Search results for: upper respiratory tract infection

Authors: Jiangbo Li, Wenqian Huang

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Early detection of fungal infection in citrus fruit is one of the major problems in the postharvest commercialization process. The automatic and nondestructive detection of infected fruits is still a challenge for the citrus industry. At present, the visual inspection of rotten citrus fruits is commonly performed by workers through the ultraviolet induction fluorescence technology or manual sorting in citrus packinghouses to remove fruit subject with fungal infection. However, the former entails a number of problems because exposing people to this kind of lighting is potentially hazardous to human health, and the latter is very inefficient. Orange is used as a research object. This study would focus on this problem and proposed an effective method based on Vis-NIR hyperspectral imaging in the wavelength range of 400-1000 nm with a spectroscopic resolution of 2.8 nm. In this work, three normalization approaches are applied prior to analysis to reduce the effect of sample curvature on spectral profiles, and it is found that mean normalization was the most effective pretreatment for decreasing spectral variability due to curvature. Then, principal component analysis (PCA) was applied to a dataset composing of average spectra from decayed and normal tissue to reduce the dimensionality of data and observe the ability of Vis-NIR hyper-spectra to discriminate data from two classes. In this case, it was observed that normal and decayed spectra were separable along the resultant first principal component (PC1) axis. Subsequently, five wavelengths (band) centered at 577, 702, 751, 808, and 923 nm were selected as the characteristic wavelengths by analyzing the loadings of PC1. A multispectral combination image was generated based on five selected characteristic wavelength images. Based on the obtained multispectral combination image, the intensity slicing pseudocolor image processing method is used to generate a 2-D visual classification image that would enhance the contrast between normal and decayed tissue. Finally, an image segmentation algorithm for detection of decayed fruit was developed based on the pseudocolor image coupled with a simple thresholding method. For the investigated 238 independent set samples including infected fruits infected by Penicillium digitatum and normal fruits, the total success rate is 100% and 97.5%, respectively, and, the proposed algorithm also used to identify the orange infected by penicillium italicum with a 100% identification accuracy, indicating that the proposed multispectral algorithm here is an effective method and it is potential to be applied in citrus industry.

Keywords: citrus fruit, early rotten, fungal infection, hyperspectral imaging

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Authors: Alieh Farshbaf

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Introduction: Current anti-retroviral drugs have some restrictions for treatment of HIV-1 infection. The efficacy of retroviral drugs is not same in different infected patients and the virus rebound from latent reservoirs after stopping them. Recently, the engineering of stem cells and gene therapy provide new approaches to eliminate some drug problems by induction of resistance to HIV-1. Literature review: Up to now, AIDS-restriction genes (ARGs) were suitable candidate for gene and cell therapies, such as cc-chemokine receptor-5 (CCR5). In this manner, CCR5 provide effective cure in Berlin and Boston patients by inducing of HIV-1 resistance with allogeneic stem cell transplantation. It is showed that Zinc Finger Nuclease (ZFN) could induce HIV-1 resistance in stem cells of infected patients by homologous recombination or non-end joining mechanism and eliminate virus loading after returning the modified cells. Then, gene modification by HIV restriction factors, as TRIM5, introduced another gene candidate for HIV by interfering in infection process. These gene modifications/editing provided by stem cell futures that improve treatment in refractory disease such as HIV-1. Conclusion: Although stem cell transplantation has some complications, but in compare to retro-viral drugs demonstrated effective cure by elimination of virus loading. On the other hand, gene therapy is cost-effective for an infected patient than retroviral drugs payment in a person life-long. The results of umbilical cord blood stem cell transplantation showed that gene and cell therapy will be applied easier than previous treatment of AIDS with high efficacy.

Keywords: stem cell, AIDS, gene modification, cell engineering

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Authors: K. Sekulová, M. Šimon

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This article focuses on upper-extremity musculoskeletal disorders risk assessment model at workplace. In this model are used risk factors that are responsible for musculoskeletal system damage. Based on statistic calculations the model is able to define what risk of MSD threatens workers who are under risk factors. The model is also able to say how MSD risk would decrease if these risk factors are eliminated.

Keywords: ergonomics, musculoskeletal disorders, occupational diseases, risk factors

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Authors: Jeremy Schultz, Kelly Bricker

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Ecotourism has proven itself to be a forerunner in the advancement of environmental conservation all the while supporting cultural tradition, uniqueness, and pride among indigenous communities. Successful private-community partnerships associated with ecotourism operations are vital to the overall prosperity of both the businesses and the local communities. Such accomplishments can be seen through numerous livelihood goals including income, food security, health, reduced vulnerability, governance, and empowerment. Private-community partnerships also support global initiatives such as the sustainable development goals and sustainable development frameworks including those proposed by the United Nations World Tourism Organization (WTO). Understanding such partnerships assists not only large organizations such as the WTO, but it also benefits smaller ecotourism operators and entrepreneurs who are trying to achieve their sustainable tourism development goals. This study examined the partnership between an ecotourism company (Rivers Fiji) and two rural villages located in Fiji’s Upper Navua Conservation Area. Focus groups were conducted in each village. Observation journals were also used to record conversations outside of the focus groups. Data were thematically organized and analyzed to offer researcher interpretations and understandings. This research supported the notion that respectful and emboldening partnerships between communities and private enterprise are vital to the composition of successful ecotourism operations that support sustainable development protocol. Understanding these partnerships can assist in shaping future ecotourism development and re-molding existing businesses. This study has offered an example of a thriving partnership through community input and critical researcher analysis. Research has identified six contributing factors to successful ecotourism partnerships, and this study provides additional support to that framework.

Keywords: community partnerships, conservation areas, ecotourism, Fiji, sustainability

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Authors: Patcharawalai Jaisamut, Kamonthip Wiwattanawongsa, Ruedeekorn Wiwattanapatapee

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Supersaturation of drugs in the gastrointestinal tract is one approach to increase the absorption of poorly water-soluble drugs. The stabilization of a supersaturated state was achieved by adding precipitation inhibitors that may act through a variety of mechanisms.In this study, the effect of the natural gums, acacia, gelatin, pectin and tragacanth on curcumin supersaturation in simulated gastric fluid (SGF) (pH 1.2), fasted state simulated gastric fluid (FaSSGF) (pH 1.6), and simulated intestinal fluid (SIF) (pH 6.8)was investigated. The results indicated that all natural gums significantly increased the curcum insolubility (about 1.2-6-fold)when compared to the absence of gum, and assisted in maintaining the supersaturated drug solution. Among the tested gums, pectin at 3% w/w was the best precipitation inhibitor with a significant increase in the degree of supersaturation about 3-fold in SGF, 2.4-fold in FaSSGF and 2-fold in SIF.

