switch from therapy with Stribild®. In one further patient with VF two novel mutations in the RT-gene were observed when compared to historical genotypic test result (V106I/M and M184V), whereby it is not clear whether they evolved during or already before the switch to Stribild®. Conclusions: Effectiveness of Stribild® for treatment-naïve patients was consistent with data obtained in clinical trials. The safety and tolerability profile as well as resistance development confirmed clinical efficacy of Stribild® in a daily practice setting.

Keywords: ART, HIV, integrase inhibitor, stribild

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Authors: Elena Ionescu, Elena Iacob, Marie-Louise Ionescu, Carmen Elena Tebrencu, Oana Teodora Ciuperca

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Echinacea species (Asteraceae) has received a global attention because it is widely used for treatment of cold, flu and upper respiratory tract infections. Echinacea species contain a great variety of chemical components that contribute to their activity. The most important components responsible for the biological activity are those with high molecular-weight such as polysaccharides, polyacetylenes, highly unsaturated alkamides and caffeic acid derivatives. The principal factors that may influence the chemical composition of Echinacea include the species and the part of plant used (aerial parts or roots ). In recent years the market for Echinacea has grown rapidly and also the cases of adultery/replacement especially for Echinacea root. The identification of presence or absence of same biomarkers provide information for safe use of Echinacea species in food supplements industry. The aim of the study was the preliminary evaluation and fingerprinting by UV-VISIBLE spectroscopy of biomarkers in terms of content in phenolic derivatives of some Echinacea species (E. purpurea, E. angustifolia and E. pallida) for identification and authentication of the species. The steps of the study were: (1) samples (extracts) preparation from Echinacea species (non-hydrolyzed and hydrolyzed ethanol extracts); (2) samples preparation of reference substances (polyphenol acids: caftaric acid, caffeic acid, chlorogenic acid, ferulic acid; flavonoids: rutoside, hyperoside, isoquercitrin and their aglycones: quercitri, quercetol, luteolin, kaempferol and apigenin); (3) identification of specific absorption at wavelengths between 700-200 nm; (4) identify the phenolic compounds from Echinacea species based on spectral characteristics and the specific absorption; each class of compounds corresponds to a maximum absorption in the UV spectrum. The phytochemical compounds were identified at specific wavelengths between 700-200 nm. The absorption intensities were measured. The obtained results proved that ethanolic extract showed absorption peaks attributed to: phenolic compounds (free phenolic acids and phenolic acids derivatives) registrated between 220-280 nm, unsymmetrical chemical structure compounds (caffeic acid, chlorogenic acid, ferulic acid) with maximum absorption peak and absorption "shoulder" that may be due to substitution of hydroxyl or methoxy group, flavonoid compounds (in free form or glycosides) between 330-360 nm, due to the double bond in position 2,3 and carbonyl group in position 4 flavonols. UV spectra showed two major peaks of absorption (quercetin glycoside, rutin, etc.). The results obtained by UV-VIS spectroscopy has revealed the presence of phenolic derivatives such as cicoric acid (240 nm), caftaric acid (329 nm), caffeic acid (240 nm), rutoside (205 nm), quercetin (255 nm), luteolin (235 nm) in all three species of Echinacea. The echinacoside is absent. This profile mentioned above and the absence of phenolic compound echinacoside leads to the conclusion that species harvested as Echinacea angustifolia and Echinacea pallida are Echinacea purpurea also; It can be said that preliminary fingerprinting of Echinacea species through correspondence with the phenolic derivatives profile can be achieved by UV-VIS spectroscopic investigation, which is an adequate technique for preliminary identification and authentication of Echinacea in medicinal herbs.

Keywords: Echinacea species, Fingerprinting, Phenolic compounds, UV-VIS spectroscopy

Procedia PDF Downloads 239

Authors: Sushmita Roy Chowdhury, Steve Holding, Sujoy Khan

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The immunogenic responses of the lung towards the fungus Aspergillus fumigatus may range from invasive aspergillosis in the immunocompromised, fungal ball or infection within a cavity in the lung in those with structural lung lesions, or allergic bronchopulmonary aspergillosis (ABPA). Patients with asthma or cystic fibrosis are particularly predisposed to ABPA. There are consensus guidelines that have established criteria for diagnosis of ABPA, but uncertainty remains on the serologic cut-off values that would increase the diagnostic specificity of ABPA. We retrospectively analyzed 80 patients with severe asthma and evidence of peripheral blood eosinophilia ( > 500) over the last 3 years who underwent all serologic tests to exclude ABPA. Total IgE, specific IgE and specific IgG levels against Aspergillus fumigatus were measured using ImmunoCAP Phadia-100 (Thermo Fisher Scientific, Sweden). The Modified ISHAM working group 2013 criteria (obligate criteria: asthma or cystic fibrosis, total IgE > 1000 IU/ml or > 417 kU/L and positive specific IgE Aspergillus fumigatus or skin test positivity; with ≥ 2 of peripheral eosinophilia, positive specific IgG Aspergillus fumigatus and consistent radiographic opacities) was used in the clinical workup for the final diagnosis of ABPA. Patients were divided into 3 groups - definite, possible, and no evidence of ABPA. Specific IgG Aspergillus fumigatus levels were not used to assign the patients into any of the groups. Of 80 patients (males 48, females 32; mean age 53.9 years ± SD 15.8) selected for the analysis, there were 30 patients who had positive specific IgE against Aspergillus fumigatus (37.5%). 13 patients fulfilled the Modified ISHAM working group 2013 criteria of ABPA (‘definite’), while 15 patients were ‘possible’ ABPA and 52 did not fulfill the criteria (not ABPA). As IgE levels were not normally distributed, median levels were used in the analysis. Median total IgE levels of patients with definite and possible ABPA were 2144 kU/L and 2597 kU/L respectively (non-significant), while median specific IgE Aspergillus fumigatus at 4.35 kUA/L and 1.47 kUA/L respectively were significantly different (comparison of standard deviations F-statistic 3.2267, significance level p=0.040). Mean levels of IgG anti-Aspergillus fumigatus in the three groups (definite, possible and no evidence of ABPA) were compared using ANOVA (Statgraphics Centurion Professional XV, Statpoint Inc). Mean levels of IgG anti-Aspergillus fumigatus (Gm3) in definite ABPA was 125.17 mgA/L ( ± SD 54.84, with 95%CI 92.03-158.32), while mean Gm3 levels in possible and no ABPA were 18.61 mgA/L and 30.05 mgA/L respectively. ANOVA showed a significant difference between the definite group and the other groups (p < 0.001). This was confirmed using multiple range tests (Fisher's least significant difference procedure). There was no significant difference between the possible ABPA and not ABPA groups (p > 0.05). The study showed that a sizeable proportion of patients with asthma are sensitized to Aspergillus fumigatus in this part of India. A higher cut-off value of Gm3 ≥ 80 mgA/L provides a higher serologic specificity towards definite ABPA. Long-term studies would provide us more information if those patients with 'possible' APBA and positive Gm3 later develop clear ABPA, and are different from the Gm3 negative group in this respect. Serologic testing with clear defined cut-offs are a valuable adjunct in the diagnosis of ABPA.

