Classy non-wovens based on animate L. gasseri-inanimate poly(vinyl alcohol): upstream application in food engineering.

We explored electrospinning as a feasible and practicable mode for encapsulation and stabilization of Lactobacillus gasseri. The utilized nanocomposite was prepared using sol-gel composed of animate L. gasseri and inanimate PVA. The objective was to examine the ability of electrospinning method to protect functional properties of probiotic L. gasseri. The PVA was used as an encapsulation matrix as it is biocompatible and hydrophilic in nature thus facilitate an easy revival of bacteria. The characterization of as-spun bioproduct was done by energy-dispersive X-ray spectrometer, SEM, and TEM, whereas thermal behavior was analyzed by thermogravimetry. The viability was confirmed by traditional pour plate method and fluorescence microscopy. Furthermore, to test whether the functionality of L. gasseri was affected, the encapsulated L. gasseri were fed to mouse for colonization. Our results pointed out that encapsulated bacteria were viable for months, and their metabolism was not affected by immobilization; thus, they could be used in food engineering and trade.

Antibacterial activity and interaction mechanism of electrospun zinc-doped titania nanofibers.

In this study, a biological evaluation of the antimicrobial activity of Zn-doped titania nanofibers was carried out using Escherichia coli ATCC 52922 (Gram negative) and Staphylococcus aureus ATCC 29231 (Gram positive) as model organisms. The utilized Zn-doped titania nanofibers were prepared by the electrospinning of a sol-gel composed of zinc nitrate, titanium isopropoxide, and polyvinyl acetate; the obtained electrospun nanofibers were vacuum dried at 80°C and then calcined at 600°C. The physicochemical properties of the synthesized nanofibers were determined by X-ray diffraction pattern, field emission scanning electron microscopy, energy-dispersive X-ray spectroscopy, electron probe microanalysis, thermogravimetry, and transmission electron microscopy (TEM). The antibacterial activity and the acting mechanism of Zn-doped titania nanofibers against bacteria were investigated by calculation of minimum inhibitory concentration and analyzing the morphology of the bacterial cells following the treatment with nanofibers solution. Our investigations reveal that the lowest concentration of Zn-doped titania nanofibers solution inhibiting the growth of S. aureus ATCC 29231 and E. coli ATCC 52922 strains is found to be 0.4 and 1.6 µg/ml, respectively. Furthermore, Bio-TEM analysis demonstrated that the exposure of the selected microbial strains to the nanofibers led to disruption of the cell membranes and leakage of the cytoplasm. In conclusion, the combined results suggested doping promotes antimicrobial effect; synthesized nanofibers possess a very large surface-to-volume ratio and may damage the structure of the bacterial cell membrane, as well as depress the activity of the membranous enzymes which cause bacteria to die in due course.

Controlled synthesis of Mn2O3 nanowires by hydrothermal method and their bactericidal and cytotoxic impact: a promising future material.

Mn2O3 nanowires with diameter ~70 nm were synthesized by a simple hydrothermal method using Mn(II) nitrate as precursor. X-ray diffraction, field emission scanning electron microscopy, transmission electron microscopy (TEM), and energy dispersive X-ray spectroscopy techniques were employed to study structural features and chemical composition of the synthesized nanowires. A biological evaluation of the antimicrobial activity and cytotoxicity of Mn2O3 nanowires was carried out using Escherichia coli and mouse myoblast C2C12 cells as model organism and cell lines, respectively. The antibacterial activity and the acting mechanism of Mn2O3 nanowires were investigated by using growth inhibition studies and analyzing the morphology of the bacterial cells following the treatment with nanowires. These results suggest that the pH is critical factor affecting the morphology and production of the Mn2O3 nanowires. Method developed in the present study provided optimum production of Mn2O3 nanowires at pH ~ 9. The Mn2O3 nanowires showed significant antibacterial activity against the E. coli strain, and the lowest concentration of Mn2O3 nanowires solution inhibiting the growth of E. coli was found to be 12.5 µg/ml. TEM analysis demonstrated that the exposure of the selected microbial strains to the nanowires led to disruption of the cell membranes and leakage of the internal contents. Furthermore, the cytotoxicity results showed that the inhibition of C2C12 increases with the increase in concentration of Mn2O3 nanowires. Our results for the first time highlight the cytotoxic and bactericidal potential of Mn2O3 nanowires which can be utilized for various biomedical applications.

