Identification of Natural Compounds as Inhibitors of Pyruvate Kinase M2 for Cancer Treatment.

The reliance of tumor cells on aerobic glycolysis is one of the emerging hallmarks of cancer. Pyruvate kinase M2 (PKM2), an important enzyme of glycolytic pathway, is highly expressed in a number of cancer cells. Tumor cells heavily depend on PKM2 to fulfill their divergent energetic and biosynthetic requirements, suggesting it as novel drug target for cancer therapies. Based on this context, we performed enzymatic-assay-based screening of the in-house phenolic compounds library for the identification of PKM2 inhibitors. This screening identified silibinin, curcumin, resveratrol, and ellagic acid as potential inhibitors of PKM2 with IC50 values of 0.91 µM, 1.12 µM, 3.07 µM, and 4.20 µM respectively. For the determination of Ki constants and the inhibition type of hit compounds, Lineweaver-Burk graphs were plotted. Silibinin and ellagic acid performed the competitive inhibition of PKM2 with Ki constants of 0.61 µM and 5.06 µM, while curcumin and resveratrol were identified as non-competitive inhibitors of PKM2 with Ki constants of 1.20 µM and 7.34 µM. The in silico screening of phenolic compounds against three binding sites of PKM2 provided insight into the binding pattern and functionally important amino residues of PKM2. Further, the evaluation of cytotoxicity via MTT assay demonstrated ellagic acid as potent inhibitor of cancer cell growth (IC50 = 20 µM). These results present ellagic acid, silibinin, curcumin, and resveratrol as inhibitors of PKM2 to interrogate metabolic reprogramming in cancer cells. This study has also provided the foundation for further research to validate the potential of identified bioactive entities for PKM2 targeted-cancer therapies.

In silico-based identification of phytochemicals as novel human phosphoglycerate mutase 1 (PGAM1) inhibitors for cancer therapy.

Targeting cancer-specific metabolic and mitochondrial remodeling has emerged as a novel and selective strategy for cancer therapy during recent years. Phosphoglycerate Mutase 1 (PGAM1) is an important glycolytic enzyme that catalyzes the conversion of 3-phosphoglycerate to 2-phosphoglycerate and plays a critical role in cancer progression by coordinating glycolysis and biosynthesis. PGAM1 has been reported to be over expressed in a variety of cancer types and its inhibition results in decreased tumor growth and metastasis. Recently, there has been a growing interest in identification and characterization of novel PGAM1 inhibitors for the treatment of cancer. In the current study, in silico tools were used to find out natural inhibitors of PGAM1. For docking studies, a database of 5006 phytochemicals were docked against PGAM1, using MOE software in order to identify the compounds which show better binding affinities than PGMI-004A. Out of 5006 compounds screened, eight compounds (1,3-cyclopentanedione, glyflavanone B, 6-demethoxytangeretin, gnaphaliin, lantalucratin A and -(-) morelensin, abyssinin II and monotesone-A) showed significant binding affinity with PGAMI active site. Further, the eight selected compounds were evaluated for different pharmacokinetics parameters using admetSAR, the obtained results demonstrated that none of these hit compounds violated Lipinski's drug rule of 5 and all the hit compounds possess favorable ADMET properties. This study has unveiled the potential of phytochemicals that could serve as probable lead candidates for the development of PGAM1 inhibitors as anti-cancer agents.

Eriocalyxin B induces apoptosis in human triple negative breast cancer cells via inhibiting STAT3 activation and mitochondrial dysfunction.

Eriocalyxin B (EriB), a potent ent-kaurene extracted from Isodon eriocalyx, has turned up as novel anti-cancer agent during recent years against a range of cancer types. TNBC (Triple negative breast cancer) is highly aggressive breast cancer, which is resistant towards current therapeutics due to absence of drug targets. Here, we have probed the molecular mechanism of EriB-induced apoptosis in TNBC (MDA-MB231) cells to check whether its anticancer activity is mediated by modulation of STAT3 and NF-Ï°B. EriB induced apoptosis in MDA-MB231 cells via inhibiting NF-Ï°Bp65, STAT3 phosphorylation, increasing Bax/Bcl-2 ratio, MMP dissipation, and activation of caspase-3. These results provide a rationale for further in vivo investigations on EriB, which might also prove to be a potential drug candidate for developing novel therapeutics against TNBC.