Keywords: allergic bronchopulmonary aspergillosis, Aspergillus fumigatus, asthma, IgE level

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Authors: Francis Gyapong, Denis Yar

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The use of cell phones has become an indispensable tool in the hospital's settings. Cell phones are used in hospitals without restrictions regardless of their unknown microbial load. However, the indiscriminate use of mobile devices, especially at health facilities, can act as a vehicle for transmitting pathogenic bacteria and other microorganisms. These potential pathogens become exogenous sources of infection for the patients and are also a potential health hazard for self and as well as family members. These are a growing problem in many health care institutions. Innovations in mobile communication have led to better patient care in diabetes, asthma, and increased in vaccine uptake via SMS. Notwithstanding, the use of cell phones can be a great potential source for nosocomial infections. Many studies reported heavy microbial contamination of cell phones among healthcare workers and communities. However, limited studies have been reported in our region on bacterial contamination on cell phones among healthcare workers. This study assessed microbial contamination of cell phones of health care workers (HCWs) at the Mampong Municipal Government Hospital (MMGH), Ghana. A cross-sectional design was used to characterize bacterial microflora on cell phones of HCWs at the MMGH. A total of thirty-five (35) swab samples of cell phones of HCWs at the Laboratory, Dental Unit, Children’s Ward, Theater and Male ward were randomly collected for laboratory examinations. A suspension of the swab samples was each streak on blood and MacConkey agar and incubated at 37℃ for 48 hours. Bacterial isolates were identified using appropriate laboratory and biochemical tests. Kirby-Bauer disc diffusion method was used to determine the antimicrobial sensitivity tests of the isolates. Data analysis was performed using SPSS version 16. All mobile phones sampled were contaminated with one or more bacterial isolates. Cell phones from the Male ward, Dental Unit, Laboratory, Theatre and Children’s ward had at least three different bacterial isolates; 85.7%, 71.4%, 57.1% and 28.6% for both Theater and Children’s ward respectively. Bacterial contaminants identified were Staphylococcus epidermidis (37%), Staphylococcus aureus (26%), E. coli (20%), Bacillus spp. (11%) and Klebsiella spp. (6 %). Except for the Children ward, E. coli was isolated at all study sites and predominant (42.9%) at the Dental Unit while Klebsiella spp. (28.6%) was only isolated at the Children’s ward. Antibiotic sensitivity testing of Staphylococcus aureus indicated that they were highly sensitive to cephalexin (89%) tetracycline (80%), gentamycin (75%), lincomycin (70%), ciprofloxacin (67%) and highly resistant to ampicillin (75%). Some of these bacteria isolated are potential pathogens and their presence on cell phones of HCWs could be transmitted to patients and their families. Hence strict hand washing before and after every contact with patient and phone be enforced to reduce the risk of nosocomial infections.

Keywords: mobile phones, bacterial contamination, patients, MMGH

Procedia PDF Downloads 82

Authors: Aboli Gore, Aritra Das, Sunil Sonthalia, Tanmay Mahapatra, Sridhar Srikantiah, Hemant Shah

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AMANAT is an initiative, taken in collaboration with the Government of Bihar, aimed at improving the Quality of Maternal and Neonatal care services at Bihar’s public health facilities – those offering either the Basic Emergency Obstetric and Neonatal care (BEmONC) or Comprehensive Emergency Obstetric and Neonatal care (CEmONC) services. The effectiveness of this program is evaluated by conducting cross-sectional assessments at the concerned facilities prior to (baseline) and following completion (endline) of intervention. Direct Observation of Delivery (DOD) methodology is employed for carrying out the baseline and endline assessments – through which key obstetric and neonatal care practices among the Health Care Providers (especially the nurses) are assessed quantitatively by specially trained nursing professionals. Assessment of vitals prior to delivery improved during all three phases of BEmONC and all four phases of CEmONC training with statistically significant improvement noted in: i) pulse measurement in BEmONC phase 2 (9% to 68%), 3 (4% to 57%) & 4 (14% to 59%) and CEmONC phase 2 (7% to 72%) and 3 (0% to 64%); ii) blood pressure measurement in BEmONC phase 2 (27% to 84%), 3 (21% to 76%) & 4 (36% to 71%) and CEmONC phase 2 (23% to 76%) and 3 (2% to 70%); iii) fetal heart rate measurement in BEmONC phase 2 (10% to 72%), 3 (11% to 77%) & 4 (13% to 64%) and CEmONC phase 1 (24% to 38%), 2 (14% to 82%) and 3 (1% to 73%); and iv) abdominal examination in BEmONC phase 2 (14% to 59%), 3 (3% to 59%) & 4 (6% to 56%) and CEmONC phase 1 (0% to 24%), 2 (7% to 62%) & 3 (0% to 62%). Regarding infection control, wearing of apron, mask and cap by the delivery conductors improved significantly in all BEmONC phases. Similarly, the practice of handwashing improved in all BEmONC and CEmONC phases. Even on disaggregation, the handwashing showed significant improvement in all phases but CEmONC phase-4. Not only the positive practices related to handwashing improved but also negative practices such as turning off the tap with bare hands declined significantly in the aforementioned phases. Significant decline was also noted in negative maternal care practices such as application of fundal pressure for hastening the delivery process and administration of oxytocin prior to delivery. One of the notable achievement of AMANAT is an improvement in active management of the third stage of labor (AMTSL). The overall AMTSL (including administration of oxytocin or other uterotonics uterotonic in proper dose, route and time along with controlled cord traction and uterine massage) improved in all phases of BEmONC and CEmONC mentoring. Another key area of improvement, across phases, was in proper cutting/clamping of the umbilical cord. AMANAT mentoring also led to improvement in important immediate newborn care practices such as initiation of skin-to-skin care and timely initiation of breastfeeding. The next phase of the mentoring program seeks to institutionalize mentoring across the state that could potentially perpetuate improvement with minimal external intervention.

Keywords: capacity building, nurse-mentoring, quality of care, pregnancy, newborn care