Keywords: curcumin, solubility, supersaturation, precipitation inhibitor

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Authors: W. Mohammed Saeed, A. J. Mcbain, S. M. Cruickshank, C. A. O’Neill

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In the gut, probiotics have been shown to protect epithelial cells from pathogenic bacteria through a number of mechanisms: 1-Increasing epithelial barrier function, 2-Modulation of the immune response especially innate immune response, 3-Inhibition of pathogen adherence and down regulation of virulence factors. Since probiotics have positive impacts on the gut, their potential effects on other body tissues, such as skin have begun to be investigated. The purpose of this project is to characterize the potential of probiotic bacteria lysate as therapeutic agent for preventing or reducing the S. aureus infection. Normal human primary keratinocytes (KCs) were exposed to S. aureus (106/ml) in the presence or absence of L. rhamnosus GG lysate (extracted from 108cfu/ml). The viability of the KCs was measured after 24 hours using a trypan blue exclusion assay. When KCs were treated with S aureus alone, only 25% of the KCs remained viable at 24 hours post infection. However, in the presence of L. rhamnosus GG lysate the viability of pathogen infected KCs increased to 58% (p=0.008, n=3). Furthermore, when KCs co-exposed, pre- exposed or post-exposed to L. rhamnosus GG lysate, the viability of the KCs increased to ≈60%, the L. rhamnosus GG lysate was afforded equal protection in different conditions. These data suggests that two possible separate mechanisms are involved in the protective effects of L. rhamnosus GG such as reducing S. aureus growth, or inhibiting of pathogenic adhesion. Interestingly, a lysate of L rhamnosus GG provided significant reduction in S. aureus growth and adhesion of S. aureus that being viable following 24 hours incubation with S aureus. Therefore, a series of Liquid Chromatography (RP-LC) methods were adopted to partially purify the lysate in combination with functional assays to elucidate in which fractions the efficacious molecules were contained. In addition, the Mass Spectrometry-based protein sequencing was used to identify putative proteins in the fractions. The data presented from purification process demonstrated that L. rhamnosus GG lysate has the potential to protect keratinocytes from the toxic effects of the skin pathogen, S. aureus. Three potential mechanisms were identified: inhibition of pathogen growth; competitive exclusion; and displacement of the pathogen from keratinocyte binding sites. In this study, ‘moonlight’ proteins were identified in the current study’s MS/MS data for L. rhamnosus GG lysate, which could elucidate the ability of lysate in the competitive exclusion and displacement of S. aureus from keratinocyte binding sites. Taken together, it can be speculated that L. rhamnosus GG lysate utilizes different mechanisms to protect keratinocytes from S. aureus toxicity. The present study indicates that the proteinaceous substances are involved in anti-adhesion activity. This is achieved by displacing the pathogen and preventing the severity of pathogen infection and the moonlight proteins might be involved in inhibiting the adhesion of pathogens.

Keywords: lysate, fractions, adhesion, L. rhamnosus GG, S. aureus toxicity

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Authors: Worawanna Panyakotkaew, Falan Srisuriyachai

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Cyclic steam injection (CSI) is a thermal recovery technique performed by injecting periodically heated steam into heavy oil reservoir. Oil viscosity is substantially reduced by means of heat transferred from steam. Together with gas pressurization, oil recovery is greatly improved. Nevertheless, prediction of effectiveness of the process is difficult when reservoir contains degree of heterogeneity. Therefore, study of heterogeneity together with interest reservoir properties must be evaluated prior to field implementation. In this study, thermal reservoir simulation program is utilized. Reservoir model is firstly constructed as multi-layered with coarsening upward sequence. The highest permeability is located on top layer with descending of permeability values in lower layers. Steam is injected from two wells located diagonally in quarter five-spot pattern. Heavy oil is produced by adjusting operating parameters including soaking period and steam quality. After selecting the best conditions for both parameters yielding the highest oil recovery, effects of degree of heterogeneity (represented by Lorenz coefficient), vertical permeability and permeability sequence are evaluated. Surprisingly, simulation results show that reservoir heterogeneity yields benefits on CSI technique. Increasing of reservoir heterogeneity impoverishes permeability distribution. High permeability contrast results in steam intruding in upper layers. Once temperature is cool down during back flow period, condense water percolates downward, resulting in high oil saturation on top layers. Gas saturation appears on top after while, causing better propagation of steam in the following cycle due to high compressibility of gas. Large steam chamber therefore covers most of the area in upper zone. Oil recovery reaches approximately 60% which is of about 20% higher than case of heterogeneous reservoir. Vertical permeability exhibits benefits on CSI. Expansion of steam chamber occurs within shorter time from upper to lower zone. For fining upward permeability sequence where permeability values are reversed from the previous case, steam does not override to top layers due to low permeability. Propagation of steam chamber occurs in middle of reservoir where permeability is high enough. Rate of oil recovery is slower compared to coarsening upward case due to lower permeability at the location where propagation of steam chamber occurs. Even CSI technique produces oil quite slowly in early cycles, once steam chamber is formed deep in the reservoir, heat is delivered to formation quickly in latter cycles. Since reservoir heterogeneity is unavoidable, a thorough understanding of its effect must be considered. This study shows that CSI technique might be one of the compatible solutions for highly heterogeneous reservoir. This competitive technique also shows benefit in terms of heat consumption as steam is injected periodically.

Keywords: cyclic steam injection, heterogeneity, reservoir simulation, thermal recovery

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Authors: Samar Mousa, Ryan Kerstein, Mohanad Adam

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Regional anaesthesia has been associated with favourable outcomes in patients undergoing a wide range of surgeries. Beneficial effects have been demonstrated in terms of postoperative respiratory and cardiovascular endpoints, 7-day survival, time to ambulation and hospital discharge, and postoperative analgesia. Our project aimed at assessing the regional anaesthesia practice in the plastic surgery department of Buckinghamshire trust and finding out ways to improve the service in collaboration with the anaesthesia team. It is a retrospective study associated with a questionnaire filled out by plastic surgeons and anaesthetists to get the full picture behind the numbers. The study period was between 1/3/2022 and 23/5/2022 (12 weeks). The operative notes of all patients who had an operation under plastic surgery, whether emergency or elective, were reviewed. The criteria of suitable candidates for the regional block were put by the consultant anaesthetists as follows: age above 16, single surgical site (arm, forearm, leg, foot), no drug allergy, no pre-existing neuropathy, no bleeding disorders, not on ant-coagulation, no infection to the site of the block. For 12 weeks, 1061 operations were performed by plastic surgeons. Local cases were excluded leaving 319 cases. Of the 319, 102 patients were suitable candidates for regional block after applying the previously mentioned criteria. However, only seven patients had their operations under the regional block, and the rest had general anaesthesia that could have been easily avoided. An online questionnaire was filled out by both plastic surgeons and anaesthetists of different training levels to find out the reasons behind the obvious preference for general over regional anaesthesia, even if this was against the patients’ interest. The questionnaire included the following points: training level, time taken to give GA or RA, factors that influence the decision, percentage of RA candidates that had GA, reasons behind this percentage, recommendations. Forty-four clinicians filled out the questionnaire, among which were 23 plastic surgeons and 21 anaesthetists. As regards the training level, there were 21 consultants, 4 associate specialists, 9 registrars, and 10 senior house officers. The actual percentage of patients who were good candidates for RA but had GA instead is 93%. The replies estimated this percentage as between 10-30%. 29% of the respondents thought that this percentage is because of surgeons’ preference to have GA rather than RA for their operations without medical support for the decision. 37% of the replies thought that anaesthetists prefer giving GA even if the patient is a suitable candidate for RA. 22.6% of the replies thought that patients refused to have RA, and 11.3% had other causes. The recommendations were in 5 main accesses, which are protocols and pathways for regional blocks, more training opportunities for anaesthetists on regional blocks, providing a separate block room in the hospital, better communication between surgeons and anaesthetists, patient education about the benefits of regional blocks.

