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Objectives: The main objective was to evaluate the outcome of laparoscopic cholecystectomy in terms of intra-operative complications and the rate and reasons of conversion to open cholecystectomy. Study Design: Descriptive Study. Setting: Hayatabad Medical Complex, Peshawar. Period: June 2018 to May 2019. Material & Methods: After taking consent of Hospital ethical & research committee, patients admitted with clinical diagnosis of cholelithiasis and chronic cholecystitis, confirmed by abdominal ultrasound, undergoing laparoscopic cholecystectomy fulfilling inclusion criteria were selected. Results: A total of 150 were included in the study. Mean age was 39.2yrs with female to male ratio of 9.75:1. Laparoscopic cholecystectomy was successfully accomplished in 98% cases. In 2% (3 patients) converted cases the most common cause of conversion observed was dense adhesions in the calots triangle. Intra-operative complications were noted in 1.4% patients, those included bile duct injury and leakage from the gallbladder bed. However other complications such as bowel injury, blood vessel injury, and post operative hemorrhage did not occur. Overall morbidity was 1.4% with no mortality. Conclusion: Laparoscopic cholecystectomy is a safe and effective procedure in our setup to the accepted standards, as evident by the national and international studies. And it can be accomplished with minimal morbidity and low rate of conversion with the increasing surgeon’s experience.

The widespread deficiency of iron (Fe) and sulfur (S) is becoming a global concern. The underlying mechanisms regulating Fe and S sensing and signaling have not been well understood. We investigated the crosstalk between Fe and S using mutants impaired in Fe homeostasis, sulfate assimilation, and glutathione (GSH) biosynthesis. We showed that chlorosis symptoms induced by Fe deficiency were not directly related to the endogenous GSH levels. We found dynamic crosstalk between Fe and S networks and more interestingly observed that the upregulated expression of IRT1 and FRO2 under S deficiency in Col-0 was missing in the cad2-1 mutant background, which suggests that under S deficiency, the expression of IRT1 and FRO2 was directly or indirectly dependent on GSH. Interestingly, the bottleneck in sulfite reduction led to a constitutively higher IRT1 expression in the sir1-1 mutant. While the high-affinity sulfate transporter (Sultr1;2) was upregulated under Fe deficiency in the roots, the low-affinity sulfate transporters (Sultr2;1, and Sultr2;2) were down-regulated in the shoots of Col-0 seedlings. Moreover, the expression analysis of some of the key players in the Fe–S cluster assembly revealed that the expression of the so-called Fe donor in mitochondria (AtFH) and S mobilizer of group II cysteine desulfurase in plastids (AtNFS2) were upregulated under Fe deficiency in Col-0. Our qPCR data and ChIP-qPCR experiments suggested that the expression of AtFH is likely under the transcriptional regulation of the central transcription factor FIT.

Selenium is an essential trace element required for seleno-protein synthesis in many eukaryotic cells excluding higher plants. However, a substantial fraction of organically bound selenide in human nutrition is directly or indirectly derived from plants, which assimilate inorganic selenium into organic seleno-compounds. In humans, selenium deficiency is associated with several health disorders Despite its importance for human health, selenium assimilation and metabolism is barely understood in plants. Here, we analyzed the impact of the two dominant forms of soil-available selenium, selenite and selenate, on plant development and selenium partitioning in plants. We found that the reference plant Arabidopsis thaliana discriminated between selenate and selenite application. In contrast to selenite, selenate was predominantly deposited in leaves. This explicit deposition of selenate caused chlorosis and impaired plant morphology, which was not observed upon selenite application. However, only selenate triggered the accumulation of the macronutrient sulfur, the sister element of selenium in the oxygen group. To understand the oxidation state-specific toxicity mechanisms for selenium in plants, we quantified the impact of selenate and selenite on the redox environment in the plastids and the cytosol in a time-resolved manner. Surprisingly, we found that selenite first caused the oxidation of the plastid-localized glutathione pool and had a marginal impact on the redox state of the cytosolic glutathione pool, specifically in roots. In contrast, selenate application caused more vigorous oxidation of the cytosolic glutathione pool but also impaired the plastidic redox environment. In agreement with the predominant deposition in leaves, the selenate-induced oxidation of both glutathione pools was more pronounced in leaves than in roots. Our results demonstrate that Se-species dependent differences in Se partitioning substantially contribute to whole plant Se toxicity and that these Se species have subcellular compartment-specific impacts on the glutathione redox buffer that correlate with toxicity symptoms.

