Method optimization and validation for the routine analysis of multi-class pesticide residues in Kinnow Mandarin and fruit quality evaluation.

The present study describes the selection of a sensitive multi-residue method that can be used for the routine testing of pesticides in Kinnow Mandarin. The citrate-buffered QuEChERS extraction followed by primary secondary amines and C18 clean-up was found suitable for the analysis of fifty four pesticides. The limit of quantification for the selected pesticides was lower than maximum residue limits (MRLs) set by European Union, Codex Alimentarius Commission (CAC), and twelve other countries. The method's accuracy ranged from 74.4 to 112% and expanded uncertainty ranged from 7.5 to 49.6%. The validated method was applied to Kinnow Mandarin samples, collected from 22 export units of district Sargodha, Pakistan. Almost 27% of the samples (n = 22) were exceeding the CAC-MRLs. The index of quality for residues (IqR), for 64% of the samples, was considered adequate. The study indicates the need for regular monitoring to protect public health and ensure safe and consistent trade.

Antihyaluronidase and Alkaline Phosphatase (ALP) Activities of Medicinal Plants to Combat Echis carinatus Venom-Induced Toxicities.

Snakebite is one of the most neglected diseases of developing countries. Deaths due to snakebite envenoming are quite high in Pakistan, and many deaths are caused by Echis carinatus envenomation. Traditional use of medicinal plants against snakebites is a common practice in Pakistan due to countless benefits. The current study was performed with the objective to evaluate eighteen Pakistani medicinal plants inhibitory potential against hyaluronidase and alkaline phosphatase enzymes of Pakistani Echis carinatus venom. Hyaluronidase activity (0.2-1.6 mg/0.1 mL) and alkaline phosphatase activity (0.1-0.8 mg/0.1 mL) were measured in dose-dependent manner. Crude methanolic extracts of medicinal plants were used for in vitro investigation of their inhibitory activity against toxic enzymes. All active plants were fractioned using different solvents and were again analyzed for inhibitory activity of same enzymes. Results indicated all plants were able to neutralize hyaluronidase that Swertia chirayita (Roxb. ex Flem.) Karst., Terminalia arjuna Wight and Arn, Rubia cordifolia Thumb., and Matthiola incana (L.) R.Br. inhibited maximum hyaluronidase activity equivalent to standard reference (p > 0.5). Pakistani medicinal plants are dense with natural neutralizing metabolites and other active phytochemicals which could inhibit hyaluronidase activity of Pakistani Echis carinatus venom. Further advanced studies at molecular level could lead us to an alternative for envenoming of Pakistani Echis carinatus venom.

Anti-5'-Nucleotidases (5'-ND) and Acetylcholinesterase (AChE) Activities of Medicinal Plants to Combat Echis carinatus Venom-Induced Toxicities.

Echis carinatus is one of the highly venomous snakes of Pakistan that is responsible for numerous cases of envenomation and deaths. In Pakistan, medicinal plants are commonly used traditionally for snakebite treatment because of their low cost and easy availability in comparison with antivenom. The current research is aimed at evaluating the inhibitory activity of Pakistani medicinal plants against acetylcholinesterase and 5'-nucleotidases present in Echis carinatus venom. Acetylcholinesterase and 5'-nucleotidase enzymatic assays were performed at different venom concentrations to check the activity of these enzymes. Methanolic extracts from different parts of plants were used for in vitro determination of their inhibitory activity against 5'-nucleotidases in snake venom. Active methanolic extracts were subsequently fractioned using different solvents, and these fractions were also assessed for their anti-5'-nucleotidase activity. Results of this study exhibited that Eugenia jambolana Willd. ex O. Berg, Rubia cordifolia L., Trichodesma indicum (L.) R. Br., Calotropis procera (Wild.) R. Br., Curcuma longa L., and Fagonia arabica L. were able to significantly (p > 0.5) neutralize the 5'-nucleotidase activity by 88%, 86%, 86%, 85%, 83.7%, and 83%, respectively, compared with a standard antidote (snake venom antiserum). Thus, this study indicates that these plants possess the potential to neutralize one of the toxic enzymatic components of Echis carinatus venom and hence can help to augment the future efforts of developing alternative therapy for the management of snakebites.

Synthesis, characterization and docking studies of amide ligands as anti-leishmanial agents.

