Unveiling the Chemistry and Synthetic Potential of Catalytic Cycloaddition Reaction of Allenes: A Review.

Allenes with two carbon-carbon double bonds belong to a unique class of unsaturated hydrocarbons. The central carbon atom of allene is sp hybridized and forms two σ-bonds and two π-bonds with two terminal sp2 hybridized carbon atoms. The chemistry of allenes has been well documented over the last decades. They are more reactive than alkenes due to higher strain and exhibit significant axial chirality, thus playing a vital role in asymmetric synthesis. Over a variety of organic transformations, allenes specifically undergo classical metal catalyzed cycloaddition reactions to obtain chemo-, regio- and stereoselective cycloadducts. This review briefly describes different types of annulations including [2+2], [2+2+1], [3+2], [2+2+2], [4+2], [5+2], [6+2] cycloadditions using titanium, cobalt, rhodium, nickel, palladium, platinum, gold and phosphine catalyzed reactions along with a mechanistic study of some highlighted protocols. The synthetic applications of these reactions towards the synthesis of natural products such as aristeromycin, ent-[3]-ladderanol, waihoensene(-)-vindoline and (+)-4-epi-vindoline have also been described.

Targeting the effect of sofosbuvir on selective oncogenes expression level of hepatocellular carcinoma Ras/Raf/MEK/ERK pathway in Huh7 cell line.

Direct acting antiviral agents are emerging line of treatment to eradicate Hepatitis C virus. Recent controversy over whether direct acting antiviral agents increase rate of hepatocellular carcinoma in HCV patients or prevent it, has increased the need to elaborate underlying mechanisms on molecular basis. This work was aimed to investigate the effect of sofosbuvir on the expression of selected oncogenes from the Ras/Raf/MEK/ERK pathway in Huh7 cell line. Results found concrete molecular evidence that sofosbuvir has significantly altered the expression of selected genes when huh7 cell line was treated with sofosbuvir. Nine genes related to HCC were found to be affected by sofosbuvir in a mixed effect manner. The relative expression of growth factors (VEGF, PDGFRB and HGF) was increased in sofosbuvir treated cell lines. The kinase family genes H-RAS, B-RAF, MET except MAPK1 were downregulated. Similarly, DUSP1 was upregulated and SPRY2 was slightly downregulated; both were negative feedback inhibitors of ERK signalling cascade. Sofosbuvir upregulated the growth factors and MAPK1 which suggests it to be a carcinogen. The downregulation of kinases and upregulation of DUSP1 make it an anticancer drug. Hence, the results from this study are important to prove that sofosbuvir neither reduce nor induce hepatocellular carcinoma.

Identification of phytochemical inhibitors of SARS-CoV-2 protease 3CLpro from selected medicinal plants as per molecular docking, bond energies and amino acid binding energies.

Recent worldwide outbreak of SARS-COV-2 pandemic has increased the thirst to discover and introduce antiviral drugs to combat it. The bioactive compounds from plant sources, especially terpenoid have protease inhibition activities so these may be much effective for the control of viral epidemics and may reduce the burden on health care system worldwide. Present study aims the use of terpenoid from selected plant source through bioinformatics tools for the inhibition of SARS-COV-2. This study is based on descriptive analysis. The Protein Data Bank and PubChem database were used for the analysis of SARS-COV-2 protease and plant source terpenoids. Molecular docking by using molegro virtual docker (MVD) software was carried out. The findings of study are based on the inhibitory actions of different plant sourced terpenoid against SARS-COV-2. As per the available resources and complementary analysis these phytochemicals have capacity to inhibit 3CLpro protease. The study reports that (3,3-dimethylally) isoflavone (Glycine max), licoleafol (Glycyrrhiza uralensis), myricitrin (Myrica cerifera), thymoquinone (Nigella sativa), bilobalide, ginkgolide A (Ginkgo biloba), Salvinorin A (Salvia divinorum), citral (Backhousia citriodora) and prephenazine (drug) showed high activity against SARS-COV-2 protease 3CLpro. The drug like and ADMET properties revealed that these compounds can safely be used as drugs. Cross structural analysis by using bioinformatics study concludes that these plant source terpenoid compounds can be effectively used as antiprotease drugs for SARS-COV-2 in future.