The widespread deficiency of iron (Fe) and sulfur (S) is becoming a global concern. The underlying mechanisms regulating Fe and S sensing and signaling have not been well understood. We investigated the crosstalk between Fe and S using mutants impaired in Fe homeostasis, sulfate assimilation, and glutathione (GSH) biosynthesis. We showed that chlorosis symptoms induced by Fe deficiency were not directly related to the endogenous GSH levels. We found dynamic crosstalk between Fe and S networks and more interestingly observed that the upregulated expression of

For the protection of human health, clean and safe drinking water supply is important. In connection with this, the drinking and irrigation water of village Darra Pezu and its neighboring areas of district Lakki Marwat, Pakistan was evaluated for different physical and chemical constraints including pH, electrical conductivity (EC), alkalinity, turbidity, hardness, total dissolved solids (TDS) and total suspended solids (TSS). The standard analytical procedures were adopted for the study of each of these parameter. The obtained results of each parameter were compared to the standard permissible limits given by the National Drinking Water Quality Standard (NDWQS) as well as World Health Organization (WHO) standards for drinking water and were found to be according to the safe limits prescribed by NDWQS and WHO except well water. The two well water samples of Wanda Sharbat and Wanda Gulmir have high pH, turbidity, EC, hardness, alkalinity, TDS, TSS values than that given by WHO and NDWQS. It is because the well water is open to atmosphere so due to which there is contamination by chemicals pollutants and high growth of algae. Overall, water from all the sources is reported irrigation while well water of the area is not fit for drinking and needs the government attention.

Serine acetyltransferase (SAT) (EC 2.3.1.30) is the rate-limiting enzyme of cysteine (Cys) biosynthesis, providing the decisive precursor for the ubiquitous defense thiol glutathione (GSH). Together with O-acetylserine (thiol) lyase (OAS-TL; EC 2.5.1.47) SAT generates Cys in the cytosol, plastids, and mitochondria of vascular plants. The current study aimed to overproduce Cys and GSH for enhanced stress tolerance via overexpression of the feedback-insensitive isoform of serine acetyltransferase from tobacco, i.e., NtSAT4. Constitutive overexpression of NtSAT4 in Brassica napus resulted in the 2.6-fold-4-fold higher SAT activity in different subcellular compartment-specific lines. This higher SAT activity led to a 2.5-fold-3.5-fold higher steady-state level of free Cys and 2.2-fold-5.3-fold elevated level of GSH in leaves compared with nontransformed plants. Among the compartment-specific lines, the mitochondrial targeted NtSAT4 overexpressor line M-182 showed the highest levels of Cys (3.5-fold) and GSH (5.3-fold) compared with wild-type plants. Overexpression of NtSAT4 conferred a physiological advantage in terms of enhanced tolerance against oxidative stress with hydrogen peroxide and the heavy metal cadmium (Cd). The NtSAT4 overexpressor lines showed a significantly higher amount of iron (Fe) translocation from roots to shoots compared with nontransformed plants. Overall, these results suggest that overexpression of NtSAT4 is a promising approach to creating plants with tolerance to heavy metals and oxidative stress and, in addition, may potentially improve plant nutrition in terms of enhanced Fe translocation from roots to shoots.

