The presence of heavy metals especially arsenic and lead in drinking water above the normal level (WHO standards) has threatened the use of ground water as major source of drinking water. According to different reports, majority of water wells in urban areas of Pakistan have arsenic level above the WHO recommended level of 10 ppb. In addition high level of lead is also reported especially in areas which are close to highways. Both arsenic and lead pose serious health threat to mankind as they disrupt the function of many important tissues and organs of the body. The acute and chronic arsenic exposure results in damage to digestive, cardiovascular and nervous system and also leads to many malignancies. Lead also leads to many abnormalities and especially effect nervous system. In addition, it mainly affects skin and kidney and disrupts their normal function. Lead is also possible human carcinogen and leads to malignancies of lung and stomach etc. Stem cells are type of undifferentiated cells that have the potential of self-renewal and differentiation into specialized cells. On the basis of their origin and differentiation potential they are categorized as embryonic stem cells (ESCs) and adult stem cells. Mesenchyaml stem cells (MSCs) are adult stem cells that have differentiation potential into adipocytes, chondrocytes and osteoblasts etc. In this dissertation, the effect of arsenic and lead has been described on the MSCs and differentiation of MSCs into osteoblasts at molecular level. MSCs were isolated from mice (mMSCs) and rat (rMSCs) bone marrow by simply flushing them out, and from human (hMSCs) bone marrow by density gradient centrifugation and growing them on the basis of plastic adherence. The adult MSCs (AMSCs) were isolated by enzymatic digestion of human adipose tissue whereas human fibroblasts (hFib) were isolated by explant and enzymatic digestion. The stemness of hMSCs was checked by colony forming unit assay. The doubling time of all the cells lines was checked by growing them in 6 well plates for 5 days and counting the cells everyday by hemocytometer. The doubling time of mMSCs, rMSCs, hMSCs and AMSCs was 44.32, 47.93, 60.79 and 58.59 h, respectively. The cytotoxicity assays were done by neutral red method for all the cell lines, in which 5 x 103 cells were exposed to arsenic (0-10 μg/ml) and lead (0-100 μg/ml) for a total period of 6, 12, 24 and 48 h, LC50 values were calculated. The LC50 values of arsenic against mMSCs were 2.4, 2.8, 3.4 and 3.6 μg/ml on exposure to arsenic for 6,12, 24 and 48 h, respectively, while for all other cell lines mixed results were obtained. The LC50 values for arsenic obtained were 2.5, 2.8, 3.5 and 3.5 μg/ml for rMSCs, 8, 7, 5, and 2.5 μg/ml for hMSCs, 7.6, 4.6, 4.1 and 4.8 μg/ml for AMSCs, and 9, 4, 3.8 and 3 μg/ml for hFib on exposure to arsenic for 6, 12, 24 and 48 h, respectively. The maximum reduction in proliferation of cells on exposure to lead was observed in mouse and rat MSCs. In the case of mMSCs the percentage reduction was 43, 30, 46 and 24 while it was 43, 32, 48 and 30 in the case of rMSCs. The percentage reduction was 16, 23, 28 and 6 in hMSCs, 21, 14, 12 and 16 in AMSCs and 26, 37, 32 and 28 in hFib on exposure to lead for 6, 12, 24 and 48 h, respectively. The effect of metals on morphology of cells was observed under microscope and on exposure to arsenic, cells became round in shape and were detached from plate surface, whereas in the presence of lead the effect was not as drastic as that of the arsenic and attached cells were observed even on exposure to 100 μg/ml lead. The genotoxic effects of arsenic was investigated by comet assay in which all the cell types were exposed to arsenic (1 μg/ml) and lead (10 μg/ml) for 12 h and then slides were prepared and genotoxity was checked by observing different comet parameters (Comet length, height, area, intensity, head diameter, tail length, tail area, % DNA in tail, tail moment, % DNA in head). The comet assay results showed that there was maximum DNA damage on exposure