Procedia PDF Downloads 139

Authors: W. S. Thulasitha, N. Umasuthan, G. I. Godahewa, Jehee Lee

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In fish, innate immune defense is the first immune response against microbial pathogens which consists of several antimicrobial components. Galectins are one of the carbohydrate binding lectins that have the ability to identify pathogen by recognition of pathogen associated molecular patterns. Galectins play a vital role in the regulation of innate and adaptive immune responses. Rock bream Oplegnathus fasciatus is one of the most important cultured species in Korea and Japan. Considering the losses due to microbial pathogens, present study was carried out to understand the molecular and functional characteristics of a galectin in normal and pathogenic conditions, which could help to establish an understanding about immunological components of rock bream. Complete cDNA of rock bream galectin like protein B (rbGal like B) was identified from the cDNA library, and the in silico analysis was carried out using bioinformatic tools. Genomic structure was derived from the BAC library by sequencing a specific clone and using Spidey. Full length of rbGal like B (contig14775) cDNA containing 517 nucleotides was identified from the cDNA library which comprised of 435 bp in the open reading frame encoding a deduced protein composed of 145 amino acids. The molecular mass of putative protein was predicted as 16.14 kDa with an isoelectric point of 8.55. A characteristic conserved galactose binding domain was located from 12 to 145 amino acids. Genomic structure of rbGal like B consisted of 4 exons and 3 introns. Moreover, pairwise alignment showed that rock bream rbGal like B shares highest similarity (95.9 %) and identity (91 %) with Takifugu rubripes galectin related protein B like and lowest similarity (55.5 %) and identity (32.4 %) with Homo sapiens. Multiple sequence alignment demonstrated that the galectin related protein B was conserved among vertebrates. A phylogenetic analysis revealed that rbGal like B protein clustered together with other fish homologs in fish clade. It showed closer evolutionary link with Takifugu rubripes. Tissue distribution and expression patterns of rbGal like B upon immune challenges were performed using qRT-PCR assays. Among all tested tissues, level of rbGal like B expression was significantly high in gill tissue followed by kidney, intestine, heart and spleen. Upon immune challenges, it showed an up-regulated pattern of expression with Edwardsiella tarda, rock bream irido virus and poly I:C up to 6 h post injection and up to 24 h with LPS. However, In the presence of Streptococcus iniae rbGal like B showed an up and down pattern of expression with the peak at 6 - 12 h. Results from the present study revealed the phylogenetic position and role of rbGal like B in response to microbial infection in rock bream.

Keywords: galectin like protein B, immune response, Oplegnathus fasciatus, molecular characterization

Procedia PDF Downloads 339

Authors: Drago Bratkovic, Curtis Gravance, David Ketteridge, Ravi Krishnan, Michael Imperiale

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The mucopolysaccharidoses (MPSs) are a group of lysosomal storage diseases that have a common defect in the catabolism of glycosaminoglycans (GAGs). MPS I is the most common of the MPS diseases. Manifestations of MPS I include coarsening of facial features, corneal clouding, developmental delay, short stature, skeletal manifestations, hearing loss, cardiac valve disease, hepatosplenomegaly, and umbilical and inguinal hernias. Treatments for MPS I restore or activate the missing or deficient enzyme in the case of enzyme replacement therapy (ERT) and haematopoietic stem cell transplantation (HSCT). Pentosan polysulfate sodium (PPS) is a potential treatment to improve bone and joint manifestations of MPS I. The mechanisms of action of PPS that are relevant to the treatment of MPS I are the ability to: (i) Reduce systemic and accumulated GAG, (ii) Reduce inflammatory effects via the inhibition of NF-kB, resulting in the reduction in pro-inflammatory mediators. (iii) Reduce the expression of the pain mediator nerve growth factor in osteocytes from degenerating joints. (iv) Inhibit the cartilage degrading enzymes related to joint dysfunction in MPS I. PPS is being evaluated as an adjunctive therapy to ERT and/or HSCT in an open-label, single-centre, phase 2 study. Patients are ≥ 5 years of age with a diagnosis of MPS I and previously received HSCT and/or ERT. Three white, female, patients with MPS I-Hurler, ages 14, 15, and 19 years, and one, white male patient aged 15 years are enrolled. All were diagnosed at ≤2 years of age. All patients received HSCT ≤ 6 months after diagnosis. Two of the patients were treated with ERT prior to HSCT, and 1 patient received ERT commencing 3 months prior to HSCT. Two patients received 0.75mg/kg and 2 patients received 1.5mg/kg of PPS. PPS was well tolerated at doses of 0.75 and 1.5 mg/kg to 47 weeks of continuous dosing. Of the 19 adverse events (AEs), 2 were related to PPS. One AE was moderate (pre-syncope) and 1 was mild (injection site bruising), experienced in the same patient. All AEs were reported as mild or moderate. There have been no SAEs. One subject experienced a COVID-19 infection and PPS was interrupted. The MPS I signature GAG fragments, sulfated disaccharide and UA-HNAc S, tended to decrease in 3 patients from baseline through Week 25. Week 25 GAG data are pending for the 4th patient. Overall, most biomarkers (inflammatory, cartilage degeneration, and bone turnover) evaluated in the 3 patients with 25-week assessments have indicated either no change or a reduction in levels compared to baseline. In 3 patients, there was a trend toward improvement in the 2MWT from baseline to Week 48 with > 100% increase in 1 patient (01-201). In the 3 patients that had Week 48 assessments, patients and proxies reported improvement in PGIC, including “worthwhile difference” (n=1), or “made all the difference” (n=2).

Keywords: MPS I, pentosan polysulfate sodium, clinical study, 2MWT, QoL

Procedia PDF Downloads 98

Authors: I. C. Orabueze, A. A. Amudalat, S. A. Adesegun, A. A. Usman

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Dental and associated oral diseases are increasingly affecting a considerable portion of the population and are considered some of the major causes of tooth loss, discomfort, mouth odor and loss of confidence. This study focused on the ethnobotanical survey of medicinal plants used in oral therapy and evaluation of the antimicrobial activities of methanolic extracts of two selected plants from the survey for their efficacy against dental microorganisms. The ethnobotanical survey was carried out in six herbal markets in Lagos State, Nigeria by oral interviewing and information obtained from an old family manually complied herbal medication book. Methanolic extracts of Olax subscorpioidea (stem bark) and Bridelia ferruginea (stem bark) were assayed for their antimicrobial activities against clinical oral isolates (Aspergillus fumigatus, Candida albicans, Streptococcus spp, Staphylococcus aureus, Lactobacillus acidophilus and Pseudomonas aeruginosa). In vitro microbial technique (agar well diffusion method and minimum inhibitory concentration (MIC) assay) were employed for the assay. Chlorhexidine gluconate was used as the reference drug for comparison with the extract results. And the preliminary phytochemical screening of the constituents of the plants were done. The ethnobotanical survey produced plants (28) of diverse family. Different parts of plants (seed, fruit, leaf, root, bark) were mentioned but 60% mentioned were either the stem or the bark. O. subscorpioidea showed considerable antifungal activity with zone of inhibition ranging from 2.650 – 2.000 cm against Aspergillus fumigatus but no such encouraging inhibitory activity was observed in the other assayed organisms. B. ferruginea showed antibacterial sensitivity against Streptococcus spp, Staphylococcus aureus, Lactobacillus acidophilus and Pseudomonas aeruginosa with zone of inhibitions ranging from 3.400 - 2.500, 2.250 - 1.600, 2.700 - 1.950, 2.225 – 1.525 cm respectively. The minimum inhibitory concentration of O. subscorpioidea against Aspergillus fumigatus was 51.2 mg ml-1 while that of B. ferruginea against Streptococcus spp was 0.1mg ml-1 and for Staphylococcus aureus, Lactobacillus acidophilus and Pseudomonas aeruginosa were 25.6 mg ml-1. A phytochemical analysis reveals the presence of alkaloids, saponins, cardiac glycoside, tannins, phenols and terpenoids in both plants, with steroids only in B. ferruginea. No toxicity was observed among mice given the two methanolic extracts (1000 mg Kg-1) after 21 days. The barks of both plants exhibited antimicrobial properties against periodontal diseases causing organisms assayed, thus up-holding their folkloric use in oral disorder management. Further research could be done viewing these extracts as combination therapy, checking for possible synergistic value in toothpaste and oral rinse formulations for reducing oral bacterial flora and fungi load.

