Phosphorus Accumulation in Extracellular Polymeric Substances (EPS) of Colony-Forming Cyanobacteria Challenges Imbalanced Nutrient Reduction Strategies in Eutrophic Lakes.

Extracellular polymeric substances (EPS) are crucial for cyanobacterial proliferation; however, certain queries, including how EPS affects cellular nutrient processes and what are the implications for nutrient management in lakes, are not well documented. Here, the dynamics of cyanobacterial EPS-associated phosphorus (EPS-P) were examined both in a shallow eutrophic lake (Lake Taihu, China) and in laboratory experiments with respect to nitrogen (N) and phosphorus (P) availability. Results indicated that 40-65% of the total cyanobacterial aggregate/particulate P presented as EPS-P (mainly labile P and Fe/Al-P). Phosphorus-starved cyanobacteria rapidly replenished their EPS-P pools after the P was resupplied, and the P concentration in this pool was stable for long afterward, although the environmental P concentration decreased dramatically. A low-N treatment enhanced the EPS production alongside two-fold EPS-P accumulation (particularly labile P) higher than the control. Such patterns occurred in the lake where EPS and EPS-P contents were high under N limitation. EPS-P enrichment increased the P content in cyanobacteria; subsequently, it could hold the total P concentration higher for longer and make bloom mitigation harder. The findings outline a new insight into EPS functions in the P process of cyanobacterial aggregates and encourage consideration of both N and P reductions in nutrient management.

KLF7 promotes pancreatic cancer growth and metastasis by up-regulating ISG expression and maintaining Golgi complex integrity.

Pancreatic ductal adenocarcinoma (PDAC) is an aggressive cancer with a dismal prognosis. Currently, there is no effective therapy for PDAC, and a detailed molecular and functional evaluation of PDACs is needed to identify and develop better therapeutic strategies. Here we show that the transcription factor Krüppel-like factor 7 (KLF7) is overexpressed in PDACs, and that inhibition of KLF7 blocks PDAC tumor growth and metastasis in cell culture and in mice. KLF7 expression in PDACs can be up-regulated due to activation of a MAP kinase pathway or inactivation of the tumor suppressor p53, two alterations that occur in a large majority of PDACs. ShRNA-mediated knockdown of KLF7 inhibits the expression of IFN-stimulated genes (ISGs), which are necessary for KLF7-mediated PDAC tumor growth and metastasis. KLF7 knockdown also results in the down-regulation of Discs Large MAGUK Scaffold Protein 3 (DLG3), resulting in Golgi complex fragmentation, and reduced protein glycosylation, leading to reduced secretion of cancer-promoting growth factors, such as chemokines. Genetic or pharmacologic activation of Golgi complex fragmentation blocks PDAC growth and metastasis similar to KLF7 inhibition. Our results demonstrate a therapeutically amenable, KLF7-driven pathway that promotes PDAC growth and metastasis by activating ISGs and maintaining Golgi complex integrity.

Comparative study of Amsel's criteria and Nugent scoring for diagnosis of bacterial vaginosis in a tertiary care hospital, Nepal.

BACKGROUND: The most common pathological cause of abnormal vaginal discharge in reproductive-aged women is bacterial vaginosis (BV). Amsel's criteria and Nugent scoring systems are commonly employed approaches for the diagnosis of BV. Despite the Nugent scoring system being the gold standard method for diagnosing BV, Amsel's criteria are generally preferred in clinical setup owing to the fact Nugent scoring requires considerable time and expert microscopist. This study was conducted to determine the diagnostic value of Amsel's criteria by comparing it with the Nugent scoring system. METHODS: This was a descriptive cross-sectional study conducted at Tribhuvan University Teaching Hospital, Nepal from October 2016 to September 2017. Vaginal specimens were collected from a total of 141 women presenting with abnormal vaginal discharge. The sensitivity, specificity, positive predictive value, and negative predictive value of Amsel's criteria were calculated, and each component of Amsel's criteria was compared to the Nugent scoring system. RESULTS: The sensitivity, specificity, positive predictive value, and negative predictive value of Amsel's criteria were 50%, 98.2%, 87.5%, and 88.8% respectively. The clue cells showed 100% specificity and vaginal discharge with pH > 4.5 had 89.3% sensitivity while compared with Nugent's scoring system. CONCLUSIONS: Amsel's criteria can be used as an adjunct method to Nugent scoring for the diagnosis of BV in the hands of skilled manpower in resources limited countries. The presence of clue cell and positive whiff test of Amsel's criteria shows good match with Nugent's score.

