Having a Same Type IIS Enzyme's Restriction Site on Guide RNA Sequence Does Not Affect Golden Gate (GG) Cloning and Subsequent CRISPR/Cas Mutagenesis.

Golden gate/modular cloning facilitates faster and more efficient cloning by utilizing the unique features of the type IIS restriction enzymes. However, it is known that targeted insertion of DNA fragment(s) must not include internal type IIS restriction recognition sites. In the case of cloning CRISPR constructs by using golden gate (GG) cloning, this narrows down the scope of guide RNA (gRNA) picks because the selection of a good gRNA for successful genome editing requires some obligation of fulfillment, and it is unwanted if a good gRNA candidate cannot be picked only because it has an internal type IIS restriction recognition site. In this article, we have shown that the presence of a type IIS restriction recognition site in a gRNA does not affect cloning and subsequent genome editing. After each step of GG reactions, correct insertions of gRNAs were verified by colony color and restriction digestion and were further confirmed by sequencing. Finally, the final vector containing a Cas12a nuclease and four gRNAs was used for Agrobacterium-mediated citrus cell transformation. Sequencing of PCR amplicons flanking gRNA-2 showed a substitution (C to T) mutation in transgenic plants. The knowledge derived from this study could widen the scope of GG cloning, particularly of gRNAs selection for GG-mediated cloning into CRISPR vectors.

Diversity of Vibrio parahaemolyticus in marine fishes of Bangladesh.

AIMS: To determine the occurrence, diversity, antibiotic resistance and biofilm formation of Vibrio parahaemolyticus isolated from marine fishes in Bangladesh. METHODS AND RESULTS: A total of 80 marine fishes were obtained from the local markets and examined for the presence of V. parahaemolyticus. All the isolated V. parahaemolyticus were characterized for the presence of virulence markers, thermostable direct hemolysin (TDH) or thermostable direct hemolysin related hemolysin (TRH). Isolates were serotyped and further characterized by enterobacterial repetitive intergenic consensus sequence PCR (ERIC-PCR) typing to analyse the genetic diversity. Moreover, biofilm formation and antibiotic resistance patterns were also determined. About 63·75% (51/80) of the tested marine fishes were contaminated with V. parahaemolyticus. From the contaminated fishes, 71 representatives V. parahaemolyticus were isolated and none of them harboured tdh and trh virulence genes. Nine different O-groups and seven different K-types were found by serological analysis and the dominant serotype was O5:KUT. In ERIC-PCR analysis, eight clusters (A-H) were found and the most common pattern was A (46·5%). All of the isolates were resistant to ampicillin and 78·9% of isolates were resistant to streptomycin. The highest biofilm formation was found at 37°C compared to 25°C and 4°C. CONCLUSION: Diverse V. parahaemolyticus are present in marine fishes in the local market of Bangladesh with antibiotic-resistant properties and biofilm formation capacity. SIGNIFICANCE AND IMPACT OF THE STUDY: The widespread prevalence of diverse V. parahaemolyticus in marine fishes is an issue of serious concern, and it entails careful monitoring to ascertain the safety of seafood consumers.

Clostridioides difficile ribotypes isolated from domestic environment and from patients in Bangladesh.

Clostridioides difficile infection (CDI) is an emerging but often understudied infectious disease in developing countries. This study was aimed to isolate and characterize C. difficile from shoe sole swabs and diarrheal patient's stool samples in Bangladesh. We collected 94 shoe sole swabs samples from urban communities in Dhaka and 208 diarrheal stool samples from hospitalized patients over a period of 4 months. Samples were incubated anaerobically for C. difficile growth, confirmed toxigenic, and PCR-ribotyped. Eleven of 94 (11.7%) shoe sole swabs and 4 of 208 (1.9%) stool samples were culture positive of which 9 shoe sole isolates were toxigenic. Six PCR ribotypes from the 9 toxigenic isolates were identified with ribotype F014-020 being the most common (n = 4; 44%). The recently identified ribotype 106 strain was also identified. To the best of our knowledge, this is the first report of C. difficile culture, isolation and characterization from environmental sources in Bangladesh.

Physical activity levels in Bangladeshi adults: results from STEPS survey 2010.