Songklanakarin Journal of Science and Technology, 41, 6, 1348-1355

Phytohormone abscisic acid (ABA) is the canonical trigger for stomatal closure upon abiotic stresses like drought. Soil-drying is known to facilitate root-to-shoot transport of sulfate. Remarkably, sulfate and sulfide&mdash, a downstream product of sulfate assimilation&mdash, have been independently shown to promote stomatal closure. For induction of stomatal closure, sulfate must be incorporated into cysteine, which triggers ABA biosynthesis by transcriptional activation of NCED3. Here, we apply reverse genetics to unravel if the canonical ABA signal transduction machinery is required for sulfate-induced stomata closure, and if cysteine biosynthesis is also mandatory for the induction of stomatal closure by the gasotransmitter sulfide. We provide genetic evidence for the importance of reactive oxygen species (ROS) production by the plasma membrane-localized NADPH oxidases, RBOHD, and RBOHF, during the sulfate-induced stomatal closure. In agreement with the established role of ROS as the second messenger of ABA-signaling, the SnRK2-type kinase OST1 and the protein phosphatase ABI1 are essential for sulfate-induced stomata closure. Finally, we show that sulfide fails to close stomata in a cysteine-biosynthesis depleted mutant. Our data support the hypothesis that the two mobile signals, sulfate and sulfide, induce stomatal closure by stimulating cysteine synthesis to trigger ABA production.

Understanding and controlling the processes occurring at electrode/electrolyte interface are very important in optimizing energy conversion devices. Cyclic voltammetry is a very sophisticated and accurate electroanalytical method enables us to explore the mechanism of such electrode reactions. In this work, electrochemical experiments using cyclic voltammetry were performed in aqueous KCl solution containing potassium ferricyanide, K3[Fe(CN)6] at a glassy carbon disc working electrode and the mechanism of the reactions is highlighted. The CV measurements shows that the ferricyanide [Fe(CN)6]3− reduction to ferrocyanide [Fe(CN)6]4− and the reverse oxidation process follows an outer sphere electrode reaction mechanism. Voltammetry analysis further indicates that the reaction is reversible and diffusion controlled one electron transfer electrochemical process. The peak currents due to [Fe(CN)6]4− oxidation and the peak current due to [Fe(CN)6]3− reduction increase with increase of concentration and scan rate increase. The diffusion co-efficient was determined by applying Randles-Sevcik equation. The report could be helpful for university students to understand the basics of electron transfer in redox processes and cyclic voltammetry.

The regeneration potential of the economically important plant Saccharum officinarum (Sugarcane) was investigated. Callus induction and shoot regeneration along with somatic embryogenesis were induced from bud explants incubated on Murashige and Skoog (MS)-medium supplemented with different plant growth regulators (PGRs) and white sugar. The best callus induction (83.33%) was observed on explants incubated on MS-medium plus 1.0 mg·l−1 2,4-dichlorophenoxyacetic acid (2,4-D) and 4.0 mg·l−1 2,4-D (70%) after 6 weeks of culture. Other combinations (BA, IBA, IAA, NAA and GA3) of PGRs were less effective than 2,4-D. It was observed that lower concentrations of 2,4-D induced somatic embryos in bud explants of Saccharum officinarum, whereas higher concentrations induced non-embryogenic calli. Subsequent sub-culturing of calli onto MS-medium supplemented with BA (6-benzyladenine) induced shoot organogenesis. Highest shoot induction (98%) was recorded for 2.0 mg·l−1 after 3 weeks of culture. With this conce...

Selenium (Se)-fortified broccoli (Brassica oleracea var. italica) has been proposed as a functional food for cancer prevention, based on its high glucosinolate (GSL) content and capacity for Se accumulation. However, as selenate and sulphate share the initial assimilation route, Se fertilization could interfere with sulphur metabolism and plant growth. Consequently, GSL accumulation could be compromised. To evaluate these potentially adverse effects of Se fertilization, we performed a comprehensive study on sand-grown young broccoli plants (weekly selenate applications of 0.8 µmol plant(-1) via the root) and field-grown adult broccoli plants during head formation (single foliar selenate application: 25.3 or 253 µmol plant(-1) ). The results show that under these conditions, Se application does not affect plant growth, contents of cysteine, glutathione, total GSL, glucoraphanin (major aliphatic GSL) or the expression of BoMYB28 (encoding a functionally confirmed master regulator for aliphatic GSL biosynthesis). Conversely, due to the changed expression of sulphate transporters (BoSULTR1;1, 1;2, 2;1, and 2;2), sulphate and total S contents increased in the shoot of young plants while decreasing in the root. We conclude that broccoli can be fertilized with Se without reduction in GSL content, even with Se accumulation exceeding the level recommended for human consumption.

