The presence of Toxoplasma gondii in soil, their transmission, and their influence on the small ruminants and human population: A review.

The ecosystem approach has been developed since the 1940s. An ecosystem is a community of living organism and their interaction and conjugation with abiotic factors of the environment. The ecosystem is not endemic to the aquatic environment only but, the terrestrial environment is also considered to be a part of an ecosystem. Soil act as mother role for the survival of different microorganism. The Toxoplasma gondii oocysts stay survive for a long time in the soil. This presence of these oocysts might critically enhance the success of this parasite in two ways. First, this parasite can widespread; second, it can create a lot of consequences regarding animals and their economic value. Soil contamination caused by Toxoplasma gondii Y is a significant and profound issue for animals and public health. Therefore, the current study was aimed to summarize and correlate the soil and parasite, their transmission, infection, and some aspects related to T. gondii. The small animals are pose at a high risk therefore, it was concluded that some preventive measures should be taken to keep secure itself from zoonotic diseases.

Diagnostic validity of ultrasonography in thyroid nodules.

OBJECTIVE: To determine the diagnostic validity of thyroid ultrasound in differentiating between benign and malignant thyroid nodules. METHODS: The cross-sectional study was conducted at Aga Khan University Hospital, Karachi, from August 2011 to January 2013, and comprised all patients of either gender with thyroid nodules referred for ultrasound thyroid and fine needle aspiration cytology. Ultrasonography was performed by radiologists and ultrasound parameters were assessed and compared with cytology results in all nodules. Diagnostic validity of each ultrasound feature was calculated. RESULTS: Of the 101 patients, 81(80%) were female. The overall mean age was 43±13 years (range: 15-73 years). On histocytopathology, 96(95%) nodules were benign and 5(4.9%) were malignant. The sensitivity and specificity of ultrasound features in predicting malignancy were calcification 80% and 68%; hypoechogenecity 80% and 52%; ill-defined lobulated margin 40% and 96%; solid 80% and 40%; taller than wider 50% and 63%. Each ultrasound feature had negative predictive value ranging from 95% to 98% in malignant nodules. CONCLUSIONS: Identification of calcification, hypoechogenecity and solid with ill-defined margins in a thyroid nodule on ultrasound was helpful in suspecting thyroid malignancy and warranted urgent diagnostic fine needle aspiration cytology.

Heterologous expression and characterization of mutant cellulase from indigenous strain of Aspergillus niger.

The purpose of current research work was to investigate the effect of mutagenesis on endoglucanase B activity of indigenous strain of Aspergillus niger and its heterologous expression studies in the pET28a+ vector. The physical and chemical mutagens were employed to incorporate mutations in A. niger. For determination of mutations, mRNA was isolated followed by cDNA synthesis and cellulase gene was amplified, purified and sequenced both from native and mutant A. niger. On comparison of gene sequences, it was observed that 5 nucleotide base pairs have been replaced in the mutant cellulase. The mutant recombinant enzyme showed 4.5 times higher activity (428.5 µmol/mL/min) as compared to activity of native enzyme (94 µmol/mL/min). The mutant gene was further investigated using Phyre2 and I-Tesser tools which exhibited 71% structural homology with Endoglucanase B of Thermoascus aurantiacus. The root mean square deviation (RMSD), root mean square fluctuation (RMSF), solvent accessible surface area (SASA), radius of gyration (Rg) and hydrogen bonds analysis were carried at 35°C and 50°C to explore the integrity of structure of recombinant mutant endoglucanase B which corresponded to its optimal temperature. Hydrogen bonds analysis showed more stability of recombinant mutant endoglucanase B as compared to native enzyme. Both native and mutant endoglucanase B genes were expressed in pET 28a+ and purified with nickel affinity chromatography. Theoretical masses determined through ExPaSy Protparam were found 38.7 and 38.5 kDa for native and mutant enzymes, respectively. The optimal pH and temperature values for the mutant were 5.0 and 50°C while for native these were found 4.0 and 35°C, respectively. On reacting with carboxy methyl cellulose (CMC) as substrate, the mutant enzyme exhibited less Km (0.452 mg/mL) and more Vmax (50.25 µmol/ml/min) as compared to native having 0.534 mg/mL as Km and 38.76 µmol/ml/min as Vmax. Among metal ions, Mg2+ showed maximum inducing effect (200%) on cellulase activity at 50 mM concentration followed by Ca2+ (140%) at 100 mM concentration. Hence, expression of a recombinant mutant cellulase from A. niger significantly enhanced its cellulytic potential which could be employed for further industrial applications at pilot scale.