Diagnostic evaluation of IgM ELISA and IgM Immunofluorescence assay for the diagnosis of Acute Scrub Typhus in central Nepal.

BACKGROUND: Scrub typhus is an acute febrile illness caused by the obligate intracellular bacterium, Orientia tsutsugamushi. Immunochromatography (ICT) and IgM ELISA are two of the routinely employed antibody based assays for diagnosis of Scrub typhus fever in Nepal, although the recommended gold standard diagnostic test is IgM Immunofluorescence assay (IFA). This study evaluated InBios Scrub Typhus Detect™ Immunoglobulin M (IgM) ELISA and IgM Immunofluorescence assays in single serum sample at the time of admission. METHOD: Study participants (1585 suspected cases), were enrolled based on acute febrile illness with suspected scrub typhus cases in central Nepal. Blood sample was collected from the suspected patients of scrub typhus, presenting with acute febrile illness. IgM antibody to Orientia tsusugamushi was detected by using Scrub Typhus Detect™ Kit and an in-house IgM IFA. The IFA assay was performed with the Gilliam, Karp, Kato strains and O. chuto antigens following the ARRL protocol. RESULT: Statistical analysis of IgM ELISA results when compared to reference test, IgM IFA results demonstrated the following characteristics, sensitivity 84.0% (95%CI: 79.73-87.68%), specificity 94.82% (95% CI: 93.43-95.99%), positive likelihood ratio 16.21% (95% CI: 12.71-20.67%), negative likelihood ratio 0.17% (95% CI: 0.13-0.21%), disease prevalence 22.08% (95% CI: 20.06 -24.21%), positive predictive value 82.12% (95% CI: 78.28-85.42%) and negative predictive value 95.44% (95% CI: 94.27-96.38%) respectively. CONCLUSION: Although IgM IFA is considered the gold standard test for the diagnosis of scrub typhus cases, it is relatively expensive, requires trained personal and a microscope with fluorescence filters. Scrub typhus IgM ELISA may be the best alternative test and possible viable option for resource limited endemic countries like Nepal.

Bacteria on Medical Professionals' White Coats in a University Hospital.

The transient contamination of medical professional's attires including white coats is one of the major vehicles for the horizontal transmission of microorganisms in the hospital environment. This study was carried out to determine the degree of contamination by bacterial agents on the white coats in a tertiary care hospital in Nepal. Sterilized uniforms with fabric patches of 10 cm × 15 cm size attached to the right and left pockets were distributed to 12 nurses of six different wards of a teaching hospital at the beginning of their work shift. Worn coats were collected at the end of the shifts and the patches were subjected for total bacterial count and identification of selected bacterial pathogens, as prioritized by the World Health Organization (WHO). Fifty percent of the sampled swatches were found to be contaminated by pathogenic bacteria. The average colony growth per square inch of the patch was 524 and 857 during first and second workdays, respectively, indicating an increase of 63.6% in colony counts. The pathogens detected on patches were Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, and Acinetobacter sp. Additional bacteria identified included Bacillus sp., Micrococcus sp., and coagulase-negative staphylococci (CoNS). The nurses working in the maternity department had their white coats highly contaminated with bacteria. On the other hand, the least bacterial contamination was recorded from the nurses of the surgery ward. One S. aureus isolate from the maternity ward was resistant to methicillin. This study showed that pathogens belonging to the WHO list of critical priority and high priority have been isolated from white coats of nurses, thus posing the risk of transmission to patients. White coats must be worn, maintained, and washed properly to reduce bacterial contamination load and to prevent cross-contamination of potential superbugs. The practice of wearing white coats outside the healthcare zone should be strictly discouraged.

