Cucumis sativus and Cucurbita maxima extract attenuate diabetes-induced hepatic and pancreatic injury in a rat model.

Diabetes is usually associated with oxidative stress that causes hepatic and pancreatic tissue injury. This work was carried out to evaluate the effect of Cucumis sativus and Cucurbita maxima methanol extracts on the streptozotocin-induced diabetic hepatic and pancreatic injury in rats. Diabetes was induced in seven equal groups of rats by a single intraperitoneal injection of streptozotocin (40 mg/kg), in addition to the non-diabetic control group. Two diabetic groups were treated with Cucumis sativus methanol extract and two were treated with Cucurbita maxima, each at 200 and 400 mg/kg for 21 days after streptozotocin-induced diabetes. Another diabetic group was treated with both Cucumis sativus and Cucurbita maxima at 200 mg/kg of each. Another group was treated with metformin (200 mg/kg orally). The plant extracts normalized serum liver enzymes activities, oxidative stress markers, and restored serum proteins and lipid profile. They also significantly reduced blood sugar to values comparable to non-diabetic rats. The hypoglycemic effect is also confirmed by the improvement of the immunohistochemical expression of insulin in ß-cells of islets of Langerhans. Hepatic and pancreatic protection was also confirmed by the improvement of the histopathological picture as compared to STZ-diabetic rats. The GC-MS analysis revealed the presence of 35 and 34 compounds in the methanol extract of cucumber and pumpkin, respectively. Finally, the methanol extract of cucumber and pumpkin could be beneficial acting synergistically in the protection of the liver and pancreas against diabetes-induced tissue damage.

Current trends of Hepatitis C virus genotypes and associated risk factors in hemophilia patients in Pakistan

@# Hemophilia is a rare bleeding disorder that needs plasma or clotting factor concentrate transfusion. Therefore chances of blood-borne pathogens like HCV transmission increase due to high prevalence in healthy donors. This study was aimed to determine the prevalence of HCV genotypes and associated risk factors in hemophilia patients of Khyber Pakhtunkhwa, Pakistan. Blood samples and data were collected from 672 hemophiliacs after proper consent obtained from each patient. Samples were analyzed for anti-HCV, HCV RNA and HCV genotype/s detection. Of the total, 22.32% (150) were anti-HCV positive, of which HCV RNA was detected in 18.45% (124) individuals. HCV genotype 3a was found with significantly higher prevalence (p<0.05) (19.35%) as compared to 2a (16.13%) and 1a (12.90%). HCV-3b and HCV-4 were found each in 3.22% samples. Dual infection of genotypes was found in 22.58% of individuals and 22.58% HCV RNA positive sampels were not typed. A total of 572 (85.12%) subjects had hemophilia A and 100 (14.88%) had hemophilia B. In hemophiliacs A the most dominant genotype was 3a (19.27%) while in hemophilia B, genotype 1a was prevalent (26.67%). Whole blood and plasma transfusion were observed as the main risk factors of HCV. It is concluded that HCV genotype 3a and 2a are prevalent in hemophilia patients of Khyber Pakhtunkhwa Pakistan and the main risk factor observed was an unscreened whole blood transfusion.

Current trends of Hepatitis C virus genotypes and associated risk factors in hemophilia patients in Pakistan.

Hemophilia is a rare bleeding disorder that needs plasma or clotting factor concentrate transfusion. Therefore chances of blood-borne pathogens like HCV transmission increase due to high prevalence in healthy donors. This study was aimed to determine the prevalence of HCV genotypes and associated risk factors in hemophilia patients of Khyber Pakhtunkhwa, Pakistan. Blood samples and data were collected from 672 hemophiliacs after proper consent obtained from each patient. Samples were analyzed for anti-HCV, HCV RNA and HCV genotype/s detection. Of the total, 22.32% (150) were anti-HCV positive, of which HCV RNA was detected in 18.45% (124) individuals. HCV genotype 3a was found with significantly higher prevalence (p<0.05) (19.35%) as compared to 2a (16.13%) and 1a (12.90%). HCV-3b and HCV-4 were found each in 3.22% samples. Dual infection of genotypes was found in 22.58% of individuals and 22.58% HCV RNA positive sampels were not typed. A total of 572 (85.12%) subjects had hemophilia A and 100 (14.88%) had hemophilia B. In hemophiliacs A the most dominant genotype was 3a (19.27%) while in hemophilia B, genotype 1a was prevalent (26.67%). Whole blood and plasma transfusion were observed as the main risk factors of HCV. It is concluded that HCV genotype 3a and 2a are prevalent in hemophilia patients of Khyber Pakhtunkhwa Pakistan and the main risk factor observed was an unscreened whole blood transfusion.