This research was conducted to determine inbreeding depression in S1 maize lines for reproductive and yield related traits, at NWFP Agricultural University, Peshawar, Pakistan, in two successive seasons during 2002. Maize variety"Pahari" was selfed for one generation and 63 S1 lines were evaluated using "lattice square design" with two replications along with the parental population. Days to mid silking and mid pollen shed (anthesis), plant height and ear height (cm), and 200 grain weight (g) showed inbreeding depression with varying degrees. Severe inbreeding depression was observed for yield with the average of 709 kg ha-1. Inbreeding depression for 200 grains weight averaged 19 g, while for plant height and ear height it was calculated as 33 and 16 cm. Inbreeding depression for days to mid silking and pollen shed was lower as compared to morphological and yield components. Silking and pollen shed were delayed by 1.8 and 1.5 days on the average, respectively, as compared to the parental variety. Yield was significantly correlated with all maturity and morphological characters. Maturity characters were positively and significantly correlated with each other, showing good synchronization in maturity characters, while they were negatively and significantly correlated with yield components.

The proximate composition, mineral constituents and amino acid profile of four important legumes (chickpea, lentil, cowpea and green pea) were studied in order to evaluate their nutritional performance. Significant (P < 0.05) variations existed among the legumes with respect to their proximate composition, mineral constituent and amino acid profile. Lentil was found to be a good source of protein, while cowpea was good in ash among the grain legumes tested. All four types of legumes were also better suppliers of mineral matter, particularly potassium, phosphorus, calcium, copper, iron, and zinc. However, the concentrations of various mineral constituents was not in good nutritional balance. It was concluded that the four legumes tested were rich in lysine, leucine and arginine and can fulfil the essential amino acid requirement of human diet except for S-containing amino acids and tryptophan. In order to make good, the deficiency of certain essential amino acids in legume protein, they must be supplemented with other vegetables, meat and dairy products (e.g., Whey, yogurt).

The experiment was conducted at NWFP Agricultural University, Peshawar to study heterosis and inbreeding depression in 7×7 half diallel crosses of sunflower. The planted materials consisted of parental inbred lines, their F1 hybrids and F2 populations using randomized complete block design with three replications. Data were recorded on yield and other important agronomic characters. Significant genetic differences were observed among the parents, their F1 hybrids and F2 populations for all the characters under study. Yield and leaf area showed highly significant heterosis in F1 hybrids ranging from 102 to 309% and46.3 to 163.9%, respectively, while inbreeding depression in the F2 population ranged from 17-71% and -9.7-43%, respectively. Leaves per plant showed low level of heterosis in F1 hybrids (-0.9 to 39.7%), whereas the effect of inbreeding depression in F2 population was comparatively high (1.1 to 22.2%) for this character. The parent RHA-822 proved itself to be a good general combiner by making higher contribution towards heterosis both in F1 hybrids and in F2 populations.

Selenium (Se)-fortified broccoli (Brassica oleracea var. italica) has been proposed as a functional food for cancer prevention, based on its high glucosinolate (GSL) content and capacity for Se accumulation. However, as selenate and sulphate share the initial assimilation route, Se fertilization could interfere with sulphur metabolism and plant growth. Consequently, GSL accumulation could be compromised. To evaluate these potentially adverse effects of Se fertilization, we performed a comprehensive study on sand-grown young broccoli plants (weekly selenate applications of 0.8 µmol plant(-1) via the root) and field-grown adult broccoli plants during head formation (single foliar selenate application: 25.3 or 253 µmol plant(-1) ). The results show that under these conditions, Se application does not affect plant growth, contents of cysteine, glutathione, total GSL, glucoraphanin (major aliphatic GSL) or the expression of BoMYB28 (encoding a functionally confirmed master regulator for aliphatic GSL biosynthesis). Conversely, due to the changed expression of sulphate transporters (BoSULTR1;1, 1;2, 2;1, and 2;2), sulphate and total S contents increased in the shoot of young plants while decreasing in the root. We conclude that broccoli can be fertilized with Se without reduction in GSL content, even with Se accumulation exceeding the level recommended for human consumption.