Microelectrophoretic study of environmentally induced DNA damage in fish and its use for early toxicity screening of freshwater bodies.

This study investigates the potential of the comet and micronucleus assays of fish DNA as a means of screening the toxicity of aquatic environments. Catla catla and Cirrhinus mrigala collected from the River Chenab in Pakistan were used as a case study for the application of comet and micronucleus techniques. Comet and micronucleus assays were used to compare DNA damage in C. catla and C. mrigala collected from polluted areas of the River Chenab and farmed fish. Atomic absorption spectrophotometry showed an acute level of toxicity from Cd, Cu, Mn, Zn, Pb, Cr, Sn, and Hg in river water. Comet assay showed significant (p < 0.05) DNA damage in C. catla representing 17.33 ± 2.42, 11.53 ± 2.14, and 14.17% DNA in the comet tail, averaged from three sites of the polluted area of the river. Tail moment was observed as 10.06 ± 2.71, 3.11 ± 0.74, and 14.70 ± 1.89, while olive moment was 8.85 ± 1.84, 3.83 ± 0.76, and 7.11 ± 0.73, respectively. Highly significant (p < 0.01) damage was reported in C. mrigala as 37.29 ± 2.51, 34.96 ± 2.53, and 38.80 ± 2.42% DNA in comet tail, tail moment was 23.48 ± 3.90, 19.78 ± 4.26, and 14.30 ± 1.82, and olive moment was 16.22 ± 2.04, 13.83 ± 1.96, and10.99 ± 0.90. Significant (p < 0.05) differences were observed in genotoxicity between farmed and polluted area fish. Micronucleus assay showed a similar picture of significant difference in respect to single and double micronucleus induction: i.e., 23.20 ± 4.19 and 2.80 ± 1.07‰ in C. catla and 44.80 ± 3.73 and 06.20 ± 0.97‰, respectively, in C. mrigala. Nuclear abnormalities were found as 6.00 ± 0.84 and 09.60 ± 1.72/thousand cells, respectively, in both species. The results of this study suggest that these novel fish DNA damage assays can be used as an expedient toxicity screening for aquatic environments.

Assessment of pesticide residues in flesh of Catla catla from Ravi River, Pakistan.

The levels of dichlorodiphenyltrichloroethane (DDT), dichlorodiphenyldichloroethylene (DDE), endosulfan, endosulfan sulfate, carbofuran, and cartap which were estimated in the flesh of Catla catla sampled from ten sites of Ravi River between its stretches from Shahdara to Head Balloki were studied to know the level of contamination of the selected pesticides by GC-ECD method. All fish samples were found contaminated with different concentrations of DDT, DDE, endosulfan, and carbofuran; however, DDT and DDE concentrations were more than the maximum residue limits (MRLs) about food standards, while endosulfan sulfate and cartap were not detected. Pesticide concentrations in the fish flesh were ranged from 3.240 to 3.389 for DDT, 2.290 to 2.460 for DDE, 0.112 to 0.136 for endosulfan, and 0.260 to 0.370 µg g(-1) for carbofuran. The findings revealed that the pesticide concentrations in the fish flesh decreased in the order: DDT > DDE > carbofuran > endosulfan. After Degh fall and After Hudiara nulla fall river sampling sites were found severely contaminated. It is proposed that a constant monitoring programs are needed to be initiated to overcome the present alarming situation.

Pesticides in the River Ravi and its tributaries between its stretches from Shahdara to Balloki Headworks, Punjab-Pakistan.

The present study investigated the presence and potent source of pesticides and specific pesticide bearing effluent release points on the River Ravi, between the Shahdara and Balloki Headworks. Pakistan has banned the use of organochlorine and nitrogen containing pesticides, yet no attention has been paid to the continuous monitoring and assessment of these banned pesticides to make sure that they are not being used in practice. Levels of selected organochlorine and nitrogen containing pesticide residues were assessed in water collected from 18 sampling sites on the River Ravi and its tributaries using a gas chromatograph equipped with an electron capture detector (GC-ECD). All water samples were found to be contaminated with varying concentrations of pesticide residues. However, levels of pesticides were below the tolerance limits suggested in national and international standards. Pesticide concentrations in the water of river sites ranged from 0.034 to 0.045 microg/L for DDT, 0.033 to 0.046 microg/L for DDE, 0.108 to 0.123 microg/L for endosulfan and 0.028 to 0.040 microg/L for carbofuran. In tributaries, pesticide concentrations ranged from 0.0468 to 0.0685 microg/L for DDT, 0.0390 to 0.0637 microg/L for DDE, 0.111 to 0.147 microg/L for endosulfan and 0.0396 to 0.0631 microg/L for carbofuran. The results show pesticide concentrations in river water in the order: endosulfan > DDE > DDT > carbofuran. Pesticide concentrations in tributary waters decreased in the order: endosulfan > DDT> DDE > carbofuran. After Degh Fall and After Hudiara Nulla Fall river sampling sites were severely contaminated while, among the tributaries, Degh Fall and Hudiara Drain were severely contaminated with DDT, DDE, endosulfan and carbofuran. Constant monitoring programs should to be initiated to reform the present situation.