Acute gastroenteritis associated with Rotavirus A among children less than 5 years of age in Nepal.

BACKGROUND: Rotavirus gastroenteritis is a major public health problem in Nepal. This study was conducted to obtain information associated with Rotavirus gastroenteritis and to perform genotyping of Rotavirus A. METHODS: Hospital based cross sectional study was conducted from January to December 2017 among children less than 5 years of age attending Kanti Children's Hospital and Tribhuvan University Teaching Hospital. Rotavirus A antigen detection was performed by Enzyme Linked Immunosorbent Assay (ELISA) using ProSpecT Rotavirus Microplate Assay. Rotavirus A positive strains were further confirmed by genotyping using Reverse-Transcription Polymerase Chain Reaction (RT-PCR). RESULTS: A total of 1074 stool samples were collected, of them 770 were hospitalized, and 304 were non-hospitalized cases. Rotavirus A infection was found in 28% of children with infection rate higher in hospitalized (34%) than in non-hospitalized (14%) children. Rotavirus A detection was higher in male (31%) than in female (24%), but this was statistically not significant (p > 0.05). Rotavirus A positivity was higher in children of age group 0-23 months, this result was statistically not significant (p > 0.05) with higher frequency found in the months of November, December, January, February and March (p < 0.05). On the basis of molecular analysis of Rotavirus A genotyping, G12P[6] (46.39%) was found to be the predominant followed by G1P[8] (35.05%), G3P[8] (7.21%) and G1P[6] (5.15%) while 4.12% was mixed infection and 1.03% was partially typed (p < 0.05). CONCLUSION: Rotavirus A infection occurred throughout the year, but the infection was significantly higher during the month of March. The higher frequency of rotavirus infection was observed among children of age group 0-23 months; however this was not found to be statistically significant. In this study, G12P[6] is predominant genotype observed. The results of genotyping are essential for the introduction of Rotavirus vaccine in Nepal.

Effects of Phosphorus on Interspecific Competition between two cell-size Cyanobacteria: Synechococcus sp. and Microcystis aeruginosa.

Pico-cyanobacteria and micro-cyanobacteria coexist ubiquitously in many lakes. Differences in cell size and abilities to utilize nutrients may influence their distribution patterns. In this study, Synechococcus sp. and Microcystis aeruginosa were chosen as pico- and micro-cyanobacteria, respectively. Gradient phosphorus treatments (0.002, 0.01, 0.05, and 0.25 mg P L-1) were designed in mono- and co-cultures. Growth curves were recorded and fitted by the Monod equation. Moreover, the interspecific competition was analyzed by the Lotka-Volterra model. When mono-cultured in lower P conditions (≤ 0.01 mg P L-1), Synechococcus sp. obtained much higher biomass than M. aeruginosa. But, M. aeruginosa grew faster than Synechococcus sp. in higher P groups (≥ 0.05 mg P L-1) (p < 0.05). Synechococcus sp. has abilities to thrive in low-phosphorus environments, whereas M. aeruginosa favored high-phosphorus conditions. In co-cultures, Synechococcus sp. strongly inhibited M. aeruginosa at each P treatment.

Comparison of Four Quantitative Techniques for Monitoring Microalgae Disruption by Low-Frequency Ultrasound and Acoustic Energy Efficiency.

Ultrasound has been regarded as an environmental friendly technology to utilize microalgae biomass and control algal blooms. In this study, four quantitative techniques, including cell counting, optical density of algal suspension, pigments release, and protein release, were performed on three species of microalgae ( M. aeruginosa, C. pyrenoidosa, and C. reinhardtii) to develop effective techniques for rapid monitoring of cell disruption and to optimize the acoustic energy efficiency. Results showed optical density of algal suspensions was not an optimal indicator to monitor cell disruption, although it is a common technique for determining cell concentration in microbial cultures. Instead, an accurate and reliable technique was to determine the release of intracellular pigments (absorbance peaks of supernatant) for indicating cell rupture. The protein released during sonication could also be a useful indicator if it is the component of interest. A fitted power functional model showed a strong relationship between cell disruption and energy consumption ( R2 > 0.87). This model could provide an effective approach to directly compare the energy efficiency of ultrasound in different systems or with varying microalgae species. This study provides valuable information for microalgae utilization and the treatment of algal blooms by ultrasound, so as to achieve energy conservation and environmental safety.