The role of sulfite reductase (SiR) in assimilatory reduction of inorganic sulfate to sulfide has long been regarded as insignificant for control of flux in this pathway. Two independent Arabidopsis thaliana T-DNA insertion lines (sir1-1 and sir1-2), each with an insertion in the promoter region of SiR, were isolated. sir1-2 seedlings had 14% SiR transcript levels compared with the wild type and were early seedling lethal. sir1-1 seedlings had 44% SiR transcript levels and were viable but strongly retarded in growth. In mature leaves of sir1-1 plants, the levels of SiR transcript, protein, and enzymatic activity ranged between 17 and 28% compared with the wild type. The 28-fold decrease of incorporation of 35S label into Cys, glutathione, and protein in sir1-1 showed that the decreased activity of SiR generated a severe bottleneck in the assimilatory sulfate reduction pathway. Root sulfate uptake was strongly enhanced, and steady state levels of most of the sulfur-related metabolites, as well as the expression of many primary metabolism genes, were changed in leaves of sir1-1. Hexose and starch contents were decreased, while free amino acids increased. Inorganic carbon, nitrogen, and sulfur composition was also severely altered, demonstrating strong perturbations in metabolism that differed markedly from known sulfate deficiency responses. The results support that SiR is the only gene with this function in the Arabidopsis genome, that optimal activity of SiR is essential for normal growth, and that its downregulation causes severe adaptive reactions of primary and secondary metabolism.

Reductive assimilation of inorganic sulfate to sulfide is an essential metabolic process in higher plants for the synthesis of cysteine and all downstream compounds containing reduced sulfur in the cell. Sulfite reductase (SiR) palys a central role in the assimilatory sulfate reduction pathway by catalyzing the reduction of sulfite to sulfide. An Arabidopsis T-DNA insertion line (sir1-1) with an insertion in the promoter region of SiR was isolated in order to address the exact role of SiR in vivo. Detailed characterization of sir1-1 revealed that homozygous sir1-1 plants are viable, but severely affected in growth. Homozygous sir1-1 plants flower and set viable seeds, albeit later than wild-type plants grown under the same conditions. Evaluation of SiR transcript levels in the leaves of sir1-1 plants revealed that the mRNA was down-regulated to about 50% of wild-type level. Consequently, the amount of SiR protein and the SiR activity were reduced in the same manner. The significant differences between the leaves of sir1-1 and Col-0 plants for most of the sulfur-containing and other related compounds such as cysteine, O-acetylserine (OAS), sulfate, nitrate, total glucosinolates, total carbon (C), nitrogen (N), and sulfur (S) suggests strong perturbations in the entire metabolism of sir1-1 plants. A reduction of approximately 25.6-fold and 32.7-fold in the incorporation of 35S label into cysteine and GSH fractions, respectively, of sir1-1 leaves compared to wild-type plants was observed, suggesting the fact that the activity of SiR generates a severe bottleneck in the sulfur assimilation pathway. Investigations of the transcript levels through microarray analysis revealed that the expression of many genes related to sulfur metabolism was altered in response to reduced sulfide synthesis. Out of 920 selected genes related to sulfur metabolism, the expression of 67 genes in the leaves and 180 genes in the roots of sir1-1, were significantly up- or down-regulated compared to wild-type. The high affinity sulfate transporters, sulfate transporter 1;1 (SULTR 1;1) and sulfate transporter 1;2 (SULTR 1;2) showed a significant up-regulation in the roots of sir1-1 compared to Col-0. The up-regulation of the high affinity sulfate transporters in the roots of sir1-1 suggests that instead of steady-state sulfate levels, the amount of reduced sulfur present in the cell, likely forms the signal for their their induction. The preliminary results for a second T-DNA insertion line (sir1-2) strongly indicate that an insertion more closer to the gene, in the promoter region of SiR causes early seedling lethality. All results point towards the exclusiveness of SiR for sufite reduction and that its optimal activity is essential for the normal growth of Arabidopsis plants. Treatment of different Arabidopsis lines with selenate, which is quite similar to sulfate, caused an increase in the total sulfur and selenium contents of the plants. This might happen due to the up-regulation of sulfate transporters which might eventually lead to an increase in total sulfur and selenium due to elevated sulfate/selenate contents.