Keywords: regional anaesthesia, regional block, plastic surgery, general anaesthesia

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Authors: Sangyeop Shin, D. C. M. Kulatunga, S. H. S. Dananjaya, Chamilani Nikapitiya, Jehee Lee, Mahanama De Zoysa

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Saprolegniasis is one of the most devastating fungal diseases in freshwater fish which is caused by species in the genus Saprolegnia including Saprolegnia parasitica. In this study, we isolated the strain of S. parasitica from diseased rainbow trout in Korea. Morphological and molecular based identification confirmed that isolated fungi belong to the member of S. parasitica, supported by its typical fungal features including cotton-like whitish mycelium, zoospores (primary and secondary) and phylogenetic analysis with internal transcribed spacer (ITS) region. Pathogenicity of isolated S. parasitica was developed in embryo, larvae, juvenile and adult zebrafish as a disease model. Up regulation of host genes encoding ZfTnf-α, Zfc-Rel, ZfIl-12, ZfLyz-c, Zfβ-def, and ZfHsp-70 was identified in zebrafish larvae after experimental challenge of S. parasitica showing the host immune responses against the S. parasitica. Survival of the juveniles upon fungal infection might be due to the increased immune protection in the host. Investigation of antifungal susceptibility of S. parasitica with natural lawsone (2-hydroxy-1,4-naphthoquinone) revealed the minimum inhibitory concentration (MIC) and percentage inhibition of radial growth (PIRG %) as 200 µg/mL and 31.8%, respectively. Lawsone was able to change the membrane permeability, and cause irreversible damage and disintegration to the cellular membranes of S. parasitica which might have effect on fungi growth inhibition. Moreover, the mycelium exposed to lawsone (MIC level) changed the transcriptional responses of S. parasitica genes. Overall results indicate that lawsone could be a potential and novel anti-S. parasitica agent for controlling S. parasitica infection.

Keywords: host-pathogen interactions, lawsone, rainbow trout, Saprolegnia parasitica, Saprolegniasis, zebrafish

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Authors: Meryam Labyad, Karima Fakiri, Widad Lahmini, Ghizlane Draiss, Mohamed Bouskraoui, Nadia Ouzennou

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Background: Asthma is the most common chronic disease in children, affecting nearly 235 million people worldwide (OMS). In Morocco, asthma is much more common in children than in adults; the prevalence rate in children between 13 and 14 years of age is 20%.1 This pathology is marked by high morbidity, a significant impact on the quality of life and development of children 2 This requires a rigorous management strategy in order to achieve clinical control and reduce any risk to the patient 3 A search for aggravating factors is mandatory if a child has difficulty maintaining good asthma control. The objective of the present study is to describe asthma control during this confinement period in children aged 4 to 11 years followed by a pneumo-paediatric consultation. For children whose asthma is not controlled, a search for associations with promoting factors and adherence to treatment is also among the objectives of the study. Knowing the level of asthma control and influencing factors is a therapeutic priority in order to reduce hospitalizations and emergency care use. Objective: To assess asthma control and determine the factors influencing asthma levels in children with asthma during confinement following the COVID 19 pandemic. Method: Prospective cross-sectional study by questionnaire and structured interview among 66 asthmatic children followed in pediatric pneumology consultation at the CHU MED VI of Marrakech from 13/06/2020 to 13/07/2020, asthma control was assessed by the Childhood Asthma Control Test (C-ACT). Results: 66 children and their parents were included (mean age is 7.5 years), asthma was associated with allergic rhinitis (13.5% of cases), conjunctivitis (9% of cases), eczema (12% of cases), occurrence of infection (10.5% of cases). The period of confinement was marked by a decrease in the number of asthma attacks translated by a decrease in the number of emergency room visits (7.5%) of these asthmatic children, control was well controlled in 71% of the children, this control was significantly associated with good adherence to treatment (p<0.001), no infection (p<0.001) and no conjunctivitis (p=002) or rhinitis (p<0.001). This improvement in asthma control during confinement can be explained by the measures taken in the Kingdom to prevent the spread of COVID 19 (school closures, reduction in industrial activity, fewer means of transport, etc.), leading to a decrease in children's exposure to triggers, which justifies the decrease in the number of children having had an infection, allergic rhinitis or conjunctivitis during this period. In addition, the close monitoring of parents resulted in better therapeutic adherence (42.4% were fully observant). Confinement was positively perceived by 68% of the parents; this perception is significantly associated with the level of asthma control (p<0.001). Conclusion: Maintaining good control can be achieved through improved therapeutic adherence and avoidance of triggers, both of which were achieved during the containment period following the VIDOC pandemic 19.

Keywords: Asthma, control , COVID-19 , children

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Authors: Dimitra V. Peristeri, Hussameldin M. Nour, Amiya Ahsan, Sameh Abogabal, Krishna K. Singh, Muhammad Shafique Sajid

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Introduction: The use of optimal skin antiseptic agents for the prevention of surgical site infection (SSI) is of critical importance, especially during abdominal surgical procedures. Alcohol-based chlorhexidine gluconate (CHG) and aqueous-based povidone-iodine (PVI) are the two most common skin antiseptics used nowadays. The objective of this article is to evaluate the effectiveness of alcohol-based CHG versus aqueous-based PVI used for skin preparation before abdominal surgery to reduce SSIs. Methods: Standard medical databases such as MEDLINE, Embase, Pubmed, and Cochrane Library were searched to find randomised, controlled trials (RCTs) comparing alcohol-based CHG skin preparation versus aqueous-based PVI in patients undergoing abdominal surgery. The combined outcomes of SSIs were calculated using an odds ratio (OR) with 95% confidence intervals (95% CI). All data were analysed using Review Manager (RevMan) Software 5.4, and the meta-analysis was performed with a random effect model analysis. Results: A total of 11 studies, all RCTs, were included (n= 12072 participants), recruiting adult patients undergoing abdominal surgery. In the random effect model analysis, the use of alcohol-based CHG in patients undergoing abdominal surgery was associated with a reduced risk of SSI compared to aqueous-based PVI (OR: 0.84; 95% CI [0.74, 0.96], z= 2.61, p= 0.009). Conclusion: Alcohol-based CHG may be more effective for preventing the risk of SSI compared to aqueous-based PVI agents in abdominal surgery. The conclusion of this meta-analysis may add a guiding value to reinforce current clinical practice guidelines.