Methods to quantify gait rehabilitation following lower limb fractures.

Lower limb fragility fractures require long-term rehabilitation and are also very expensive to treat. Clinically, early weight bearing and walking stability were reported as key measures of fracture restoration. This study introduces methods to numerically quantify these performance indices for a range of ankle and knee joint fractures. As a follow-up of initial treatment, experimental data was collected using force plates from 367 subjects divided into seven groups: ankle fracture (AF), lower leg ankle fracture (AL), calcaneus foot fracture (CF), knee tibia fracture (KF), knee patella fracture (KP), kneecap rupture (KR), and normal limb (NL). For each joint, data was analysed to evaluate intralimb and interlimb weight-bearing and walking stability for all fracture conditions. These thresholds were statistically compared with normal subjects. Some advantages of evaluating fracture restoration indices over the others include:•to quantify fracture restoration (weight-bearing, walking stability, and gait symmetry) using minimum setup and signal requirements.•to provide comprehensive tools to assess and overcome fracture-associated complications through a detailed preview of fractured limb functionality during subphases of a gait cycle.•in clinical research, such assessments are important as a reference to evaluate existing or new rehabilitative interventions.

Biomonitoring of heavy metals and their association with DNA damage in Indian peafowl (Pavo cristatus) under captivity.

Environmental pollution and changing climatic conditions are likely to damage biodiversity not only on organismal level but on molecular level as well. The aim of the present study was to find the concentration of heavy metals in soil, water, feed, feathers, and blood and association of heavy metals with DNA damage of P. cristatus. The results showed that lead (Pb) was significantly (P < 0.01) higher in soil and cadmium (Cd) was significantly (P < 0.01) higher in soil and water. Chromium (Cr), zinc (Zn), nickel (Ni), and cobalt (Co) were significant (P < 0.01) in feed. Manganese (Mn) was significantly (P < 0.01) higher in feed and soil (surface). In addition, Pb and Cd concentrations were significant (P < 0.01) in feathers while Cr and Zn concentrations were significantly (P < 0.01) higher in feces. Nickel was significantly (P < 0.01) higher in feathers and eggshell while Mn and Co concentrations were significantly (P < 0.01) higher in blood and feces, respectively. Furthermore, significant positive correlation between Pb (rs = 0.75; P < 0.05) and Cd (rs = 0.67; P < 0.05) concentrations in blood with tail DNA was found. It was concluded that heavy metals exist in the soil, water, feathers, and blood and have association with DNA damage of P. cristatus.

Hepatoprotective effects of almond shells against carbon tetrachloride induced liver injury in albino rats.

Liver injury is a prevalent pathological process that can give rise to conditions such as fatty liver, cirrhosis, fibrosis, and even cancer. It has been observed that plants and natural products possess significant protective effects against liver injury. Current study was performed to investigate the efficacy of almonds shell against carbon tetrachloride (CCl4) induced hepatotoxicity in rat model. As almonds shell contain a large variety of phenolic and flavonoid compounds, which are largely associated with antioxidant and hepatoprotective properties. For this purpose, screening of small-scale library of twenty plant extracts was performed for evaluation of antioxidant potential by DPPH assay. The data revealed that almonds shell extract (ASEE) exhibited potent antioxidant activity. This potent extract was further evaluated for hepatoprotective activity in in vivo rat model on 30 rats, divided into 6 groups of 5 rats each. On 29th day all rats were sacrificed and blood serum was collected for further analysis. Liver tissues were also preserved in formalin for histopathology. The results demonstrated that ASEE displayed a protective effect on liver function tests (LFT), renal function tests (RFT), and lipid biomarkers in comparison to the CCl4 group. The histological data also unveiled a substantial safeguarding impact on liver damage, characterized by a reduction in apoptosis, diminished liver hemorrhage, and decreased accumulation of cellular debris. The data indicates that ethanolic extract from almond shells possesses hepatoprotective potential, suggesting its viability as an alternative source for hepatoprotective drug development after pre-clinical research.