Genetic engineering of Lactococcus lactis to produce an amylase inhibitor for development of an anti-diabetes biodrug.

Diabetes is known as a multifactorial disease. The treatment of diabetes is complicated due to its inherent pathophysiological factors related to the disease. One of the complications of diabetes is postprandial hyperglycemia. Glucosidase inhibitors, particularly ?-amylase inhibitors can help manage postprandial hyperglycemia. The low molecular weight inhibitor of ? -amylases called PAMI (peptide amylase inhibitor) inhibits the ? -amylase. In this study we cloned this amylase blocker PAMI in Lactococcus lactis. Using this Lactococcus lactis expressing the PAMI, we prepared yogurt and fed it to diabetic mice models. There was decrease in the blood glucose level after 20 days of oral administration of the yogurt. This product be used as a biodrug in maintaining the blood glucose level in diabetic patients.

Impact of Crohn's disease during pregnancy on children with attention deficit hyperactivity disorder: A review.

BACKGROUND: Iron deficiency anemia (IDA) is a common complication of inflammatory bowel disease (IBD) in pregnant women. Although studies have shown that certain maternal autoimmune diseases are associated with attention deficit hyperactivity disorder (ADHD) in children, no studies have found a relationship specifically between and IDA in pregnant women with IBD and ADHD in their children. This review aims identify a relationship between maternal Crohn's disease (CD) with IDA and ADHD in children. MATERIALS AND METHODS: A review of existing literature was conducted using PubMed to search for articles on pregnant women with CD and IDA and children with ADHD. The studies included nested case-control studies, cohort studies, cross-sectional studies, case-control studies, and literature reviews published from 2012 to 2021. RESULTS: Among 876 articles generated, 11 studies were chosen for this review. Inclusion criteria consisted of no animal studies, meta-analysis or systematic reviews followed by ADHD related topics and maternal CD and IDA. The findings show that ADHD in progeny may be attributed to maternal CD. Although no studies have shown that IDA in mothers with CD is related to ADHD in offspring, several studies have shown a positive correlation between maternal IDA and ADHD in offspring. Some studies suggest inflammation in IBD during pregnancy can inflame the central nervous system, leading to ADHD in offspring. CONCLUSION: IDA is a prevalent complication in CD, and inadequate iron levels are associated with neurodevelopmental problems, such as ADHD. Iron therapy for pregnant mothers diagnosed with CD is suggested to prevent ADHD in offspring.

Comparative Study of Serum Lipid Profiles in Nepalese Cancer Patients Attending a Tertiary Care Hospital

Significant efforts have been made to study cancer at the biochemical and cellular level and identify factors associated with progression. The aim of this hospital based randomized comparative study at the Nepalese Army Institute of Health science hospital was to assess factors in 52 people diagnosed with different types of cancer and 56 normal control persons. Fasting blood samples were analyzed for serum total cholesterol (TC), high density lipoprotein (HDL), triglycerides (TG) and low density lipoprotein (LDL). We found that biochemical parameter TC, TG, VLDL (very low density lipoprotein), LDL and HDL were significantly different in the cancer patients and healthy controls. Levels of TC, TG, LDL, HDL and VLDL were higher in the age group below 50 and that of TG was found to be higher in women than men. Our results indicate that TC, TG and HDL are increased, while LDL and VLDL are lowered in cancer patients. Our study provides clues to risk factors associated with life style, eating habits, and exercise regimens. Monitoring of these parameters with aging is recommended.

A Pilot Study on Screening of BRCA1 Mutations (185delAG, 1294del40) in Nepalese Breast Cancer Patients.