to arsenic whereas the DNA fragmentation was less in the case of lead. No DNA damage was observed in the case of control samples. Both arsenic and lead proved to have genotoxic effects on all the cell lines used in the present study. For differentiation of mMSCs, cells were exposed to 50 μg/ml ascorbic acid, 10 mM β-glycerophosphate and 1 x 10-7 M dexamethasone. Alkaline phosphatase (ALP) activity, total protein and expression profile of proteins was analyzed at different time intervals. Alizarin Red S staining was done after 21 days in the differentiation medium. The differentiation of mMSCs was done both in the presence and absence of dexamethasone. There was more than 3 fold increase in ALP activity in differentiating cells compared with the control cells. The control cells had greater concentration of total protein and more number of cells compared to differentiating cells. Dexamethasone significantly enhanced the proliferation of cells. After 21 days of growth, both in the presence and absence of dexamethasone, there was mineral deposition and differentiated cells stained red when treated with Alizarin Red while control cells didn’t stain red. SDS-PAGE did not show any difference in expression of protein in control and differentiating cells. The proliferation of cells was greatly reduced when hMSCs were grown in the differentiation medium with arsenic (100 ng/ml) and lead (1μg/ml). The ALP activity increased gradually in the differentiation medium with or without metals compared to control cells. The difference in ALP activity was very less among the differentiating cells and metal exposed differentiating cells but it was high compared to control cells. The differentiating cells stained red with Alizarin Red but in the presence of arsenic and lead, the cells did not stain red. On crystal violet staining the morphology of cells changed from spindle to oval in the differentiation medium. In the presence of arsenic, the morphology of cells was totally changed. Dendrites like out growth were observed (blebbing phenomenon), while in the presence of lead the morphology of cells was comparable to differentiated cells. The expression of two marker genes of stemness, Oct4 and HLA-DR alpha were detected in the stem cells but were absent in the control cells (fibroblasts). Beside that the expression of differentiation markers, osteocalcin, osteopontin, osteonectin, Runx2, BMP2, ALP and collagen were found to increase in the differentiation medium with and without lead, whereas the expression of BMP2 and Runx2 remained the same. The effect of arsenic could not be detected due to high cytotoxicity and genotoxicity. The cells died rapidly and good quality RNA could not be isolated for further analysis. No change was observed in protein pattern of cells in the differentiating medium with and without metals. The expression of apoptosis related genes including Bax, Bcl-2, c-myc and P53 were monitored in the cells exposed to 100ng/ml arsenic and 1 μg/ml lead for 12 h. The expression of P53 gene only was detected in arsenic exposed cells which indicated that arsenic triggers the apoptosis pathway of cells and results in death of cells. To conclude, MSCs were isolated from a different source on the basis of their adherence potential to plastic surfaces and differentiated into osteoblasts using standard differentiation protocol. mMSCs, could be differentiated into osteoblasts like cells even in the absence of dexamethasone, which is a key component of osteoblast differentiation medium. Arsenic and lead proved to have both cytotoxic and genotoxic effects on all the cell types used but arsenic proved to be much more toxic than lead. In addition, both arsenic and lead have adverse effects on the morphology of cells. Longer exposure to arsenic resulted in blebbing outgrowth of plasma membrane. The exposure of differentiating cells to arsenic and lead at 1 μg/ml and 10 μg/ml respectively did not significantly affect the expression of differentiation marker genes. At molecular level both metals severely affected the differentiation process so the exposure of these metals especially in early age can result in several bone abnormalities. According to reports from environmental protection department (EPD), the level of arsenic and lead is increasing in drinking water all over the country so there is great need to figure out sources of water contamination and proper measures should be taken to reduce the exposure of people to these heavy metals." xml:lang="en_US