Molecular characterization and expression of cyclic nucleotide gated ion channels 19 and 20 in Arabidopsis thaliana for their potential role in salt stress.

Cyclic nucleotide gated ion channels (CNGCs) in plants have very important role in signaling and development. The study reports role of CNGC19 and CNGC20 in salt stress in A. thaliana. In-silico, genome wide analysis showed that CNGC19 and CNGC20 are related to salt stress with maximum expression after 6 h in A. thaliana. The position of inserted T-DNA was determined (in-vivo) through TAIL-PCR for activation tagged mutants of CNGC19 and CNGC20 under salt stress. The expression of AtCNGC19 and AtCNGC20 after cloning under 35S promoter of expression vectors pBCH1 and pEarleyGate100 was determined in A. thaliana by real-time PCR analysis. Genome wide analysis showed that AtCNGC11 had lowest and AtCNGC20 highest molecular weight as well as number of amino acid residues. In-vivo expression of AtCNGC19 and AtCNGC20 was enhanced through T-DNA insertion and 35S promoter in over-expressed plants under high salt concentration. AtCNGC19 was activated twice in control and about five times under 150 mM NaCl stress level, and expression value was also higher than AtCNGC20. Phenotypically, over-expressed plants and calli were healthier while knock-out plants and calli showed retarded growth under salinity stress. The study provides new insight for the role of AtCNGC19 and AtCNGC20 under salt stress regulation in A. thaliana and will be helpful for improvement of crop plants for salt stress to combat food shortage and security.

Exploration of Carica papaya bioactive compounds as potential inhibitors of dengue NS2B, NS3 and NS5 protease.

Current outbreak of dengue has shown serious health concerns in Pakistan. The present study reports the anti-dengue potential of Carica papaya natural compounds. The leaves of C. papaya have previously shown promising results in cure of Dengue fever. The aim of this project is to find specific bioactive compounds by computational screening and biological activities of C. papaya against serine NS2B, NS3 and NS5 proteases of dengue virus. Docking study resulted in the screening of nine bioactive compounds having highest docking scores. However, three compounds namely epigallocatchin, catechin and protocatechuric acid had the strongest binding affinity with the active residues i.e., Ser135, His51 and Asp75 of dengue virus serine proteases. Results also indicated that the extract of C. papaya was a strong antimicrobial and antioxidant agent. It is concluded that the C. papaya compounds can be commercially applied for medical formulations against dengue virus.

Pharmacokinetics profile of serum and cellular Sofosbuvir along with its concentration effect analysis in HCV patients receiving Sofosbuvir and Ribavirin.

Sofosbuvir along with ribavirin is being widely used for treatment of HCV in Pakistan but it may show delayed response and reoccurrence of disease in some cases. The aim of the study was to investigate pharmacokinetics and concentration effect analysis of sofosbuvir. HCV patients (n=100) received 400 mg sofosbuvir along with low dose or weight based ribavirin (400 mg). Nonlinear mixed effects modeling (NONMEM) and unpaired t-test were used for the association of concentrations and treatment outcomes. Average day 10 sofosbuvir metabolite BM 331007 concentration was higher in patients having haemoglobin nadir value <10 g/dl compared to the patients having heamoglobin nadir value >10 g/dl (5.34 versus 4.87 pmol/106 cells; p=0.03). The average concentration trends of GS331007 at day 10 was towards being higher in the patients achieved sustained virologic response (SVR) as compare to the patients relapsed (5.19 versus 4.86 pmol/106 cells; p=0.05). Sofosbuvir (GS331007) thresholds concentration (suggested at day 10 through receiver operating characteristic curve) was 5.4 pmol/106 cells for SVR (p=0.05) and haemoglobin nadir cells was 6.3 pmol/106 with sensitivity and specificity of >60%. Dosing simulations shows that 400 mg sofosbuvir twice daily produce day 10 concentration range of 5.4 to 6.7 pmol/106 cells. The range of therapeutic values was identified for HCV patients receiving sofosbuvir in combination with ribavirin for 24 weeks, suggesting a potential pharmaceutical basis for individualized therapeutic dosing.

