A study of toxicity and differential gene expression in murine liver following exposure to anti-malarial drugs: amodiaquine and sulphadoxine-pyrimethamine

BACKGROUND: Amodiaquine (AQ) along with sulphadoxine-pyrimethamine (SP) offers effective and cheaper treatment against chloroquine-resistant falciparum malaria in many parts of sub-Saharan Africa. Considering the previous history of hepatitis, agranulocytosis and neutrocytopenia associated with AQ monotherapy, it becomes imperative to study the toxicity of co-administration of AQ and SP. In this study, toxicity and resulting global differential gene expression was analyzed following exposure to these drugs in experimental Swiss mice. METHODS: The conventional markers of toxicity in serum, oxidative stress parameters in tissue homogenates, histology of liver and alterations in global transcriptomic expression were evaluated to study the toxic effects of AQ and SP in isolation and in combination. RESULTS: The combination therapy of AQ and SP results in more pronounced hepatotoxicity as revealed by elevated level of serum ALT, AST with respect to their individual drug exposure regimen. Furthermore, alterations in the activity of major antioxidant enzymes (glutathione peroxidase, superoxide dismutase, catalase, glutathione reductase), indicating the development of oxidative stress, was more significant in AQ+SP combination therapy. cDNA microarray results too showed considerably more perturbed gene expression following combination therapy of AQ and SP as compared to their individual drug treatment. Moreover, a set of genes were identified whose expression pattern can be further investigated for identifying a good biomarker for potential anti-malarial hepatotoxicity. CONCLUSION: These observations clearly indicate AQ+SP combination therapy is hepatotoxic in experimental Swiss mice. Microarray results provide a considerable number of potential biomarkers of anti-malarial drug toxicity. These findings hence will be useful for future drug toxicity studies, albeit implications of this study in clinical conditions need to be monitored with cautions

Random amplified polymorphic DNA (RAPD) markers reveal genetic diversity in bael (Aegle marmelos Correa) genotypes of Andaman Islands, India

The present study evaluated genetic variability of superior bael genotypes collected from different parts of Andaman Islands, India using fruit characters and random amplified polymorphic DNA (RAPD) markers. Genomic DNA extracted from leaf material using cetyl trimethyl ammonium bromide (CTAB) method was subjected to polymerase chain reaction (PCR) with 12 polymorphic primers. A total of 476 polymorphic loci were identified with mean value of 39.66 bands per primer and 63.99% polymorphism. Application of unweighted pair group method using arithmetic average cluster analysis generated three genotypic groups. ‘Bael-5’ and ‘Bael-8’ were most similar genotypes whereas ‘Bael-7’ and ‘Bael-1’ were extreme divergent. The clusters based on molecular data were not in agreement with the morphological traits in most of the cases as low level of correlation was observed between the classification methods based on fruit characteristics and RAPD markers. The bael genotypes were found to have considerable genetic variability, demonstrating the importance of RAPD markers to analyse each genotype in a collection in order to efficiently maintain the germplasm collection for genetic improvement of bael.Keywords: Aegle, Bael, random amplified polymorphic DNA (RAPD) and genetic diversity.African Journal of Biotechnology Vol. 12(42), pp. 6055-606