BACKGROUND: Breast cancer is the second most common malignancy among Nepalese women, accounting for 60% of the total cancer cases in females. Women diagnosed with germline mutations in BRCA1 like 185delAG, 1294del40 develop breast and/or ovarian cancer with a lifelong likelihood of up to 85% whereas presence of a mutation increases the risk for mutations to occur in other genes. The major objective of this study was to find the prevalence of these mutations in Nepalese cancer patients. MATERIALS AND METHODS: This prospective study was carried out at two cancer hospitals in the Kathmandu valley over a period of 11 months. Irrespective of age group and stage of canceran appropriate amount of blood was withdrawn from 50 breast cancer patients and 20 controls. DNA was extracted manually and subjected to PCR using primers for 185delAG and 1294del40 mutations. PCR products were then digested with restriction enzyme (DdeII) followed by electrophoresis. RESULTS: Prevalence of 185delAG in reference breast cancer patients was found to be 4/50 (8%) but no 1294del40 was apparent. CONCLUSIONS: Several mutations occurring in different exons of BRCA1 as well as mutations in other genes like BRCA2, for example, should also be taken in account.

Role of hyperinsulinemia in increased risk of prostate cancer: a case control study from Kathmandu Valley.

AIM: To investigate the effect of hyperglycemia and hyperinsulinemia on prostate cancer risk. MATERIALS AND METHODS: This hospital based study was carried out using data retrieved from the register maintained in the Department of Biochemistry of a tertiary care hospital of Kathmandu, Nepal between 31st December, 2011 and 31st October, 2013. The variables collected were age, serum cholesterol, serum calcium, PSA, fasting blood glucose, serum insulin. Analysis was performed by descriptive statistics and testing of hypothesis using Excel 2003, R 2.8.0, Statistical Package for the Social Sciences (SPSS) for Windows Version 16.0 (SPSS Inc; Chicago, IL, USA) and the EPI Info 3.5.1 Windows Version. RESULTS: Of the total 125 subjects enrolled in our present study, 25 cases were of PCa and 100 were healthy controls. The mean value of fasting plasma glucose was 95.5 mg/dl in cases of prostatic carcinoma and the mean value of fasting plasma insulin was 5.78 µU/ml (p value: 0.0001*). The fasting insulin levels µU/ml were categorized into the different ranges starting from ≤2.75, >2.75 to ≤4.10, >4.10 to ≤6.10, >6.10µU/ml. The maximum number of cases of prostatic carcinoma of fasting insulin levels falls in range of >6.10µU/ml. The highest insulin levels (>6.10µU/ml) were seen to be associated with an 2.55 fold risk of prostatic carcinoma when compared with fasting insulin levels of (<2.75 µU/ml). CONCLUSIONS: Elevated fasting levels of serum insulin appear to be associated with a higher risk of prostate cancer.

Diabetes mellitus and renal cell carcinoma--a hospital based study from Kathmandu Valley.

OBJECTIVE: To diagnose renal cell carcinoma at early stages and for better prognosis , the main objective of our current study was to understand any association with diabetes with relation to age, gender, history of disease, diabetic laboratory parameters, tumor size and grade. MATERIALS AND METHODS: This hospital based study was carried out using data retrieved from the register maintained in the Department of Biochemistry of Nepalese Army Institute of Health Sciences between 1st December, 2011 and 31st May, 2012. The variables collected were age, gender, HbA1c, serum creatinine, fasting blood glucose. One way ANOVA was applied to examine statistical significance of differences between groups. The LSD post hoc test was used for the comparison of means of case groups. RESULTS: Of the total 140 cases of renal cell carcinoma, 79 patients were also suffering from diabetes mellitus. The number of females (47) was more in diabetic RCC patients when compared to males (32). Significance was observed in levels of serum creatinine for tumor size >10 cm (0.0001*). The highest value of glycated hemoglobin (8.9%) and fasting blood sugar(148.3mg/dl)in cases of renal cell carcinoma along with diabetes mellitus was found in tumour size of 1-5 cm. CONCLUSION: Diabetes mellitus has independent prognostic significance in RCC in relation to tumour size and grade.

Association of type II diabetes mellitus with hepatocellular carcinoma occurrence--a case control study from Kathmandu Valley.