Aim of this study was to synthesize new inhibitors on the basis of active site of aspartic protease enzyme and to evaluate their intended biological activity. A3D model of an enzyme was generated via homology modeling and series of novel amide ligands were synthesized by using a short high yield process, subsequently, analyzed in-silico and in-vitro anti-leishmanial activities. Characterization and identification was accomplished via NMR (H1& C13), infrared and mass spectroscopic techniques. Among all compound (4) was found to show significant activity (IC50 58±0.01) against Leishmania major (L. major) species. Furthermore, docking studies confirmed the inhibition of a targeted enzyme that supported the interaction of potent compound (4) with key residues (aspartic protease) via hydrogen bonds. Present study conferred about novel compound (4) as a promising compound to antagonize L. major activities in future.

Plastic driven pollution in Pakistan: the first evidence of environmental exposure to microplastic in sediments and water of Rawal Lake.

Microplastic pollution is as an emerging environmental threat. Focus of most of the current researches has been on microplastics in the marine environment. However, there is limited information for microplastic availability in the fresh water, especially in Pakistan. The current study was conducted with the objective of investigating the microplastic presence and concentration in the surface water and sediments of the Rawal Lake, in the capital city of Pakistan. The average microplastic abundance for water and sediments was 0.142 items/0.1 L and 1.04 items/0.01 kg, respectively. Results indicated that the fibers and fragments were the most dominant types of microplastics. The dominant colors were blue, red, black, and transparent. FTIR analysis of visible microplastic particles displayed a greater similarity with polyethylene, polypropylene, polyesters, polyethylene terephthalate, and polyvinyl chloride because of the appearance of characteristic peaks of these polymers. The study also revealed greater concentration of microplastics in the sediments as compared with water of the Rawal Lake. High population density surrounding lake, improper waste disposal, tourism, and recreational activities may be the major reasons for the microplastic contamination of the lake.

Simultaneous mitigation of cadmium and drought stress in wheat by soil application of iron nanoparticles.

Excess amount of cadmium (Cd) in arable soils and shortage of good quality water are the major abiotic factors affecting the crop yield which needs immediate solution to feed the increasing population worldwide. Recently, nanoparticles (NPs) are widely used in various industries including agriculture which is due to the unique properties of NPs. Among NPs, iron (Fe) NPs might be used to alleviate the abiotic stresses in crops but limited informations are available in the literature about the role of Fe-NPs in crops under metal stress. The present study was designed to highlight the efficiency of Fe-NPs on Cd accumulation in Cd and drought-stressed wheat. Wheat plants were grown in Cd-contaminated soil after the supply of different levels of Fe-NPs and two water regimes were introduced in the soil in latter growth stages of the plants. Cadmium and drought stress negatively affected the wheat photosynthesis, yield and caused oxidative stress in leaves with excess accumulation of Cd in grains and other plant tissues. The NPs improved the photosynthesis, yield, Fe concentrations and diminished the Cd concentrations in tissues. The NPs alleviated the oxidative stress in leaves and the efficiency depends on the NPs concentrations applied in the soil. The results obtained indicated that Fe-NPs may be employed aiming to get wheat grains with excess Fe and decreased Cd contents. However, field investigations with various sizes, shapes and levels of NPs are needed before final recommendations to the farmers.

Effects of silicon nanoparticles on growth and physiology of wheat in cadmium contaminated soil under different soil moisture levels.

Soil degradation with different stress conditions like accumulation of cadmium (Cd) contents in soil and drought stress has become one of the most dangerous issues that obstruct the sustainable agriculture production. Silicon nanoparticles (Si NPs) play beneficial roles in combating various biotic and abiotic stresses but their role under combined metal and drought stress is not studied. A pot study was designed to determine the effect of Si NPs on wheat (Triticum aestivum L.) growth and uptake of Cd grown in Cd contaminated soil with different water levels under ambient conditions. Four different levels of Si NPs (0, 25, 50, and 100 mg/kg) were applied in the soil before 1 week of wheat sowing and two water levels (70% and 35% soil water-holding capacity) were introduced after 50 days of seed sowing for the remaining growth period. The lowest biomass, yield, and photosynthesis were observed in the control plants while oxidative stress and the highest Cd concentrations in shoots, roots, and grains were observed in the control plants, and the drought stress further enhanced this effect on the plants. The Si NPs treatments improved the plant growth indicators and photosynthesis, and reduced the Cd concentrations in wheat tissues, especially in grains either without or with drought stress. The Si NPs reduced the oxidative stress in leaves as was indicated by the reduced production of hydrogen peroxide, electrolyte leakage, and malondialdehyde contents, and increase in superoxide dismutase and peroxidase activities. The improvement in wheat growth and a reduction in oxidative stress and Cd concentration in tissues were dependent on the levels of Si NPs and the effect was the highest with the highest level of NPs used.

