Assessment of heavy metals and associated oxidative stress in occupationally exposed workers from Bannu and Karak Districts in Pakistan.

Heavy metals (HMs) are extensively found in occupationally exposed miners and industrial workers, which may cause serious health-related problems to the large workforce. In order to evaluate the impact of these toxic pollutants, we have investigated the effect of cadmium (Cd), chromium (Cr), copper (Cu), and lead (Pb) concentration on exposed workers of mining, and woolen textile mill and compared the findings with unexposed individuals. From each category like exposed workers (mining, and woolen mill textile site) and unexposed individuals, 50 blood samples were taken. The occurrence of HMs in a sample was investigated through atomic absorption spectrometry while the oxidative stress marker malondialdehyde (MDA) and antioxidant enzyme statuses such as superoxide dismutase (SOD) and catalase (CAT) were analyzed in exposed and control samples. The results showed significant (p < 0.05) variation in Cd, Cr, Cu, and Pb levels in exposed and control samples. The concentration of Cd in the blood of WMWs, KMWs, and control group was 5.75, 3.89, and 0.42 µg/dL, respectively. On the other hand, the concentration of Pb in the blood of WMWs, MWs, and control was 32.34, 24.39, and 0.39 µg/dL while the concentrations of Cr and Cu in the blood of WMWs, MWs, and control group were 11.61 and 104.14 µg/dL, 4.21 and 113.21 µg/dL, 0.32 and 65.53 µg/dL, respectively. An increase in MDA was recorded in the exposed workers' group as compared to control subjects, whereas SOD and CAT activities decreased. Meanwhile, MDA was significantly and positively (p < 0.01) correlated with HMs, while negative significant correlations were found among HMs with SOD and CAT.

Screening of various Brassica species for phytoremediation of heavy metals-contaminated soil of Lakki Marwat, Pakistan.

Natural resources, especially agrarian soils, have been much contaminated with various pollutants including heavy metals since industrial revolution, so it is pertinent to utilize green technology, the so-called phytoremediation technology for reclamation of heavy metal-contaminated soils. A pot experiment was conducted to screen four different species (Brassica juncea, Brassica napus, Brassica rapa, Brassica campestris) of Brassicaceae family for the remediation of HMs contaminated soil of Lakki Marwat city, Pakistan, irrigated with municipal wastewater. Plants were analyzed for various morpho-physiological, biochemical, and phytoextraction factors like bioaccumulation (BAF) and translocation factor (TF). Results showed maximum morpho-physiological responses including seed germination, chlorophyll content, root fresh and dry weights, and shoot fresh and dry weights in B. juncea followed by B. napus, B. campestris, and B. rapa. Plant biochemical analyses of antioxidant enzymes including superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) also exhibited maximum activity in B. juncea followed by B. napus, B. campestris, and B. rapa, respectively, in both control and contaminated soils. Conversely, plant oxidative stress markers including malondialdehyde (MDA) and hydrogen peroxide (H2O2) showed maximum contents in B. rapa followed by B. campestris, B. napus, and B. juncea in both soils. Plant bioconcentration factors i.e. BAF and TF measured for all species in both soils confirmed that B. juncea accumulated maximum heavy metals. Similarly, enhanced phytoextraction capacity was noticed for all Brassica species in decreasing order i.e. B. napus > B. campestris > B. rapa. Hence, all the results confirmed that B. juncea excelled and can be recommended for phytoremediation purpose in soils of study area.

Multivariate geo-statistical perspective: evaluation of agricultural soil contaminated by industrial estate's effluents.

Present study was carried out to explore heavy metals pollution and potential ecological risk factors associated with agriculture soil irrigated with industrial effluents of Hayatabad industrial estate, Peshawar (HIEP) and Gadoon industrial estate, Sawabi (GIES) of Khyber-Pakhtunkhwa, Pakistan through multivariate geo-statistical tools. Diverse statistical tools like cluster analysis (HC) and principal component analysis (PCA), along with geo-statistical approaches were applied to highlight the geogenic and anthropogenic sources of pollution. The results indicated that concentration of heavy metals in target areas was significantly higher than control. Both soils had significant to moderate enrichment of heavy metals, while Gadoon soil had moderate to considerable ecological risk factor. The geo accumulation indices (Igeo) tendency for heavy metals in both target soil (Gadoon and Hayatabad) were the same. The Pb concentration of both target areas falls in the extremely severely polluted category because of the excessive presence of industries producing lead-containing products. Hence, this study indicated that the majority of toxic heavy metals contributed to soil pollution in the studied areas are coming from industrial and commercial activities.