OBJECTIVES: Physical inactivity is an established risk factor for non-communicable diseases (NCD) and identified as the major public health concern worldwide. However, nationally representative and internationally comparable data on physical activity (PA) are lacking in Bangladesh. The objective of this paper was to determine nationally representative prevalence of PA levels among Bangladeshi adults. STUDY DESIGN: Cross-sectional survey. METHODS: Data, on PA for this paper, were analysed from the NCD risk factors survey 2010 in Bangladesh. A standardized approach known as STEPS (STEPSwise approach to Surveillance for NCD risk factors) was followed for this survey. A total of 9275 adults (aged ≥ 25 years) were interviewed. Data on PA were processed and analysed according to Global Physical Activity Questionnaire (GPAQ) version 2 analysis framework. RESULTS: Of total 9275 respondents 4312 were men and 4963 women with a mean age of 42.4 (±13.5) years. Median MET-minutes of total PA in a typical week was double in rural areas (3360) than urban (1680) areas. The overall country wide prevalence of low PA was 34.5% (95% confidence interval, 33.5-35.5), urban 37.7% (36.3-39.1) and rural 31.6% (30.3-32.9). Women in general were more inactive (women, 53.6% [52.2-55.0], men 15.4% [14.9-17.1]). The main contributions to total PA were from work (urban 47.0%, rural 61.0%), and active commuting (38.0%, 30.0%) domains. Leisure-time PA represented only a small proportion (15.0%, 9.0%). CONCLUSIONS: Insufficient physical activity is highly prevalent among the Bangladeshi adult population. Promoting overall PA at leisure-time and commuting considering country context can be feasible options with special attention to the women.

Effects of Intensive versus Non-Intensive Physical Therapy on Children with Cerebral Palsy.

Cerebral Palsy (CP) is one of the most common causes of all childhood disorders. There are tone, posture and movements difficulty due to non-progressive damage to the immature brain in CP. The hallmark of CP is a disability in the development of gross motor function (GMF). The influence of gross motor development on fine motor development is more important in early developmental period, specially under three years old and in children with CP. Various therapeutic interventions have been used in the management of GMF development. Among them physical therapy is the most common intervention in CP and is usually a component of mandated programs. Physical therapy means physical stimulations in the form of various therapeutic exercises, touch, massage, limbs and trunk movement, balancing and coordination training, gait and ambulation training, cognitive stimulation as well as speech, language and occupational therapy. Our study focused to see the effect by short term intensive versus non-intensive physical therapy on children GMF development by using gross motor function measure (GMFM) Score sheet, GMFM-88, version 1.0. Study provides the information that physical therapy intervention is effective in GMF development and intensive interventions are more effective in children with spastic CP than non-intensive one. Study also inform that the more early treatment the more effective result.

Effects of Feeding Rate and Water Temperature on Growth and Body Composition of Juvenile Korean Rockfish, Sebastes schlegeli (Hilgendorf 1880).

Three feeding trials were conducted to evaluate the effects of feeding rate and water temperature on growth and body composition of juvenile Korean rockfish, Sebastes schlegeli rearing at 3 different water temperatures. A total of 270 fish (each experiment) individual body weight (BW) averaging 16±0.3 g (mean±SD) were fed a commercial diet for 4 wk at 16°C, 20°C, and 24°C. At each temperature, triplicate tanks were assigned to one of 6 feeding rates: 1.5%, 2.5%, 2.8%, 3.1%, 3.4%, and satiation (3.7% BW/d) at 16°C, 1.9%, 2.9%, 3.2%, 3.5%, 3.8% and satiation (4.1% BW/d) at 20°C and 1.7%, 2.7%, 3.0%, 3.3%, 3.6%, and satiation (3.9% BW/d) at 24°C water temperature. Weight gains of fish in satiation and 3.4% groups at 16°C, in satiation and 3.8% groups at 20°C and in satiation and 3.6% groups at 24°C were significantly higher than those of fish in the other treatments (p<0.05). A broken line regression analysis of weight gain indicated that optimum feeding rates of juvenile Korean rockfish were 3.41% at 16°C, 3.75% at 20°C and 3.34% at 24°C water temperature. Results of the present study indicate that the optimum feeding rate could be >3.1% but <3.41% at 16°C, >3.5% but <3.75% at 20°C and >3.0% but <3.34% at 24°C. As we expected results suggest that fish performed better at 20°C than 16°C or 24°C water temperature and the optimum feeding rate could be 3.1% BW/d to 3.7% BW/d in 16 g of juvenile Korean rockfish.