Evaluation of 99mTc-sulfadiazine as Bacillus microorganisms infection imaging agent using animal model.

Bacterial infection is one of the vital sources of morbidity and mortality. The development of single photon emission computed tomography (SPECT) radiotracer agents using antibiotics, for targeting in-vivo bacteria, helps in antibiotic dose calibration, targeted infection therapy and reduction in mortality rate. The aim of this study was to appraised 99mTc-labeling sulfadiazine as a radiopharmaceutical for bacillus infections imaging. Radiolabeling of sulfadiazine with technetium-99m was carried out by subsequent addition of 1.5 mL aqueous solution of sulfadiazine (1mg/mL), 120µg stannous tartrate, gentistic acid as stabilizing agent and 185 MBq normal saline solution of 99mTcO4-1 (pertechnetate) at pH = 5. The reaction mixture was incubated for 40 min at room temperature with light stirring. The quality control analysis (ITLC-SG and paper chromatography analysis) revealed ~ 98% labeling yield. Biodistribution and scintigraphic study was carried using bacillus bacterial infection induced New Zealand white rabbits. Due to the ease of 99mTc-sulfadiazine conjugation method, high labeling efficiency, shelf stability (>95% up to 6h), blood serum stability (~90% up to 6h) and high uptake in the infected muscle (T/NT =2.21 at 1H), 99mTc-SDZ could be used as radiopharmaceutical of choice for further pre-clinical and clinical studies.

Zn-doped SiO2 nanoparticles preparation and characterization under the effect of various solvents: Antibacterial, antifungal and photocatlytic performance evaluation.

The biological profiling (antibacterial and antifungal) and photocatalytic activity (PCA) was evaluated of newly synthesized Zinc-doped SiO2 nanoparticles (Zn-d-SiO2 NPs) under the effect of different solvents (n-hexane, acetonitrile, isoamyl alchol). Antibacterial potential against Bacillus subtilis and Escherichia coli, and antifungal activities against Candida parapsilosis and Aspergilus niger were investigated by the Disc diffusion method. The advanced techniques including Scanning Electron Microscopy (SEM), Fourier Transform Infrared Spectroscopy (FT-IR), Transmission Electron Microscopy (TEM), X-ray diffraction (XRD), Thermo-gravimeteric Analysis (TGA), Atomic forced microscopy (AFM) and Dynamic Light Scattering (DLS) were used for the characterization of Zn-d-SiO2 NPs. FT-IR characterization confirmed the presence of metal bond SiO2, while XRD showed the hexagonal structure with 7.93 nm average particle size of Zn-d-SiO2 NPs. SEM and TEM analysis revealed the variation in morphology of NPs prepared in different solvents. The Zn-d-SiO2 NPs prepared in the presence of acetonitrile showed highest antibacterial activities (6% and 4.5%) followed by isoamyl alchol (3% and 2.4%) and n-hexane (2%and 1%) against B. subtilis and E. coli, respectively. The antifungal activities were recorded in following order; n-hexane (0.34% and 0.43%) < isoamyl alchol (0.27% and 0.19%) < acetonitrle (0.21% and 0.17%) against C. parapsilosis and A. niger, respectively. The PCA of Zn-d-SiO2 NPs was evaluated by degrading Crystal Violet (CV) dye under solar light irradiation and up to 85.5% dye degradation was achived for 120 min irradiation. Results revealed that the solvents have a significant effect on physicochemical properties and Zinc-doped SiO2 NPs showed promising PCA and antimicrobial activities.

Vibrio fischeri bioluminescence inhibition assay for ecotoxicity assessment: A review.