Novel route of tannic acid biotransformation and their effect on major biopolymer synthesis in Azotobacter sp. SSB81.

AIMS: To examine tannic acid (TA) utilization capacity by nitrogen-fixing bacteria, Azotobacter sp. SSB81, and identify the intermediate products during biotransformation. Another aim of this work is to investigate the effects of TA on major biopolymers like extracellular polysaccharide (EPS) and polyhydroxybutyrate (PHB) synthesis. METHODS AND RESULTS: Tannic acid utilization and tolerance capacity of the strain was determined according to CLSI method. Intermediate products were identified using high-performance liquid chromatography, LC-MS/MS and (1) H NMR analysis. Intermediates were quantified by multiple reactions monitoring using LC-MS/MS. The strain was able to tolerate a high level of TA and utilized through enzymatic system. Growth of Azotobacter in TA-supplemented medium was characterized by an extended lag phase and decreased growth rate. Presence of TA catalytic enzymes as tannase, polyphenol oxidase (PPO) and phenol decarboxylase was confirmed in cell lysate using their specific substrates. PPO activity was more prominent in TA-supplemented mineral medium after 48 h of growth when gallic to ellagic acid (EA) reversible reaction was remarkable. Phase contrast and scanning electron microscopic analysis revealed elongated and irregular size of Azotobacter cells in response to TA. (1) H NMR analysis indicated that TA was transformed into gallic acid (GA), EA and pyrogallol. Biopolymer (EPS and PHB) production was decreased several folds in the presence of TA compared with cells grown in only glucose medium. CONCLUSIONS: This is the first evidence on the biotransformation of TA by Azotobacter and also elevated level of EA production from gallotannins. Azotobacter has developed the mechanism to utilize TA for their carbon and energy source. SIGNIFICANCE AND IMPACT OF THE STUDY: The widespread occurrence and exploitation of Azotobacter sp. strain SSB81 in agricultural and forest soil have an additional advantage to utilize the soil-accumulated TA and detoxifies the allelopathic effect of constant accumulated TA in soil.

Determination of some significant batch culture conditions affecting acetyl-xylan esterase production by Penicillium notatum NRRL-1249.

BACKGROUND: Acetyl-xylan esterase (AXE, EC 3.1.1.72) hydrolyses acetate group from the linear chain of xylopyranose residues bound by ß-1,4-linkage. The enzyme finds commercial applications in bio-bleaching of wood pulp, treating animal feed to increase digestibility, processing food to increase clarification and converting lignocellulosics to feedstock and fuel. In the present study, we report on the production of an extracellular AXE from Penicillium notatum NRRL-1249 by solid state fermentation (SSF). RESULTS: Wheat bran at a level of 10 g (with 4 cm bed height) was optimized as the basal substrate for AXE production. An increase in enzyme activity was observed when 7.5 ml of mineral salt solution (MSS) containing 0.1% KH(2)PO(4), 0.05% KCl, 0.05% MgSO(4).7H(2)O, 0.3% NaNO(3), 0.001% FeSO(4).2H(2)O and 0.1% (v/w) Tween-80 as an initial moisture content was used. Various nitrogen sources including ammonium sulphate, urea, peptone and yeast extract were compared for enzyme production. Maximal enzyme activity of 760 U/g was accomplished which was found to be highly significant (p ≤ 0.05). A noticeable enhancement in enzyme activity was observed when the process parameters including incubation period (48 h), initial pH (5), 0.2% (w/w) urea as nitrogen source and 0.5% (v/w) Tween-80 as a stimulator were further optimized using a 2-factorial Plackett-Burman design. CONCLUSION: From the results it is clear that an overall improvement of more than 35% in terms of net enzyme activity was achieved compared to previously reported studies. This is perhaps the first report dealing with the use of P. notatum for AXE production under batch culture SSF. The Plackett-Burman model terms were found highly significant (HS), suggesting the potential commercial utility of the culture used (df = 3, LSD = 0.126).