Advances in the application of functional nanomaterial and cold plasma for the fresh-keeping active packaging of meat.

The most recent advancements in food science and technology include cold sterilization of food and fresh-keeping packaging. Active packaging technology has received much interest due to the photocatalytic activity (PCA) of functional nanoparticles, including titanium dioxide (TiO2) and ferric oxide (Fe2O3). However, there are still significant concerns about the toxicity and safety of these functional nanoparticles. This review emphasizes the bacteriostatic and fresh-keeping properties of functional nanoparticles as well as their packaging strategies using the ultraviolet photo-catalysis effect. High-voltage electric field cold plasma (HVEF-CP) is the most innovative method of cold-sterilizing food. HVEF-CP sterilizes by producing photoelectrons, ions, and active free radicals on food media, which come into contact with the bacteria's surface and destroy their cells. Next, this review also assesses the photocatalytic activity and bacteriostasis kinetics of nanosized TiO2 and Fe2O3 in poultry, beef, and lamb. In addition, this review also emphasizes the importance of exploiting the complex interaction processes between TiO2 and Fe2O3, along with dietary components and their utilization in the fresh meat industry.

Identification of Macrolepiota procera extract as a novel G6PD inhibitor for the treatment of lung cancer.

Tumor metabolism, an emerging hallmark of cancer, is characterized by aberrant expression of enzymes from various metabolic pathways including glycolysis and PPP (pentose phosphate pathway). Glucose 6 phosphate dehydrogenase (G6PD) and 6-phosphogluconate dehydrogenase (6PGD), oxidative carboxylases of PPP, have been reported to accomplish different biosynthetic and energy requirements of cancer cells. G6PD and 6PGD have been proposed as potential therapeutic targets for cancer therapy during recent years due to their overexpression in various cancers. Here, we have employed enzymatic assay based screening using in-house G6PD and 6PGD assay protocols for the identification of mushroom extracts which could inhibit G6PD or 6PGD enzymatic activity for implications in cancer therapy. For the fulfillment of the objectives of present study, nine edible mushrooms were subjected to green extraction for preparation of ethanolic extracts. 6xhis-G6PD and pET-28a-h6PGD plasmids were expressed in BL21-DE3 E. coli cells for the expression and purification of protein of interests. Using purified proteins, in house enzymatic assay protocols were established. The preliminary screening identified two extracts (Macrolepiota procera and Terfezia boudieri) as potent and selective G6PD inhibitors, while no extract was found highly active against 6PGD. Further, evaluation of anticancer potential of mushroom extracts against lung cancer cells revealed Macrolepiota procera as potential inhibitor of cancer cell proliferation with IC50 value of 6.18 µg/ml. Finally, screening of M. procera-derived compounds against G6PD via molecular docking has identified paraben, quercetin and syringic acid as virtual hit compounds possessing good binding affinity with G6PD. The result of present study provides novel findings for possible mechanism of action of M. procera extract against A549 via G6PD inhibition suggesting that M. procera might be of therapeutic interest for lung cancer treatment.

Detecting Aquatic Pollution Using Histological Investigations of the Gills, Liver, Kidney, and Muscles of Oreochromis niloticus.

The present study aimed to determine the degree of changes in the histological architecture of the liver, gills, kidneys, and muscles of fish Oreochromis niloticus collected from different polluted river sites. Fish samples collected from the Faisalabad Fish Hatchery and upstream of Chakbandi drain acted as a control. Necrosis, hemorrhage, and epithelial hyperplasia were observed in the gills of fish inhabiting the river downstream of the Chakbandi drain entrance. Liver tissues were found to be affected by vacuolated cytoplasm, bile duct proliferation, melanomacrophages, and necrosis. In kidney tissues, shrinkage of the renal cortex, necrosis, and destructive renal tubules were observed. Histopathology of muscles indicates the presence of hypertrophy and swollen myofibers. In contrast, upstream specimens of fish exhibited mild tissue alterations. Histopathology of gills tissue showed vacuolization. Liver tissues indicated the presence of hypertrophy and more frequent Kupffer cells than usual. The vacuolation was also observed in kidney tissues. Muscle tissues expressed splitting of muscle fibres and degeneration in muscle bundles. However, sections of tissues collected from farmed fish have normal morphology and no anomalies. The histopathological assessment indicated various cellular, biochemical, and histological changes in response to the contamination in the vicinity of the fish.