Vibrio fischeri bioluminescence inhibition bioassay (VFBIA) has been widely applied for the monitoring of toxicity on account of multiple advantages encompassing shorter test duration, sensitive, cost-effective and ease of operation. Moreover, this bioassay found to be equally applicable to all types of matrices (organic & inorganic compounds, metals, wastewater, river water, sewage sludge, landfill leachate, herbicides, treated wastewater etc.) for toxicity monitoring. This review highlights the apparent significance of Vibrio fischeri bioluminescence inhibition assay for ecotoxicological screening and evaluation of diverse chemical substances toxicity profile. The biochemical and genetic basis of the bioluminescence assay and its regulatory mechanism have been concisely discussed. The basic test protocol with ongoing improvements, widespread applications, typical advantages and probable limitations of the assay have been overviewed. The sensitivity of VFBIA and toxicity bioassays has also been compared.

Healing potential of Adiantum capillus-veneris L. plant extract on bisphenol A-induced hepatic toxicity in male albino rats.

Bisphenol A (BPA) is a widely used environmental pollutant in the production of plastics but causes hepatotoxicity in mammals. In the present study, we studied the BPA-induced oxidative stress in rats and ameliorative potential of Adiantum capillus-veneris L. plant. It was concluded that the BPA can reduce the body and liver weight, increase in biochemical levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST), total bilirubin, and disturb the normal hepatic physiology, histology, and metabolism. Additionally, liver histology shows hepatic necrosis, congestion, and vacuolization in exposed individuals. In contrast, simultaneous exposure of A. capillus-veneris and BPA showed declining trend in serum biomarker levels and normal histopathological structures. We conclude that the A. capillus-veneris plant is antioxidant in nature and can reduce the BPA-induced toxicity. These findings are very helpful to understand the BPA-induced hepatic toxicity and ameliorative potential of A. capillus-veneris plant and are of great importance in risk assessment of xenobiotics.

Sorafenib tosylate, Ribavirn and Sofosbuvir combination therapy for HCV virus infected patients with decompensated liver cancer.

Hepatitis C is the most common health problem worldwide and is major cause of death due to proliferation of hepatocellular carcinoma. The medicines available for HCV treatment overcome up-to 95% complications of HCV. However, liver cancer needs some additional care. Normally Sorafenib tosylate 200 mg is recommended for liver cancer. There is no such trial in which this drug could effectively be used in combination of direct acting antivirals for HCV. The study was conducted for HCV patients (n=30) with liver cancer having decompensated stage. Combination of Sorafenib tosylate, Ribavirn and Sofosbuvir were used for the pharmacokinetics of these medicines. Child pugh score less then 7 (CP A) in adults during treatment phase (received 12 weeks of Sorafenib tosylate 200 mg, Ribavirn and Sofosbuvir 400 mg once daily) have no side effect while child pugh score 7-9 (CP B) have evidence of hypertension. The main efficiency end point sustained virology response with overcoming liver cancer as well in 12 weeks after end treatment (SVR-LLC 12). Mean pharmacokinetic exposure to Sorafenib tosylate 200 mg, Ribavirn and Sofosbuvir at week 8th was 2.1, 1.5,1.2 times greater in CP B than in CP A. Adverse effects (AEs) were observed in 12 out of 30 patients but not severe as lethal for life. Treatment with Sorafenib tosylate, Ribavirn and Sofosbuvir for twelve weeks was harmless and well accepted, 100 % patients achieve (SVR LLC 12) with 10-fold cure rate more than previous ones. The combination therapy of Sorafenib tosylate, Ribavirn and Sofosbuvir was found helpful for the management of decompensated liver cancer.

Acidic and enzymatic saccharification of waste agricultural biomass for biotechnological production of xylitol.

BACKGROUND: The plant biomass and agro-industrial wastes show great potential for their use as attractive low cost substrates in biotechnological processes. Wheat straw and corn cob as hemicellulosic substrates were acid hydrolyzed and enzymatically saccharified for high xylose production. The hydrolysate was concentrated and fermented by using Saccharomyces cerevisiae and Kluyveromyces for production of xylitol. RESULTS: Acid hydrolysis of wheat straw and corn cob in combination with enzymatic hydrolysis showed great potential for production of free sugars from these substrates. Kluyveromyces produced maximum xylitol from acid treated wheat straw residues with enzymatic saccharification. The percentage xylitol yield was 89.807 g/L and volumetric productivity of 0.019 g/L/h. Kluyveromyces also produced maximum xylitol from corn cob acid hydrolyzed liquor with xylitol yield 87.716 g/L and volumetric productivity 0.018 g/L/h. CONCLUSION: Plant and agro-industrial biomass can be used as a carbohydrate source for the production of xylitol and ethanol after microbial fermentation. This study revealed that wheat straw acid and enzyme hydrolyzed residue proved to be best raw material for production of xylitol with S. cerevisiae. The xylitol produced can be utilized in pharmaceuticals after purification on industrial scale as pharmaceutical purposes.