Optimization and Detergent Compatibility of Protease Produced from Aspergillus oryzae by Utilizing Agro Wastes.

The biotechnological process called solid-state fermentation (SSF) was applied for hyper production of protease by using a fungal strain called Aspergillus oryzae. From screening of 9 different local substrates (peanut shell, wheat bran, guava leaves, sugar cane bagasse, rice polish, wheat straw, corn straw, reed grass, and rice straw), peanut shells serve as the best substrates for protease production under optimized cultured conditions. The varying physiochemical parameters such as pH (2-9.5), temperature (30-52 °C), incubation time (1-10 days), inoculum size (1-8 mL), moisture level (20-125%), and substrate concentration (1-7 g) were optimized by response surface methodology (RSM). The highest activity of protease was recorded to be 1101.778 U/mL at 660 nm using peanut shell was optimum at pH 8, temperature 52 °C, incubation time 8 days, inoculum size 2 mL, moisture level 20%, and substrate concentration 2 g. The crude form of enzymes produced were further purified through ammonium sulfate precipitation, dialysis, and gel filtration chromatography. Then, purified enzymes were characterized at different pH, temperature, and incubation time. For characterization of purified protease, pH, temperature, and incubation time were 8, 52 °C, and 8 days for peanut shell and was done by one factor at a time method. Hence, isolated enzymes were alkaline in nature, i.e., alkaline proteases. Then, protease produced from peanut shells was applied to locally available detergents to increase their catalytic activity for strain removal. At last, the final results were interpreted in the form of 3D surface and contour plots using Microsoft Excel 2013 and Minitab 17 software. In conclusion, the utilization of A. oryzae and peanut shell as the substrate in the biotechnological process of SSF demonstrated successful hyper production of alkaline protease. The optimized conditions resulted in high enzyme activity and showcased the potential application of the isolated enzymes in improving the catalytic activity of locally available detergents.

Thermal and hydraulic performance of ZnO/EG based nanofluids in mini tubes of different diameters: An experimental investigation.

The present experimental study investigates the thermal and hydraulic performance of Ethylene Glycol (EG)-based ZnO nanofluids (NFs) in circular minichannel test sections, each of 330 mm in length and 1.0-2.0 mm inner diameters. The experiments were conducted under steady-state constant heat flux and laminar flow conditions. The stable ZnO/EG-based NFs were synthesized using a standard two-step method in varying nanoparticles (NPs) loadings (0.012-0.048 wt%). The morphological characteristics, crystal structure, and specific surface area (SSA) showed that the NPs were sized in nm, possessing excellent crystal structure and enhanced surface area. Thermal conductivity (TC) and viscosity (VC) of the NFs were examined in the 20-60 °C temperature range. Both TC and VC possessed an increasing trend with the rise in concentration of the NPs. However, with the temperature rise, TC increased while the VC decreased and vice versa. The highest enhancements in TC and VC were 14.38 % and 15.22 %, respectively, at 40 °C and 0.048 wt% of NPs loading. The highest enrichment recorded in the local and average heat transfer coefficient (HTC) were 14.80 % and 13.48% in a minichannel with 1.0 mm inner diameter, respectively. It was directly proportional to the NPs loading and volume flow rate of the NFs. The friction factor was also directly proportional to the test section's inner cross-sectional area, while the pressure gradient showed an inverse behavior. An inverse relationship was recorded for the volume flow rate of the NFs and vice versa. Maximum friction factor and the pressure drop for all three minichannel test sections were recorded as 34.58 % and 32.16 %, respectively. The well-known Shah correlation predicted the local and average HTC within ±15.0 %, while the friction factor and the pressure gradient were well predicted by the Darcy correlation within the ±10.0 % range.

Screening of Chitinolytic Microfungi and Optimization of Parameters for Hyperproduction of Chitinase Through Solid-State Fermentation Technique.