LC-ESI-QTOF/MS characterization of antimicrobial compounds with their action mode extracted from vine tea (Ampelopsis grossedentata) leaves.

Vine tea (Ampelopsis grossedentata) is a tea plant cultivated south of the Chinese Yangtze River. It has anti-inflammatory properties and is used to normalize blood circulation and detoxification. The leaves of vine tea are the most abundant source of flavonoids, such as dihydromyricetin and myricetin. However, as the main bioactive flavonoid in vine tea, dihydromyricetin was the main focus of previous research. This study aimed to explore the antibacterial activities of vine tea against selected foodborne pathogens. The antimicrobial activity of vine tea extract was evaluated by the agar well diffusion method. Cell membrane integrity and bactericidal kinetics, along with physical damage to the cell membrane, were also observed. The extract was analyzed using a high-performance liquid chromatography-diode array detector (HPLC-DAD), and the results were confirmed using a modified version of a previously published method that combined liquid chromatography and electrospray-ionized quadrupole time-of-flight mass spectrometry (LC-ESI-QTOF/MS). Cell membrane integrity and bactericidal kinetics were determined by releasing intracellular material in suspension and monitoring it at 260 nm using an ultraviolet (UV) spectrophotometer. A scanning electron microscope (SEM) was used to detect morphological alterations and physical damage to the cell membrane. Six compounds were isolated successfully: (1) myricetin (C15H10O8), (2) myricetin 3-O-rhamnoside (C21H20O12), (3) 5,7,8,3,4-pentahydroxyisoflavone (C15H10O7), (4) dihydroquercetin (C15H12O7), (5) 6,8-dihydroxykaempferol (C15H10O8), and (6) ellagic acid glucoside (C20H16O13). Among these bioactive compounds, C15H10O7 was found to have vigorous antimicrobial activity against Bacillus cereus (AS11846) and Staphylococcus aureus (CMCCB26003). A dose-dependent bactericidal kinetics with a higher degree of absorbance at optical density 260 (OD260) was observed when the bacterial suspension was incubated with C15H10O7 for 8 h. Furthermore, a scanning electron microscope study revealed physical damage to the cell membrane. In addition, the action mode of C15H10O7 was on the cell wall of the target microorganism. Together, these results suggest that C15H10O7 has vigorous antimicrobial activity and can be used as a potent antimicrobial agent in the food processing industry.

Chirality of the biomolecules enhanced its stereospecific action of dihydromyricetin enantiomers.

The present study explores the effect of chirality of the biological macromolecules, its functional aspects, and its interaction with other food components. Dihydromyricetin (DHM) is a natural novel flavonol isolated from the vine tea (Ampelopsis grossedentata) leaves. However, limited progress in enantiopure separation methods of such compounds hinder in the development of enantiopure functional studies. This study is an attempt to develop a simple, accurate, and sensitive extraction method for the separation of the enantiopure DHM from vine tea leaves. In addition, the identification and purity of the extracted enantiopure (-)-DHM were further determined by the proton nuclear magnetic resonance (1H-NMR) and the carbon nuclear magnetic resonance (13C-NMR). The study further evaluates the antimicrobial activity of isolated (-)-DHM in comparison with racemate (+)-DHM, against selected foodborne pathogens, whereas the action mode of enantiopure (-)-DHM to increase the integrity and permeability of the bacterial cell membrane was visualized by confocal laser scanning microscopy using green fluorescence nucleic acid dye (SYTO-9) and propidium iodide (PI). Moreover, the morphological changes in the bacterial cell structure were observed through field emission scanning electron microscope. During analyzing the cell morphology of B. cereus (AS11846), it was confirmed that enantiopure (-)-DHM could increase the cell permeability that leads to the released of internal cell constituents and, thus, causes cell death. Therefore, the present study provides an insight into the advancement of enantiopure isolation along with its antimicrobial effect which could be served as an effective approach of biosafety.