Physiological, ultrastructural, biochemical, and molecular responses of glandless cotton to hexavalent chromium (Cr6+) exposure.

Glandless cotton can be grown to obtain cotton seeds free of toxic gossypol for use as both food and feed. However, they are not grown normally due to their lesser productivity and higher susceptibility to biotic stress. Great attention has been paid to biotic stresses rather than abiotic stresses on glandless cotton. Chromium (Cr) is a common pollutant of soil and considered a serious threat to plants due to its adverse effects on different functions. Although numerous studies are available on the toxicity of Cr6+ in various plants. However, its adverse effects and mechanism of toxicity in glandless cotton can seldom be found in the literature. This study examined the Cr6+ effect on glandless cotton in comparison to glanded cotton. Four pairs of glanded and glandless cotton near-isogenic lines (NILs) were exposed to different doses (0, 10, 50, and 100 µM/L) of Cr6+ for seven days, and biochemical, physiological, molecular, and ultrastructure changes were observed, which were significantly affected by Cr6+ at high concentrations in all NILs. The effect of Cr6+ on ionic contents shows the same trend in glanded and glandless NILs except for manganese (Mn2+) that show inhibition in glandless (ZMS-12w and Coker-312w) and enhance in the glanded NIL (ZMS-17). The gene expression of superoxide dismutase (SOD) and peroxidase (POD) revealed similar trends as enzyme activities in glandless NILs. The principal component analysis (PCA) and Agglomerative hierarchical clustering (AHC) results of all NILs from morpho-physiological traits, cluster ZMS-16, and ZMS-17 into Cr6+ sensitive group. While the glandless NILs have the potential to cope with the Cr toxicity by increasing the antioxidant enzyme activity and their gene expression. This study also revealed that Cr6+ tolerance in cotton is genotypic and has an independent mechanism in the root that not related to low gossypol.

Reduced Glutathione Protects Subcellular Compartments From Pb-Induced ROS Injury in Leaves and Roots of Upland Cotton (Gossypium hirsutum L.).

Heavy metals-based changes in the plants and their alleviation through eco-friendly agents including reduced glutathione (GSH) have been widely studied. In the present experiment, we tested the alleviatory role of reduced glutathione (GSH) in seedlings of upland cotton cultivar, TM-1 under lead (Pb) toxicity. Plants were grown in the Hoagland solution containing Pb (0 µM), Pb (500 µM), GSH (50 µM), and GSH + Pb (50 µM + 500 µM). Lead exposure exacerbated hydrogen peroxide (H2O2) and hydroxyl radical (OH•) levels, induced lipid peroxidation (MDA), and decreased the activities of catalase (CAT) and ascorbate peroxidase (APX) in the terminal and median leaves of 28-days old cotton seedlings stressed for 10 days. However, in the primary and secondary roots, CAT activity was increased but APX decreased. Similarly, peroxidase (POD) and superoxide dismutase (SOD) activities were enhanced in the median leaves but a declining trend was observed in the terminal leaves, primary roots and secondary roots. Glutathione reductase (GR) activity, ascorbic acid (AsA) contents and GSH concentrations were increased in all parts except AsA in the median leaves. Transmission electron micrographs of Pb-treated plants exhibited deformed cell wall and cell membrane, disfigured chloroplasts and irregularly shaped mitochondria in the terminal and median leaves. Further, cell membrane, mitochondria, nucleus and other cell organelles in root cells were severely affected by the Pb. Thus their identification was little bit difficult through ultramicroscopy. External GSH stabilized leaf and root ultramorphology by stabilizing cell membranes, stimulating formation of multivesicular body vesicles, and by maintaining structural integrity of other organelles. Evidently, GSH played major alleviatory role against Pb toxicity in upland cotton.

Use of various biomarkers to explore the effects of GSM and GSM-like radiations on flowering plants.