"رسالہ " عربی لفظ ہے ۔جس کا معنی ترسیل کا آلہ ہے ۔ اردو رسائل نے اردو زبان و ادب کی ترویج و ترقی میں گراں قدر خدمات سرانجام دی ہیں ۔ ادبی رسالہ میں ادب کی مختلف ادبی جہات مثلاً شاعری،ناول ،افسانہ کو موضوع بنایا گیا ہے ۔ زمانہ قدیم سے عہدجدید تک اردوادبی رسائل کی اہمیت سے انحراف نہیں کیا جاسکتا ۔دنیا کےکسی بھی زبان کے ادب کے فروغ میں رسائل اہم کردار ادا کرتے ہیں ۔ جہاں اردو رسائل نے ہماریروایات و اقدار،تہذیب وتمدن،کلچر،ثقافت میں اہم کردار ادا کیا ہے وہیں ادب کے فروغ میں بھی کلیدی حثیت رکھتے ہیں ۔ اردو ادبی رسائل ہمارےتنقیدی نظریات و معیاری تخلیقات اور فکر و فن کو موضوع سخن بناتے ہیں اور ادب کی ترویج و ترقی میں اہم کردار ادا کرتے ہیں ۔ اردو رسائل نےادب کے مختلف موضوعات کو اپنے دامن میں جگہ دی ہے۔اورادبی رسائل خصوصی ادبی شمارے یا ادبی رسائل نمبرز بھی شائع کرتے ہیں جن میں افسانہ نمبر ، ناول نمبر ، غالب نمبر ، شاعری نمبر ، اقبال نمبر ، غزل نمبر،مرثیہ نمبر ،نعت نمبر اور ناولٹ نمبراور خصوصی شخصیت نمبر بھی جاری کئے ہیں ۔ اردو رسائل و جرائدادب کے ترجمان ہیں اردو ادبی ورسائل وجرائد انسانی جذبات و احساسات کی بھر پور عکاسی کرتے ہیں۔ ادبی رسائل ادبی صحافت کے میدان میں بھی بہت اہمیت کے حامل ہیں ۔ان ادبی رسائل میں ہفتہ وار، ماہنامہ ، سہ ماہی ، ششماہی ، اور سالانہ مجلے بھی ہوتے ہیں کیوں کہ ان کی اشاعت کی مدت مختلف ہوتی ہے ۔ یہ ۔سیاسی ، سماجی ،مذہبی اور ادبی شعور کو اجاگر کرتے ہیں ۔
In every period, hadith experts have contributed to the preservation of the Hadith. Imam Malik composed M’ūaṭa to safeguard the hadith, and Imam al-Bukhari afterwards turned to al-Mu'tah for assistance. The research methodology of this paper is an analytical study of the reliance of Imām Bukhārī on Al-M’ūaṭa Imam Mālik in Ṣaḥiḥ Bukhārī. In our article, we came to the conclusion that Imam Bukhari recounted a total of six hundred sixty-eight narrations from Imam Malik in his Sahih, of which six hundred four are scriptural narrations that he took from ten versions of Al- M’ūaṭa. As a result, these narrations make up more than ninety percent of all narrations. Contrasted with in his Sahih, Imam Muslim narrates three hundred eighty-nine narrations from Imam Malik, three hundred twelve of which are scriptural narrations that he has derived from eight of narrations of al M’ūaṭa's. These narrations so account for more than eighty percent of all narrations.
To examine the prevalence of aflatoxins in poultry feeds and to substantiate the effects of aflatoxin B1 on broilers’ performance, a study was conducted on commercial poultry feeds and their ingredients during 2006- 2007. A total of 216 samples comprised of wheat, maize, rice, cotton seed meal, broiler starter and finisher rations were collected from local poultry markets of Peshawar, Swat and D. I. Khan regions of North West Frontier Province (NWFP) of Pakistan. Sampling was carried out during the winter (December-February), spring (March-May), summer (June-August) and autumn (September-November) seasons of the year 2006/2007. It was found that water activity (aw) of samples collected from Peshawar, Swat and D.I. Khan regions ranged from 0.500 to 0.834 aw, 0.408 to 0.815 aw, and 0.554 to 0.747 aw, respectively. Seasonal variation significantly affected water activity of the samples. Moisture sorption isotherms of the samples showed that all the commodities were hygroscopic in nature. The dominant fungal genera isolated from the feeds were Penicillium spp (25.45 %), Aspergillus spp (19.01%), Rhizophus spp (14.32%), Mucor spp (10.52%), Fusarium spp (10.69%), and Eurotium spp (9.46%) across all the three regions. Total fungal viable count ranged from 6.45 to 26.69 x 103, 6.80 to 16.90 x 103 and 6.43 to 25.58 x 103 