Prognostic value of quantitative-PCR versus serology for detection of CMV in pre- and post- transplantation patients.

Two hundred-twenty six pre- and post- kidney transplantation patients were screened for CMV. They were categorized into three groups I: Eighty-five dialysis patients suffering from chronic renal failure, II: Sixty -two end stage renal disease patients prepared for transplantation, and III: Seventy-nine post-kidney transplant patients under immunosuppressive and cytotoxic therapy. The last group was subdivided according to clinical presentation into asymptomatic, mild, and invasive CMV disease subgroups. A control group comprised of fifty-two apparently healthy kidney and blood donors were also included. Serum and plasma samples were utilized for detection of anti-CMV antibodies by EIA, and determination of CMV viral load by quantitative-PCR respectively. QT-PCR results revealed high viral load; (4000-10.000 copy/ml) in 16 (100%) kidney recipient with invasive CMV disease, While only 4 (25%) of them were positive for IgM anti-CMV. Another ten patients with symptoms of CMV syndrome were found CMV-PCR positive with low viral load (436-3070 copy/ml) and all were negative for IgM anti-CMV. On the other hand, no CMV viral load was detectable among end-stage renal disease patients, although 6/62 (9.7%) were positive for IgM anti-CMV. All subjects were positive for IgG anti-CMV. It is concluded that QT-PCR rather than serology should be used for monitoring of CMV infection in pre- and post-transplantation patient.

First Report of Citrus bent leaf viroid and Citrus dwarfing viroid from Citrus in Punjab, Pakistan.

Pakistan is among the top 10 citrus-producing countries of the world and the leader in Kinnow mandarin production with production concentrated in the province of Punjab, which produces more than 96% of the total citrus crop. To evaluate the presence and distribution of citrus viroids in this area, 34 samples were collected in September 2008 from citrus orchards in the Sargodha, Bhalwal, and Faisalabad areas of Punjab, including 15 'Mosambi' and two 'Bloodred' sweet oranges (Citrus sinensis), eight 'Kinnow' and four 'Feutrell Early' mandarins (C. reticulata), three 'Jatti Khatti' rough lemon (C. jambhiri), and two grapefruit (C. paradisi), which showed stunting, bark scaling, and cracking symptoms on the rootstock which was either citrange (Poncirus trifoliata × C. sinensis) or sweet lime (C. limetta). Infected budwood from these trees was grafted onto indicator plants of Arizona 861-S-1 'Etrog citron' (C. medica) budded on rough lemon rootstock, and after 3 months, the citron showed typical viroid symptoms of mild epinasty and leaf roll with 23 of the 34 samples. A one-step multiplex reverse transcription (RT)-PCR assay (3) was used to detect simultaneously Citrus exocortis viroid (CEVd), Citrus bent leaf viroid (CBLVd), Hop stunt viroid (HSVd), Citrus dwarfing viroid (CDVd), and Citrus bark cracking viroid (CBCVd). On the basis of amplification of the appropriately sized DNA, CEVd, CBLVd, HSVd, and CDVd were detected in 12, 8, 31, and 17 samples, respectively, whereas CBCVd was not detected. Twenty-three of 34 infected samples harbored more than one viroid species and one had four viroids. Budwood from 11 trees did not induce viroid symptoms on Etrog citron. Two of these trees were infected with CBLVd only and nine with HSVd only. Four primer pairs were used to amplify the full sequences of CEVd, CBLVd, HSVd, and CDVd by RT-PCR (2), which were cloned by standard methods. Sequences of three cDNA clones each of CEVd (Nos. FJ773253, FJ773254, and FJ773255), CBLVd (Nos. FJ773262, FJ773263, and FJ773267), HSVd (Nos. FJ773268, FJ773269, and FJ773271), and CDVd (Nos. FJ773274, FJ773275, and FJ773276) were deposited in GenBank. BLAST analysis showed that these nucleotide sequences had greater than 97% nucleotide identity to the most similar genome sequences in GenBank. One of the HSVd sequences, FJ773271, presented the cachexia determinants. To our knowledge, this is the first report of CBLVd and CDVd in Pakistan (1). These results indicate the need for proper indexing of mother trees and a virus-free propagation scheme to create healthy budwood sources in Pakistan. References: (1) M. Arif et al. Pak. J. Bot. 37:407, 2005. (2) L. Bernard and N. Duran-Vila. Mol. Cell. Probes 20:105, 2006. (3) X. F. Wang et al. Eur. J. Plant. Pathol, 124:175, 2009.