OBJECTIVES: To find and document clinico-hematological features of bone marrow aspirates inpatients presenting with various forms of cytopenia(s) at Saidu Teaching Hospitals Swat.STUDY DESIGN: Descriptive study.PLACE AND DURATION OF STUDY: This study was carried out over a period of one year (1stJanuary 2013 to 31st December 2013) at Pathology department, Saidu Medical College, Saidu Sharif,Swat.METHODS AND MATERIALS: Total 200 patients of both genders and all age groups withcytopenias referred from Saidu Teaching Hospital for bone marrow cytology were included for bonemarrow aspiration and assessment.RESULTS: Two hundred patients underwent bone marrow sampling procedure. Patient’s age rangedfrom 6 months to 90 years. There were 56% male and 44% female at the ratio of 1.5:1. The mostcommon non malignant hematological condition was megaloblastic anemia 28%, mixed nutritionaldeficiency anemia 16%, hypoplasia 8%, hemolytic anemia 7%, iron deficiency anemia was present in6% cases, visceral leishmaniasis 4%, thrombocytopenia due to any cause (immune thrombocytopenicpurpura (ITP)) 2%, anemia of chronic illness was present in 2% cases, malaria and sepsis leading tocytopenia were 1% each. Malignant neoplasms in our study were 50 cases (25%), which were comprisedof; ALL 10%, AML 5%, CML 4%, MDS 3%, multiple myeloma 1.5% and lymphoproliferativedisorders in 1% cases.CONCLUSION: This study showed the pattern of morphological features on bone marrow aspiration,which was found to be different from pattern seen in European and Middle East regions and similar tothat found in Asian subcontinent.KEY WORDS: Cytopenias, pancytopenia, bicytopenia, anemia, thrombocytopenia, megaloblasticanemia, acute leukemia, myelodysplastic syndrome.

Objectives: The objective of the study was to determine the frequency of HELLPsyndrome in preeclampsiaand eclampsia patients presented to saidu teaching hospital Swat. Methodology: This was a descriptive, cross sectional study. It was conducted in the department ofgynaecology and obstetrics of Saidu Teaching Hospital Swat. All pregnant women attending antenatal clinicduring T1 Jan 2007 to 31s' Dec 2009 were included in the study. Women after 20 weeks of gestation withpreeclampsia requiring admission in obstetric ward for monitoring and controlling blood pressure and allwho were admitted in labour room with severe preeclampsia and eclampsia were included while womenwith gestational hypertension but no proteinurea were excluded. Non probability purposive samplingtechnique was used in the study. Result: During three years time period, 16722 pregnant women were registered, out of which 537 werediagnosed having preeclampsia, 3.21% (n = 16722) . While 39 patients among 537, were diagnosed withHELLPsyndrome, 7.26% (n = 537) in limited and 0.227% (n = 16722) in the overall sample population. Outof total 39 patients 38 were antenatal 1 was post-natal. About 69.25% women were between 30 to 40 weeksof gestation and 30.7% were less than 30 weeks of gestation. In our study the frequency of HELLPsyndromewas high between 26-30 years (38.46%) and in multigravidas (46.15%). Conclusion: The study revealed that although in spite of backward area, lack of education and health careawareness with special reference to mother and child health the incidence of HELLP syndrome is less thanthe internationally reported data, but this may be due the fact that all patients with eclampsia and preeclampsia do not reached hospital and treated at home due to superstitious believes. A community basedintervention is needed to improve health awareness and education. All patients with preeclampsia andeclampsia needs to be investigated for HELLP syndrome and those with HELLP syndrome should beprovide the best possible care at the appropriate time. Key words: Pre-Eclampsia, Eclampsia, HELLPsyndrome