Molecular and functional diversity in Capsicum landraces of Andaman Islands

The present study analyzed the diversity in 26 landraces of Capsicum from Andaman Islands using 20 morphological, 16 biochemical and 10 DNA markers. Significant differences were observed in tested landraces and 16 reference genotypes from mainland India. Biochemical markers grouped all the genotypes into eight clusters with inter-cluster distance of 0.5 to 1.9 while seven quantified morphological traits divided the test genotypes into three major clusters and seven sub-clusters with 0.1 to 1.6 inter-cluster distance value. The random amplified polymorphic DNA (RAPD) and inter simple sequence repeat (ISSR) markers assured the genetic nature of diversity in landraces. The similarity matrix from RAPD and ISSR markers revealed 48% diversity among 42 genotypes with polymorphism information content (PIC) values of 0.43 and 0.41, respectively. The correspondence in morphological and biochemical markers indicates their interdependence for observed traits. However, poor correlation between DNA profiles and functional markers suggest further screening of more number of markers.The study identified phytochemical rich landraces CA-334, SPG-7, CARI-1 and CCB-2. The information will be useful in chemo-taxonomic foot-printing of Capsicum landraces and devising apposite conservation and utilization strategies.Keywords: Capsicum, landraces, functional diversity, chemo-taxonomic diversity, DNA markersAfrican Journal of Biotechnology Vol. 12(39), pp. 5729-573

Mucinous tubular and spindle cell carcinoma of the kidney

Mucinous tubular and spindle cell carcinoma (MTSCC) is a unique neoplasm attributing to less than 1% of all renal cell carcinoma (RCC). The median age is 6th decade with a significant female preponderance.1 These tumors classically consist of tightly packed, elongated, and anastomosing tubules which merge with bland spindle cells in a myxoid stroma in variable proportions. Tubules with tufting or small papillae and foci of foamy macrophages may occur. The tumor cells are low-grade and may show clear cytoplasm or oncocytic changes with rare mitosis. Immunohistochemically, PAX8, CK7, AMACR, and CD10 are positive in this tumor.1,2 Though it is a morphological diagnosis, it can be challenging to differentiate from a solid variant of papillary RCC, sarcomatoid RCC, or myoid-predominant angiomyolipoma. Immunohistochemistry may be of little help due to overlapping profiles.3 Copy number analyses can help establish a diagnosis in challenging cases or core biopsies since these are associated with multiple chromosomal losses involving chromosomes 1, 4, 6, 8, 9, 13, 14, 15, and 22. Novel biomarkers like VSTM2A overexpression are also emerging, which can be detected by RNA in-situ hybridization. Recurrent Hippo pathway aberrations have been defined as the molecular signature of MTSCC with increased nuclear YAP1 protein expression.4 Adverse features such as necrosis, solid growth, single file infiltration, sarcomatoid transformation, lymphovascular invasion, and increased mitoses are indicators of metastatic disease. Although it has indolent behavior, rare cases with classic morphology have been seen to develop metastases.5-6 We describe gross and histopathological findings of mucinous tubular and spindle cell carcinoma in a 65-year-old male patient. He presented with right abdominal pain for one month. The pain was dull and mild to moderate in intensity. On contrast-enhanced ultrasound, a well-defined, smoothly marginated heterogeneous hyperechoic lesion measuring 5.5x6cm was noted at the upper pole of the right kidney. The lesion was hyper enhancing compared to the rest of the renal parenchyma. Based on the radiological findings, possibilities of chromophobe RCC and oncocytoma were considered. The abdominal computed tomography (CT) revealed a hyperdense lesion in the upper pole of the right kidney with a relatively well-defined margin measuring 63x65x69mm with no evidence of significant post-contrast enhancement in the either arterial, venous or delayed phase. The patient underwent a right radical nephrectomy. On gross examination, a well-encapsulated mass was seen wholly occupying the upper pole of the kidney measuring 6.9x6.5x5.5cm. The cut surface was variegated with grey-white, firm areas admixed with mucinous and hemorrhagic foci (Figure 1A). No capsular breach or perinephric fat extension was noted. The renal pelvis, sinus, and renal vessels were free of tumor. Figure 1 A - gross nephrectomy specimen showing a large, well-demarcated tumor involving the upper pole of the kidney measuring 6.9x6.5x5.5cm. The cut surface is variegated and appears grey-white and firm, along with intervening mucinous and hemorrhagic areas (scale bar = 5 cm); B - microscopically, the tumor is well-circumscribed with sharp demarcation from adjacent normal renal parenchyma (H&E; 40x); C - the dominant epithelial component is seen against the background stroma containing significant extracellular mucin (H&E; 100x); D - higher magnification showing tightly packed anastomosing tubules lined by low-grade cuboidal cells with cytoplasmic vacuoles against mucinous background (H&E; 400x).: Microscopically, the tumor was well-demarcated from the adjacent native renal parenchyma (Figure 1B). It was composed of the dominant epithelial element against the background of extracellular mucinous stroma (Figure 1C). The epithelial component contained tightly packed, elongated, tufted, and anastomosing tubules lined by low-grade cuboidal cells and are seen merging with bland spindle cells (Figure 1D). The nuclei display fine vesicular chromatin, inconspicuous nucleoli, and moderate cytoplasm with vacuolations. Mitotic figures are infrequent (<1/10HPF). No necrosis, lymphovascular invasion, or high-grade transformation was found. However, foamy macrophage collections and lymphoid aggregates are admixed. Cholesterol clefts with focal foreign body giant cell response indicated the long-standing nature of the tumor. Based on the gross and microscopy findings, the diagnosis of mucinous tubular and spindle cell carcinoma [(pT1bpNx; American Joint Committee on Cancer (AJCC) staging manual; 8th edition)] was rendered