Influence of Combined Effect of Ultra-Sonication and High-Voltage Cold Plasma Treatment on Quality Parameters of Carrot Juice.

Influence of the combined effect of ultra-sonication (US) and high-voltage cold plasma treatment (HVCP) on the quality parameters of fresh carrot juice has been studied. During the treatment of ultra-sonication, carrot juice was subjected to a 0.5 inch probe for 3 min by adjusting the pulse duration 5 s on and off at 20 kHz frequency, amplitude level 80%. The ultrasound intensity was measured by using a thermocouple and was 46 Wcm-2. The temperature was maintained at 10 °C by an automatic control unit. During the treatment of HVCP, carrot juice was then subjected to dielectric barrier discharge (DBD) plasma discharge at 70 kV voltage for 4 min. Significant increases were observed when HVCP treated carrot juice was tested against total carotenoids, lycopene, and lutein when compared to the control treatments. Moreover, this increase was raised to its highest in all pigments, chlorogenic acid, sugar contents, and mineral profile, as the results of ultra-sonication when combined with high voltage atmospheric cold plasma (US-HVCP). Whereas, a significant decreased was observed in Mg, total plate count, yeast, and mold after US-HVCP treatment. Furthermore, results indicated that the combined effect of US-HVCP treatment has improved the quality and led to a higher concentration of lycopene, lutein, chlorogenic acid, and mineral compounds (Na, K, and P). Therefore, the findings of the current study suggested that US-HVCP treatment is a novel combined technique that could provide better quality and more stability during the processing of carrot juice with better physicochemical properties and bio-available nutrients, so this novel processing technique could serve as an alternative to traditional processes.

Exploring the Potential of High-Voltage Electric Field Cold Plasma (HVCP) Using a Dielectric Barrier Discharge (DBD) as a Plasma Source on the Quality Parameters of Carrot Juice.

The main aim of the current investigation was to contemplate the impact of high-voltage electric field cold plasma (HVCP) on different quality characteristics (enzymes, microbial activities, coloring pigments, ascorbic acid, polyphenolic compounds, °Brix, acidity, and color index) of carrot juice in correlation with thermal processing. A carrot juice (250 mL) sample sealed in pre-sterilized food-grade bottles, which placed between two dielectric quartz plates for HVCP treatment. The gap between the plates was 30 mm, and a stable and uniform plasma dielectric barrier discharge (DBD) generated for 3 and 4 min at 60, 70, and 80 kV. Air was used as a working gas during the DBD-based plasma treatment. The observed rise in temperature was 2-5 °C during the HVCP treatment. A water bath was used to carry out thermal treatment (100 °C for 5 min). The maximum inactivation of enzymes and microorganisms was achieved with thermal treatment and then with HVCP treatment at 70 kV for 4 min. However, maximum retention of coloring compounds, ascorbic acid, total phenols, flavonoids, and tannins was found following HVCP (70 kV for 4 min) treatment compared to thermal treatment. The °Brix, pH, and acidity remained unchanged irrespective of treatments. These findings suggest that HVCP treatment at 70 kV for 4 min may be a good alternative to thermal treatment, and it may successfully be applied in carrot juice production, resulting in reduced enzymes, lower microbial activity, and improved bioactive compounds. The prospects of overcoming the existing conventional physical and chemical methods for sterilization make it a novel and more economical technique to maintain food's natural nourishment, composition, appearance, structure, and freshness.

Genetic diversity in ex-situ conserved Lens culinaris for botanical descriptors, biochemical and molecular markers and identification of landraces from indigenous genetic resources of Pakistan.

Lentil, one of the oldest legumes was investigated for diversity based on botanical descriptors, total seed proteins, isozymes and random amplified polymorphic DNA (RAPD) markers. About one fourth of accessions were heterogeneous for botanical descriptors and a seed protein profile. The germplasm collected from the province of Baluchistan revealed the prevalence of indigenous landraces as high diversity was observed for all of the techniques. Diversity explored through various techniques revealed validity irrespective of the sample size or geographic pattern, RAPD being the best choice for investigating both inter- and intra-accession variation in lentil. Although all of the techniques were able to resolve genetic diversity in lentil, isozymes and seed proteins gave low levels of genetic diversity, suggesting that more investigation into isozymes of specific proteins is required. RAPD is the best option for determining inter- and intra-accession variation, and will be required to extend germplasms and primers to continue the study of botanical descriptors.