Keywords: antimicrobial activities, Bridelia ferruginea, dental disinfection, methanolic extract, Olax subscorpioidea, ethnobotanical survey

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Authors: Aurora Sheehy, Caitlin Prince

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Within current orthodox medical protocols, trauma and mental health issues are deemed to reside within the realm of cognitive or psychological therapists and are marginalised in these areas, in part due to limited drugs option available, mostly manipulating neurotransmitters or sedating patients to reduce symptoms. By contrast, this research presents examples from the clinical practice of how trauma can be assessed and treated physiologically. Adverse Childhood Experiences (ACEs) are a tally of different types of abuse and neglect. It has been used as a measurable and reliable predictor of the likelihood of the development of autoimmune disease. It is a direct way to demonstrate reliably the health impact of traumatic life experiences. A second assessment tool is Allostatic Load, which refers to the cumulative effects that chronic stress has on mental and physical health. It records the decline of an individual’s physiological capacity to cope with their experience. It uses a specific grouping of serum testing and physical measures. It includes an assessment of neuroendocrine, cardiovascular, immune and metabolic systems. Allostatic load demonstrates the health impact that trauma has throughout the body. It forms part of an initial intake assessment in clinical practice and could also be used in research to evaluate treatment. Examining medicinal plants for their physiological, neurological and somatic effects through the lens of Polyvagal theory offers new opportunities for trauma treatments. In situations where Polyvagal theory recommends activities and exercises to enable parasympathetic activation, many herbs that affect Effector Memory T (TEM) cells also enact these responses. Traditional or Indigenous European herbs show the potential to support the polyvagal tone, through multiple mechanisms. As the ventral vagal nerve reaches almost every major organ, plants that have actions on these tissues can be understood via their polyvagal actions, such as monoterpenes as agents to improve respiratory vagal tone, cyanogenic glycosides to reset polyvagal tone, volatile oils rich in phenyl methyl esters improve both sympathetic and parasympathetic tone, bitters activate gut function and can strongly promote parasympathetic regulation. Auricular Acupuncture uses a system of somatotopic mapping of the auricular surface overlaid with an image of an inverted foetus with each body organ and system featured. Given that the concha of the auricle is the only place on the body where the Vagus Nerve neurons reach the surface of the skin, several investigators have evaluated non-invasive, transcutaneous electrical nerve stimulation (TENS) at auricular points. Drawn from an interdisciplinary evidence base and developed through clinical practice, these assessment and treatment tools are examples of practitioners in the field innovating out of necessity for the best outcomes for patients. This paper draws on case studies to direct future research.

Keywords: polyvagal, auricular acupuncture, trauma, herbs

Procedia PDF Downloads 66

Authors: Tomasz Borowski, Dawid Sołoducha, Agata Markowska-Szczupak, Aneta Wesołowska, Marian Kordas, Rafał Rakoczy

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Essential oils (EOs) are fragrant volatile oils obtained from plants. These are used for cooking (for flavor and aroma), cleaning, beauty (e.g., rosemary essential oil is used to promote hair growth), health (e.g. thyme essential oil cures arthritis, normalizes blood pressure, reduces stress on the heart, cures chest infection and cough) and in the food industry as preservatives and antioxidants. Rosemary and thyme essential oils are considered the most eminent herbs based on their history and medicinal properties. They possess a wide range of activity against different types of bacteria and fungi compared with the other oils in both in vitro and in vivo studies. However, traditional uses of EOs are limited due to rosemary and thyme oils in high concentrations can be toxic. In light of the accessible data, the following hypothesis was put forward: Low frequency rotating magnetic field (RMF) increases the antimicrobial potential of EOs. The aim of this work was to investigate the antimicrobial activity of commercial Salvia Rosmarinus L. and Thymus vulgaris L. essential oil from Polish company Avicenna-Oil under Rotating Magnetic Field (RMF) at f = 25 Hz. The self-constructed reactor (MAP) was applied for this study. The chemical composition of oils was determined by gas chromatography coupled with mass spectrometry (GC-MS). Model bacteria Escherichia coli K12 (ATCC 25922) was used. Minimum inhibitory concentrations (MIC) against E. coli were determined for the essential oils. Tested oils in very small concentrations were prepared (from 1 to 3 drops of essential oils per 3 mL working suspensions). From the results of disc diffusion assay and MIC tests, it can be concluded that thyme oil had the highest antibacterial activity against E. coli. Moreover, the study indicates the exposition to the RMF, as compared to the unexposed controls causing an increase in the efficacy of antibacterial properties of tested oils. The extended radiation exposure to RMF at the frequency f= 25 Hz beyond 160 minutes resulted in a significant increase in antibacterial potential against E. coli. Bacteria were killed within 40 minutes in thyme oil in lower tested concentration (1 drop of essential oils per 3 mL working suspension). Rapid decrease (>3 log) of bacteria number was observed with rosemary oil within 100 minutes (in concentration 3 drops of essential oils per 3 mL working suspension). Thus, a method for improving the antimicrobial performance of essential oil in low concentrations was developed. However, it still remains to be investigated how bacteria get killed by the EOs treated by an electromagnetic field. The possible mechanisms relies on alteration in the permeability of ionic channels in ionic channels in the bacterial cell walls that transport in the cells was proposed. For further studies, it is proposed to examine other types of essential oils and other antibiotic-resistant bacteria (ARB), which are causing a serious concern throughout the world.

Keywords: rotating magnetic field, rosemary, thyme, essential oils, Escherichia coli

Procedia PDF Downloads 144

Authors: Chee Wee Gan, Anthony Herr Cheun Ng, Yee Shan Wong, Subbu Venkatraman, Lynne Hsueh Yee Lim

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Otitis media with effusion (OME) is the leading cause of hearing loss in children worldwide. Surgery to insert grommet tube into the eardrum is usually indicated for OME unresponsive to antimicrobial therapy. It is the most common surgery for children. However, current commercially available grommet tubes are non-bioresorbable, not drug-treated, with unpredictable duration of retention on the eardrum to ventilate middle ear. Their functionality is impaired when clogged or chronically infected, requiring additional surgery to remove/reinsert grommet tubes. We envisaged that a novel fully bioresorbable grommet tube with sustained antibiotic release technology could address these drawbacks. In this study, drug-loaded bioresorbable poly(L-lactide-co-ε-caprolactone)(PLC) copolymer grommet tubes were fabricated by microinjection moulding technique. In vitro drug release and degradation model of PLC tubes were studied. Antibacterial property was evaluated by incubating PLC tubes with P. aeruginosa broth. Surface morphology was analyzed using scanning electron microscopy. A preliminary animal study was conducted using guinea pigs as an in vivo model to evaluate PLC tubes with and without drug, with commercial Mini Shah grommet tube as comparison. Our in vitro data showed sustained drug release over 3 months. All PLC tubes revealed exponential degradation profiles over time. Modeling predicted loss of tube functionality in water to be approximately 14 weeks and 17 weeks for PLC with and without drug, respectively. Generally, PLC tubes had less bacteria adherence, which were attributed to the much smoother tube surfaces compared to Mini Shah. Antibiotic from PLC tube further made bacteria adherence on surface negligible. They showed neither inflammation nor otorrhea after 18 weeks post-insertion in the eardrums of guinea pigs, but had demonstrated severe degree of bioresorption. Histology confirmed the new PLC tubes were biocompatible. Analyses on the PLC tubes in the eardrums showed bioresorption profiles close to our in vitro degradation models. The bioresorbable antibiotic-loaded grommet tubes showed good predictability in functionality. The smooth surface and sustained release technology reduced the risk of tube infection. Tube functional duration of 18 weeks allowed sufficient ventilation period to treat OME. Our ongoing studies include modifying the surface properties with protein coating, optimizing the drug dosage in the tubes to enhance their performances, evaluating their functional outcome on hearing after full resoption of grommet tube and healing of eardrums, and developing animal model with OME to further validate our in vitro models.