This study is intended for the production of chitinase enzyme from locally isolated fungal strains. Out of 10 isolated fungal strains from district Gujrat, Punjab, Pakistan, Aspergillus terreus SB3 (accession number ON738571) was found with maximum chitinolytic potential (80.8 U/mL/min). By applying central composite design (CCD) through response surface methodology (RSM) under solid-state fermentation (SSF), eight nutritional and physical parameters were optimized. Among these, temperature, substrate concentration, and pH were found as significant factors toward chitinase production in the first phase. Moisture and nitrogen source were found as significant factors during second phase of chitinase production. The effect of incubation period, inoculum size, and magnesium source was observed as non-significant. The chitinase activity was successfully enhanced more than 2 folds up to 198.5 U/mL/min at optimized conditions of 35 °C temperature, 4.5 pH, 20 g substrate concentration, 4-day incubation period, 55% moisture content, 4.5 mL inoculum size, 0.25 g ammonium sulfate, and 0.30 g magnesium sulfate using RSM design. It was also found that Ganoderma lucidum (bracket fungus) has more potential to be used for the production of chitinase compared to fish scales. The present study exhibited Aspergillus terreus SB3 (ON738571) as a potential indigenous strain capable for hyperproduction of chitinase through cheap fermentation technology that might be employed for the eradication of chitin-based sea waste to remove the marine pollution.

Incorporation of zinc sulfide nanoparticles, Acinetobacter pittii and Bacillus velezensis to improve tomato plant growth, biochemical attributes and resistance against Rhizoctoniasolani.

Green nanobiotechnology and beneficial bacterial strains as biofertilizers are crucial in agriculture to achieve food security. Both these strategies have been individually studied in improving plant resistance against phytopathogens along with enhancing plant productivity. Therefore, objective of this study was to explore the eco-friendly and cost-effective approach of utilizing plant growth promoting and disease suppressing bacterial strains and nanoparticles, individually as well as in combination, as bio-stimulants to improve plant growth, antioxidant defense system, nutrition and yield of tomato. A pot experiment was conducted to investigate the zinc sulfide nanoparticles (ZnS NPs) synthesized by using Jacaranda mimosifolia flower extracts (JFE), Acinetobacter pittii and Bacillus velezensis either individually or in combinations to check their potential against Rhizoctonia solani in tomato to suppress root rot infection and improve growth and yield. Among all the combinations the JFE-ZnS NPs + B. velezensis compared to untreated infected plants showed minimum disease incidence and maximum significant protection (66%) against R. solani instigated root rot that was followed by JFE-ZnS NPs + A. pittii and individual application of JFE-ZnS NPs by 58%. The same treatment showed maximum significant increase in plant fresh and dry biomass. B. velezensis significantly increased the photosynthetic pigments when applied individually. However, JFE-ZnS NPs alone and in mixed treatments with B. velezensis efficiently improved total soluble protein, sugar and phenolic contents. The same interactive application of JFE-ZnS NPs + B. velezensis improved the tomato plant nutrition (silicon (Si), magnesium (Mg), calcium (Ca) and potassium (K)) and redox quenching status by improving the activity of antioxidant defense enzymes. Overall, the interactive use of JFE-ZnS NPs with A. pittii and B. velezensis very appropriately prepared the host plant to fight against the negative effects of root rot pathogen in tomato. Advancements in interactively investigating the nanoparticles with beneficial plant growth promoting bacterial strains importantly can contribute in resolving the challenges of food security. According to our information, this is a pioneer report for implying JFE-ZnS NPs in synergism with A. pittii and B. velezensis to hinder the root rot in tomatoes.

UV filter ethylhexyl salicylate affects cardiovascular development by disrupting lipid metabolism in zebrafish embryos.

Ethylhexyl salicylate (EHS) is an organic UV filter commonly used in sunscreens to protect people from the UV radiation. The widespread use of EHS will enter the aquatic environment along with human activities. EHS readily accumulates in adipose tissue as a lipophilic compound, but its toxic effects on lipid metabolism and cardiovascular system of aquatic organisms have not been studied. This study investigated the effects of EHS on lipid metabolism and cardiovascular development during zebrafish embryogenesis. The results showed that EHS caused defects such as pericardial edema, cardiovascular dysplasia, lipid deposition, ischemia, and apoptosis in zebrafish embryos. In addition, qPCR and whole-mount in situ hybridization (WISH) results indicated that EHS treatment significantly altered the expression of genes related to cardiovascular development, lipid metabolism, erythropoiesis, and apoptosis. The hypolipidemic drug rosiglitazone was able to alleviate the cardiovascular defects caused by EHS, indicating that EHS affected cardiovascular development by disrupting lipid metabolism. In addition, severe ischemia caused by cardiovascular abnormalities and apoptosis were observed in the EHS-treated embryos, which was likely to be the main cause of embryonic mortality. In conclusion, this study shows that EHS has toxic effects on lipid metabolism and cardiovascular formation. Our findings provide new evidence for assessing UV filter EHS toxicity and contribute to raising awareness of the safety risks of EHS.