OBJECTIVE: To assess associations of Type II DM with hepatocellular carcinoma occurrence in Nepal. MATERIALS AND METHODS: This case control study was carried out using data retrieved from the register maintained in the Department of Biochemistry of Nepalese Army Institute of Health Sciences between 1st January, 2012, and 31st August, 2012. The variables collected were age, gender, HbA1c. All biochemical parameters were analyzed in the Central Laboratory of our hospital by standard validated methods. One way ANOVA was used to examine the statistical significant difference between groups with the LSD post-hoc test for comparison of means of case groups. Odds ratios (OR) were calculated using simple logistic-regression analysis. RESULTS: Etiological factors for HCC were HBV, HCV, alcohol and cryptogenic cirrhosis. The highest age group belonged to the etiological category of HCV with a mean of 71.9 ± 3.6 (CI 69.3, 74.5) years and the lowest age group to the etiological category of HBV with 61.7 ± 5.3(CI 57.9, 65.5) years. The main imperative basis of HCC in present study was HCV (39.5%) and second most significant cause of HCC was alcohol (26%). Glycated hemoglobin was found to be more in males with HCC (7.9%) as compared to females (7.3%). The percentage of Type II diabetes mellitus was greater in HCC patients when compared to controls. This difference was statistically significant with an odd ratio of 4.63 (p<0.001). CONCLUSION: Type II DM influences incidence, risk of recurrence, overall survival, and treatment-related complications in HCC patients.

Des-gamma-carboxyprothrombin for early identification and prognosis of hepatocellular carcinoma--a case control study from western Nepal.

OBJECTIVE: To assess the diagnostic and prognostic value of AFP and des-gamma-carboxyprothrombin (DCP) in combination and alone for hepatocellular carcinoma. MATERIALS AND METHODS: A case control study carried out in the Department of Biochemistry of Manipal College of Medical Sciences, Pokhara, Nepal between 1st January 2010 and 31st December 2011. The variables collected were age, gender, BMI, total proteins, albumin, AST, ALT, total bilirubin, DCP, AFP. Approval for the study was obtained from the institutional research ethical committee. Estimation of AFP was performed by ELISA reader for all cases. Analysis was done using descriptive statistics and confidence interval (CI). The data was analyzed using Excel 2003, R 2.8.0 Statistical Package for the Social Sciences (SPSS) for Windows Version 16.0 (SPSS Inc; Chicago, IL, USA) and the EPI Info 3.5.1 Windows Version. RESULTS: The mean age of HCC cases was 53.6±14.93 yrs. The percentage of females was less than males in both cases (23%) and controls (29%). The specificity of DCP reached 100% when its values was equal or greater than 150 (MAU/ml) for 0, 3, 6, 9, 12 months preceding the diagnosis of HCC. Similarly, the specificity for AFP was also nearly 100% when its value was equal or greater than 200 ng/ml 0, 3, 6, 9, 12 months earlier to the finding of HCC. The specificity of DCP (≥40 MAU/mL) and AFP(≥20 ng/mL) in combination was 93%, 97%, 95%, 96%, 97% in respect to 0, 3, 6, 9, 12 months prior to the diagnosis of HCC. CONCLUSION: The combination of both DCP and AFP will improve the finding of initial HCC and the sensitivity of these markers was utmost at the time of HCC identification and noticeably lesser at former time points.

Serum amyloid a as an independent prognostic factor for renal cell carcinoma--a hospital based study from the Western region of Nepal.

OBJECTIVE: The objective of our present study was to assess the role of serum amyloid A (SAA) in stages and prognosis of renal cell carcinoma. MATERIAL AND METHODS: It was a hospital based retrospective study carried out in the Department of Medicine and Biochemistry of Manipal Teaching Hospital, Pokhara, Nepal between 1st January 2008 and 31st December 2011. The variables collected were SAA, CRP. Approval for the study was obtained from the institutional research ethical committee. Quantitative analysis of human SAA and C-reactive protein (CRP) was performed by radial immune diffusion (RID) assay for all cases. RESULTS: Of the 422 total cases of renal cell carcinoma, 218 patients had normal and 204 abnormal SAA. SAA levels were grossly elevated in T3 stage (122.3±SD35.7) when compared to the mean for the T2 stage (84.2±SD24.4) (p value: 0.0001). Similarly, SAA levels were grossly elevated in M1 stage (190.0±SD12.7) when compared to the M0 stage (160.9±SD24.8) (p: 0.0001). There was no significant association with elevated CRP levels (209.1±SD22.7, normal 199.0±SD19.5) . CONCLUSION: The validity of SAA in serum as being of independent prognostic significance in RCC was demonstrated with higher levels in advanced stage disease.