Negative effects of bisphenol A on testicular functions in albino rats and their abolitions with Tribulus terristeris L

ABSTRACT This study was conducted to find out the ameliorative properties of Tribulus terristeris L (TT) on BPA induced spermatotoxicity in male albino rats. Mature male albino rats were divided into five groups, Group A was taken as control for comparison group, whereas the other four groups namely B(vehicle control), C (toxic), D (preventive control) and Group E (amelioration group) received distilled water, olive oil, BPA, TT, and (TT + BPA) respectively. Macroscopic results revealed decreased body weight of rats, weight of testes, and the relative tissue weight index (RTWI) in BPA induced group. Hormonal (testosterone) assay results revealed the decreased values of BPA treated group. Microscopic examination of testis of BPA treated rats showed reduction in leydig cells, decreased diameter of seminiferous tubules and low values of Johnsen's scoring. Histological examination showed discontinuity and irregularity of basement membrane and sloughing of the germinal cell linage. Group E showed the body weights of rats, weight of testes, RTWI, and increased, while reduced level of testosterone, reduced number of Leydig cells, decreased diameter of seminiferous tubules and low values of Johnsen's scoring were restored near to normal. These results demonstrate that TT might be beneficial in combating the spermatotoxicity, induced by BPA.

Bisphenol A exposure and healing effects of Adiantum capillus-veneris L. plant extract (APE) in bisphenol A-induced reproductive toxicity in albino rats.

The current study presents the bisphenol A exposure and the ameliorative effects of Adiantum capillus-veneris on testicular toxicity induced by bisphenol A. Adult male albino rats were divided into five groups of five animals each: A (control), B (vehicle control), C (toxic), D (protective), and E (ameliorative) were served distilled water, olive oil, bisphenol A (BPA) at 100 mg/kg body weight, A. capillus-veneris plant extract at 25 mg/kg body weight, and BPA + A. capillus-veneris, respectively. All of the doses were administered orally for 15 days, and the rats were then sacrificed. Blood samples for the testosterone assay and both testes were collected for histological examination. The body weight, paired testes weight, relative tissue weight index, Johnsen scoring of tubules, and level of serum testosterone decreased in BPA-treated rats. Similarly, histological examination of the testes in BPA-treated animals revealed a lower number of Leydig cells, an irregular basement membrane, sloughing of germinal layers, vacuolization, a lower number of spermatocytes, and debris in the lumen. However, co-administration of A. capillus-veneris with BPA increased the total antioxidative capacity (330.82 ± 22.46 µmol/mg protein) of the testes and restored the serum testosterone level (1.70 ng/ml); histological features showed restoration in the stages of spermatogenesis. Conclusively, A. capillus-veneris plant extract overcomes the estrogenic effects of BPA on the reproductive system of rats and protects rats' testes against BPA-induced injury/damage via an antioxidative mechanism that appears to be conciliated.

Chemical Composition and in-Vitro Evaluation of the Antimicrobial and Antioxidant Activities of Essential Oils Extracted from Seven Eucalyptus Species.