Keywords: skin preparation, surgical site infection, chlorhexidine, skin antiseptics

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Authors: Luciano Tarantino, Emanuele Balzano, Paolo Tarantino, Riccardo Aurelio Nasto, Aurelio Nasto

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Liver transplantation (OLT) is contraindicate in Portal Vein tumor Thrombosis (PVTT) from Hepatocellular Carcinoma at hepatic hilum(pH-HCC) Surgery,Thermal ablation and chemotherapy show poorer outcomes Electrochemotherapy (ECT) has been successfully used in patients with pH-HCC with PVTT. We report the results of ECT as downstaging aimed to definitive cure by OLT. F.P. 53 years HBV related Cirrhosis Child-Pugh B7 class; EGDS F2 aesophageal Varices. Diabetes. April 2016 : Enhanced Computed Tomography (CT) detected HCC(n.3 nodules in VII-VIII-VI;diameter range=25 cm) and PVTT of right portal vein. The patient was considered ineligible for OLT. May 2016: first ablation session with percutaneous Radiofrequency-ablation(RFA) of 3 HCC-nodules . August 2016: second ablation session with ECT of PVTT. CT october 2016: disappearance of PVTT and patent right portal vein. No intraparenchymal recurrence. CT march 2017: No recurrence in portal vein and in the left lobe. local recurrence in the VII-VIII segments. May 2017 : transarterial chemoembolization (TACE) of right lobe recurrences. CT October 2017: patent right portal vein. No recurrence. The patient was reconsidered for OLT. He underwent OLT in April 2018. At 36-months follow-up , no intrahepatic recurrence of HCC occurred. March 2021: enhanced CT and PET/CT detected a single small nodule (1.5 cm) uptaking tracer in the left upper pulmonary lobe, no hepatic recurrence . CT-guided FNB showed metastasis from HCC . June 2021: left lung upper lobectomy . At the current time the patient is alive and recurrence-free at 64 months follow-up. ECT Could be aneffective technique as pre-OLT dowstaging in HCC with PVTT.

Keywords: liver tumor ablation, interventional ultrasound, electrochemotherapy, liver transplantation

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Authors: Hasmithaa Neha, Atul Kumar Patidar, Girish Ch Kothyari

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The northward convergence of the Indian plate has a dominating influence over the structural and geomorphic development of the Himalayan region. The highly deformed and complex stratigraphy in the area arises from a confluence of exogenic and endogenetic geological processes. This region frequently experiences natural hazards such as debris flows, flash floods, avalanches, landslides, and earthquakes due to its harsh and steep topography and fragile rock formations. Therefore, remote sensing technique-based examination and real-time monitoring of tectonically sensitive regions may provide crucial early warnings and invaluable data for effective hazard mitigation strategies. In order to identify unusual changes in the river gradients, the current study demonstrates a spatial quantitative geomorphic analysis of the upper Alaknanda River basin, Uttarakhand Himalaya, India, using gradient length anomaly analysis (GLAA). This basin is highly vulnerable to ground creeping and landslides due to the presence of active faults/thrusts, toe-cutting of slopes for road widening, development of heavy engineering projects on the highly sheared bedrock, and periodic earthquakes. The intersecting joint sets developed in the bedrocks have formed wedges that have facilitated the recurrence of several landslides. The main objective of current research is to identify abnormal gradient lengths, indicating potential landslide-prone zones. High-resolution digital elevation data and geospatial techniques are used to perform this analysis. The results of GLAA are corroborated with the historical landslide events and ultimately used for the generation of landslide susceptibility maps of the current study area. The preliminary results indicate that approximately 3.97% of the basin is stable, while about 8.54% is classified as moderately stable and suitable for human habitation. However, roughly 19.89% fall within the zone of moderate vulnerability, 38.06% are classified as vulnerable, and 29% fall within the highly vulnerable zones, posing risks for geohazards, including landslides, glacial avalanches, and earthquakes. This research provides valuable insights into the spatial distribution of landslide-prone areas. It offers a basis for implementing proactive measures for landslide risk reduction, including land-use planning, early warning systems, and infrastructure development techniques.

Keywords: landslide vulnerability, geohazard, GLA, upper Alaknanda Basin, Uttarakhand Himalaya

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Authors: Sara A. Al-Jaberi, Salma Ben-Salem, Meriam Messedi, Fatma Ayadi, Lihadh Al-Gazali, Bassam R. Ali

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Background: The chemokine receptor components play crucial roles in the immune system and some of them serve as co-receptors for the HIV virus. Several studies have documented those variants in chemokine receptors are correlated with susceptibility and resistance to infection with HIV virus. For example, mutations in the chemokine receptor 5 gene (CCR5) resulting in loss-of-function (such as the homozygous CCR5Δ32) confer high degree of resistance to HIV infection. Heterozygotes for these variants exhibit slow progression to AIDS. The prevalence of CCR5 polymorphisms varies among ethnic and geographical groups. For example, the CCR5 Δ32 variant is present in 10–15% of north Europeans but is rarely encountered among Africans. This study aims to identify the prevalence of some CCR5 variants in two geographically distant Arab populations (namely Emiratis and Tunisians). Methodology: The prevalence of CCR5 gene variants including CCR5Δ32, FS299, C101X, A29S and C178R has been determined using PCR and direct DNA sequencing. A total of 403 unrelated healthy individuals (253 Emiratis and 150 Tunisians) were genotyped for the CCR5Δ32 variant using PCR amplification and gel electrophoresis. In addition, 200 Emiratis have been screened for other SNPs using Sanger DNA sequencing. Results: Among Emiratis, the allele frequency of the CCR5Δ32 variant has been found to be 0.002. In addition, two variants L55Q and A159 were found at a frequency of 0.002.Moreover, the prevalence of the CCR5Δ32 variant in Tunisians was estimated to be 0.013 which is relatively higher than its frequency in Emiratis but lower than Europeans. Conclusion: We conclude that the allele frequency of the most critical CCR5 polymorphism (Δ32) is extremely low among Emiratis compared to other Arabs and North Europeans. In addition, very low allele frequencies of other CCR5 polymorphisms have been detected among Emiratis.