Keywords: bioresorbable polymer, drug release, grommet tube, guinea pigs, otitis media with effusion

Procedia PDF Downloads 437

Authors: Massimiliano Biagini, Marco Spinsanti, Gabriella De Angelis, Sara Tomei, Ilaria Ferlenghi, Maria Scarselli, Alessia Biolchi, Alessandro Muzzi, Brunella Brunelli, Silvana Savino, Marzia M. Giuliani, Isabel Delany, Paolo Costantino, Rino Rappuoli, Vega Masignani, Nathalie Norais

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The gram-negative bacterium Neisseria meningitidis serogroup B (MenB) is an exclusively human pathogen representing the major cause of meningitides and severe sepsis in infants and children but also in young adults. This pathogen is usually present in the 30% of healthy population that act as a reservoir, spreading it through saliva and respiratory fluids during coughing, sneezing, kissing. Among surface-exposed protein components of this diplococcus, factor H binding protein is a lipoprotein proved to be a protective antigen used as a component of the recently licensed Bexsero vaccine. fHbp is a highly variable meningococcal protein: to reflect its remarkable sequence variability, it has been classified in three variants (or two subfamilies), and with poor cross-protection among the different variants. Furthermore, the level of fHbp expression varies significantly among strains, and this has also been considered an important factor for predicting MenB strain susceptibility to anti-fHbp antisera. Different methods have been used to assess fHbp expression on meningococcal strains, however, all these methods use anti-fHbp antibodies, and for this reason, the results are affected by the different affinity that antibodies can have to different antigenic variants. To overcome the limitations of an antibody-based quantification, we developed a quantitative Mass Spectrometry (MS) approach. Selected Reaction Monitoring (SRM) recently emerged as a powerful MS tool for detecting and quantifying proteins in complex mixtures. SRM is based on the targeted detection of ProteoTypicPeptides (PTPs), which are unique signatures of a protein that can be easily detected and quantified by MS. This approach, proven to be highly sensitive, quantitatively accurate and highly reproducible, was used to quantify the absolute amount of fHbp antigen in total extracts derived from 105 clinical isolates, evenly distributed among the three main variant groups and selected to be representative of the fHbp circulating subvariants around the world. We extended the study at the genetic level investigating the correlation between the differential level of expression and polymorphisms present within the genes and their promoter sequences. The implications of fHbp expression on the susceptibility of the strain to killing by anti-fHbp antisera are also presented. To date this is the first comprehensive fHbp expression profiling in a large panel of Neisseria meningitidis clinical isolates driven by an antibody-independent MS-based methodology, opening the door to new applications in vaccine coverage prediction and reinforcing the molecular understanding of released vaccines.

Keywords: quantitative mass spectrometry, Neisseria meningitidis, vaccines, bexsero, molecular epidemiology

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Authors: Benjamin Osei Tutu, Sharon Annison

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On 21^st November, 2016, an outbreak of foodborne illness occurred after a buffet lunch served during a stakeholders’ consultation meeting held in Accra. An investigation was conducted to characterise the affected people, determine the etiologic food, the source of contamination and the etiologic agent and to implement appropriate public health measures to prevent future occurrences. A retrospective cohort study was conducted via telephone interviews, using a structured questionnaire developed from the buffet menu. A case was defined as any person suffering from symptoms of foodborne illness e.g. diarrhoea and/or abdominal cramps after eating food served during the stakeholder consultation meeting in Accra on 21^st November, 2016. The exposure status of all the members of the cohort was assessed by taking the food history of each respondent during the telephone interview. The data obtained was analysed using Epi Info 7. An environmental risk assessment was conducted to ascertain the source of the food contamination. Risks of foodborne infection from the foods eaten were determined using attack rates and odds ratios. Data was obtained from 54 people who consumed food served during the stakeholders’ meeting. Out of this population, 44 people reported with symptoms of food poisoning representing 81.45% (overall attack rate). The peak incubation period was seven hours with a minimum and maximum incubation periods of four and 17 hours, respectively. The commonly reported symptoms were diarrhoea (97.73%, 43/44), vomiting (84.09%, 37/44) and abdominal cramps (75.00%, 33/44). From the incubation period, duration of illness and the symptoms, toxin-mediated food poisoning was suspected. The environmental risk assessment of the implicated catering facility indicated a lack of time/temperature control, inadequate knowledge on food safety among workers and sanitation issues. Limited number of food samples was received for microbiological analysis. Multivariate analysis indicated that illness was significantly associated with the consumption of the snacks served (OR 14.78, P < 0.001). No stool and blood or samples of etiologic food were available for organism isolation; however, the suspected etiologic agent was Staphylococcus aureus or Clostridium perfringens. The outbreak could probably be due to the consumption of unwholesome snack (tuna sandwich or chicken. The contamination and/or growth of the etiologic agent in the snack may be due to the breakdown in cleanliness, time/temperature control and good food handling practices. Training of food handlers in basic food hygiene and safety is recommended.

Keywords: Accra, buffet, conference, C. perfringens, cohort study, food poisoning, gastroenteritis, office workers, Staphylococcus aureus

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Authors: S. Fata, M. A. Tokat, N. Bagardi, B. Yilmaz