Eucalyptus is well reputed for its use as medicinal plant around the globe. The present study was planned to evaluate chemical composition, antimicrobial and antioxidant activity of the essential oils (EOs) extracted from seven Eucalyptus species frequently found in South East Asia (Pakistan). EOs from Eucalyptus citriodora, Eucalyptus melanophloia, Eucalyptus crebra, Eucalyptus tereticornis, Eucalyptus globulus, Eucalyptus camaldulensis and Eucalyptus microtheca were extracted from leaves through hydrodistillation. The chemical composition of the EOs was determined through GC-MS-FID analysis. The study revealed presence of 31 compounds in E. citriodora and E. melanophloia, 27 compounds in E. crebra, 24 compounds in E. tereticornis, 10 compounds in E. globulus, 13 compounds in E. camaldulensis and 12 compounds in E. microtheca. 1,8-Cineole (56.5%), α-pinene (31.4%), citrinyl acetate (13.3%), eugenol (11.8%) and terpenene-4-ol (10.2%) were the highest principal components in these EOs. E. citriodora exhibited the highest antimicrobial activity against the five microbial species tested (Staphylococcus aureus, Bacillus subtilis, Escherichia coli, Aspergillus niger and Rhizopus solani). Gram positive bacteria were found more sensitive than Gram negative bacteria to all EOs. The diphenyl-1-picrylhydazyl (DPPH) radical scavenging activity and percentage inhibition of linoleic acid oxidation were highest in E. citriodora (82.1% and 83.8%, respectively) followed by E. camaldulensis (81.9% and 83.3%, respectively). The great variation in chemical composition of EOs from Eucalyptus, highlight its potential for medicinal and nutraceutical applications.

PROTECTIVE EFFECT OF SACCHARUM OFFICINARUM L. (SUGAR CANE) JUICE ON ISONIAZID INDUCED HEPATOTOXICITY IN MALE ALBINO MICE.

BACKGROUND: Isoniazid (INH) is the drug of choice for treatment of tuberculosis (TB) and it is a well-known-cause of acute clinical liver injury which can be severe and sometimes fatal. The study was designed to investigate the effects of Saccharum officinarum L. juice on oxidative liver injury due to INH in mice. METHODS: This was a laboratory based experimental study. Thirty mice were divided into three groups, containing 10 mice each. Group A being the control, group B and C were experimental and were treated orally with INH 100 mg/kg per day and INH 100 mg/kg per day plus Saccharum officinarum L. juice 15 ml/ kg per day respectively for a period of 30 days. Blood samples were taken at 30th day by cardiac puncture under anaesthesia and liver in each was taken out for microscopic examination. RESULTS: INH treated mice showed; rise in serum ALT, AST, ALP and total bilirubin levels (Mean?SEM), while group C mice treated with Saccharum officinarum L. juice significantly decreased the levels of these biochemical parameters. The histopathological examination of groups A showed normal liver structure which was deranged in (INH) group B, whereas group C showed significant recovery in histological structure. Saccharum officinarum L. constituents, especially flavanoids and anthocyanins have strong antioxidant properties which provides hepatoprotection against oxidative liver injury produced by INH. CONCLUSION: INH-induced liver injury is associated with oxidative stress, and co-administration of Saccharum officinarum L. juice (15 ml/Kg bw) may reduce this damage effectively in mice.

Evaluating Insects as Bioindicators of Heavy Metal Contamination and Accumulation near Industrial Area of Gujrat, Pakistan.

To study the accumulation and contamination of heavy metals (i.e., Cd, Cr, Cu, Ni, and Zn) in soil, air, and water, few insect species were assayed as ecological indicators. Study area comes under industrial zone of district Gujrat of Punjab, Pakistan. Insects used as bioindicators included a libellulid dragonfly (Crocothemis servilia), an acridid grasshopper (Oxya hyla hyla), and a nymphalid butterfly (Danaus chrysippus) near industrial zone of Gujrat. Accumulation of Cd was highest in insect species followed by Cu, Cr, Zn, and Ni at p < 0.05. Hierarchical cluster analysis (HACA) was carried out to study metal accumulation level in all insects. Correlation and regression analysis confirmed HACA observations and declared concentration of heavy metals above permissible limits. Metal concentrations in insects were significantly higher near industries and nallahs in Gujrat and relatively higher concentrations of metals were found in Orthoptera than Odonata and Lepidoptera. The total metal concentrations in insects were pointed significantly higher at sites S3 (Mid of HalsiNala), S9 (End of HalsiNala), and S1 (Start of HalsiNala), whereas lowest value was detected at site S6 (Kalra Khasa) located far from industrial area. HACA indicates that these insect groups are potential indicators of metal contamination and can be used in biomonitoring.