Effects of Low-Frequency Ultrasound on Microcystis aeruginosa from Cell Inactivation to Disruption.

Ultrasound can be used to induce cell resonance and cavitation to inhibit cyanobacterial growth, but it can also lead to increase in dissolved nutrients because of cell disruption. This study investigated the process from cell inactivation to disruption of Microcystis aeruginosa. Algal cells were sonicated (at 35 kHz) under various intensities and durations. Results showed that chlorophyll a content and Fv/Fm values decreased slightly within the first 5 min. Superoxide dismutase activity was stimulated and its peak value appeared at the fifth minute. After 20 min, considerable number of ruptured cells were observed and the concentrations of dissolved nitrogen and phosphorus increased rapidly. Finally, ammonia and nitrate merely composed a small portion of dissolved nitrogen. This study demonstrated that excessive ultrasound treatment can significantly rupture algal cells and lead to the release of cellular inclusions, which may cause ecological issues or public health problems. Based on our findings, ultrasonic intensity controlled at 0.035 W/mL and applied for a duration of 20 min delivers the optimal result in effectively inhibiting physiological activities of Microcystis aeruginosa without marked cell disruption. This will ultimately help to achieve algal control, while conserving energy and preserving the environment and human health.

Molecular epidemiology of Rotavirus causing diarrhea among children less than five years of age visiting national level children hospitals, Nepal.

BACKGROUND: Rotaviruses are the major cause of diarrhea among the infants and young children all over the world causing over 500,000 deaths and 2.4 million hospitalizations each year. In Nepal Rotavirus infection positivity rates ranges from 17.0 to 39.0% among children less than 5 years. However, little is known about the molecular genotypes of Rotavirus prevailing. The objective of this study was to estimate the burden of Rotavirus gastroenteritis and determine the genotypes of Rotavirus among children less than 5 years. METHODS: The cross sectional study was conducted from January to November 2014 among children less than 5 years old visiting Kanti Children's Hospital and Tribhuvan University Teaching Hospital. Rotavirus antigen detection was performed by Enzyme Linked Immunosorbent Assay (ELISA) using ProSpecT Rotavirus Microplate Assay. Among the Rotavirus antigen positive samples, 59 samples were used for Rotavirus RNA extraction. Multiplex PCR was performed to identify G type comprising G1-G4, G8-G10 and G12 and P type comprising P[4], P[6], P[8], P[9], P[10], and P[11]. RESULTS: A total of 717 diarrheal stool samples were collected from patients ranging from 10 days to 59 months of age. Rotavirus antigen positive was found among (N = 164)22.9% of patients. The highest number of the diarrhea was seen in January. Molecular analysis of Rotavirus genotypes revealed that the predominant G-Type was G12 (36%) followed by G9 (31%), G1 (21%), G2 (8.6%). The predominant P- type was P6 (32.8%) followed by P8 (31%), P10 (14.8%), P4 (14.8%). A total of seven G/P type combinations were identified the most common being G12P [6] (35.8%), G1P [8] (15.1%), G9P [8] (15.1%). CONCLUSION: Rotavirus diarrhea is, mostly affecting children from 7 to 24 months in Nepal, mostly occurring in winter. The circulating genotypes in the country are found to be primarily unusual genotypes and predominance of G12P[6]. It is recommended to conduct genotyping of Rotavirus on large samples before starting vaccination in the country.

Monomethyl Auristatin E Phosphate Inhibits Human Prostate Cancer Growth.