Outbreak investigation of NDM-producing Burkholderia cepacia causing neonatal sepsis in Pakistan.

Aim: To investigate the outbreak of Burkholderia cepacia complex (BCC), mortality, antimicrobial resistance and associated risk factors in the neonatal intensive care unit. Method: Eighteen blood culture samples from neonates and twenty swab samples from different neonatal intensive care unit surfaces were collected. The VITEK 2 was used to confirm the isolates and generate the antibiogram. PCR was used to identify blaNDM. Results: Eighteen samples tested positive for BCC, and 10/18 (55.5%) of the neonates died. 13/18 (72%) of the neonates had late-onset neonatal sepsis, and 10/18 (55%) had low birth weight. Resistance to minocycline and chloramphenicol was 100%, 72.2% to meropenem; 72.2% NDM gene was found in neonates and was 20% from the environment. Conclusion: Outbreak of NDM-producing BCC resulting in high neonatal mortality in NICU.

Neonatal septicemia, or blood poisoning, is a dangerous illness in newborns. It is caused by bacteria or other infections entering the blood and spreading. Pregnancy, labor, delivery and exposure after birth can result in infection of the newborn. Neonatal septicemia kills 700,000 babies worldwide, mostly in low- and middle-income countries. Burkholderia cepacia complex bacteria can cause infections in people with weaker immune systems or other disorders. They are particularly dangerous in hospitals, as they can cause chronic lung problems. This study collected blood samples from newborns with blood poisoning. Most samples that contained Burkholderia cepacia complex were not susceptible to drugs. Four of the newborns carried the same bacteria, indicating that hospital staff should practice hand washing and equipment and environmental cleaning to prevent the spread of the bacteria.

Farmer's knowledge and suggested approaches for controlling aflatoxin contamination of raw milk in Pakistan.

Monitoring of aflatoxin levels in milk is often complicated in developing countries due to the dominance of informal markets channeling milk in raw form. Farmer's awareness and voluntary participation in aflatoxin mitigation can be critical in such scenarios. Therefore, the present study was conducted to understand the perceptions of dairy farmers about aflatoxins and link it with aflatoxin mitigation programs on milk in Pakistan. Information was collected from 450 peri-urban dairy farmers in seven cities using questionnaires. Majority (77.9%) of the farmers were aware of the negative impact of moldy feed on animal health. However, only 40.6% of the farmers were aware of the transferability of the toxins from moldy feed to milk. The farmers had almost no awareness of aflatoxins as 95% never heard of the term. After receiving an onsite briefing on effects of the toxin on animal and human health, and its transferability to milk, 98.3% farmers showed willingness to buy aflatoxin-safe feedstuffs, while 88.5% showed willingness to control aflatoxin in milk. Around half of the farmers considered aflatoxin control programs as affordable. On average, farmers agreed to pay 10.1% higher price for aflatoxin certified oilseed cakes. Availability of feedstuffs certified of low aflatoxin content was suggested by 22% of the participants as the critical step in reducing aflatoxins in milk. Other important suggestions included; subsidy on quality feeds (18%), raising awareness (18%), and legislation and monitoring (16%). The present results suggest that the current practice of milk monitoring in the country can yield desirable results only if it is coupled with feed certification programs ensuing availability of aflatoxin-safe feeds. Further, awareness can positively impact participation of producers in aflatoxin control programs. In this regard, awareness about effects of aflatoxins on animal health was found to be a more powerful trigger of voluntary control compared with the awareness of the toxin's transferability to milk.

Neonatal AKI profile using KDIGO guidelines: A cohort study in tertiary care hospital ICU of Lahore, Pakistan.