Serum Electrolyte Status of Patients with Acute Stroke Admitted in a Tertiary Care Hospital.

Stroke, usually a focal rather than global neurological deficit resulting from vascular origin with sudden onset, may be with cerebral infarction or intracerebral haemorrhage. It results in brain oedema following vascular injury and electrolyte imbalance. A descriptive cross sectional study was carried out in the Department of Medicine, Mymensingh Medical College Hospital, Mymensingh, Bangladesh during March 2016 to May 2018 to assess the electrolyte levels among 220 purposively selected patients with stroke confirmed by CT scan. Data were collected by the principal investigator himself by using interview schedule and case record form after attaining consent. Blood samples were collected from the patients to carry out biochemical and haematological tests and to assess serum electrolyte levels. Data were cross-checked for completeness, consistency and relevancy, and were analyzed by computer software SPSS 20.0. Age was significantly higher in haemorrhagic stroke (64.88±13.00 years) than ischaemic stroke (60.92±13.96 years). Male (55.91%) were predominant than female (44.09%). One hundred nineteen (54.09%) patients had ischaemic stroke and 101(45.91%) patients had haemorrhagic stroke. The serum concentration of Na+, K+, Cl- and HCO3- were measured during acute period of stroke. Imbalance in serum Sodium, Chloride, Potassium and Bicarbonate level were observed in 37.27%, 29.55%, 23.18% and 6.36% patients respectively. Hyponatremia, hypokalemia, hypochloremia and acidosis were most common electrolyte imbalance in both ischaemic and haemorrhagic strokes. In ischaemic stroke hyponatremia was 35.29%, hypernatremia was 3.36%, hypokalemia 19.33%, hyperkalemia 0.84%, hypochloraemia 30.25%, hyperchloraemia 3.36%, acidosis was in 6.72% and alkalosdis in 1.68% patients while in haemorrhagic stroke hyponatremia 33.66%, hypernatremia 1.98%, hypokalaemia 22.77% hyperkalemia 3.96%, hypochloremia 19.80%, hyperchloraemia 4.95%, acidosis 2.97% and alkalosis was in 0.99% of patients. Mortality was more in hyponatremic, hypokalemic and in hypochloremic patients.

Multidrug-resistant Escherichia coli isolated from patients and surrounding hospital environments in Bangladesh: A molecular approach for the determination of pathogenicity and resistance.

Extended spectrum ß-lactamase producing Escherichia coli (ESBL E. coli) is a primary concern for hospital and community healthcare settings, often linked to an increased incidence of nosocomial infections. This study investigated the characteristics of ESBL E. coli isolated from hospital environments and clinical samples. In total, 117 ESBL E. coli isolates were obtained. The isolates were subjected to molecular analysis for the presence of resistance and virulence genes, antibiotic susceptibility testing, quantitative adherence assay, ERIC-PCR for phylogenetic analysis and whole genome sequencing of four highly drug resistant isolates. Out of the 117 isolates, 68.4% were positive for blaCTX-M, 39.3% for blaTEM, 30.8% for blaNDM-1, 13.7% for blaOXA and 1.7% for blaSHV gene. Upon screening for diarrheagenic genes, no isolates were found to harbour any of the tested genes. In the case of extraintestinal pathogenic E. coli (ExPEC) virulence factors, 7.6%, 11%, 5.9%, 4.3% and 21.2% of isolates harbored the focG, kpsMII, sfaS, afa and iutA genes, respectively. At a temperature of 25°C, 14.5% of isolates exhibited strong biofilm formation with 21.4% and 28.2% exhibiting moderate and weak biofilm formation respectively, whereas 35.9% were non-biofilm formers. On the other hand at 37°C, 2.6% of isolates showed strong biofilm formation with 3.4% and 31.6% showing moderate and weak biofilm formation respectively, whereas, 62.4% were non-biofilm formers. Regarding antibiotic susceptibility testing, all isolates were found to be multidrug-resistant (MDR), with 30 isolates being highly drug resistant. ERIC-PCR resulted in 12 clusters, with cluster E-10 containing the maximum number of isolates. Hierarchical clustering and correlation analysis revealed associations between environmental and clinical isolates, indicating likely transmission and dissemination from the hospital environment to the patients. The whole genome sequencing of four highly drug resistant ExPEC isolates showed the presence of various antimicrobial resistance genes, virulence factors and mobile genetic elements, with isolates harbouring the plasmid incompatibility group IncF (FII, FIB, FIA). The sequenced isolates were identified as human pathogens with a 93.3% average score. This study suggests that ESBL producing E. coli are prevalent in the healthcare settings of Bangladesh, acting as a potential reservoir for AMR bacteria. This information may have a profound effect on treatment, and improvements in public healthcare policies are a necessity to combat the increased incidences of hospital-acquired infections in the country.