Erupt of malaria, dengue and chikungunya in Pakistan: Recent insights about prevalence, diagnosis and treatment.

Malaria, dengue and chikungunya are the most rampant mosquito-borne infections predominantly in Pakistan. They pose a serious threat and cause a havoc for the victims owing to the life threatening signs and symptoms marked with elevated morbidity and mortality rate. It seems hard to discriminate due to common indications, consequently, deserves appropriate diagnosis prior elevated toll of death. Present article encompasses depth insights about their prevalence, diagnosis and clinical manifestation if erupt in the pandemic. However, host-vector-host cycle is the root cause of transmission and diverse mosquito species confer dissimilar infections. Indeed these infections are seasonal but other factors like flood, open irrigation channels, immense agricultural land, rich fauna and water reservoirs can't be overlooked. Dire need was felt to acknowledge and aware the public about local transmission, vector control, entomologic, research resources, diagnosis and advancement in healthcare system to alleviate them absolutely in future.

Evaluation of different Pakistani medicinal plants for inhibitory potential against Echis carinatus induced Phospholipase A2 toxicity.

Medicinal plants of Pakistan are known for their curative properties against snake bite as rural people have been using natural herbs for such injuries for hundreds to thousands of years. People of rural areas of Pakistan are prone to snakebite, and on the whole death due to snakebite has been increasing worldwide. The objective of this study was to test the neutralizing potential of 17 Pakistani medicinal plant extracts against phospholipase A2 activity in Echis carinatus venom. Plant material was extracted by simple maceration and fractionation of active plant extracts. Venom was collected by manual massage of the venom glands. The PLA2 enzymatic assay was performed to map out the venomous activity of Echis carinatus envenomation. Snake venom released fatty acids at different concentrations (0.1-5 mg/ml) of venom in a dose-dependent manner. Reduction of pH by 01 correlated with 133 µmol of fatty acids released at 5mg/ml of venom. All plants extract inhibited PLA2 activity, however, Curcuma longa, Citrullus colocynthis and Rubia cordifolia inhibited maximum of PLA2 activity (⁓78%) comparable to the standard antidote (p>0.5). Medicinal plants possess secondary metabolites and many active compounds that may have neutralizing or inhibiting properties against the PLA2 activity of Echis venom. Further studies such as compound analysis could provide an alternative against snakebites injuries resulting from Echis carinatus venom.

Air pollution tolerance index of plants around brick kilns in Rawalpindi, Pakistan.

In any contaminated environment, the sensitive plant species can serve as bio-indicator of air pollution while tolerant plant species can act as a sink for air pollutants. Air pollution tolerance index (APTI) is an important tool to screen out plants based on their tolerance or sensitivity level to different air pollutants. The present study was aimed to identify the sensitive and tolerant plant species in the vicinity of brick kilns in the Rawalpindi city. To determine the susceptibility level of the selected plant species, four biochemical parameters, ascorbic acid, total chlorophyll content, relative water content and pH of leaf extract, were assessed and APTI was calculated. Plant sampling was carried out with increasing distance of 100, 300 and 500 m around three brick kiln sites and APTI values were calculated by following the standard methods. The results of the study revealed that among nine studied plant species, Calotropis procera (APTI = 20.05) and Althernanthera pungens (APTI = 17.13) were found to be the most tolerant species, whereas Malva neglecta (APTI = 8.83) was found to be the most sensitive species. Inconsistent trend of variations was seen in the APTI values at each site. The present study suggested that the most tolerant species, C. procera and A. pungens, could be grown in the vicinity of such pollution sources as a remedial measure of brick kiln pollution.

Effects of ozone on crops in north-west Pakistan.

Although ozone is well-documented to reduce crop yields in the densely populated Indo-Gangetic Plain, there is little knowledge of its effects in other parts of south Asia. We surveyed crops close to the city of Peshawar, in north-west Pakistan, for visible injury, linking this to passive measurements of ozone concentrations. Foliar injury was found on potato, onion and cotton when mean monthly ozone concentrations exceeded 45 ppb. The symptoms on onion were reproduced in ozone fumigation experiments, which also showed that daytime ozone concentrations of 60 ppb significantly reduce the growth of a major Pakistani onion variety. Aphid infestation on spinach was also reduced at these elevated ozone concentrations. The ozone concentrations measured in April-May in Peshawar, and used in the fumigation experiment, are comparable to those that have been modelled to occur over many parts of south Asia, where ozone may be a significant threat to sensitive crops.