Background and objective: Acute kidney injury (AKI) was observed in sick neonates and was associated with poor outcomes. Our cohort represents the neonatal characteristics of those diagnosed with AKI using Kidney Disease: Improved Global Outcome (KDIGO) guidelines. Methodology: A cohort study was conducted in the NICU of FMH from June 2019 to May 2021. Data were collected on a proforma. All continuous variables were not normally distributed and expressed as the median and interquartile range. Categorical variables were analyzed by proportional differences with the Pearson chi-square test or Fisher's exact tests. A multinomial logistic regression model was used to explore the independent risk factors for AKI. Time to the event (death) and the cohort's survival curves were plotted using the Cox proportional hazard model. Results: AKI occurred in 473 (37.6%) neonates. The risk factors of AKI were outborn birth [adjusted odds ratio (AOR): 3.987, 95% confidence interval (CI): 2.564-6.200, p: 0.000], birth asphyxia (AOR: 3.567, 95% CI: 2.093-6.080, p: 0.000), inotropic agent (AOR: 2.060, 95% CI: 1.436-2.957, p: 0.000), antenatal steroids (AOR: 1.721, 95% CI: 1.213-2.443, p: 0.002), central lines (AOR: 1.630, 95% CI: 1.155-2.298, p: 0.005) and intraventricular hemorrhage (IVH)/intracranial hemorrhage/disseminated intravascular coagulopathy (AOR: 1.580, 95% CI: 1.119-2.231, p: 0.009). AKI significantly increases the duration of stay and mortality rates by 16.5% vs. 3.9% in neonates with normal renal function (p < 0.001). Conclusion: About one-third of critically sick neonates had AKI. Significant risk factors for AKI were outborn birth, asphyxia inotropic agents, necrotizing enterocolitis, antenatal steroids central lines, and IVH. AKI is associated with an increased length of stay and increased mortality.

Combined ability of salicylic acid and spermidine to mitigate the individual and interactive effects of drought and chromium stress in maize (Zea mays L.).

Application of the growth regulator salicylic acid (SA) and the polyamine spermidine (Spd) can be used to manage various plant abiotic stresses. We aimed to evaluate the sole and combined effects of SA and Spd on maize (Zea mays) under individual and combined drought and chromium (Cr) stress. Drought, Cr, and drought + Cr treatments caused oxidative stress by inducing higher production of reactive oxygen species (H2O2, O2-), enhanced malondialdehyde content and increased relative membrane permeability. Increased oxidative stress and higher Cr uptake in the host plant reduced the content of carotenoids, other photosynthetic pigments and protein, and changed carbohydrate metabolism. Combined drought + Cr stress was more damaging for the growth of maize plants than the individual stresses. Exogenous treatments of SA and Spd alleviated the adverse effects of drought and Cr toxicity, reflected by accumulations of osmolytes, antioxidants and endogenous polyamines. Single applications of Spd (0.1 mM) increased plant height, shoot fresh weight, leaf area, above-ground dry matter accumulation and polyamine content under drought, Cr, and drought + Cr stress conditions. However, the combined treatment SA + Spd (0.25 mM + 0.05 mM) was more effective in increasing protein and water contents, photosynthetic pigments, and carotenoids. The same treatment increased Cr tolerance in the maize plants by decreasing uptake of this heavy metal from root to shoot. The SA + Spd treatment also decreased oxidative stress by promoting antioxidant enzyme activities, and enhanced levels of proline, soluble sugars, and carbohydrate contents under individual and combined stress conditions. Results indicate that the combined half-dose application of SA + Spd may be utilized to boost the tolerance in maize under individual as well as combined drought and Cr stress conditions.

Induction of defense-related enzymes and enhanced disease resistance in maize against Fusarium verticillioides by seed treatment with Jacaranda mimosifolia formulations.