Trauma in surgery and its incidence, circumstances as well as clinical consequences.

This study was conducted from 10.02.08 to 23.05.09 in general surgery wards of Chittagong Medical College Hospital, Bangladesh with multi-factorial views of clinical aspects and an alarming model image in such relation was depicted. 188 (72.9 %) patients in this study out of 258 were male it was and found that different types of trauma had been relatively higher in 51-70 yrs (35.1 %) age group followed by 31-50 yrs (28.7 %) age group in case of male subjects. On the contrary, in female, those were quite higher in 11-30 yrs (31.4 %) age group followed by 51-70 yrs (24.8 %) age group. As for the mortality, this study suggest that it was significantly higher in case of female subjects (31.4 %) (22) in relation to male subjects (18.1 %) (34). But to our knowledge probably it has no clinical significance at all. Rather, it may have socio-economic associations, as for instance- traditional negligence to female in developing countries like Bangladesh. Another remarkable observation that was clear in this study was variation in different levels of morbidity. In male, 30.9 % patients had morbidity from 1 week to 1 month; followed by 28.7 % patients with morbidity less than 1 week, whereas, in case of female, it was 25.7 % and 20 %, respectively. With regard to morbidity over 1 month 23.4 % male and 22.9 % female were found in this group and in case of both male and female, the so called p-values were quite significant. This study also reflects that majority of trauma occurred due to road traffic accidents (RTAs) (24.5 %) followed by natural disasters (21.3 %) and assault (17 %) in case of male, whereas, in female 34.3 % trauma occurred by assault and homicidal attempts followed by 20 % due to natural disaster. One of the most remarkable associations of traumatic patients was that 82.3 % (28) of all deaths occurred in the first 24 hours of injury in male and on the other hand, it was 72.7 % (16) in case of female patients (Tab. 3, Fig. 7, Ref. 31).

Psychiatric Morbidity in Irritable Bowel Syndrome.

Psychiatric morbidity is frequent among outpatients with Irritable Bowel Syndrome (IBS) attending gastroenterology clinics. The lifetime prevalence of major depressive and anxiety disorders in IBS patients are up to 76% and 54% respectively. The point prevalence of Major depressive disorder and Anxiety disorders in IBS patients are ranged 9.6 to 54% and from 11.1 to 52.4% respectively. IBS is not usually life-threatening but it has a significant impact on quality of life. So, screening for psychiatric disorders in IBS patients attending Gastroenterology clinics is worthwhile. The objective of this study was to evaluate psychiatric morbidity among the patients with IBS. This cross sectional comparative study was conducted in the Department of Psychiatry, Sylhet MAG Osmani Medical College Hospital, Sylhet, Bangladesh during the period from January 2012 to June 2012. Inclusion criteria were patients of irritable bowel syndrome fulfilling the Rome-III criteria, age between 18 to 60 years irrespective of sex. Patients with any red flag sign, serious cognitive impairment such as dementia, schizophrenia, mania, delusional disorder, prior history of abdominal surgery and other chronic diseases were excluded. One hundred and ten patients with IBS were selected according to inclusion and exclusion criteria and categorized as IBS group. Another 110 age and sex matched subjects were selected as control group. The mean±SD age of the patients was 34.3±12.2 years which was almost identical to control group (p=0.820). Most of the patients (66.4%) were below the age of 40 years. There were 64 (58.2%) male and 46 (41.8%) female which was similar to control group (p=0.643). Psychiatric co-morbidity was most frequent in IBS patients (56.4%) as compared to control group (16.4%) p value <0.001. The most common psychiatric disorders were Anxiety disorders (22.7%), followed by major depressive disorder (19.1%) and somatization disorder (14.5%). The results of this study suggest that a significant number of patients with IBS suffer from psychiatric disorders than control, which are similar to the IBS group in age and gender.