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Nowadays, various traditional practices are used throughout the world with aim to improve fertility. Various traditional remedies, acupuncture, religious practices such as sacrifice are frequently used. Studies often evaluate the traditional practices used by the women. But the use of this non-medical practice by couples and specific application reasons of this methods has been less investigated. The aim of this study was to evaluate the common used non-medical practices and determine perceived benefits by couples with fertility problems in Turkey. This is a descriptive study. Research data were collected between May-July 2016, in Izmir Ege Birth Education and Research Hospital Assisted Reproduction Clinic, from 151 couples with fertility problem. Personal Information Form and Non-Medical Practices Used for Fertility Evaluation Form was used. Number 'GOA 2649' permission letter from Dokuz Eylul University Non-Invasive Research Ethics Board, permission letter from the institution and the written consent from participants has been received to carry out the study. In the evaluation of the data, frequencies and proportions analysis were used. The average age of women participating in the study was 32.87, the 35.8% were high school graduates, 60.3% were housewife and the 58.9% lived in city. The 30.5% of husbands were high school graduates, the 96.7% were employed and the 60.9% lived in city. The 78.1% of couples lived as a nuclear family, the average marriage year was 7.58, in 33.8% the fertility problem stems from women, 42.4% of them received a diagnosis for 1-2 years, 35.1% were being treated for 1-2 years. The 35.8% of women reported use of non-medical applications. The 24.4% of women used figs, onion cure, hacemat, locust, bee-pollen milk, the 18.2% used herbs, the 13.1% vowed, the 12.1% went to the tomb, the 10.1% did not bath a few days after the embryo transfer, the 9.1% used thermal water baths, the 5.0% manually corrected the womb, the 5.0% printed amulets by Hodja, the 3.0% went to the Hodja/pilgrims. Among the perceived benefits of using non-medical practices; facilitate pregnancy and implantation, improve oocyte quality were the most recently expressed. Women said that they often used herbs to develop follicles, did not bath after embryo transfer with aim to provide implantation, and used thermal waters to get rid of the infection. Compared to women, only the 25.8% of men used the non-medical practice. The 52.1% reported that they used peanuts, hacemat, locust, bee-pollen milk, the 14.9% used herbs, the 12.8% vowed, the 10.1% went to the tomb, the 10.1% used thermal water baths. Improve sperm number, motility and quality were the most expected benefits. Men said that they often used herbs to improve sperm number, used peanuts, hacemat, locust, bee-pollen milk to improve sperm motility and quality. Couples in Turkey often use non-medical practices to deal with fertility problems. Some of the practices considered as useful can adversely affect health. Healthcare providers should evaluate the use of non-medical practices and should inform if the application is known adverse effects on health.

Keywords: fertility, couples, non-medical practice, perceived benefit

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Authors: Surachart Koyadun

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Introduction: Plasmodium knowlesi (P. knowlesi) malaria is a zoonotic disease that is classified as type 5 of human malaria. Commonly found in macaques (Macaca fascicularis) and (Macaca nemestrina), P. knowlesi is capable of resulting in both uncomplicated and severe malaria in humans. Situation of P. knowlesi malaria in Phang-Nga province for the past 3 years from 2020 – 2022 revealed no case report in 2020, however, a total of 14 cases had been reported in 2021 - 2022. This research aimed to 1) study the epidemiology of P. knowlesi, 2) examine the clinical manifestations of P. knowlesi patients, 3) analyze the ecology and entomology of P. knowlesi, and 4) analyze the diagnosis and treatment of P. knowlesi. Method: This research was a retrospective descriptive study/case report. The study was conducted in 14 patients with P. knowlesi malaria between 2021 and 2022 in 4 districts of Phang-Nga Province, Thailand including Thapput, Kapong, Takuapa and Khuraburi. Results: The study subjects of P. knowlesi malaria were all males. Most of them were working age groups as farmers and worked in forest or plantation areas. All had no history of blood transfusions. Most of the patients did not use mosquito nets and had a history of camping in the forest prior to the onset of fever. An analysis of all 14 sources of infection unveiled the area is home to macaques, and that area has detected Anopheles mosquito, which is the carrier of the disease. Majority of them got sick in the dry season of Thailand (December-April). The main symptoms brought to the hospital were fever, chills, headache, body aches. Laboratory findings on the first day of diagnosis were as follows: The white blood cell count was found within the normal range. In the proportion of white blood cells, eosinophils were found to be slightly higher than normal. Slight anemia was found on early examination. The platelet count was found to be below normal in all cases. Severely low platelet count (2,000 cells/mm3) was found in severe cases with multiple complications. No patient was found dead but 85.7% of complications were found, with acute renal failure being the most common. Patients with delayed diagnosis and treatment of malaria (inaccurate diagnosis or late access to the hospital) had the highest severity and complications than those who had seen the doctor since the first 3-4 days of illness or the screening of symptoms and risk history by the malaria clinic staff at vector-borne disease control unit. Conclusion and Recommendation: P. knowlesi malaria is an emerging infectious disease transmitted from animals to humans. There are challenges in epidemiology, entomology, ecology for effective surveillance, prevention and control. Early diagnosis and treatment would reduce complications and prevent death.

Keywords: malaria, plasmodium knowlesi, epidemiology, ecology, entomology, diagnosis, treatment

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Authors: Sara R. Knigge, Sugat R. Tuladhar, Alexander Becker, Tobias Schilling, Birgit Glasmacher

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Commercially available mechanical or biological heart valve prostheses both tend to fail long-term due to thrombosis, calcific degeneration, infection, or immunogenic rejection. Moreover, these prostheses are non-viable and do not grow with the patients, which is a problem for young patients. As a result, patients often need to undergo redo-operations. Tissue-engineered (TE) heart valves based on degradable electrospun fiber scaffolds represent a promising approach to overcome these limitations. Such scaffolds need sufficient mechanical properties to withstand the hydrodynamic stress of intracardiac hemodynamics. Additionally, the scaffolds should be colonized by autologous or homologous cells to facilitate the in vivo remodeling of the scaffolds to a viable structure. This study investigates how process parameters of electrospinning and degradation affect the mechanical properties of electrospun scaffolds made of FDA-approved, biodegradable polymer polycaprolactone (PCL). Fiber mats were produced from a PCL/tetrafluoroethylene solution by electrospinning. The e-spinning process was varied in terms of scaffold thickness, fiber diameter, fiber orientation, and fiber interconnectivity. The morphology of the fiber mats was characterized with a scanning electron microscope (SEM). The mats were degraded in different solutions (cell culture media, SBF, PBS and 10 M NaOH-Solution). At different time points of degradation (2, 4 and 6 weeks), tensile and cyclic loading tests were performed. Fresh porcine pericardium and heart valves served as a control for the mechanical assessment. The progression of polymer degradation was quantified by SEM and differential scanning calorimetry (DSC). Primary Human aortic endothelial cells (HAECs) and Human induced pluripotent stem cell-derived endothelial cells (iPSC-ECs) were seeded on the fiber mats to investigate the cell colonization potential. The results showed that both the electrospinning parameters and the degradation significantly influenced the mechanical properties. Especially the fiber orientation has a considerable impact and leads to a pronounced anisotropic behavior of the scaffold. Preliminary results showed that the polymer became strongly more brittle over time. However, the embrittlement can initially only be detected in the mechanical test. In the SEM and DSC investigations, neither morphological nor thermodynamic changes are significantly detectable. Live/Dead staining and SEM imaging of the cell-seeded scaffolds showed that HAECs and iPSC-ECs were able to grow on the surface of the polymer. In summary, this study's results indicate a promising approach to the development of a TE aortic heart valve based on an electrospun scaffold.