Pollutant fate and spatio-temporal variation and degree of sedimentation of industrial- and municipal wastes in Chakbandi drain and River Chenab.

This study's objective was to assess a seasonal impact of industrial and sewage waste disposal on water quality of the river upstream Trimu Head. Considering the significance of the river, drain wastewater was analyzed during the summer and the winter seasons from pre-determined locations. Water quality parameters were recorded higher than the maximum permissible limits prescribed by WHO for freshwater bodies. Level of these Physio-chemical variables was higher in the winter due to the least amount of water from domestic sewage. Although some of these parameters indicated sedimentation hitherto the water quality of River Chenab was found very poor due to the pollution bestowed by tributary waste water from drains. Findings of this investigation suggest the importance of continuous monitoring and assessment to improve the water quality of the river. This study provides a baseline data which may be compared to assess any further deterioration in the water quality and may also be used to plan future monitoring and required restoration of habitat for the safe supply of fish to the population of this region.

Fish eco-genotoxicology: Comet and micronucleus assay in fish erythrocytes as in situ biomarker of freshwater pollution.

Owing to white meat production Labeo rohita have vast economic importance, but its population has been reduced drastically in River Chenab due to pollution. Atomic absorption spectrophotometry showed a merciless toxicity level of Cd, Cu, Mn, Zn, Pb, Cr, Sn and Hg. Comet assay results indicated significant (p < .05) DNA fragmentation in Labeo rohita as 42.21 ±â€¯2.06%, 31.26 ±â€¯2.41% and 21.84 ±â€¯2.21% DNA in comet tail, tail moment as 17.71 ±â€¯1.79, 10.30 ±â€¯1.78 and 7.81 ±â€¯1.56, olive moment as 13.58 ±â€¯1.306, 8.10 ±â€¯1.04 and 5.88 ±â€¯0.06, respectively, from three different polluted sites on the river. Micronucleus assay showed similar findings of single micronucleus induction (MN) as 50.00 ±â€¯6.30‰, double MN 14.40 ±â€¯2.56‰, while nuclear abnormalities (NA) were found as 150.00 ±â€¯2.92‰. These higher frequencies of MN induction and NA were found to be the cause of reduction of 96% of the population of this fish species in an experimental area of the River Chenab. This fish species has been found near extinction through the length of the river Chenab and few specimens in rainy seasons if restored by flood, may die in sugarcane mill season. Due to sweeping extinction Labeo rohita showed the highest sensitivity for pollution and could be used as bioindicator and DNA fragmentation in this column feeder fish species as a biomarker of the pollution load in freshwater bodies.

Study on impact of habitat degradation on proximate composition and amino acid profile of Indian major carps from different habitats.

This investigation is aimed to study an impact of habitat degradation on proximate composition and amino acid (AAs) profile of Catla catla, Labeo rohita and Cirrhinus mrigala collected from polluted, non-polluted area (upstream) and a commercial fish farm. The amino acid profile was estimated by the amino acid analyzer. C. catla collected from the polluted environment had highest lipid, protein and ash contents (12.04 ±â€¯0.01, 13.45 ±â€¯0.01 and 0.93 ±â€¯0.03%, respectively). The high protein content (14.73 ±â€¯0.01 and 14.12 ±â€¯0. 01%) was recorded in C. catla procured from non-polluted (upstream) wild habitat of River Chenab and controlled commercial fish farm. Farmed fish species showed comparatively higher moisture contents followed by upstream and polluted area fishes. C. mrigala showed significant differences in amino acid and proximate composition collected from a polluted site of the river Chenab. C. catla collected from non-polluted site of the river showed an excellent nutrient profile, followed by L. rohita (wild and farmed) and C. mrigala (polluted area), respectively. All fishes from the polluted areas of the River Chenab indicated a significant decrease in the concentration of some AAs when compared to farmed and wild (upstream) major carps. Omitting of some important AAs was also observed in the meat of fish harvested from polluted habitat of this river. C. mrigala and L. rohita exhibited a significant increase in the concentration of some of non-essential amino acids such as cysteine in their meat. The results indicated that wild fish (upstream) and farmed fish species had highest protein contents and amino acid profile and hence appeared to be the best for human consumption. The proximate composition and AAs profiles of fish harvested from the polluted area of the river clearly indicated that efforts shall be made for the restoration of habitat to continue the requirement of high quality fish meat at a low cost to the human population.