Pattern of Tubercular Uveitis in Active Pulmonary Tuberculosis.

Ocular tuberculosis is an extra-pulmonary form of systemic Tuberculosis (TB). It is rarely found concomitant with active pulmonary tuberculosis (PTB). The aim of this prospective observational study was to evaluate the pattern of tubercular uveitis (TBU) in the patients with active PTB who attended in our Uvea clinic at NIO&H from July 2018 to December 2020. Active PTB patients who had uveitis consistent with TBU and TBU patients who were confirmed as active lung lesion were included in the study. Chest X-ray, TST, IGRA (QuantiFERON-TB Gold Test), RT PCR of aqueous fluid for tubercular bacillus, Gene Xpert TB test of sputum were done to confirm the diagnosis of ocular and primary TB. If the induration is 15 mm of TST was considered patient in this study. Among 33 patients of active PTB, 4 had TBU (12.12%). Among 48 patients of TBU, 5 had active PTB (10.41%). 13 eyes of 9 patients were affected. Mean age at presentation was 41.8 years. There were anterior uveitis in 23.00% eyes, posterior uveitis in 69% eyes and panuveitis in 7.00% eyes. Choroidal granuloma was the most common presentation of posterior uveitis (46.00%). There were more than 4 times chance to develop choroidal granuloma in patients with active PTB than extrapulmonary TB (Χ²=4.53, <0.05), TBU patients with choroidal granuloma should be evaluated meticulously for active PTB and active PTB patients should be evaluated routinely by ophthalmologist for the search of TBU.

Healthcare Facilities as Potential Reservoirs of Antimicrobial Resistant Klebsiella pneumoniae: An Emerging Concern to Public Health in Bangladesh.

The emergence of virulent extended spectrum ß-lactamase producing Klebsiella pneumoniae (ESBL-KP) including carbapenem-resistant Klebsiella pneumoniae (CRKP) in hospital-acquired infections has resulted in significant morbidity and mortality worldwide. We investigated the antibiotic resistance and virulence factors associated with ESBL-KP and CRKP in tertiary care hospitals in Bangladesh and explored their ability to form biofilm. A total of 67 ESBL-KP were isolated from 285 Klebsiella pneumoniae isolates from environmental and patient samples from January 2019 to April 2019. For ESBL-KP isolates, molecular typing was carried out using enterobacterial repetitive intergenic consensus polymerase chain reaction (ERIC-PCR), antibiotic susceptibility testing, PCR for virulence and drug-resistant genes, and biofilm assays were also performed. All 67 isolates were multidrug-resistant (MDR) to different antibiotics at high levels and 42 isolates were also carbapenem-resistant. The most common ß-lactam resistance gene was blaCTX-M-1 (91%), followed by blaTEM (76.1%), blaSHV (68.7%), blaOXA-1 (29.9%), blaGES (14.9%), blaCTX-M-9 (11.9%), and blaCTX-M-2 (4.5%). The carbapenemase genes blaKPC (55.2%), blaIMP (28.4%), blaVIM (14.9%), blaNDM-1 (13.4%), and blaOXA-48 (10.4%) and virulence-associated genes such as fimH (71.6%), ugeF (58.2%), wabG (56.7%), ureA (47.8%) and kfuBC (28.4%) were also detected. About 96.2% of the environmental and 100% of the patient isolates were able to form biofilms. ERIC-PCR-based genotyping and hierarchical clustering of K. pneumoniae isolates revealed an association between environmental and patient samples, indicating clonal association with possible transmission of antimicrobial resistance genes. Our findings can help in improving patient care and infection control, and the development of public health policies related to hospital-acquired infections.

Consequences of abdominal adenocarcinoma in post-menopausal woman in relation to surgical and non-surgical management.