Since last decade, GSM (Global System for Mobile Communication) technology has evidently revolutionized our digital world. It uses electromagnetic frequency radiations (EMFr), ranging 850-1900 MHz, and is being composed of three main units (i.e., mobile station, access and core networks). GSM technology has significant impact on our daily life as revealed by increased number of mobile users in the world over. The main goal of the present review is to critically revisit the available literature regarding the responses of various flowering plant species towards GSM and GSM-like radiations using physiological, biochemical, molecular and cytological markers using in vitro approaches. Different monocots (tomato, onion, wheat and maize etc.) and dicots (pulses, mustard and flax) have been studied using both GSM mobile phone and GSM simulators. Different studies revealed overall reductions in germination, root-shoot lengths, dry weight, in both dose and time-dependent manners. However, there could be found incline in various parameters at biochemical and molecular levels. Furthermore, there could be found disturbances at cytological levels upon exposure of roots of onion to EMFr radiations. The overall literature review shows the negative effects of GSM and GSM-like radiations on targeted plant species. In order to alleviate the stressful effects of EMFr radiations on plants, in vivo studies need to be done using various cost-effective approaches such as use of biochar and various organic amendments.

Ethnomedicinal Cichorium intybus Seed Extracts: An Impending Preparation against Multidrug Resistant Bacterial Pathogens.

BACKGROUND: The present study was undertaken to analyze the phytochemical content and biological activity of Cichorium intybus seeds traditionally used in Charsadda, Pakistan against multidrug resistant (MDR) bacterial pathogens. OBJECTIVES: This study explored the qualitative and quantitative antibacterial potential of C. intybus. Further qualitative analysis of phytochemical content was performed. METHODS: Cichorium intybus seed extracts were prepared in aqueous, chloroform, ethanol, and hexane separately. RESULTS: All the extracts of C. intybus seeds were screened for antibacterial activity and phytochemical content. Cichorium intybus seed extract showed considerable activity against MDR pathogenic bacteria. In the well diffusion method, aqueous extracts showed a higher zone of inhibition against Pseudomonas aeruginosa (16 mm ± 0.7 mm) and Acinetobacter baumannii (13 mm ± 0.5 mm), whereas chloroform, ethanol, and hexane extracts showed activity against P. aeruginosa (11 mm ± 0.3 mm, 12 mm ± 0.5 mm, and 11 mm ± 0 mm, respectively) as compared to Imipenem, a broad spectrum antibiotic. Minimum inhibitory concentration and minimum bactericidal concentration values for aqueous and ethanol extracts indicate that they were more effective against MDR bacteria. Phytochemical analysis revealed that aqueous and ethanol extracts were rich in alkaloids, carbohydrates, gallotannins, and triterpenoids, whereas chloroform and hexane extracts were more concentrated with phenolics, pseudotannins, saponins, and tannins. Cichorium intybus seed extract demonstrated potential activity against MDR human pathogenic bacteria. CONCLUSIONS: The undertaken study has for the first time reported the effects of C. intybus seed extracts against MDR bacterial pathogens. Findings of the current study will be helpful for further elucidation of bioactive molecules for therapeutic use against MDR bacterial pathogens.

Biosorption of heavy metals by Pseudomonas species isolated from sugar industry.

Heavy metal-resistant bacteria can be efficient bioremediators of metals and may provide an alternative or additional method to conventional methods of metal removal. In this study, 10 bacterial isolates were isolated from soil samples of a sugar industry, located at Peshawar, Pakistan. Morphological, physiological, and biochemical characteristics of these isolates were observed. Sequence analysis (16S ribosomal RNA) revealed that isolated strains were closely related to the species belonging to the genera Pseudomonas, Arthrobacter, Exiguobacterium, Citrobacter, and Enterobacter Bacterial isolates were resistant with a minimum inhibitory concentration (500-900 ppm) to lead ion (Pb(2+)), (500-600 ppm) nickel ion (Ni(2+)), (500-800 ppm) copper ion (Cu(2+)), and (600-800 ppm) chromium ion (Cr(3+)) in solid media. Furthermore, biosorption of metals proved considerable removal of heavy metals by isolated metal-resistant strains. Pseudomonas sp. reduced 37% (Pb(2+)), 32% (Ni(2+)), 29% (Cu(2+)), and 32% (Cr(3+)) and was thus found to be most effective, whereas Enterobacter sp. reduced 19% (Pb(2+)), 7% (Ni(2+)), 14% (Cu(2+)), and 21% (Cr(3+)) and was found to be least effective. While average reduction of Pb(2+), Ni(2+), Cu(2+), and Cr(3+) by Citrobacter sp. was found to be 24%, 18%, 23%, and 27%, respectively, among recognized species. This study revealed that Pseudomonas sp. may provide a new microbial community that can be used for enhanced remediation of contaminated environment.