Metabolic changes enhance the cardiovascular risk with differentiated thyroid carcinoma--a case control study from Manipal Teaching Hospital of Nepal.

OBJECTIVE: To evaluate several metabolic changes in patients with differentiated thyroid carcinoma (DTC ) which enhance cardiovascular risk in the western region of Nepal. MATERIALS AND METHODS: This hospital based case control study was carried out using data retrieved from the register maintained in the Department of Biochemistry of the Manipal Teaching Hospital, Pokhara, Nepal between 1st January, 2009 and 31st December, 2011. The variables collected were age, gender, BMI, glucose, insulin, HbA1C, CRP, fibrinogen, total cholesterol, triglycerides, HDL, LDL, VLDL, f-T3, f-T4, TSH. One way ANOVA was used to examine statistical significance of differences between groups, along with the Post Hoc test LSD for comparison of means. RESULTS: fT3 values were markedly raised in DTC cases (5.7±SD1.4) when compared to controls (2.2±SD0.9). Similarly, fT4 values were also moderately raised in cases of DTC (4.9±SD1.3 and 1.7 ±SD0.9). In contrast, TSH values were lowered in DTC cases (0.39±SD0.4) when compared to controls (4.2 ±SD 1.4). Mean blood glucose levels were decreased while insulin was increased and HDL reduced (39.5±SD4.7 as compared to the control 43.1±SD2.2). CONCLUSION: Cardiovascular risk may be aggravated by insulin resistance, a hypercoagulable state, and an atherogenic lipid profile in patients with differentiated thyroid cancer.

Electrospun nylon-6 spider-net like nanofiber mat containing TiO(2) nanoparticles: a multifunctional nanocomposite textile material.

In this study, electrospun nylon-6 spider-net like nanofiber mats containing TiO(2) nanoparticles (TiO(2) NPs) were successfully prepared. The nanofiber mats containing TiO(2) NPs were characterized by SEM, FE-SEM, TEM, XRD, TGA and EDX analyses. The results revealed that fibers in two distinct sizes (nano and subnano scale) were obtained with the addition of a small amount of TiO(2) NPs. In low TiO(2) content nanocomposite mats, these nanofiber weaves were found uniformly loaded with TiO(2) NPs on their wall. The presence of a small amount of TiO(2) NPs in nylon-6 solution was found to improve the hydrophilicity (antifouling effect), mechanical strength, antimicrobial and UV protecting ability of electrospun mats. The resultant nylon-6/TiO(2) antimicrobial spider-net like composite mat with antifouling effect may be a potential candidate for future water filter applications, and its improved mechanical strength and UV blocking ability will also make it a potential candidate for protective clothing.

Photocatalytic and antibacterial properties of a TiO2/nylon-6 electrospun nanocomposite mat containing silver nanoparticles.

Silver-impregnated TiO(2)/nylon-6 nanocomposite mats exhibit excellent characteristics as a filter media with good photocatalytic and antibacterial properties and durability for repeated use. Silver nanoparticles (NPs) were successfully embedded in electrospun TiO(2)/nylon-6 composite nanofibers through the photocatalytic reduction of silver nitrate solution under UV-light irradiation. TiO(2) NPs present in nylon-6 solution were able to cause the formation of a high aspect ratio spider-wave-like structure during electrospinning and facilitated the UV photoreduction of AgNO(3) to Ag. TEM images, UV-visible and XRD spectra confirmed that monodisperse Ag NPs (approximately 4 nm in size) were deposited selectively upon the TiO(2) NPs of the prepared nanocomposite mat. The antibacterial property of a TiO(2)/nylon-6 composite mat loaded with Ag NPs was tested against Escherichia coli, and the photoactive property was tested against methylene blue. All of the results showed that TiO(2)/nylon-6 nanocomposite mats loaded with Ag NPs are more effective than composite mats without Ag NPs. The prepared material has potential as an economically friendly photocatalyst and water filter media because it allows the NPs to be reused.