BACKGROUND: Bone metastasis from primary prostate cancer leads to markedly diminished quality of life with poor long-term survival. Bone seeking treatment options are limited with adverse consequences on rapidly proliferating tissues such as bone marrow. In the present study, we seek to determine the bone-enriching capabilities of monomethyl auristatin E phosphate (MMAEp), a derivative of the potent antimitotic monomethyl auristatin E (MMAE). METHODS: The in vitro actions and mechanisms of cytotoxicity were assessed using cell viability, immunofluorescence, flow cytometry, and Western blot analysis. In vivo efficacy was determined using an intratibial xenograft mouse model of human prostate cancer and live animal imaging. RESULTS: The half maximal inhibitory concentration (IC50) of MMAE and MMAEp was determined to be approximately 2 and 48 nM, respectively, in PC-3 and C4-2B cell lines. MMAEp retained the mechanism of action of MMAE in reducing microtubule polymerization and stalling cell cycle progression at the G2/M transition. MMAEp was able to bind hydroxyapatite in in vitro assays. MMAEp significantly reduced intratibial tumor growth compared to the vehicle control treatment. CONCLUSIONS: MMAEp is an antimitotic compound that binds to calcium ions in the bone and inhibits prostate tumor growth in the bone. Prostate 76:1420-1430, 2016. © 2016 Wiley Periodicals, Inc.

The alkyllysophospholipid edelfosine enhances TRAIL-mediated apoptosis in gastric cancer cells through death receptor 5 and the mitochondrial pathway.

The ether phospholipid edelfosine is the prototype of a group of synthetic antitumor alkyllysophospholipid (ALP) compounds that exert pro-apoptotic effects in various types of cancer cells through cell type-dependent mechanisms. In this study, we examined the antitumor effect of edelfosine in human gastric cancer cells. Edelfosine decreased cell viability and induced autophagic death at a moderate concentration (~30 µM), whereas it induced apoptotic cell death at concentrations over 30 µM. Interestingly, low concentrations of edelfosine (5-10 µM) effectively enhanced recombinant human tumor necrosis factor (TNF)-related apoptosis-inducing ligand (rhTRAIL/TNFSF10)-induced apoptosis and clonogenicity in gastric cancer cells, including TRAIL-resistant AGS cells. Edelfosine upregulated the protein level of death receptor 5 (DR5/TNFRSF10B) and/or increased DR5 upregulation in lipid rafts. In addition, edelfosine-mediated rhTRAIL sensitization was regulated by the DR5 pathway. Edelfosine also activated p38MAPK (MAPK14), and edelfosine-mediated rhTRAIL sensitization was partially regulated by a p38-mediated decrease in mitochondrial membrane potential. This study suggests a novel therapeutic strategy targeting gastric cancer cells by using the combination of edelfosine and TRAIL.

IL-17 and insulin/IGF1 enhance adhesion of prostate cancer cells to vascular endothelial cells through CD44-VCAM-1 interaction.

BACKGROUND: Extravasation is a critical step in cancer metastasis, in which adhesion of intravascular cancer cells to the vascular endothelial cells is controlled by cell surface adhesion molecules. The role of interleukin-17 (IL-17), insulin, and insulin-like growth factor 1 (IGF1) in adhesion of prostate cancer cells to the vascular endothelial cells is unknown, which is the subject of the present study. METHODS: Human umbilical vein endothelial cells (HUVECs) and human prostate cancer cell lines (PC-3, DU-145, LNCaP, and C4-2B) were analyzed for expression of vascular cell adhesion molecule 1 (VCAM-1), integrins, and cluster of differentiation 44 (CD44) using flow cytometry and Western blot analysis. The effects of IL-17, insulin, and IGF1 on VCAM-1 expression and adhesion of prostate cancer cells to HUVECs were examined. The interaction of VCAM-1 and CD44 was assessed using immunoprecipitation assays. RESULTS: Insulin and IGF1 acted with IL-17 to increase VCAM-1 expression in HUVECs. PC-3, DU-145, LNCaP, and C4-2B cells expressed ß1 integrin but not α4 integrin. CD44 was expressed by PC-3 and DU-145 cells but not by LNCaP or C4-2B cells. When HUVECs were treated with IL-17, insulin or IGF1, particularly with a combination of IL-17 and insulin (or IGF1), adhesion of PC-3 and DU-145 cells to HUVECs was significantly increased. In contrast, adhesion of LNCaP and C4-2B cells to HUVECs was not affected by treatment of HUVECs with IL-17 and/or insulin/IGF1. CD44 expressed in PC-3 cells physically bound to VCAM-1 expressed in HUVECs. CONCLUSIONS: CD44-VCAM-1 interaction mediates the adhesion between prostate cancer cells and HUVECs. IL-17 and insulin/IGF1 enhance adhesion of prostate cancer cells to vascular endothelial cells through increasing VCAM-1 expression in the vascular endothelial cells. These findings suggest that IL-17 may act with insulin/IGF1 to promote prostate cancer metastasis.