Fusarium verticillioides is an important fungal pathogen of maize, causing stalk rot and severely affecting crop production. The aim of this study was to characterize the protective effects of formulations based on Jacaranda mimosifolia leaf extracts against F. verticillioides in maize. We compared different seed treatments comprising J. mimosifolia extracts, chemical fungicide (mefenoxam) and salicylic acid to modulate the defense system of maize host plants. Both aqueous and methanolic leaf extracts of J. mimosifolia (1.2% w/v) resulted in 96-97% inhibition of mycelial growth of F. verticillioides. While a full-dose (1.2%) extract of J. mimosifolia provided significant protective effects on maize plants compared to the inoculated control, a half-dose (0.6% w/v) application of J. mimosifolia in combination with half-strength mefenoxam was the most effective treatment in reducing stalk rot disease in pot and field experiments. The same seed treatment significantly upregulated the expression of genes in the leaves encoding chitinase, glucanase, lipid transfer protein, and pathogenesis-related proteins PR-1, PR-5 and PR-10, 72 h after inoculation. This treatment also induced the activities of peroxidase, polyphenol oxidase, protease, acid invertase, chitinase and phenylalanine ammonia lyase. We conclude that seed pre-treatment with J. mimosifolia extract with half-strength chemical mefenoxam is a promising approach for the management of stalk rot in maize.

Pre-pregnancy maternal BMI as predictor of neonatal birth weight.

INTRODUCTION: BMI is a tool to measure maternal nutritional status. Maternal malnutrition is frequently reported health problem especially during child bearing age and effects neonatal birth weight. AIM: To determine relationship between prepregnancy maternal BMI and neonatal birth weight. METHODS AND MATERIAL: Prospective, cross sectional study conducted in Fatima Memorial Hospital, Lahore, Pakistan over a period of 1 year including 2766 mother-neonate pairs. All full term, live born neonates of both gender in early neonatal period (<72 hours) with documented maternal pre-pregnancy and/or first trimester BMI were enrolled. Data analysis using SPSS version 20, was performed. RESULTS: Data analysis of 2766 mother-neonates pairs showed that there were 32.9% overweight and 16.5% obese mothers. More than two third of all overweight and obese mothers were of age group between 26-35 years. Diabetes mellitus, hypertension, medical illness, uterine malformations and caesarean mode of delivery were more prevalent in obese mothers as 22.8%, 10.1%, 13.2%, 2.6% and 75.4% respectively. Mean birth weight, length and OFC increased with increasing maternal BMI. Comparing for normal weight mothers, underweight mothers were at increased risk of low birth weight (p< 0.01) and low risk of macrosomic neonates (p<0.01). However overweight and obese mothers were comparable to normal weight mothers for delivering macrosomic neonates (p 0.89 and p 0.66 respectively). CONCLUSIONS: Our study highlights that direct relationship exists between maternal BMI and neonatal birth weight.

GC-MS analysis, antimicrobial, antioxidant, antilipoxygenase and cytotoxic activities of Jacaranda mimosifolia methanol leaf extracts and fractions.

Jacaranda mimosifolia trees are grown in frost-free regions globally. The aim of this study was to evaluate the methanol crude extract and various fractions of increasing polarity of J. mimosifolia leaves for bioactive metabolites, as well as antimicrobial, antioxidant and anticancer activities. The anti-inflammatory potential of the various fractions of J. mimosifolia leaf extract was studied via the lipoxygenase (LOX) inhibitory assay. Methanol crude extract (ME), derived fractions extracted with chloroform (CF) and ethyl acetate (EAF), and residual aqueous extract (AE) of dried J. mimosifolia leaves were assayed for polyphenolic compounds, their antioxidant, antimicrobial and lipoxygenase (LOX) inhibitory activities, and anticancer properties. Polyphenolic compounds were determined via HPLC while phytochemicals (total phenolics, flavonoids, tannins and ortho-diphenol contents), antioxidant activities (DPPH, hydrogen peroxideperoxide, hydroxyl and superoxide radical anions) and LOX were measured via spectrophotometry. Methanol extracts and various fractions were evaluated for antibacterial activities against Bacillus subtilis, Klebsiella pneumonia, Pseudomonas aeruginosa and Staphylococcus aureus. Antifungal potential of the fractions was tested against three species: Aspergillus flavus, Aspergillus fumigatus and Fusarium oxysporum. The highest values for total phenolic content (TPC), total flavonoid content (TFC), flavonols, tannins and ortho-diphenols were in the ME, followed by CF > EAF > AE. ME also had the highest antioxidant activity with EC50 values 48±1.3, 45±2.4, 42±1.3 and 46±1.3 µg/mL based on the DPPH, hydrogen peroxide, hydroxyl radical and superoxide radical assays, respectively. TPC and TFC showed a significant, strong and positive correlation with the values for each of these antioxidant activities. ME exhibited anti-inflammatory potential based on its LOX inhibitory activity (IC50 = 1.3 µg/mL). ME also had the maximum antibacterial and antifungal potential, followed by EAF > CF > AE. Furthermore, ME showed the strongest cytotoxic effect (EC50 = 10.7 and 17.3 µg/mL) against human hormone-dependent prostate carcinoma (LnCaP) and human lung carcinoma (LU-1) cell lines, respectively. Bioactive compounds present in leaf methanol extracts of J. mimosifolia were identified using gas chromatography-mass spectrometry (GC-MS). Fifteen compounds were identified including phenolic and alcoholic compounds, as well as fatty acids. Our results suggest that J. mimosifolia leaves are a good source of natural products with antioxidant, anti-inflammatory and anti-cancer properties for potential therapeutic, nutraceutical and functional food applications.