Keywords: chemokine receptors, CCR5Δ32, CCR5 polymorphisms, Emiratis, Arab populations

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Authors: Tabassum Yasmin, Hamid Pahlevan

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HIV/Hepatitis C co-infection treatments have evolved substantially and they have similar sustained virologic response rates as those of Hepatitis C monoinfected population. There are a few studies on therapy of patients with dual genotypes, especially in HIV/Hepatic C coinfected group. Most studies portrayed case reports of dual genotype chronic Hepatitis C coinfection treatment with Sofosbuvir/Ledipasvir and Ribavirin. A 79-year-old male with a history of HIV on Truvada and Isentress had chronic Hepatitis C with 1a and 2 genotypes. The patient has a history of alcohol intake for 40 years but recently stopped drinking alcohol. He has a history of intravenous drug use in the past and currently is not using any recreational drugs. Patient has Fibro score of 0.7 with Metavir score F2 to F4. AFP is 3.2. The HCV RNA is 493,034 IU/ML. The HBV viral DNA is < 1.30 (not detected). The CD4 is 687CU/MM. The FIB 4 is 3.34 with APRI index 0.717. The HIV viral load is 101 copies/ML. MRI abdomen did not show any liver abnormality. Fixed dose Sofosbuvir/Ledipasvir was used for therapy without Ribavirin. He tolerated medication except for some minor gastrointestinal side effects like abdominal bloating. He demonstrated 100% adherence rate. Patient completed 12 weeks of therapy. HCV RNA was undetectable at 4 and 12 weeks. He achieved SVR at week 12 and subsequently had undetectable RNA for 2 years. Dual genotype prevalence in chronic hepatitis C population is rare, especially in HIV/hepatic coinfection. Our case demonstrates that dual genotypic cases can still be successfully treated with Direct Acting Antiviral agents. The newer agents for therapy for pan genotypes were not available at the time the patient was being treated. We demonstrated that dual agent therapy was still able to maintain SVR in our patient.

Keywords: HIV/Hepatitis C, SVR (sustained virologic response), DAA (direct active antiviral agents, dual genotype

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Authors: F. Pal-Fam, S. Keszthelyi

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Phytosanitary condition of sunflower (Helianthus annuus L.) was endangered by several phytopathogenic agents, mainly microfungi, such as Sclerotinia sclerotiorum, Diaporthe helianthi, Plasmopara halstedtii, Macrophomina phaseolina and so on. There are more agrotechnical and chemical technologies against them, for instance, tolerant hybrids, crop rotations and eventually several in-crop chemical treatments. There are different fungicide treatment methods in sunflower in Hungarian agricultural practice in the quest of obtaining healthy and economic plant products. Besides, there are many choices of useable active ingredients in Hungarian sunflower protection. This study carried out into the examination of the effect of five different fungicide active substances (found on the market) and three different application modes (early; late; and early and late treatments) in a total number of 9 sample plots, 0.1 ha each other. Five successive vegetation periods have been investigated in long term, between 2013 and 2017. The treatments were: 1)untreated control; 2) boscalid and dimoxystrobin late treatment (July); 3) boscalid and dimoxystrobin early treatment (June); 4) picoxystrobin and cyproconazole early treatment; 5) picoxystrobin and cymoxanil and famoxadone early treatment; 6) picoxystrobin and cyproconazole early; cymoxanil and famoxadone late treatments; 7) picoxystrobin and cyproconazole early; picoxystrobin and cymoxanil and famoxadone late treatments; 8) trifloxystrobin and cyproconazole early treatment; and 9) trifloxystrobin and cyproconazole both early and late treatments. Due to the very different yearly weather conditions different phytopathogenic fungi were dominant in the particular years: Diaporthe and Alternaria in 2013; Alternaria and Sclerotinia in 2014 and 2015; Alternaria, Sclerotinia and Diaporthe in 2016; and Alternaria in 2017. As a result of treatments ‘infection frequency’ and ‘infestation rate’ showed a significant decrease compared to the control plot. There were no significant differences between the efficacies of the different fungicide mixes; all were almost the same effective against the phytopathogenic fungi. The most dangerous Sclerotinia infection was practically eliminated in all of the treatments. Among the single treatments, the late treatment realised in July was the less efficient, followed by the early treatments effectuated in June. The most efficient was the double treatments realised in both June and July, resulting 70-80% decrease of the infection frequency, respectively 75-90% decrease of the infestation rate, comparing with the control plot in the particular years. The lowest yield quantity was observed in the control plot, followed by the late single treatment. The yield of the early single treatments was higher, while the double treatments showed the highest yield quantities (18.3-22.5% higher than the control plot in particular years). In total, according to our five years investigation, the most effective application mode is the double in-crop treatment per vegetation time, which is reflected by the yield surplus.

Keywords: fungicides, treatments, phytopathogens, sunflower

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Authors: Nurgül Şenol, Melda Azman

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In this study, gastrointestinal immunohistochemistry in the Vimba vimba and the localization of CCK/gastrin were determined. Although there are a number of studies which relate to the gastrointestinal histochemistry and the localization of the peptides, a literature research in this field revealed that no histochemical or immunohistochemical study covering also the species had been found in our country. In this research, species will be provided from Vimba vimba located in Eğirdir lake. Stomach samples and intestinal samples of these fish will be exposed to routine histological tissue process, embedded in paraffin blocks, and 5-6 μ -thick sections will be taken. Using the PAP (Peroxidase anti-peroxidase) method, localization of the peptides CCK/gastrin was to be found. The densities of peptides of this species were compared, and then the findings obtained were to be evaluated through the statistical analysis methods (SPSS). Endocrine cells reactive to gastrin/CCK antiserum were demonstrated in the stomach and intestinal mucosa. There is a significant difference between gastrin and CCK when compared to regions.

Keywords: CCK, gastrin, immunoreactivity, vimba vimba

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Authors: Aschalew Cherie Workneh, K. S. Hari Prasad, Chandra Shekhar Prasad Ojha

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Due to the increase in water scarcity, the crop water stress index (CWSI) is receiving significant attention these days, especially in arid and semiarid regions, for quantifying water stress and effective irrigation scheduling. Nowadays, machine learning techniques such as neural networks are being widely used to determine CWSI. In the present study, the performance of two artificial neural networks, namely, Self-Organizing Maps (SOM) and Feed Forward-Back Propagation Artificial Neural Networks (FF-BP-ANN), are compared while determining the CWSI of rice crop. Irrigation field experiments with varying degrees of irrigation were conducted at the irrigation field laboratory of the Indian Institute of Technology, Roorkee, during the growing season of the rice crop. The CWSI of rice was computed empirically by measuring key meteorological variables (relative humidity, air temperature, wind speed, and canopy temperature) and crop parameters (crop height and root depth). The empirically computed CWSI was compared with SOM and FF-BP-ANN predicted CWSI. The upper and lower CWSI baselines are computed using multiple regression analysis. The regression analysis showed that the lower CWSI baseline for rice is a function of crop height (h), air vapor pressure deficit (AVPD), and wind speed (u), whereas the upper CWSI baseline is a function of crop height (h) and wind speed (u). The performance of SOM and FF-BP-ANN were compared by computing Nash-Sutcliffe efficiency (NSE), index of agreement (d), root mean squared error (RMSE), and coefficient of correlation (R²). It is found that FF-BP-ANN performs better than SOM while predicting the CWSI of rice crops.