Performance of potato (Solanum tuberosum L.) varieties with and without straw-mulch at Shankharapur, Kathmandu, Nepal

The suitability of crop varieties in a particular location determines the productivity of crops. A field experiment was conducted from February to May 2020 at Shankharapur, Kathmandu, Nepal to evaluate the growth and productivity of potato varieties with and without mulch. Five potato varieties (Cardinal, Janakdev, Khumal Bikas, MS- 42.3, Panauti Local) were evaluated under straw-mulch and no-mulch conditions. The treatments were arranged in two factorial Randomized Complete Block Design (RCBD) with 3 replications. The results revealed that the highest tuber yield was produced by Janakdev (34.29 t ha-1) followed by Panauti Local (29.52 t ha-1). Similarly, straw-mulch gave higher yield (25.9 t ha-1) as compared with no mulch condition (21.73 t ha-1). The higher tuber yield in the Janakdev and Panauti Local was because of higher tuber weight per hill, higher number and weight of medium size (50 -100 g) and oversize (> 100 g) tuber per hill. Similarly, higher yield in straw mulch was due to higher number and weight of tuber per hill. Higher gross return, net return was observed and the benefit cost ratioin varieties Janakdev (4.08) and Panauti Local (3.53) and rice straw mulch (2.88). Therefore, potato varieties Janakdev and Panauti Local with straw mulch are most suitable for Kathmandu like climate for improving productivity and profitability

Diversity of Morinda citrifolia L. in Andaman and Nicobar Islands (India) assessed through morphological and DNA markers

Morinda citrifolia L. is an important plant species for traditional medicine systems which also has immense scope in pharmaceutical industry. The aim of this study was to assess the genetic diversity of the 33 accessions using morphological traits and molecular markers. The accessions showed great variation among morphological parameters. The germplasm showed great variation for fruit weight which ranged from 60 to 125 g. JGH-5, GAH-2, HD-6, CHLD-17 and HBAY-11A from Andaman Islands were found to be big fruited while TRA-1 and TRA-2 from Nicobar Islands were found as small fruited accessions, suggesting difference in collections from Andaman Islands and Nicobar Islands. The extent of diversity in accessions was also supported by polymerase chain reaction (PCR) analysis using random amplified polymorphic DNA (RAPD) and inter-simple sequence repeats (ISSR) markers which grouped M. citrifolia collections into two major clusters with 74% similarity. The study revealed variation in M. citrifolia populations from two different geographical isolated islands and identified promising accessions for agronomical parameters for breeding and industrial use.Key words: Morinda citrifolia L., DNA markers, morphological markers, genetic diversity, Andaman and Nicobar Islands