Detection of biofilm production and antibiotic resistance pattern in clinical isolates from indwelling medical devices.

Microbial biofilms pose great threat for patients requiring indwelling medical devices (IMDs) as it is difficult to remove them. It is, therefore, crucial to follow an appropriate method for the detection of biofilms. The present study focuses on detection of biofilm formation among the isolates from IMDs. We also aimed to explore the antibiogram of biofilm producers. This prospective analysis included 65 prosthetic samples. After isolation and identification of bacteria following standard methodology, antibiogram of the isolates were produced following Kirby-Bauer disc diffusion method. Detection of biofilms was done by tube adherence (TA), Congo red agar and tissue culture plate (TCP) methods. Out of 67 clinical isolates from IMDs, TCP detected 31 (46.3 %) biofilm producers and 36 (53.7 %) biofilm non-producers. Klebsiella pneumoniae, Pseudomonas aeruginosa and Burkholderia cepacia complex were found to be the most frequent biofilm producers. The TA method correlated well with the TCP method for biofilm detection. Higher antibiotic resistance was observed in biofilm producers than in biofilm non-producers. The most effective antibiotics for biofilm producing Gram-positive isolates were Vancomycin and Tigecycline, and that for biofilm producing Gram-negative isolates were Polymyxin-B, Colistin Sulphate and Tigecycline. Nearly 46 % of the isolates were found to be biofilm producers. The antibiotic susceptibility pattern in the present study showed Amoxicillin to be an ineffective drug for isolates from the IMDs. For the detection of biofilm production, TA method can be an economical and effective alternative to TCP method.

Aminomethylphosphonic acid and methoxyacetic acid induce apoptosis in prostate cancer cells.

Aminomethylphosphonic acid (AMPA) and its parent compound herbicide glyphosate are analogs to glycine, which have been reported to inhibit proliferation and promote apoptosis of cancer cells, but not normal cells. Methoxyacetic acid (MAA) is the active metabolite of ester phthalates widely used in industry as gelling, viscosity and stabilizer; its exposure is associated with developmental and reproductive toxicities in both rodents and humans. MAA has been reported to suppress prostate cancer cell growth by inducing growth arrest and apoptosis. However, it is unknown whether AMPA and MAA can inhibit cancer cell growth. In this study, we found that AMPA and MAA inhibited cell growth in prostate cancer cell lines (LNCaP, C4-2B, PC-3 and DU-145) through induction of apoptosis and cell cycle arrest at the G1 phase. Importantly, the AMPA-induced apoptosis was potentiated with the addition of MAA, which was due to downregulation of the anti-apoptotic gene baculoviral inhibitor of apoptosis protein repeat containing 2 (BIRC2), leading to activation of caspases 7 and 3. These results demonstrate that the combination of AMPA and MAA can promote the apoptosis of prostate cancer cells, suggesting that they can be used as potential therapeutic drugs in the treatment of prostate cancer.

Abscisic acid signaling through LANCL2 and PPARγ induces activation of p38MAPK resulting in dormancy of prostate cancer metastatic cells.