Keywords: artificial neural networks; crop water stress index; canopy temperature, prediction capability

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Authors: Abdullah Özköse, Ahmet Tamkoç

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This study aimed to determine some plant characteristics of perennial ryegrass (Lolium perenne L.) genotypes collected from the natural flora of Ankara and correlations between these characteristics. In order to evaluate for breeding purposes according to Turkey's environmental conditions, perennial ryegrass plants collected from natural pasture of Ankara at 2004 were utilized. The collected seeds of plants were sown in pots and seedlings were prepared in greenhouse. Seedlings were transplanted to the experimental field at 50x50 cm intervals in Randomized Complete Blocks Design in 2005. Data were obtained from the observations and measurements of 568 perennial ryegrasses in 2007 and 2008. Perennial ryegrass plants’ in the spring re-growth time, color, density, growth habit, tendency to inflorescences, time of inflorescence, plant height, length of upper internode, spike length, leaf length, leaf width, leaf area, leaf shape, number of spikelets per spike, seed yield per spike, and 1000 grain weight were investigated and correlation analyses were made on the data. Correlation coefficients were estimated between all paired combinations of the traits. The yield components exhibited varying trends of association among themselves. Seed yield per spike showed significant and positive association with number of spikelets per spike, 1000 grain weight, plant height, length of upper internode, spike length, leaf length, leaf width, leaf area and color, but significant and negative association with growth habit and in the spring re-growth time spring.

Keywords: correlation, morphological traits, Lolium perenne

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Authors: Eman Helal, Ahmed M. Esmat

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Statement of problem: The development of computer-aided design/computer-aided manufacturing (CAD/CAM) resin dentures has simplified complete denture production. Most of the studies evaluated the mechanical properties of the material, but the hygienic performance of the CAD /CAM denture and their ability to maintain clean surfaces and minimize bacterial accumulation is still lacking. Purpose evaluation of the antibacterial characteristics of the 3D printed CAD/CAM denture and to compare it with the conventional heat polymerized acrylic denture base material. Methodology a total of thirty completely edentulous patients grouped randomly into two groups (Group I: Control group) received conventional heat polymerized acrylic resin complete dentures, (Group II: Test group) received 3D printed (CAD/CAM) dentures (stereolithographic PMMA), Samples of Candida albicans culture swabs were taken after 1 month and 3 months of dentures` insertion. A culture swab was obtained by scrubbing the fitting surface of the upper denture. At each time interval, three swab samples were collected from each patient and were inoculated in three individual culture media. Results: there was a significant difference in the colonization of Candida albicans to the fitting surface of the dentures between both groups (Group I: Conventional denture cases) exhibited more adhesion of Candida Albicans to the fitting surface than did (Group II: CAD/CAM cases) (P<0.05). Conclusion: 3D printed CAD/CAM complete denture showed minimal Candida adherence upon upper denture fitting compared to conventional heat-polymerized acrylic resin, which contributes to decreasing the incidence of denture stomatitis which is considered one of the most common problems among complete denture wearers.

Keywords: CAD/CAM denture, completely edentulous, elderly patients, 3D printing, antimicrobial efficiency, conventional denture, PMMA, Candida Albicans, denture stomatitis

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Authors: Paresh Patel, Harshit Patel

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Superficial fungal skin infection is among the most common skin disease. The drug administration through skin has received attention due to several advantages: Avoidance of significant pre-systemic metabolism, drug levels within the therapeutic window, drugs with short biological half-lives, decreased side effects, the non-invasive character, and very high acceptance.

Keywords: transdermal spray, ketoconazole, Eudragit® RLPO, therapeutic window

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Authors: Numfon Rakkhumkaew, Takeru Kawasaki, Makoto Fujie, Takashi Yamada

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Chlorella viruses or chloroviruses contain a gene that encodes a function for chitin synthesis, which is expressed early in viral infection to produce chitin polysaccharide, a polymer of β-1, 4-linked GlcNAc, on the outside of Chlorella cell wall. Interestingly, chlorovirus system is an eco-friendly system which converses CO2 and solar energy from the environment into useful materials. However, infected Chlorella cells are lysed at the final stage of viral infection, and this phenomenon is caused the breaking down of polysaccharide. To postpone the lysing period and prolong the synthesis of chitin polysaccharide on cells, the slow growing virus incorporated with aphidicolin treatment, an inhibitor of DNA synthesis, was investigated. In this study, a total of 25 virus isolates from water samples in Japan region were analyzed for CHS (the gene for CH synthase) gene by PCR (polymerase chain reaction). The accumulation and appearance of chitin polysaccharide on infected cells were detected by biotinylated chitin-binding proteins WGA (wheat germ agglutinin)-biotin for chitin in conjunction with avidin-Cy 2 or Cy 3 and investigated by fluorescence microscopy, observed as green or yellow fluorescence over the cell surface. Among all chlorovirus isolates, cells infected with CNF1 revealed the accumulation of chitin over the cell surface within 30 min p.i. and continued to accumulate on cells until 4 h p.i. before cell lyses which was 1.6 times longer accumulation period than cells infected with CVK2 (prototype virus). Furthermore, addition of aphidicolin could extend the chitin accumulation on cells infected with CNF1 until 8 h p.i. before cell lyses. Whereas, CVK2-infected cells treated with aphidicolin could prolong the chitin synthesis only for 6 h p.i. before cell lyses. Therefore, chitin synthesis by Chlorella-virus system could be prolonged by using slow-growing viral isolates and with aphidicolin.

Keywords: chitin, chlorovirus, Chlorella virus, aphidicolin

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Authors: Thavasimuthu Citarasu, Mariavincent Michaelbabu, Vikram Vakharia

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The rhizome of Java grass, Cyperus rotundus was extracted different organic polar and non-polar solvents and performed the in vitro antiviral and immunostimulant activities against White Spot Syndrome Virus (WSSV) and Vibrio harveyi respectively. Based on the initial screening the ethyl acetate extract of C. rotundus was strong activities and further it was purified through silica column chromatography and the fractions were screened again for antiviral and immunostimulant activity. Among the different fractions screened against the WSSV and V. harveyi, the fractions, F-III to FV had strong activities. In order to study the in vivo influence of C. rotundus, the fractions (F-III to FV) were pooled and delivered to the F. indicus through artificial feed for 30 days. After the feeding trail the experimental and control diet fed F. indicus were challenged with virulent WSSV and studied the survival, molecular diagnosis, biochemical, haematological and immunological parameters. Surprisingly, the pooled fractions (F-III to FV) incorporated diets helped to significantly (P < 0.01) suppressed viral multiplication, showed significant (P < 0.01) differences in protein and glucose levels, improved total haemocyte count (THC), coagulase activity, significantly increased (P < =0.001) prophenol oxidase and intracellular superoxide anion production compared to the control shrimps. Based on the results, C. rotundus extracts effectively suppressed WSSV multiplication and improve the immune system in F. indicus against WSSV infection and this knowledge will helps to develop novel drugs from C. rotundus against WSSV.