Efficacy of three artemisinin combination therapies for the treatment of uncomplicated Plasmodium falciparummalaria in the Republic of Congo

Background: Presented here are the results of a comparative trial on the efficacy of three artemisinin-based combinations conducted from May to October 2004; in Pool Province; Republic of Congo.Methods: The main outcome was the proportion of cases of true treatment success at day 28. Recrudescences were distinguished from re-infections by PCR analysis. A total of 298 children of 6-59 months were randomized to receive either artesunate + SP (AS+SP); artesunate + amodiaquine (AS+AQ) or artemether + lumefantrine (AL); of which 15 (5) were lost to follow-up. Results: After 28 days; there were 21/85 (25) recurrent parasitaemias in the AS+SP group; 31/97 (32) in the AS+AQ group and 13/100 (13) in the AL group. The 28-day PCR-corrected cure rate was 90.1[95CI 80.7-95.9] for AS+SP; 98.5[95CI 92.0-100] for AS+AQ and 100[95.8-100] for AL; thereby revealing a weaker response to AS+SP than to AL (p=0.003) and to AS+AQ (p=0.06). A potential bias was the fact that children treated with AL were slightly older and in better clinical condition; but logistic regression did not identify these as relevant factors. There was no significant difference between groups in fever and parasite clearance time; improvement of anaemia and gametocyte carriage at day 28. No serious adverse events were reported. Conclusions: Considering the higher efficacy of AL as compared to AS+SP and the relatively high proportion of cases with re-infections in the AS+AQ group; we conclude that AL is clinically more effective than AS+SP and AS+AQ in this area of the Republic of Congo. Implementation of the recently chosen new national first-line AS+AQ should be monitored closely

A new synthesis of oxadiazole, thiazolidinone, N-phthalimidoamino carbonyl and arylidene derivatives with potential antimicrobial activity.

Condensation of carbohydrazide derivatives Ia, b with dimethyl acetylenedicarboxylate and acetylenedicarboxylic acid yielded benzofuran derivatives II a-d. Reaction of Ib with aromatic aldehydes formed products III a-d. Treatment of compounds III a-d with mercaptoacetic acid yielded the cyclocondensation products (IVa-d). Phthalic anhydride reacted with compounds (Ia, b)to form products (Va, b). It has been found that both khellin and visnagin (VIa, b)react with aromatic aldehydes to give arylidene derivatives (VIIa-e). Condensation of diphenyl nitrilamine with 2-arylidene furochromones VII derivatives afforded cyclo-adducts (VIII a-i). The antibacterial activities of the selected compounds were tested against Staphylococcus aureus, B. subtilis, E. coli, Pseudomonas, Salmonella and Erwinia with good results.

Preparation of new Schiff bases derived from beta-enaminonitriles and hetarylcarboxaldehydes. Anti-inflammatory and ulcerogenic activities.

Schiff bases 3a-j were prepared by reacting the beta-enaminonitriles (1a,b) with the appropriate hetarylcarboxaldehyde, in boiling ethanol. Structural elucidations for the new compounds (3a-j) were based upon compatible analytical and spectroscopic results. Compounds 1a, 1b, 3a, 3c, 3d and 3i were tested for their anti-inflammatory and ulcerogenic activities.

Biosynthesis of ascorbic acid and riboflavin in radiated germinating chickpea.

Biosynthesis of ascorbic acid and riboflavin in radiated germinating chickpea seeds was studied at ambient room temperature (20-35 degrees C). Synthesis of these vitamins increased significantly with increasing germination time up to 120 hrs depending upon the treatment (P less than 0.05). Maximum amounts of ascorbic acid, 22.32 and 16.84 mg/100 g (wet weight) were found in the 0.10 kGy samples after 120 hrs of germination in tap and distilled waters respectively. Radiation dose of 0.20 kGy resulted in the development of maximum riboflavin, 11.4 and 11.0 micrograms/g (dry weight) on germination in tap and distilled waters respectively. An overall significant linear relation (r = 0.954 to 0.957) was found between vitamin biosynthesis and germination time up to 120 hrs in both the radiated and unradiated chickpea.

Effect of irradiation and germination on trypsin inhibitor and protein content of chickpea (Cicer arientinum L.).