Prostate cancer (PCa) is one the most common malignancies in men. The high incidence of bone metastasis years after primary therapy suggests that disseminated tumor cells must become dormant, but maintain their ability to proliferate in the bone marrow. Abscisic acid (ABA) is a stress response molecule best known for its regulation of seed germination, stomal opening, root shoot growth and other stress responses in plants. ABA is also synthesized by mammalian cells and has been linked to human disease. The aim of the present study was to examine the role of ABA in regulating tumor dormancy via signaling through lanthionine synthetase C­like protein 2 (LANCL2) and peroxisome proliferator activated receptor γ (PPARγ) receptors. ABA signaling in human PCa cell lines was studied using targeted gene knockdown (KD), western blotting, quantitative PCR, cell proliferation, migration, invasion and soft agar assays, as well as co­culture assays with bone marrow stromal cells. The data demonstrated that ABA signaling increased the expression of p21, p27 and p16, while inhibiting viability, migration, invasion and colony size in a reversable manner without toxicity. ABA also induced p38MAPK activation and NR2F1 signaling. Targeted gene KD of LANCL2 and PPARγ abrogated the cellular responses to ABA. Taken together, these data demonstrate that ABA may induce dormancy in PCa cell lines through LANCL2 and PPARγ signaling, and suggest novel targets to manage metastatic PCa growth.

Risk perception among residents living near industries in Godawari Municipality of Lalitpur, Nepal.

The industrial sector stands out as a significant contributor to environmental pollution. Those who reside in close proximity to industrial areas commonly harbor concerns about potential health and environmental hazards. This study aimed to find out the perception of risk and self-reported health impacts among individuals living near industries in Godawari Municipality, Lalitpur, Nepal. Conducted as a community-based cross-sectional study, it involved 270 households. Face-to-face interviews were employed, utilizing a pretested structured questionnaire. The study zone encompassed the communities of Godawari Municipality within a 3-kilometer radius of industrial sites. Specifically, stone mines, stone crushers, and brick kilns were purposefully selected, while study participants were randomly sampled using a random table. Data analysis was performed using IBM SPSS, incorporating both univariate and bivariate techniques. Among those residing near industrial zones, a mere 9.6 % reported experiencing wheezing or whistling in the past 12 months. A substantial 36.3% consistently felt stressed due to industrial activities in their vicinity. Approximately half (51.9 %) of the participants indicated that the contaminated air in the area had adverse effects on human health. Furthermore, a palpable perception of elevated risk was associated with the proximity of industries (p<0.001). Over half of the participants perceived a notable risk stemming from the presence of industries near their homes, largely due to pollutants. These individuals also disclosed various health repercussions and expressed significant apprehension regarding their future well-being in the area. The implications of these findings are substantial, particularly for local-level planning and the development of industrial sites. Addressing the concerns surrounding people's heightened perception of risk from nearby industries is pivotal in fostering harmonious coexistence and informed decision-making.

LIMK2 promotes melanoma tumor growth and metastasis through G3BP1-ESM1 pathway-mediated apoptosis inhibition.

Melanoma is the leading cause of skin cancer-related deaths, and current melanoma therapies, including targeted therapies and immunotherapies, benefit only a subset of metastatic melanoma patients due to either intrinsic or acquired resistance. LIM domain kinase 2 (LIMK2) is a serine/threonine kinase that plays an important role in the regulation of actin filament dynamics. Here, we show that LIMK2 is overexpressed in melanoma, and its genetic or pharmacological inhibition impairs melanoma tumor growth and metastasis in both cell culture and mice. To determine the mechanism by which LIMK2 promotes melanoma tumor growth and metastatic progression, we performed a phosphoproteomics analysis and identified G3BP1 as a key LIMK2 target, which mirrored the effects of LIMK2 inhibition when inhibited. To further determine the role of G3BP1 downstream of LIMK2, we knocked down the expression of G3BP1, performed RNA-seq analysis, and identified ESM1 as a downstream target of G3BP1. G3BP1 was required for ESM1 mRNA stability, and ESM1 ectopic expression rescued LIMK2 or G3BP1 inhibition-induced suppression of melanoma growth and metastatic attributes. These results collectively identify the LIMK2→G3BP1→ESM1 pathway as a facilitator of melanoma tumor growth and metastasis and document that LIMK2 is a therapeutically tractable target for melanoma therapy.

Fecal Carriage of Extended-Spectrum beta-Lactamase Producing Escherichia coli among Health Science Students.