Diagnostic utility of α-methylacyl CoA racemase (P504S) & HMWCK in morphologically difficult prostate cancer

<p>Abstract</p> <p>Background</p> <p>To evaluate the diagnostic utility of alpha-methylacyl CoA racemase (P504S) & HMWCK (34beta E12) in morphologically difficult prostate cancer.</p> <p>Methods</p> <p>A total of 1034 cases were reviewed and divided into benign (585) malignant (399) and suspicious (50). Immunohistochemistry with HMWCK and AMACR was done on the 50 suspicious cases along with controls.</p> <p>Results</p> <p>Forty nine suspicious cases were resolved by using both markers where as 1 case was resolved by further support with CD68. The original diagnosis was changed in 15 of 50 (30%) suspicious cases from benign to malignant, one case from benign to high grade PIN and in one case from malignant to benign. Change of diagnosis was seen in 17 of 50 (34%) suspicious cases with a significant p value of 0.002. The overall diagnosis was changed in 17 of 1034 cases (1.64%) of prostatic disease (p < 0.001).</p> <p>Conclusions</p> <p>A combination of HMWCK and AMACR is of great value in combating the morphologically suspicious cases and significantly increasing the diagnostic accuracy in prostate cancer. Although, in this study the sensitivity and specificity of HMWCK and AMACR were high, yet it should be used with caution, keeping in mind all their pitfalls and limitations.</p

Nitrogen uptake and economics of black rice (Oryza sativa L. indica) under different crop geometries and nitrogen management practices

Black rice&nbsp;has more antioxidants than any other rice variety.&nbsp; It is considered to have multiple benefits in human health due to the presence of different antioxidants. A field experiment was conducted during rainy season of 2015-2016 to assess the nitrogen uptake, use efficiency and economics of black rice production under different crop geometry and nitrogen (N) management practices in Rampur, Chitwan, Nepal. The experiment was laid out in strip plot design with three replications. The experiment consisted of&nbsp; treatment combination of three crop geometry (20 cm × 20 cm, 20 cm × 15 cm and 15 cm × 15 cm) in vertical plots and three nitrogen management practices (N level: 30 kg N ha-1, 60 kg N ha-1, and LCC based N-management) in horizontal plots. The results showed that the highest N uptake was recorded from closer spacing (15 cm × 15 cm) with LCC based N management. The net return and B: C ratios were higher at a closer spacing of 15 cm × 15 cm with LCC based N management and closer spacing of 15 cm × 15 cm with N application of 60 kg ha-1. The overall analysis revealed that LCC based N management under closer crop geometry (15 cm × 15 cm) was the best management practices because of high nitrogen uptake and highest monetary return with B: C ratio of 5.76

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Stability-Indicating RP-HPLC Method for Analysis of Paracetamol and Tramadol in a Pharmaceutical Dosage Form

A simple, isocratic, rapid and accurate reversed phase high performance liquid chromatography method was developed for the quantitative determination of paracetamol and tramadol in commercial medicinal tablets. The chromatographic separation was achieved on an Intersil C18 (250 mm x 4.6 mm, 5μm) column using water pH 3.4 with orthophosphoric acid: methanol (60:40, v/v) as a mobile phase, and UV detection at 228 nm. The chromatographic resolutions between paracetamol and tramadol were found greater than five. The linear range for paracetamol and tramadol were 20.8–39.0 μg/ml and 2.4–4.5 μg/ ml was obtained with correlation coefficients ≥0.999 for each analyte. The retention time were found to be 2.1 and 3.9 min for tramadol and paracetamol respectively. Paracetamol and tramadol was subjected to stress conditions (hydrolysis (acid, base) oxidation, photolysis and thermal degradation) and the stressed samples were analyzed by use of the method. The major degradation was observed in acid and minor in base, thermal, oxidation and photolysis. The forced degradation studies prove the stability indicating power of the method