Immunolocalization of aquaporin-10 in tuberculous human ileum.

OBJECTIVE: To determine the presence of AQP-10 in the ileum of patients suffering from intestinal tuberculosis. STUDY DESIGN: A cross-sectional analytical study. PLACE AND DURATION OF STUDY: Department of Anatomy, University of Health Sciences, Lahore, in year 2010. METHODOLOGY: Thirty seven paraffin embedded blocks of either surgically resected specimens or ileal biopsies with diagnosis of intestinal tuberculosis were selected from records of the histopathology departments of local hospitals. These cases were subdivided into two groups: A-1 (with tuberculous granulomatous lesions with or without epithelium) and A-2 (without tuberculous lesion lying adjacent to the lesions and having an intact epithelium). Specimens of small intestine with malignancy, Crohn's disease, inflammatory bowel disease, irritable bowel syndrome and diarrhoeal diseases caused by Rota virus, adenovirus, Salmonella, Shigella and Escherichia coli were excluded. The variables studied were the presence/absence and location of AQP-10. RESULTS: The most common clinical symptoms found in tuberculous patients were abdominal pain followed by diarrhoea. A significant association was found between AQP-10 and site of granulomas and caseation necrosis (p=0.002 and p=0.006 respectively). Absence of AQP-10 was observed in tuberculous ileum at the site of lesion with ulceration. A strong positive staining of AQP-10 was found in the intact epithelium at sites adjacent to the tuberculous lesion indicating its localization near the epithelial lining of ileum. CONCLUSION: AQP-10 was present only on the epithelial cells occurring at the luminal side of the villi and was absent in tuberculous ileum where epithelium was absent.

Biochemical analysis and investigation on the prospective applications of alkaline protease from a Bacillus cereus strain.

Proteases have prospective financial and environment-friendly applications; hence attention is focused currently on the finding of new protease producing microorganism so as to meet the requirements of industry. A thermophilic bacterial strain producing extracellular protease activity was isolated from soil and identified as Bacillus cereus by analysis of 16S rRNA. Protease production by the microorganism was improved by studying the impact of the type of nitrogen and carbon source, fermentation period, growth temperature and initial pH of the culture medium in cultivation optimization experiments. The enzyme was purified to homogeneity in two step procedure involving Sephadex G-75 and Q-Sepharose chromatography. The molecular weight of purified enzyme was found to be 58 kDa by SDS-PAGE. Protease exhibited a pH and temperature optima of 7.5 and 60°, respectively. The enzyme was active in the pH range of 6.0-9.0 and stable up to 70°C. Histological analysis of protease treated goat and cow skin pelts showed complete removal of non leather forming structures such as hair shaft, hair follicles and glandular structures. The protease showed the stain removing property from blood stained cotton cloth and found to be compatible with six commercially available detergents. The protease could release peptides from natural proteins after digestion of coagulated egg albumin and blood clot.

Quantitative and qualitative analysis of cadaveric human pinealocytes in various age groups.

OBJECTIVE: To determine age-related quantitative and qualitative changes in human pinealocytes using cadaveric material. STUDY DESIGN: Analytical cross-sectional study. PLACE AND DURATION OF STUDY: The study was conducted in the Department of Anatomy, University of Health Sciences, Lahore, from January to December 2008. METHODOLOGY: Thirty pineal glands from human cadavers ranging from 16-80 years of age were collected from mortuary of King Edward Medical University, Lahore, using purposive non-probability sampling. These were divided into three different age groups: I, II and III each between 16 to 30, 31 to 45 and 46 to 80 years of age respectively. Pinealocytes were counted; their mean diameter and that of their nuclei was calculated from a total of 30 cells per slide, using 4 µm thick H and E stained histological sections. Mean ± S.E.M. was calculated for quantitative variables. One-way ANOVA was applied to observe group mean differences among three groups. RESULTS: The number of pinealocytes decreased with aging but the difference was statistically insignificant when compared between groups (p=0.234). There was no change in size of pinealocyte soma and its nucleus (p=0.889 and 0.898 respectively). CONCLUSION: The number and size of pinealocytes, and their nuclei remained unaltered with advancing age.