BACKGROUND: The incidence of antibiotic resistance in commensal bacteria is increasing with the production of extended-spectrum beta-lactamase. Therefore, this study was conducted to understand the status of fecal carriage of such enzyme producing Escherichia coli among health science students of seven different faculties of Institute of Medicine, Tribhuvan University. METHODS: This was a cross-sectional study conducted over six months among the health science students. One stool sample collected from each student was cultured and Escherichia coli isolates were identified, antibiotic sensitivity profile was produced, and extended-spectrum beta-lactamase production was detected following Clinical and Laboratory Standards Institute guidelines. RESULTS: A total of 156 students participated in the study, and Escherichia coli was isolated from all. Out of the total 156 Escherichia coli isolates, 11.5% were extended-spectrum beta-lactamase-producers and 14.7% were multidrug-resistant. The highest rate of fecal carriage of extended-spectrum beta-lactamase-producing Escherichia coli was found among Bachelor of Medicine and Bachelor of Surgery students (17.5%) and Bachelor of Science in Medical Imaging Technology (16.7%) students. Such enzyme producing Escherichia coli was found in the range of 6.9% to 25.0% among second- to fifth-year students. A significant number of extended-spectrum beta-lactamase-producing isolates were resistant to ciprofloxacin and gentamicin, apart from other extended-spectrum beta-lactamase substrate antibiotics, when compared with non-producers. CONCLUSIONS: A high rate of extended-spectrum beta-lactamase-producing Escherichia coli was detected from the gut of healthy health science students which indicates their possible dissemination throughout the wider community resulting in potential outbreak of infections caused by such organisms.

Biofilm Formation by Pathogens Causing Ventilator-Associated Pneumonia at Intensive Care Units in a Tertiary Care Hospital: An Armor for Refuge.

BACKGROUND: Emerging threat of drug resistance among pathogens causing ventilator-associated pneumonia (VAP) has resulted in higher hospital costs, longer hospital stays, and increased hospital mortality. Biofilms in the endotracheal tube of ventilated patients act as protective shield from host immunity. They induce chronic and recurrent infections that defy common antibiotics. This study intended to determine the biofilm produced by pathogens causing VAP and their relation with drug resistance. METHODS: Bronchoalveolar lavage and deep tracheal aspirates (n = 70) were obtained from the patients mechanically ventilated for more than 48 hours in the intensive care units of Tribhuvan University Teaching Hospital, Kathmandu, and processed according to the protocol of the American Society for Microbiology (ASM). Antibiotic susceptibility testing was done following Clinical and Laboratory Standards Institute (CLSI) 2017 guidelines. Biofilm formation was determined using the microtiter plate method described by Christensen and modified by Stepanovoic et al. RESULTS: Significant microbial growth was seen in 78.6% of the total samples with 52.7% monomicrobial, 45.5% polymicrobial, and 1.8% fungal infection. Among the 71 isolates obtained, bulk was gram-negative (n = 64, 90.1%). Pseudomonas aeruginosa (31.0%) was the predominant isolate followed by Acinetobacter calcoaceticus baumannii complex (16.9%), Klebsiella pneumoniae (16.9%), Citrobacter freundii (15.5%), Staphylococcus aureus (7.0%), Escherichia coli (5.6%), Citrobacter koseri (2.8%), Enterococcus faecalis (1.4%), Burkholderia cepacia complex (1.4%), and Candida albicans (1.4%). Of the total isolates, 56.3% were biofilm producers. Multidrug-resistant (MDR) organisms, extended-spectrum ß-lactamase (ESBL), and metallo-ß-lactamase (MBL) producers were preeminent among the biofilm producers. The highest producer of biofilm was P. aeruginosa (19.7%). Among gram-negative biofilm producers, 42.2% were MDR, 21.9% were ESBL producers, and 7.8% were MBL producers. CONCLUSION: Gram-negative nonfermenter bacteria account for the bulk of nosocomial pneumonia. MDR, ESBL, and MBL production was preponderant among the biofilm producers. The rampant spread of drug resistance among biofilm producers is summoning novel interventions to combat multidrug resistance.