CFUs g-1 in samples from Peshawar, Swat and D. I. Khan regions, respectively. It was observed that total fungal count significantly varied among the substrates and different seasons. Total culturalable A. flavus and A. parasiticus in the samples from Peshawar ranged from 1.51 to 21.42 x 102 CFUs g-1 and 2.27 to 32.14 x 102 CFUs g-1, while that in Swat and D. I. Khan samples ranged from 1.51 to 42.52 x 102 CFUs g-1, 2.27 to 63.79 x 102 CFUs g-1 and 1.57 to 102.30 x 102 and 2.36 to 153.45 x 102 CFUs g-1, respectively. It was observed that 63.33% of all the isolates of Aspergillus sec Flavi were aflatoxigenic in nature while the rest were non-aflatoxigenic. All commodities were found contaminated with aflatoxins B1 (AFB1), B2 (AFB2), G1 (AFG1) and G2 (AFG2), the ranges being 13.71 to 191.65, 5.46 to 86.85, 8.55 to 167.82 and 5.14 to 89.90ng g-1, respectively in the samples from Peshawar region. Samples from Swat region had AFB1, AFB2, AFG1 and AFG2 in the range of 13.37 to 147.34, 3.63 to 22.38, 4.27 to143.33, and 1.95 to 49.72 ng g-1 whereas that of D. I. Khan region contained in the range of 6.96 to 58.31, 3.39 to 27.92, 4.41 to 24.54 and 0.02-4.46 ng g-1, respectively. The main enzymes produced by germinating conidia of A. flavus were esterase, lipase, acid phosphatase, β-glucosidase and N-acetyl-β-D- glucosaminidase, while for A. parasiticus these were alkaline phosphatase, lipase, acid phosphatase and β-fucosidase in terms of both total (μmol 4- nitrophenol min-1 g-1) and specific activity (nmol 4-nitrophenol min-1 μg-1 protein). There were significant increases in the specific activity of all these enzymes of germinating spores of A. flavus and A. parasiticus for up to 72 hrs. The total/specific activities of the enzymes produced by A. flavus were maximum at 0.99 aw, with the exception of acid phosphatase and N-acetyl-β- D-glucosaminidase at 0.94 aw. For A. parasiticus, maximum total activity occurred at 0.99 aw but specific activity was found to be higher at lower aw levels. Calcium propionate, aw and incubation time, alone and interactively, significantly affected total fungal viable count and aflatoxins production in both starter and finisher broiler rations. Minimum culturalable fungi were counted in calcium propionate added feeds at lower aw (0.85aw) level at the start of experiment which progressively increased over time, increasing aw levels and decreasing concentration of calcium propionate in the feeds. Similar trends were observed for aflatoxins (B1, B2, G1 and G2) production in both starter and finisher broiler rations. Study in vitro on A. flavus (A-2092) and A. parasiticus (PRR-2747) showed that conidia of both species were germinated on all calcium propionate (0.5 and 1%) and aws (0.996, 0.96 and 0.94 aw) treatments, however, 1% calcium propionate at 0.94aw delayed the germination process for up to 10 and 9 days in A. flavus and A. parasiticus, respectively. The growing rates of spores of both species were slower at 1% calcium propionate and 0.94 aw. Aflatoxins (B1, B2, G1 and G2) were also minimally produced by A. flavus and A. parasiticus at 1% calcium propionate dose and 0.94 aw. However, none of the treatments completely inhibited the growth and aflatoxins production by A. flavus and A. parasiticus. Broilers’ feed intake, body weight gain, feed conversion ratio, meat of carcass and dressing percentages were significantly affected by consumption of AFB1 contaminated feed. Feed intake, average body weight gain and meat of carcass were significantly reduced with increasing levels of AFB1 in the feed. On the contrary, feed conversion ratio and dressing percentages were increased as the level of aflatoxins increased in the feed. Residues of AFB1 and AFM1 were detected in both liver and muscles of chicks only when they were fed with higher level (>20ng g-1) of AFB1 in the feed. Comparatively, higher residual levels of aflatoxins B1 and M1 were observed in the liver than in the muscles showing that liver is the primary site of metabolism of aflatoxins in chicken.