Aflatoxin Contamination of Milk Produced in Peri-urban Farms of Pakistan: Prevalence and Contributory Factors.

Aflatoxin M1 contamination of milk in Pakistan, like many developing countries, is poorly understood. The present study was therefore conducted to determine AFM1 contamination of milk and its contributory factors in Pakistan. We sampled milk and feedstuffs from 450 peri-urban dairy farms in seven major cities following a cross-sectional study design. Analysis of milk using ELISA revealed high contamination with an overall average of 3164.5 ng of AFM1/L, and significant differences (p < 0.001) between cities. The milk sampled from Gilgit, in northern hilly areas, had an average AFM1 level of 92.5 ng/L. Milk from other cities had 3529.7 ng/L average contamination, with only 5.7% samples qualifying the maximum tolerable limit of 500 ng of AFM1/L. Heavy mean aflatoxin contamination was found in bakery waste (724.6 µg/kg), and cottonseed cake (600.8 µg/kg). Rest of the other feedstuffs had moderate to low mean aflatoxin contamination, ranging from 66.0 µg/kg in maize stover to 3.4 µg/kg in wheat bran. The mean aflatoxin level in commercial dairy concentrates was 32.7 µg/kg. About 80% of the total aflatoxin intake of dairy animals was contributed by cottonseed cake alone due to its high aflatoxin contamination and proportion in dairy rations. On-farm storage time of oilseed cakes varied (p < 0.01) in different cities but was not associated with aflatoxin contamination. The exceptionally high AFM1 contamination suggests that milk from peri-urban dairy farms is a serious public health threat in Pakistan. This situation can be mitigated by reducing aflatoxin contamination in cottonseed cake and promoting the use of commercial concentrates and other feedstuffs with low contamination.

Chitosan-alginate immobilized lipase based catalytic constructs: Development, characterization and potential applications.

In this study, lipase (LIP) was isolated from Aspergillus crevinus, statistically optimized and purified via ammonium sulfate fractionation (ASF), and Sephadex G-100 gel permeation chromatography. LIP was 2.26-folds purified with a specific activity of 223.60 U/mg. The molecular mass was estimated to be 60 kDa on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (5% stacking and a 12% resolving gel) (SDS-PAGE). The active LIP fraction was immobilized onto chitosan-alginate (CTS-ALG) beads developed in a uniform size, i.e., 2.0 ±â€¯0.25 mm diameter using ultrasonically dispersed 2.0% (w/v) chitosan and alginate along with 0.5% (w/v) glutaraldehyde as a macromolecular crosslinking agent. Prior to exploit for detergent compatibility and dehairing purposes, various parameters including pH, thermal, Michaelis-Menten kinetic constants and influence of organic/inorganic and metal ions on PF-LIP and CST-ALG-LIPs fractions were investigated. The immobilized fractions were optimally active and stable over a broader pH (5-11) and temperature (75 °C) as compared to the free counterpart pH (7-8) and temperature (35 °C), respectively. However, the negligible difference between the KM and Vmax values of PF-LIP i.e., 0.133 ±â€¯0.05 mg/mL and 255.0 ±â€¯11.8 U/mL/min and CST-ALG-LIPs revealed that the conformational flexibility of LIP was retained as such. Comparative to PF-LIP, the CTS-ALG-LIPs were found much stable and retained most of their activity up to 80% in the presence of inhibitory molecules. After 75 min incubation, CTS-ALG-LIP3 retained >95% activity at pH 9.0 which was reduced to 80% at pH 10.0 and 44% at pH 11.0. Among all three samples, 100% dehairing was observed when the sheepskin was dipped for 30 min in CTS-ALG-LIP3. The dehairing of leather (sheepskin) was greatly affected by CTS-ALG-LIP3 rendering its potential candidatures for leather and tannery industry.