Keywords: electrospun scaffolds, long-term polymer degradation, mechanical behavior of electrospun PCL, tissue engineered aortic heart valve

Procedia PDF Downloads 124

Authors: Itzel Amaro-Estrada, Eduardo Vergara-Rivera, Virginia Juarez-Flores, Mayra Cobaxin-Cardenas, Rosa Estela Quiroz, Jesus F. Preciado, Sergio Rodriguez-Camarillo

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Bovine anaplasmosis is an infectious, tick-borne disease caused mainly by Anaplasma marginale; typical signs include anemia, fever, abortion, weight loss, decreased milk production, jaundice, and potentially death. Sick bovine can recover when antibiotics are administered; however, it usually remains as carrier for life, being a risk of infection for susceptible cattle. Anaplasma marginale is an obligate intracellular Gram-negative bacterium with genetic composition highly diverse among geographical isolates. There are currently no vaccines fully effective against bovine anaplasmosis; therefore, the economic losses due to disease are present. Vaccine formulation became a hard task for several pathogens as Anaplasma marginale, but peptide-based vaccines are an interesting proposal way to induce specific responses. Phage-displayed peptide libraries have been proved one of the most powerful technologies for identifying specific ligands. Screening of these peptides libraries is also a tool for studying interactions between proteins or peptides. Thus, it has allowed the identification of ligands recognized by polyclonal antiserums, and it has been successful for the identification of relevant epitopes in chronic diseases and toxicological conditions. Protective immune response to bovine anaplasmosis includes high levels of immunoglobulins subclass G2 (IgG2) but not subclass IgG1. Therefore, IgG2 from the serum of protected bovine can be useful to identify ligands, which can be part of an immunogen for cattle. In this work, phage display random peptide library Ph.D. ™ -12 was incubating with IgG2 or blood sera of immunized bovines against A. marginale as targets. After three rounds of biopanning, several candidates were selected for additional analysis. Subsequently, their reactivity with sera immunized against A. marginale, as well as with positive and negative sera to A. marginale was evaluated by immunoassays. A collection of recognized peptides tested by ELISA was generated. More than three hundred phage-peptides were separately evaluated against molecules which were used during panning. At least ten different peptides sequences were determined from their nucleotide composition. In this approach, three phage-peptides were selected by their binding and affinity properties. In the case of the development of vaccines or diagnostic reagents, it is important to evaluate the immunogenic and antigenic properties of the peptides. Immunogenic in vitro and in vivo behavior of peptides will be assayed as synthetic and as phage-peptide for to determinate their vaccine potential. Acknowledgment: This work was supported by grant SEP-CONACYT 252577 given to I. Amaro-Estrada.

Keywords: bovine anaplasmosis, peptides, phage display, veterinary vaccines

Procedia PDF Downloads 126

Authors: Hakimullah Hakim, Chiharu Toyofuku, Mari Ota, Mayuko Suzuki, Miyuki Komura, Masashi Yamada, Md. Shahin Alam, Natthanan Sangsriratanakul, Dany Shoham, Kazuaki Takehara

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Disinfectants and their application are essential part of infection control strategies and enhancement of biosecurity at farms, worldwide. Alkaline agents are well known for their strong and long term antimicrobial capacities and most frequently are applied at farms for control and prevention of biological hazards. However, inadequate information regarding such materials’ capacities to inactivate pathogens and their improper applications fail farmers to achieve the mentioned goal. Thus, this requires attention to further evaluate their efficacies, under different conditions and in different ways. Here in this study we evaluated bactericidal efficacies of food additive grade of calcium hydroxide (FdCa(OH)2) powder derived from natural calcium carbonates obtained from limestone (Fine Co., Ltd., Tokyo, Japan), and bioceramic powder (BCX) derived from chicken feces at pH 13 (NMG environmental development Co., Ltd., Tokyo, Japan), for their efficacies to inactivate bacteria in feces. [Materials & Methods] Chicken feces were inoculated by 100 µl Escherichia coli and Salmonella Infantis in falcon tubes, individually, then FdCa(OH)2 or BCX powders were individually added to make final concentration of 0, 5, 10, 20 and 30% (w/w) in total weight of 0.5g, followed by properly mixing and incubating at room temperature for certain periods of time, in a dark place. Afterwards, 10 ml 1M Tris-HCl (pH 7.2) was added onto them to reduce their pH, in order to stop powders’ activities and to harvest the remained viable bacteria, whereas using normal medium or dW2 to recover bacteria increases the mixture pH, and as a result bacteria would be inactivated soon; therefore, the latter practice brings about incorrect and misleading results. Samples were then inoculated on DHL agar plates in order to calculate colony forming units (CFU)/ml of viable bacteria. [Results and Discussion] FdCa(OH)2 powder at 10% and 5% required 3 hr and 6 hr exposure times, respectively, while BCX powder at 20% concentrations required 6 hr exposure time to kill the mentioned bacteria in feces down to lower than detectable level (≤ 3.6 log10 CFU/ml). This study confirmed capacities of FdCa(OH)2 and BCX powders to inactivate bacteria in feces, and both materials are environment friendly materials, with no risk to human or animal’s health. This finding helps farmers to properly apply alkaline agents in appropriate concentrations and exposure times in their farms, in order to prevent and control infectious diseases outbreaks and to enhance biosecurity. Finally, this finding may help farmers to implement better strategies for infections control in their livestock farms.

Keywords: bacterial inactivation, bioceramic, biosecurity at livestock farms, chicken feces

Procedia PDF Downloads 420

Authors: Atin Adhikari, Sushma Kurella, Pratik Banerjee, Nabanita Mukherjee, Yamini M. Chandana Gollapudi, Bushra Shah

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Different sharp instruments, drilling machines, and high speed rotary instruments are routinely used in dental clinics during dental cleaning. Therefore, these cleaning procedures release a lot of oral microorganisms including bacteria in clinic air and may cause significant occupational bioaerosol exposure risks for dentists, dental hygienists, patients, and dental clinic employees. Two major goals of this study were to quantify volumetric airborne concentrations of bacteria and to assess overall microbial activity in this type of occupational environment. The study was conducted in several dental clinics of southern Georgia and 15 dental cleaning procedures were targeted for sampling of airborne bacteria and testing of overall microbial activity in settled dusts over clinic floors. For air sampling, a Biostage viable cascade impactor was utilized, which comprises an inlet cone, precision-drilled 400-hole impactor stage, and a base that holds an agar plate (Tryptic soy agar). A high-flow Quick-Take-30 pump connected to this impactor pulls microorganisms in air at 28.3 L/min flow rate through the holes (jets) where they are collected on the agar surface for approx. five minutes. After sampling, agar plates containing the samples were placed in an ice chest with blue ice and plates were incubated at 30±2°C for 24 to 72 h. Colonies were counted and converted to airborne concentrations (CFU/m3) followed by positive hole corrections. Most abundant bacterial colonies (selected by visual screening) were identified by PCR amplicon sequencing of 16S rRNA genes. For understanding overall microbial activity in clinic floors and estimating a general cleanliness of the clinic surfaces during or after dental cleaning procedures, ATP levels were determined in swabbed dust samples collected from 10 cm2 floor surfaces. Concentration of ATP may indicate both the cell viability and the metabolic status of settled microorganisms in this situation. An ATP measuring kit was used, which utilized standard luciferin-luciferase fluorescence reaction and a luminometer, which quantified ATP levels as relative light units (RLU). Three air and dust samples were collected during each cleaning procedure (at the beginning, during cleaning, and immediately after the procedure was completed (n = 45). Concentrations at the beginning, during, and after dental cleaning procedures were 671±525, 917±1203, and 899±823 CFU/m3, respectively for airborne bacteria and 91±101, 243±129, and 139±77 RLU/sample, respectively for ATP levels. The concentrations of bacteria were significantly higher than typical indoor residential environments. Although an increasing trend for airborne bacteria was observed during cleaning, the data collected at three different time points were not significantly different (ANOVA: p = 0.38) probably due to high standard deviations of data. The ATP levels, however, demonstrated a significant difference (ANOVA: p <0.05) in this scenario indicating significant change in microbial activity on floor surfaces during dental cleaning. The most common bacterial genera identified were: Neisseria sp., Streptococcus sp., Chryseobacterium sp., Paenisporosarcina sp., and Vibrio sp. in terms of frequencies of occurrences, respectively. The study concluded that bacterial exposure in dental clinics could be a notable occupational biohazard, and appropriate respiratory protections for the employees are urgently needed.