Keywords: antiviral drugs, cyperus rotundus, fenneropenaeus indicus, WSSV

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Authors: Rohin Rameswarapu, Sameer Valsangkar

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Background: Globally, musculoskeletal disorders are the largest single cause of work-related illnesses accounting for over 33% of all newly reported occupational illnesses. Risk factors for MSD need to be delineated to suggest means for amelioration. Material and methods: In this current cross-sectional study, the prevalence of MSDs among workers in an electrical company assembly line, the socio-demographic and job characteristics associated with MSD were obtained through a semi-structured questionnaire. A quantitative assessment of the physical risk factors through the Rapid Upper Limb Assessment (RULA) tool, and measurement of psychosocial risk factors through a Likert scale was obtained. Statistical analysis was conducted using Epi-info software and descriptive and inferential statistics including chi-square and unpaired t test were obtained. Results: A total of 263 workers consented and participated in the study. Among these workers, 200 (76%) suffered from MSD. Most of the workers were aged between 18–27 years and majority of the workers were women with 198 (75.2%) of the 263 workers being women. A chi square test was significant for association between male gender and MSD with a P value of 0.007. Among the MSD positive group, 4 (2%) had a grand score of 5, 10 (5%) had a grand score of 6 and 186 (93%) had a grand score of 7 on RULA. There were significant differences between the non-MSD and MSD group on five out of the seven psychosocial domains, namely job demand, job monotony, co-worker support, decision control and family and environment domains. Discussion: The current cross-sectional study demonstrates a high prevalence of MSD among assembly line works with inherent physical and psychosocial risk factors and recommends that not only physical risk factors, addressing psychosocial risk factors through proper ergonomic means is also essential to the well-being of the employee.

Keywords: musculoskeletal disorders, India, occupational health, Rapid Upper Limb Assessment (RULA)

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Authors: Sunghyoun Lee, Byeongkee Yu, Chanjung Lee, Yeongtaek Lim

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Oyster mushrooms are one of the ingredients that Koreans prefer. The oyster mushroom cultivation house has multiple layers in order to increase the mushroom production per unit area. However, the growing shelves in the house act as obstacles and hinder the circulation of the interior air, which leads to the difference of cultivation environment between the upper part and lower part of the growing shelves. Due to this difference of environments, growth distinction occurs according to the area of the growing shelves. It is known that minute air circulation around the mushroom cap facilitates the metabolism of mushrooms and improves its quality. This study has utilized the computational fluid dynamics (CFD) program, that is, FLUENT R16, in order to analyze the improvement of the internal environment uniformity of the oyster mushroom cultivation house. The analyzed factors are velocity distribution, temperature distribution, and humidity distribution. In order to maintain the internal environment uniformity of the oyster mushroom cultivation house, it appeared that installing circulation fan at the upper part of the working passage towards the ceiling is effective. When all the environmental control equipment – unit cooler, inlet fan, outlet fan, air circulation fan, and humidifier - operated simultaneously, the RMS figure on the growing shelves appeared as follows: velocity 28.23%, temperature 30.47%, humidity 7.88%. However, when only unit cooler and air circulation fan operated, the RMS figure on the growing shelves appeared as follows: velocity 22.28%, temperature 0.87%, humidity 0.82%. Therefore, in order to maintain the internal environment uniformity of the mushroom cultivation house, reducing the overall operating time of inlet fan, outlet fan, and humidifier is needed, and managing the internal environment with unit cooler and air circulation fan appropriately is essential.

Keywords: air circulation fan, computational fluid dynamics, multi-layered shelves cultivation, oyster mushroom cultivation house

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Authors: Yiseyon Sunday Hosu, Motebang Dominic Vincent Nakin, Elphina N. Cishe

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Universal access to clean and safe drinking water and basic sanitation is one of the targets of the 6th Sustainable Development Goals (SDGs). This paper explores the evidence-based indicators of Water Rights Acts (2013) among households in the rural communities in the Mthatha River catchment of OR Tambo District Municipality of South Africa. Daily access to minimum 25 litres/person and the factors influencing clean water access were investigated in the catchment. A total number of 420 households were surveyed in the upper, peri-urban, lower and coastal regions of Mthatha Rivier catchment. Descriptive and logistic regression analyses were conducted on the data collected from the households to elicit vital information on domestic water security among rural community dwellers. The results show that approximately 68 percent of total households surveyed have access to the required minimum 25 litre/person/day, with 66.3 percent in upper region, 76 per cent in the peri-urban, 1.1 percent in the lower and 2.3 percent in the coastal regions. Only 30 percent among the total surveyed households had access to piped water either in the house or public taps. The logistic regression showed that access to clean water was influenced by lack of water infrastructure, proximity to urban regions, daily flow of pipe-borne water, household size and distance to public taps. This paper recommends that viable integrated rural community-based water infrastructure provision strategies between NGOs and local authority and the promotion of point of use (POU) technologies to enhance better access to clean water.

Keywords: domestic water, household technology, water security, rural community

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Authors: Celine Davis

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The novel coronavirus 2019 (COVID-19) is an infectious disease caused by the virus SARS-CoV-2, which has detrimental respiratory, cardiovascular, and neurological effects impacting over one million lives in the United States. New researches has emerged indicating long-term neurologic consequences in those who survive COVID-19 infections, including more than seven million Americans and another 27 million people worldwide. These consequences include attentional deficits, memory impairments, executive function deficits and aphasia-like symptoms which fall within the purview of speech-language pathology. The National Health Interview Survey (NHIS) is a comprehensive annual survey conducted by the National Center for Health Statistics (NCHS), a branch of the Centers for Disease Control and Prevention (CDC) in the United States. The NHIS is one of the most significant sources of health-related data in the country and has been conducted since 1957. The longitudinal nature of the study allows for analysis of trends in various variables over the years, which can be essential for understanding societal changes and making treatment recommendations. This current study will utilize NHIS data from 2020-2022 which contained interview questions specifically related to COVID-19. Adult cases of individuals between the ages of 18-50 diagnosed with COVID-19 in the United States during 2020-2022 will be identified using the National Health Interview Survey (NHIS). Multiple regression analysis of self-reported data confirming COVID-19 infection status and challenges with concentration, communication, and memory will be performed. Latent class analysis will be utilized to identify subgroups in the population to indicate whether certain demographic groups have higher susceptibility to cognitive-linguistic deficits associated with COVID-19. Completion of this study will reveal whether there is an association between confirmed COVID-19 diagnosis and heightened incidence of cognitive deficits and subsequent implications, if any, on activities of daily living. This study is distinct in its aim to utilize national survey data to explore the relationship between confirmed COVID-19 diagnosis and the prevalence of cognitive-communication deficits with a secondary focus on resulting activity limitations. To the best of the author’s knowledge, this will be the first large-scale epidemiological study investigating the associations between cognitive-linguistic deficits, COVID-19 and implications on activities of daily living in the United States population. These findings will highlight the need for targeted interventions and support services to address the cognitive-communication needs of individuals recovering from COVID-19, thereby enhancing their overall well-being and functional outcomes.