The umbilical tumor as a presenting symptom must be investigated for the possibility of metastasis. In this case, it was unlikely, as there was no primary carcinoma found elsewhere. Primary peritoneal carcinoma (also termed serous surface papillary carcinoma) is a malignancy that arises primarily from peritoneal cells. The mesothelium of the peritoneum and the germinal epithelium of the ovary arise from the same embryological origin. Although primary peritoneal carcinoma has a very poor prognosis, our patient had a disease-free interval of two years post surgery. Clinically, primary peritoneal carcinoma may be difficult to distinguish from ovarian carcinoma. Compared to ovarian carcinoma, primary peritoneal carcinoma has a higher rate of abdominal distension, volume of ascites, malignant cells in the ascitic fluid, lower rate of pelvic palpable mass, and personal breast cancer history. However, primary ovarian cancer can be excluded in this case, based on both ovaries being of normal size with no tumor involvement and a histology that favors serous carcinoma. The disease is normally disseminated throughout the peritoneum. Our patient's disease was confined to the umbilicus with no evidence of dissemination. We report a rare case of adenocarcinoma confined to the umbilicus of a 64-year-old menopausal woman presented with an umbilical lump of nine years. After five years, she presented with a 6-cm irregular periumbilical mass. She agreed to undergo an excision biopsy of the mass with total abdominal hysterectomy and bilateral salpingo-oopherectomy with omentectomy. The excision biopsy showed adenocarcinoma with an appearance suggestive of metastasis from an ovarian cystadenocarcinoma. The frozen section of the tumor showed a papillary adenocarcinoma. Histology showed adenocarcinoma favoring serous cancer. It was found that tumor markers of ovarian malignancy were normal. There was no tumor seen in the ovaries, uterus and omentum. On follow-up, there was no disease recurrence. The patient has been disease-free for two years post surgery (Fig. 5, Ref. 10).

Urethral stricture in adult as a consequence of childhood hypospadias repair.

We report our experience in treating such patients. A retrospective chart review was performed from 2004 through 2007 in Urology Department of Dhaka Medical College Hospital by using the convenient type of purposive sampling. In cases undergoing perineal urethrostomy, we suggested the "side-to-side" technique, which comprised longitudinal urethrotomy and everting the mucosal and submucosal layers of the urethra to the incised skin. Graft-based urethroplasty using buccal mucosal graft, in one-stage or two-stage repairs, was the preferred choice of formal reconstruction. Patients undergoing hypospadias surgery should receive lifelong follow-up protocol to detect latent urethral strictures (Fig. 8, Ref. 12).

Citrus Cell Suspension Culture Establishment, Maintenance, Efficient Transformation and Regeneration to Complete Transgenic Plant.

Agrobacterium-mediated transformation of epicotyl segment has been used in Citrus transgenic studies. The approach suffers, however, from limitations such as occasionally seed unavailability, the low transformation efficiency of juvenile tissues and the high frequency of chimeric plants. Therefore, a suspension cell culture system was established and used to generate transgenic plants in this study to overcome the shortcomings. The embryonic calli were successfully developed from undeveloped ovules of the three cultivars used in this study, "Sweet orange"-Egyptian cultivar (Citrus sinensis), "Shatangju" (Citrus reticulata) and "W. Murcott" (Citrus reticulata), on three different solid media. Effects of media, genotypes and ages of ovules on the induction of embryonic calli were also investigated. The result showed that the ovules' age interferes with the callus production more significantly than media and genotypes. The 8 to 10 week-old ovules were found to be the best materials. A cell suspension culture system was established in an H+H liquid medium. Transgenic plants were obtained from Agrobacterium-mediated transformation of cell suspension as long as eight weeks subculture intervals. A high transformation rate (~35%) was achieved by using our systems, confirming BASTA selection and later on by PCR confirmation. The results demonstrated that transformation of cell suspension should be more useful for the generation of non-chimeric transgenic Citrus plants. It was also shown that our cell suspension culture procedure was efficient in maintaining the vigor and regeneration potential of the cells.

The Pattern of Lipid Profile in Patients with Chronic Kidney Disease.