Fulvic acid mediates chromium (Cr) tolerance in wheat (Triticum aestivum L.) through lowering of Cr uptake and improved antioxidant defense system.

Chromium (Cr) stress is one of the most adverse environmental factors that affect plant growth and food chain contamination. Fulvic acid (FA) is known to enhance the growth and production of crops, but the studies are scare regarding the application of FA on metal tolerance in plants. The effects of FA application on alleviating Cr phytotoxicity in wheat plants were investigated in a pot experiment conducted in sand- and soil-grown plants. Three Cr (0, 0.25, and 0.50 mM) treatments in the form of K2Cr2O7 were applied in both soils with or without foliar application of 1.5 mg L(-1) FA. Plants were harvested after 4 months of treatments, and data regarding growth characteristics, biomass, photosynthetic pigments, and antioxidant enzymes were recorded. FA application increased plant biomass, photosynthetic pigments, and antioxidant enzymes while it decreased Cr uptake and accumulation in plants as compared with Cr treatments alone. We conclude that FA application contributes to decreased Cr concentrations in wheat grains and could be used as an amendment when aiming for decreased metal concentration in plants.

Cadmium-induced upregulation of lipid peroxidation and reactive oxygen species caused physiological, biochemical, and ultrastructural changes in upland cotton seedlings.

Cadmium (Cd) toxicity was investigated in cotton cultivar (ZMS-49) using physiological, ultrastructural, and biochemical parameters. Biomass-based tolerance index decreased, and water contents increased at 500 µM Cd. Photosynthetic efficiency determined by chlorophyll fluorescence and photosynthetic pigments declined under Cd stress. Cd contents were more in roots than shoots. A significant decrease in nutrient levels was found in roots and stem. A significant decrease in nutrient levels was found in roots and stems. In response to Cd stress, more MDA and ROS contents were produced in leaves than in other parts of the seedlings. Total soluble proteins were reduced in all parts except in roots at 500 µM Cd. Oxidative metabolism was higher in leaves than aerial parts of the plant. There were insignificant alterations in roots and leaves ultrastructures such as a little increase in nucleoli, vacuoles, starch granules, and plastoglobuli in Cd-imposed stressful conditions. Scanning micrographs at 500 µM Cd showed a reduced number of stomata as well as near absence of closed stomata. Cd depositions were located in cell wall, vacuoles, and intracellular spaces using TEM-EDX technology. Upregulation of oxidative metabolism, less ultrastructural modification, and Cd deposition in dead parts of cells show that ZMS-49 has genetic potential to resist Cd stress, which need to be explored.

Visual rehabilitation of patients with Stargardt's disease.

OBJECTIVE: To assess the role of low vision devices in visual rehabilitation of patients with Stargardt's disease. STUDY DESIGN: Descriptive study. PLACE AND DURATION OF STUDY: The Khyber Institute of Ophthalmic Medical Sciences, Hayatabad Medical Complex, Peshawar, Pakistan, from June 2003 to June 2005. PATIENTS AND METHODS: Patients diagnosed as having Stargardt's disease and confirmed by two senior ophthalmologists clinically and after fluorescein fundus angiography and Electro-Retinogram (ERG) when needed, were included in the study. Patients with fundus pathologies other than Stargardt's disease were excluded. Each subject underwent an ophthalmic examination. Visual acuity was tested with a logarithm of the minimum angle of resolution chart; Feinbloom chart and Snellen type. Low vision assessment was performed on all individuals. SPSS version 10 was used for analysis of the data regarding the severity of visual impairment, visual acuity and devices used for aiding vision. Chi-square test was used for comparison of proportions. RESULTS: Of this cohort of 64 patients, 72% were aged between 7 to 15 years and 28% were 16 to 32 years. Using WHO low vision criteria, the percentage of visually impaired, severe visually impaired and blind (at the time of presentation) were 56.3%; 31.3% and 9.4% respectively. Among those patients, 3.1% had distance visual acuity of 6/18 or better in the better eye and 53% had normal near visual acuity of 1M (0.8 print size). Telescopes were prescribed to 53% patients for enhancement of distance visual acuity to meet their needs. CONCLUSION: Stargardt's patients respond well to magnification. Simple bifocal glasses may be used in the early stages. Visual rehabilitation can help Stargardt's patients to learn independence in their activities of daily living.