Keywords: cognition, COVID-19, language, limitations, memory, NHIS

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Authors: Nazish Badar

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Background: To investigate the epidemic resurgent wave of influenza A (H1N1) pdm09 infections during 2015-16 Influenza season(Nov,15 –Feb,16) we compared epidemiological features of influenza A (H1N1) pdm09 associated hospitalizations and deaths during this period in Pakistan. Methods: Respiratory samples were tested using CDC Real-Time RT-PCR protocols. Demographic and epidemiological data was analyzed using SPSS. Risk ratio was calculated between age groups to compare patients that were hospitalized and died due to influenza A (H1N1) pdm09 during this period. Results: A total of 1970 specimens were analyzed; influenza virus was detected in 494(25%) samples, including 458(93%) Influenza type A and 36(7%) influenza type B viruses. Amongst influenza A viruses, 351(77%) A(H1N1) pdm09 and 107(23%) were A/H3N2. Influenza A(H1N1)pdm09 peaked in January 2016 when 250(54%) of tested patients were positive. The resurgent waves increased hospitalizations due to pdmH1N1 as compared to the rest part of the year. Overall 267(76%) A(H1N1) pdm09 cases were hospitalized. Adults ≥18 years showed the highest relative risk of hospitalization (1.2). Median interval of hospitalization and symptom onset was five days for all age groups. During this period, a total of 34 laboratory-confirmed deaths associated with pandemic influenza A (H1N1) were reported out of 1970 cases, the case fatality rate was 1.72%. the male to female ratio was 2:1in reported deaths. The majority of the deaths during that period occurred in adults ≥18 years of age. Overall median age of the death cases was 42.8 years with underlying medical conditions. The median number of days between symptom onset was two days. The diagnosis upon admission in influenza-associated fatal cases was pneumonia (53%). Acute Respiratory Distress Syndrome 9 (26%), eight out of which (88%) required mechanical ventilation. Conclusions: The present resurgence of pandemic virus cannot be attributed to a single factor. The prolong cold and dry weather, possibility of drift in virus and absence of annual flu vaccination may have played an integrated role in resurfacing of pandemic virus.

Keywords: influenza A (H1N1)pdm 09, resurgence, epidemiology, Pakistan

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Authors: Maria L. G. Lourenco, Viviane Y. Hibaru, Keylla H. N. P. Pereira, Fabiana F. Souza, Joao C. P. Ferreira, Simone B. Chiacchio, Luiz H. A. Machado

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Neonatal conjunctivitis, or ophthalmia, is an infection of the conjunctiva or cornea before opening the eyelids. It is believed that immunodeficiency contributes to the development of the condition. This study aims at reporting a case of ophthalmia neonatorum in a dog, in addition to its diagnosis and treatment. A litter of five pug neonates was admitted to the Sao Paulo State University (UNESP) Veterinary Hospital, Botucatu, Sao Paulo, Brazil, with complaints of ocular secretion. The neonates were five days old. The clinical examination revealed that three newborns presented swelling in the ocular region and a purulent secretion in the medial corner of the eye that was exerting pressure on the ocular globes, which are compatible with the description of this disease. The diagnosis was made based on the clinical signs and bacterial culture of the secretion, which revealed the presence of bacteria belonging to the genus Staphylococcus sp. The laboratory assays did not reveal any alterations. The treatment was instituted gently, opening the eyelids early and cleaning the purulent ocular secretion with saline solution. An ophthalmic ointment with retinol, amino acids, methionine, and chloramphenicol (Epitezan®) was prescribed four times a day for seven days. Blood plasma (2 mL/100 g) was administered subcutaneously because bacterial infections in neonates may represent a failure in the transference of passive immunity. A more thorough cleaning of the environment was also recommended. Neonatal conjunctivitis has a simple diagnosis and treatment. If not treated early, it can evolve to adherence of the eyelids to the cornea, ulceration, and perforation of the cornea. Therefore, the prognosis is favorable as long as the condition is diagnosed early, and the treatment is instituted quickly.

Keywords: ophthalmia neonatorum, neonatal infection, puppy, newborn

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Authors: Michal Kielbik, Izabela Szulc-Kielbik, Anna Brzostek, Jaroslaw Dziadek, Magdalena Klink

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The goal of many research groups in the world is to find new components that are important for survival of mycobacteria in the host cells. Mycobacterium tuberculosis (Mtb) possesses a number of enzymes degrading cholesterol that are considered to be an important factor for its survival and persistence in host macrophages. One of them - cholesterol oxidase (ChoD), although not being essential for cholesterol degradation, is discussed as a virulence compound, however its involvement in macrophages’ response to Mtb is still not sufficiently determined. The recognition of tubercle bacilli antigens by pathogen recognition receptors is crucial for the initiation of the host innate immune response. An important receptor that has been implicated in the recognition and/or uptake of Mtb is Toll-like receptor type 2 (TLR2). Engagement of TLR2 results in the activation and phosphorylation of intracellular signaling proteins including IRAK-1 and -4, TRAF-6, which in turn leads to the activation of target kinases and transcription factors responsible for bactericidal and pro-inflammatory response of macrophages. The aim of these studies was a detailed clarification of the role of Mtb cholesterol oxidase as a virulence factor affecting the TLR2 signaling pathway in human macrophages. As human macrophages the THP-1 differentiated cells were applied. The virulent wild-type Mtb strain (H37Rv), its mutant lacking a functional copy of gene encoding cholesterol oxidase (∆choD), as well as complimented strain (∆choD–choD) were used. We tested the impact of Mtb strains on the expression of TLR2-depended signaling proteins (mRNA level, cytosolic level and phosphorylation status). The cytokine and bactericidal response of THP-1 derived macrophages infected with Mtb strains in relation to TLR2 signaling pathway dependence was also determined. We found that during the 24-hours of infection process the wild-type and complemented Mtb significantly reduced the cytosolic level and phosphorylation status of IRAK-4 and TRAF-6 proteins in macrophages, that was not observed in the case of ΔchoD mutant. Decreasement of TLR2-dependent signaling proteins, induced by wild-type Mtb, was not dependent on the activity of proteasome. Blocking of TLR2 expression, before infection, effectively prevented the induced by wild-type strain reduction of cytosolic level and phosphorylation of IRAK-4. None of the strains affected the surface expression of TLR2. The mRNA level of IRAK-4 and TRAF-6 genes were significantly increased in macrophages 24 hours post-infection with either of tested strains. However, the impact of wild-type Mtb strain on both examined genes was significantly stronger than its ΔchoD mutant. We also found that wild-type strain stimulated macrophages to release high amount of immunosuppressive IL-10, accompanied by low amount of pro-inflammatory IL-8 and bactericidal nitric oxide in comparison to mutant lacking cholesterol oxidase. The influence of wild-type Mtb on this type of macrophages' response strongly dependent on fully active IRAK-1 and IRAK-4 signaling proteins. In conclusion, Mtb using cholesterol oxidase causes the over-activation of TLR2 signaling proteins leading to the reduction of their cytosolic level and activity resulting in the modulation of macrophages response to allow its intracellular survival. Supported by grant: 2014/15/B/NZ6/01565, National Science Center, Poland

Keywords: Mycobacterium tuberculosis, cholesterol oxidase, macrophages, TLR2-dependent signaling pathway

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