Chitosan-immobilized pectinolytics with novel catalytic features and fruit juice clarification potentialities.

Biological macromolecules are primarily composed of complex polysaccharides that strengthen microbial growth for the production of industrially relevant enzymes. The presence of polysaccharides in the form of the disrupted cell wall and cell materials are among major challenges in the fruit juice industry. The breakdown of such biological macromolecules including cellulose and pectin is vital for the juices processing. In this background, pectinolytic enzymes including polygalacturonase (PG), pectin lyase (PL), and pectin methylesterase (PME) were isolated from Aspergillus ornatus, statistically optimized and purified via ammonium sulfate fractionation (ASF), dialysis, and Sephadex G-100 gel permeation chromatography. After passing through Sephadex G-100 column, PG, PL, and PME were 2.60-fold, 3.30-fold, and 4.52-fold purified with specific activities of 475.2U/mg, 557.1U/mg, and 205.7U/mg. The active PG, PL, and PME, each separately, were surface immobilized using various concentrations of chitosan and dextran polyaldehyde as a macromolecular crosslinking agent. Prior to exploit for juice clarification purposes, various parameters including pH, thermal and Michaelis-Menten kinetic constants of purified and chitosan-immobilized fractions were investigated. A considerable improvement in the pH and thermal profiles was recorded after immobilization. However, the negligible difference between the Km and Vmax values of purified free and chitosan-immobilized fractions revealed that the conformational flexibility of pectinolytics was retained as such. A significant color and turbidity reductions were recorded after 60min treatment with CTS-PG, followed by CTS-PME, and CTS-PL. It can be concluded that the clarification of apples, mango, peach, and apricot juices was greatly affected by CTS-PG, CTS-PME, and CTS-PL treatments rendering them as potential candidatures for food industry applications.

Reach the Leech: An Unusual Cause of Hematuria.

Leeches are found in fresh water as well as moist marshy tropical areas. Orifical Hirudiniasis is the presence of leech in natural human orifices. Leech have been reported in nose, oropharynx, vagina, rectum and bladder but leech per urethra is very rare. We report a case of leech in urethra causing hematuria and bleeding disorder in the form of epistaxis and impaired clotting profile after use of stream water for ablution. The case was diagnosed after a prolonged diagnostic dilemma. Asingle alive leech was recovered from the urethra after ten days with the help of forceps. The hematuria and epistaxis gradually improved over next 48 hours and the patient became asymptomatic. Natives of leech infested areas should be advised to avoid swimming in fresh water and desist from drinking and using stream water without inspection for leeches.

GenInfoGuard--a robust and distortion-free watermarking technique for genetic data.

Genetic data, in digital format, is used in different biological phenomena such as DNA translation, mRNA transcription and protein synthesis. The accuracy of these biological phenomena depend on genetic codes and all subsequent processes. To computerize the biological procedures, different domain experts are provided with the authorized access of the genetic codes; as a consequence, the ownership protection of such data is inevitable. For this purpose, watermarks serve as the proof of ownership of data. While protecting data, embedded hidden messages (watermarks) influence the genetic data; therefore, the accurate execution of the relevant processes and the overall result becomes questionable. Most of the DNA based watermarking techniques modify the genetic data and are therefore vulnerable to information loss. Distortion-free techniques make sure that no modifications occur during watermarking; however, they are fragile to malicious attacks and therefore cannot be used for ownership protection (particularly, in presence of a threat model). Therefore, there is a need for a technique that must be robust and should also prevent unwanted modifications. In this spirit, a watermarking technique with aforementioned characteristics has been proposed in this paper. The proposed technique makes sure that: (i) the ownership rights are protected by means of a robust watermark; and (ii) the integrity of genetic data is preserved. The proposed technique-GenInfoGuard-ensures its robustness through the "watermark encoding" in permuted values, and exhibits high decoding accuracy against various malicious attacks.