Proptosis: etiology and demographic patterns.

OBJECTIVE: To determine the demographic pattern of patients presenting with proptosis and its etiology. DESIGN: A cross-sectional descriptive study. PLACE AND DURATION OF STUDY: Khyber Institute of Ophthalmic Medical Sciences (KIOMS), Hayatabad Medical Complex, Peshawar from July 2001- December 2002. PATIENTS AND METHODS: Sixty cases of proptosis presenting at KIOMS were included. Thirtysix patients were male while 24 were female. They were divided into two age groups; 0-15 years (paediatric group) and above 15 years (adult group). An elaborate work up plan was formulated for all patients, which included detailed history, clinical examination (ocular, orbital and systemic), radiological and histopathological investigations. RESULTS: Neoplasms (33%) were the most common causes of proptosis in all the patients followed by orbital inflammations (23%), orbital infections (20%), structural abnormalities (12%), vascular abnormalities (7%) and trauma (5%). CONCLUSION: Neoplasms were the most common cause of proptosis in both paediatric and adult group. Proptosis was the cardinal feature and visual deterioration was present in more than half of the patients. Timely referral, early diagnosis and appropriate management can result in reduction of visual morbidity.

Clinical profile of amblyopia in Pakistani children age 3 to 14 years.

OBJECTIVE: To study the clinical profile of amblyopia in children age 3 to 14 years. DESIGN: A cohort study. PLACE AND DURATION OF STUDY: The study was conducted over a period of two years from June 2001 to June 2003 at Khyber Institute of Ophthalmic Medical Sciences, Hayatabad Medical Complex, Peshawar, Pakistan. PATIENTS AND METHODS: This study included 316 children. Visual acuity was tested with Snellen type and Lea symbols chart according to the level of cooperation of the children. Cycloplegic refraction and orthoptic assessment was performed on all children. RESULTS: One hundred and eighty-two children were between 3 to 8 years age and 134 were between 8 to 14 years. Mean age was 8 years. One hundred and twenty children had strabismic amblyopia, 136 children had anisometropic amblyopia, while 60 children had combined mechanism amblyopia (strabismus and anisometropia both). CONCLUSION: The results indicate the importance of screening school-going children for refractive error and amblyopia and the importance of a future prospective study on the magnitude, cause and treatment of amblyopia at more treatable age.

Corneal complications of vernal catarrh.

OBJECTIVE: To identify the magnitude and types of corneal complications of vernal catarrh and the extent of visual impairment caused by them. DESIGN: A cross-sectional study. PLACE AND DURATION OF STUDY: Khyber Institute of Ophthalmic Medical Sciences (KIOMS), Hayatabad Medical Complex (HMC), Peshawar, from March, 1999 to May, 2000. SUBJECTS AND METHODS: All cases of vernal catarrh who agreed to an informed consent were included in the study. Patients who were difficult to be examined by a slit lamp were excluded from the study. The diagnosis was made mainly on the basis of history and clinical examination. Slit lamp examination was done in every case together with fluorescein staining of cornea. The variables were recorded and statistically analyzed. RESULTS: Superficial punctate keratitis (45%) was the most common corneal complication. Visual impairment was more pronounced in cases with shield ulcers (14%) and corneal plaques (8%). A strong association with keratoconus (15%) was also noted. Corneal opacification (9%), hydrops (6%) and pseudogerentoxon (3%) were also found. CONCLUSION: Corneal complications in VKC are common, potentially serious and may cause marked visual impairment.