Keywords: bioaerosols, hospital hygiene, indoor air quality, occupational biohazards

Procedia PDF Downloads 294

Authors: Norihiro Kato, Yuriko Takayama

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Some gram-negative bacteria enable the simultaneous activation of gene expression involved in N-acylhomoserine lactone (AHL) dependent cell-to-cell communication system. Such regulatory system for the bacterial group behavior is termed as quorum sensing (QS) because a diffusible AHL signal can accumulate around the cell during the increase of the cell density and trigger activation of the sequential QS process. By blocking the QS, the expression of diverse genes related to infection, antibiotic production, and biofilm formation is inhibited. Conditioning of QS by regulation of the DNA-receptor-AHL interaction is a potential target for enhancing host defenses against pathogenicity. We focused on engineered application of transcriptional regulator SpnR produced in opportunistic human pathogen Serratia marcescens. The SpnR can interact with AHL signals at an N-terminal domain and also with a promoter region of a QS target gene at a C-terminal domain. As the initial process of the QS activation, the SpnR forms a complex with the AHL to enhance the expression of pig cluster; the SpnR normally acts as an activator for the expression of the QS-dependent gene. In this research, we attempt to artificially control QS by changing the role of SpnR. The QS-dependent prodigiosin production is expected to inhibit by externally added SpnR in the culture broth of AS-1 strain because the AHL concentration was kept below the threshold by AHL-SpnR complex formation. Maltose-binding protein (MBP)-tagged SpnR (MBP-SpnR) was overexpressed in Escherichia coli and purified using an affinity chromatography equipped with an amylose resin column. The specific interaction between AHL and MBP-SpnR was demonstrated by quartz crystal microbalance (QCM) sensor. AHL with amino end-group was coupled with COOH-terminated self-assembled monolayer prepared on a gold electrode of 27-MHz quartz crystal sensor using water-soluble carbodiimide. After the injection of MBP-SpnR into a cup-type sensor cell filled with the buffer solution, time course of resonant frequency change (ΔFs) was determined. A decrease of ΔFs clearly showed the uptake of MBP-SpnR onto the AHL-immobilized electrode. Furthermore, no binding affinity was observed after the heat-inactivation of MBP-SpnR at 80ºC. These results suggest that MBP-SpnR possesses a specific affinity for AHL. MBP-SpnR was added to the culture medium as an AHL trap to study inhibitory effects on intracellularly accumulated prodigiosin. With approximately 2 µM MBP-SpnR, the amount of prodigiosin induced was half that of the control without any additives. In conclusion, the function of SpnR could be switched by adding it to the cell culture. Exogenously added MBP-SpnR possesses high affinity for AHL derived from cells and acts as an inhibitor of AHL-mediated QS.

Keywords: intracellular signaling, microbial biotechnology, quorum sensing, transcriptional regulator

Procedia PDF Downloads 253

Authors: Michelle S. Pereira, Analice C. Azevedo, Julliane D. Medeiros, Ana Claudia S. Martins, Didier S. Castellano-Filho, Claudio G. Diniz, Vania L. Silva

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Vaginal microbiota is a complex ecosystem, composed by aerobic and anaerobic bacteria, living in a dynamic equilibrium. Lactobacillus spp. are predominant in vaginal ecosystem, and factors such as immunity and hormonal variations may lead to disruptions, resulting in proliferation of opportunistic pathogens. Bacterial vaginosis (BV) is a polymicrobial syndrome, caused by an increasing of anaerobic bacteria replacing Lactobacillus spp. Microorganisms such as Gardnerella vaginalis, Mycoplasma hominis, Mobiluncus spp., and Atopobium vaginae can be found in BV, which may also be associated to other infections such as by Human Papillomavirus (HPV). HPV is highly prevalent in sexually active women, and is considered a risk factor for development of cervical cancer. As long as few data is available on vaginal microbiota of women with HPV-associated cervical lesions, our objectives were to evaluate the diversity in vaginal ecosystem in these women. To all patients, clinical and socio-demographic data were collected after gynecological examination. This study was approved by the Ethics Committee from Federal University of Juiz de Fora, Minas Gerais, Brazil. Vaginal secretion and cervical scraping were collected. Gram-stained smears were evaluated to establish Nugent score for BV determination. Viral and bacterial DNA obtained was used as template for HPV genotyping (PCR) and bacterial fingerprint (REP-PCR). In total 31 patients were included (mean age 35 and 93.6% sexually active). The Nugent score showed that 38.7% were BV. From the medical records, Pap smear tests showed that 32.3% had low grade squamous epithelial lesion (LSIL), 29% had high grade squamous epithelial lesion (HSIL), 25.8% had atypical squamous cells of undetermined significance (ASC-US) and 12.9% with atypical squamous cells that would not exclude high-grade lesion (ASC-H). All participants were HPV+. HPV-16 was the most frequent (87.1%), followed by HPV-18 (61.3%). HPV-31, HPV-52 and HPV-58 were also detected. Coinfection HPV-16/HPV-18 was observed in 75%. In the 18-30 age group, HPV-16 was detected in 40%, and HPV-16/HPV-18 coinfection in 35%. HPV-16 was associated to 30% of ASC-H and 20% of HSIL patients. BV was observed in 50% of HPV-16+ participants and in 45% of HPV-16/HPV-18+. Fingerprints of bacterial communities showed clusters with low similarity suggesting high heterogeneity in vaginal microbiota within the sampled group. Overall, the data is worrisome once cervical-cancer highly risk-associated HPV-types were identified. The high microbial diversity observed may be related to the different levels of cellular lesions, and different physiological conditions of the participants (age, social behavior, education). Further prospective studies are needed to better address correlations and BV and microbial imbalance in vaginal ecosystems which would be related to the different cellular lesions in women with HPV infections. Supported by FAPEMIG, CNPq, CAPES, PPGCBIO/UFJF.

Keywords: human papillomavirus, bacterial vaginosis, bacterial diversity, cervical cancer

Procedia PDF Downloads 176