Dyslipidemia is a common problem in chronic kidney disease patients. Dyslipidemia in chronic kidney disease patients has been known to be a major risk factor of their cardiovascular disease and may contribute to progressive renal dysfunction. The result of the study might be of interest in improving preventive strategies and in management of dyslipidemia in chronic kidney disease patients. This cross sectional study was conducted to evaluate changes in lipid profile in patients with chronic kidney disease stage-3 to stage-5 patients and to correlate the biochemical abnormalities with progression of the disease in Mymensingh Medical College Hospital, Bangladesh from October, 2016 to April, 2017. In this study 200 patients were including and subjected to do complete blood count, erythrocyte sedimentation rate, random blood sugar, routine examination of urine, serum creatinine and fasting lipid profile. Two hundred (200) patients (134 males, 66 females) with the mean age were 50.5±12.43 years. 44.5% patients were in CKD stage-5, 37.5% patients were in CKD stage-4, 18% patients were in CKD stage-3. Mean value of Triglyceride (TG) was 194±47.20. Eighty nine percent (89%) patient had hyper-triglyceridemia and 11% had normal triglyceride level. It was statistically significant increased in triglyceride level (p<0.05). Mean value of High density lipoprotein cholesterol (HDL-C) was 34±6.10. Low HDL-C had in 87.5% patients, normal in 12.5% patients and was statistically significant reduction in HDL-C level (p<0.05). Low Density Lipoprotein cholesterol (LDL-C) mean was 113±35.6. High level of LDL-C had optimal/or near optimal in 47% patients, 39% patients had borderline high and 14% patients had that was not statistically significant (p>0.10). Total cholesterol (TC) mean was 212±45.3. In 38% patients had within desirable level, 62% patients had high level of Total cholesterol (TC). It was not statistically significant change (p>0.01).

Study on Alanine Aminotransferase in Patients of Type 2 Diabetes Mellitus.

Type 2 Diabetes mellitus (DM) is associated with various types of liver disorders including elevated liver enzymes. This study was carried out to observe hepatocellular injury in type 2 Diabetes mellitus as evidenced by raised Alanine aminotransferase (ALT) among the type 2 Diabetes mellitus patients and to find out association between elevated ALT and glycaemic control. This was a descriptive cross-sectional comparative study carried out in Department of Medicine, Rajshahi Medical College Hospital and Rajshahi Diabetic Association Hospital, Rajshahi, Bangladesh from July 2009 to June 2010. One hundred (100) diagnosed type 2 diabetic patients according to WHO criteria and 30 apparently healthy people of both sexes were included. Age of the study people were >40 years. All of those study population were free from taking any hepatotoxic drugs, inj. Insulin, alcohol and preexisting liver disease. Detailed History, thorough clinical examination and relevant investigations were done for exclusion. The frequency of abnormal ALT, Serum bilirubin, Aspartate aminotransferase (AST), Alkaline phosphatase, Prothrombin time and Serum albumin were 30%, 6%, 7%, 6%, 54% and 12% respectively in type 2 diabetic patients and 3.3%, 00%, 00%, 6.7%, 10% and 3.3% respectively in normal people. Serum ALT (U/L) level (mean±SD) of the type 2 diabetic patients and normal people were (39.00±24.21) and (28.26±6.67) respectively. This difference is highly significant (p<0.001). Elevated serum ALT level of type 2 diabetic patients showed positive association with increased Body mass index (BMI) (p<0.05) and poor glycaemic control (p<0.05) and fatty liver (p=0.055). So, a high proportion of patients with type 2 diabetes mellitus have raised serum Alanine aminotransferase level. Elevated serum ALT level was also associated with high BMI, poor glycaemic control and fatty liver which may be a marker of NAFLD and insulin resistance.

Somatotropin increases plasma ceramide in relation to enhanced milk yield in cows.

Recombinant bovine somatotropin (rBST) changes metabolism to spare glucose for milk synthesis in cows. Ceramides inhibit insulin responsiveness in bovine adipocytes and are associated with insulin resistance and milk production in cows. The mechanisms by which rBST supports lactation may involve ceramide. Eight multiparous lactating Holstein cows were enrolled in a 2 × 2 replicated Latin square design with 14-d periods. Cows received a single rBST injection (Posilac; Elanco Animal Health, Indianapolis, IN; 0.062 mg/kg BW) or no injection (CON). An epinephrine challenge, insulin tolerance test, and liver biopsy were performed. Somatotropin enhanced the conversion of feed nutrients into milk components and increased plasma free fatty acid (FFA) concentrations (P < 0.01). Area-under-the-curves for FFA in response to epinephrine and insulin were greater in rBST-treated cows. In response to insulin, glucose concentrations (20- and 30-min post-challenge) and insulin area-under-the-curve were higher with rBST treatment (P < 0.05, <0.10, and <0.01), suggesting insulin resistance. Somatotropin modified the plasma lipidome. For example, rBST decreased plasma di- and triacylglycerol levels (eg, DG-50:1 and TG-18:0/16:0/16:1), phosphatidylcholines and sphingomyelins (P < 0.05). Somatotropin increased plasma total and very-long-chain (C22:0-, C24:0-, C26:0-) ceramide concentrations (P < 0.01). Liver ceramide concentrations were not modified. Plasma ceramides were positively correlated with circulating FFA (r ~ 0.57; P < 0.05) and milk yield (r ~ 0.63; P < 0.05). We conclude that rBST administration modifies the bovine lipidome and increases plasma ceramide concentrations in association with increased milk production in cows.