Expression and purification of a functional human hepatitis B virus polymerase.

AIM: To identify a method for efficient large-scale purification of functional hepatitis B virus polymerase (HBV-Pol) without addition of cellular factors. METHODS: Full-length HBV-Pol (843 amino acids) tagged with 5' end Polyhistidine was expressed at a high level in an Escherichia coli (E. coli) system. Sodium dodecyl sulfate lysis buffer was utilized to dissolve insoluble HBV-Pol, and Ni-NTA resin affinity chromatography was utilized for HBV-Pol purification. Most recombinant HBV-Pol was eluted with 100 mmol/L imidazole in the presence of NP-40, a weak detergent that keeps HBV-Pol in solution. A reducing agent was utilized throughout the purification steps to keep soluble HBV-Pol from redundant disulfide bond formation. RESULTS: The large-scale production of functional intact human HBV-Pol was achieved in an E. coli expression system. Purified HBV-Pol showed stable reverse transcriptase activity and DNA polymerase activity. The purified protein was of high purity and had stable reverse transcriptase activity. CONCLUSION: Large-scale production of HBV-Pol in pure form should facilitate crystallization and detailed analysis of the structure and mechanism of HBV-Pol. Ability of this purification approach to obtain human HBV-Pol in an enzymatically active form should be helpful for development of drugs for treatment of chronic hepatitis B.

Epidemiology and etiopathogenesis of urinary calculi in western Nepal (Pokhara).

Urinary stone disease is recorded in the literature from the dawn of the history and has spared no segment of society irrespective of age, gender, occupation and socio-economic status. It is still termed as "Refractory Disease" as complete medical management to prevent occurrence or recurrence is not so far available. We conducted a preliminary survey from the Manipal Teaching Hospital and carried out urinalyses to ascertain risk factors in the local population. This preliminary survey indicates the prevalence of stone disease is in moderate zone. Hyperoxaluria is an important risk factor in more than one fourth of the stone formers; and hypernatriuria is distinctly most common potentiating risk factor.

Spectrum of stones composition: a chemical analysis of renal stones of patients visiting NMCTH.

A general observation of clinicians suggests that the prevalence ofurolithiasis is fairly high in Kathmandu but so far no systematic study has been undertaken here to explore the etiopathogenesis of disease in this region. In this preliminary communication, we present herewith the qualitative composition of 47 renal stones collected from surgical patients admitted to NMCTH over a period of 13 months (July 2005 to July 2006). All stones were of mixed type. Calcium was present in all stones. Oxalate, phosphate and uric acid were present in 95.7%, 87.2% and 34.0% patients respectively. The probable composition, as construed from analysis, suggests that calcium oxalate stones are predominant. Strikingly, the prevalence was very high in e"20 yrs age group.

Epidemiology and etiopathogenesis of urinary calculi in western Nepal (Pokhara).

Urinary stone disease is recorded in the literature from the dawn of the history and has spared no segment of society irrespective of age, gender, occupation and socio-economic status. It is still termed as "Refractory Disease" as complete medical management to prevent occurrence or recurrence is not so far available. We conducted a preliminary survey from the Manipal Teaching Hospital and carried out urinalyses to ascertain risk factors in the local population. This preliminary survey indicates the prevalence of stone disease is in moderate zone. Hyperoxaluria is an important risk factor in more than one fourth of the stone formers; and hypernatriuria is distinctly most common potentiating risk factor.

Spectrum of stones composition: a chemical analysis of renal stones of patients visiting NMCTH.

A general observation of clinicians suggests that the prevalence ofurolithiasis is fairly high in Kathmandu but so far no systematic study has been undertaken here to explore the etiopathogenesis of disease in this region. In this preliminary communication, we present herewith the qualitative composition of 47 renal stones collected from surgical patients admitted to NMCTH over a period of 13 months (July 2005 to July 2006). All stones were of mixed type. Calcium was present in all stones. Oxalate, phosphate and uric acid were present in 95.7%, 87.2% and 34.0% patients respectively. The probable composition, as construed from analysis, suggests that calcium oxalate stones are predominant. Strikingly, the prevalence was very high in e"20 yrs age group.