Effect of irradiation (0.05-0.20 kGy) and subsequent germination on trypsin inhibitor activity (TIA) and protein content of chickpea, was studied. The results revealed a significant linear relation (r = 0.981 to -0.992) between the loss of TIA and germination time, and the rate of TIA destruction increased with irradiation dose (p less than 0.05). Maximum destruction (43.8%) of the TIA occurred on germination for 120 hr of 0.20 kGy sample (from 330.0 to 185.3 TIU/g). Initially protein content was 21.7% and the value significantly increased to maximum levels of 27.5% and 27.9% in distilled and tap water respectively during germination for 120 hr of 0.10 kGy sample (p less than 0.05). Protein contents were not affected by irradiation.

Comparative facilitated transport of oxygen.

Numerical solutions were obtained for a model of facilitated transport of O2. The dependence of the facilitated flow on the concentrations of O2 at the two boundaries of the transport path was studied. The numerical values of the parameters correspond to the adult Ascaris lumbricoides and to vertebrate red striated muscle. A global control principle is formulated. This states that for every fixed O2 concentration at the low concentration boundary there exists an O2 concentration at the high concentration boundary for which the facilitated flow is maximum. The collection of these maxima makes possible the existence of a global adjustment of the facilitated transport in contradistinction to the mere presence of a local maximum. The ranges of the pairs of boundary O2 concentrations thus defined and for which the facilitated flow is within 70% of its attainable maximum were found to coincide with the physiological ranges of boundary O2 concentrations for the Ascaris and vertebrate striated muscle. This phenomenon has the character of a graded compensatory mechanism to hypoxia. It is an intrinsic property of the carrier transport system and does not depend on sensors for hypoxia.

Synthesis and antimicrobial activity of some benzofuran derivatives.

The Friedel-Craft's acylation of 2,3-diphenyl-5-methylbenzofuran 1a with succinic anhydride gives beta-(2,3-diphenyl-5-methylbenzofuranoyl)propionic acid 1b. Acid 1b reacts with hydrazines yielding the corresponding tetrahydropyridazinones 2. 4,5-Dihydro-1,2-oxazin-6H-6-one 6 is formed by reaction of 1b with hydroxylamine hydrochloride. The reaction of the pyridazinone 2a with dialkyl sulfates, ethyl chloroacetate, Grignard reagents, amines in presence of formaldehyde (Mannich reaction), o-amino-phenol and thiourea has been investigated. The antimicrobial activities of compounds 1a-c, 2a-c, e, f, 4a, b and 5a, b as well as 7b are studied.

Facilitated transport of oxygen in the presence of membranes in the diffusion path.

Most of the experimental observations on facilitated transport have been done with millipore filters, and all the theoretical studies have assumed homogeneous spatial properties. In striated muscle there exist membranes that may impede the diffusion of the carrier myoglobin. In this paper a theoretical study is undertaken to analyze the transport in the presence of membranes in the diffusion path. For the numerical computations physiologically relevant values of the parameters were chosen. The numerical results indicate that the presence of membranes tends to decrease the facilitation. For the nonlinear chemical kinetics of the reaction of oxygen with the carrier, this decrement also depends on the location of the membranes. At the higher oxygen concentration side of each membrane the flow of combined oxygen is transferred to the flow of dissolved oxygen. The reverse process occurs at the lower concentration side. Jump discontinuities of the concentration of the oxygen-carrier compound at each membrane are associated with these transfers. The decrement of facilitation is due to the cumulative effect of these jump discontinuities.

Behaviour of hepatitis B antigen in bilharzial patients infected with HBs positive viral hepatitis.

A TOTAL of 204 cases with acute viral hepatitis were studied: 93 bilharzial and 111 non-bilharzial. This study is concerned with the 106 cases showing positive HBS Ag test. Follow up of the cases after discharge from the Fever Hospital was carried out by repeated clinical and laboratory examinations 3 montly in the first year and 6 monthly subsequently. The bilharzial and non-bilharzial patients were found to be equally susceptible to HBS Ag positive hepatitis (p=0.25). However, antigenaemia tended to last longer as a whole in the bilharzial patients (p=0. 005) and was not affected by successful specific antibilharzial treatment. It seems that bilharzial infection predisposes to "retention" of HBs antigen, after subsidance of the acute attack, thus increasing the volume of reservoir for HB virus in our rural community.