Characterization of Pathogenic Vibrio parahaemolyticus Isolated From Fish Aquaculture of the Southwest Coastal Area of Bangladesh.

Vibrio parahaemolyticus is a major foodborne pathogen responsible for significant economic losses in aquaculture and a threat to human health. Here, we explored the incidence, virulence potential, and diversity of V. parahaemolyticus isolates from aquaculture farms in Bangladesh. We examined a total of 216 water, sediment, Oreochromis niloticus (tilapia), Labeo rohita (rui), and Penaeus monodon (shrimp) samples from the aquaculture system where 60.2% (130/216) samples were positive for V. parahaemolyticus. Furthermore, we identified 323 V. parahaemolyticus strains from contaminated samples, 17 of which were found positive for trh, a virulence gene. Four isolates out of the 17 obtained were able to accumulate fluid in the rabbit ileal loop assay. The correlation between the contamination of V. parahaemolyticus and environmental factors was determined by Pearson correlation. The temperature and salinity were significantly correlated (positive) with the incidence of V. parahaemolyticus. Most of the pathogenic isolates (94.1%) were found resistant to ampicillin and amoxicillin. O8: KUT was the predominant serotype of the potentially pathogenic isolates. ERIC-PCR reveals genetic variation and relatedness among the pathogenic isolates. Therefore, this region-specific study establishes the incidence of potential infection with V. parahaemolyticus from the consumption of tilapia, rui, and shrimp raised in farms in Satkhira, Bangladesh, and the basis for developing strategies to reduce the risk for diseases and economic burden.

ESBL Producing Escherichia coli in Faecal Sludge Treatment Plants: An Invisible Threat to Public Health in Rohingya Camps, Cox's Bazar, Bangladesh.

Introduction: Human faecal sludge contains diverse harmful microorganisms, making it hazardous to the environment and public health if it is discharged untreated. Faecal sludge is one of the major sources of E. coli that can produce extended-spectrum ß-lactamases (ESBLs). Objective: This study aimed to investigate the prevalence and molecular characterization of ESBL-producing E. coli in faecal sludge samples collected from faecal sludge treatment plants (FSTPs) in Rohingya camps, Bangladesh. Methods: ESBL producing E. coli were screened by cultural as well as molecular methods and further characterized for their major ESBL genes, plasmid profiles, pathotypes, antibiotic resistance patterns, conjugation ability, and genetic similarity. Results: Of 296 isolates, 180 were phenotypically positive for ESBL. All the isolates, except one, contained at least one ESBL gene that was tested (blaCTX-M-1, blaCTX-M-2, blaCTX-M-8, blaCTX-M-9, blaCTX-M-15, blaCTX-M-25, blaTEM , and blaSHV ). From plasmid profiling, it was observed that plasmids of 1-211 MDa were found in 84% (151/180) of the isolates. Besides, 13% (24/180) of the isolates possessed diarrhoeagenic virulence genes. From the remaining isolates, around 51% (79/156) harbored at least one virulence gene that is associated with the extraintestinal pathogenicity of E. coli. Moreover, 4% (3/156) of the isolates were detected to be potential extraintestinal pathogenic E. coli (ExPEC) strains. Additionally, all the diarrhoeagenic and ExPEC strains showed resistance to three or more antibiotic groups which indicate their multidrug-resistant potential. ERIC-PCR differentiated these pathogenic isolates into seven clusters. In addition to this, 16 out of 35 tested isolates transferred plasmids of 32-112 MDa to E. coli J53 recipient strain. Conclusion: The present study implies that the faecal sludge samples examined here could be a potential origin for spreading MDR pathogenic ESBL-producing E. coli. The exposure of Rohingya individuals, living in overcrowded camps, to these organisms poses a severe threat to their health.