Comparison of knowledge and perception about adverse drug reaction reporting and pharmacovigilance between pharmacy and medical students

Background: Adverse drug responses are serious public health issues worldwide. Pharmacovigilance (PV) is an important part of the healthcare system since it assesses, monitors, and discovers medication interactions and their consequences in humans. Objective were to assess knowledge and perception about adverse drug reactions (ADR) and PV among pharmacy and medical students. Methods: A cross-sectional study was conducted for pre-final year and final-year of pharmacy and medical students. Study conducted through semi structured questionnaires and a total 607 participants were recruited in this study. Results: A total 607 participants of pharmacy (52.5%) and medical (47.4%) from pre final (50.2%), final year (49.9%) students participated in this study. Pharmacy students had a significantly better overall knowledge of ADRs than medical students (p<0.05). The perception of pharmacy students regarding ADRs reporting and PV were significantly more than those medical student’s (p<0.05). In comparison to pharmacy students, a lack of knowledge of where and how to report ADRs was the main barrier that medical students perceived to ADR reporting (0.02). Conclusions: This study shows that compared to medical students, pharmacy students had better awareness of ADR reporting and shown good perceptions of PV. To enhance the knowledge of PV among healthcare students, an instructional intervention ought to be implemented.

AN ANALYSIS OF PRESCRIPTION PATTERN, ADHERENCE TO PHARMACOTHERAPY AND QUALITY OF LIFE IN PEMPHIGUS VULGARIS PATIENTS

Objective: Pemphigus Vulgaris [PV] is a chronic autoimmune disease. Corticosteroids and Immunosuppressants are the main line of treatment. The aim of this study was to evaluate the prescription pattern and patient adherence behavior with therapy. We determined the association between patient adherence and quality of life in patients with pemphigus vulgaris.Methods: A prospective observational study was conducted from February 2020 to May 2021. The age and sex of patients who were diagnosed with PV within 1 mo period and confirmative histopathological findings in Biopsy were included in the study. A total of 35 patients were analyzed and patients were reviewed for medication adherence and quality of life by using MMAS-8 and WHOQOL respectively, monthly till 3 mo after initiation of therapy. Statistical analysis was done by using Epi Info version 7.1.5.Results: Out of 140 prescriptions, vitamin C and zinc were the most common drug prescribed in 129 prescriptions followed by prednisolone prescribed in 128 prescriptions. The average number of drugs per encounter was 8.67. 97.56 % of drugs were prescribed by generic names. No significant improvement was seen in the adherence behavior of patients from 1st follow-up to 3rd follow-up. It was observed that physical, psychological, and environmental domains have significant associations with medication adherence in all 3 follow-up visits.Conclusion: It was concluded that Polypharmacy and inappropriate use of medicines may decrease the adherence behavior of patients to the therapy. Low adherence to pharmacotherapy affects the quality of life in PV patients. This study is beneficial for raising awareness about treatment adherence and also encouraging the development of appropriate interventions.

Exercise-induced chronic fatigue impairs working memory in rats via basal ganglia output nuclei / 神经解剖学杂志

Objective:To investigate possible neuromodulatory mechanisms involved in the involvement of parvalbu-min(PV)expression in the basal ganglia output nuclei,entopeduncular nucleus(EPN)and substantia nigra pars etic-ulata(SNr),in exercise-induced chronic fatigue impairs working memory capacity.Methods:Male SD rats were divid-ed into control group and Fatigue group by random number method,and a three-stage incremental load treadmill training program was selected to establish a chronic exhaustion exercise-induced fatigue rat model.The working memory ability of rats was assessed by the Y-maze autonomous alternation experiment.Immunohistochemical staining was used to ob-serve the expression of parvalbumin(PV)positive neurons and cysteine aspartate-specific protease-3(caspase-3)in EPN and SNr of rats.Results:The accuracy of voluntary alternation in the fatigue group was obviously lower than that in control group(P＜0.05).The results of immunohistochemical staining showed that the density of PV positive neu-rons and the degree of positive fiber staining in EPN and SNr in the fatigue group were obviously lower than those in the control group(P＜0.05,P＜0.01).The number of caspase-3 positive cells per unit area of EPN and SNr in the fa-tigue group was obviously higher than that in the control group(P＜0.05,P＜0.01).Conclusion:The mechanism of impairing working memory in rats caused by exercise-induced chronic fatigue may be related to the apoptosis of PV posi-tive neurons in EPN and SNr.

The role of iron-uptake factor PiuB in pathogenicity of soybean pathogen Xanthomonas axonopodis pv. glycines / 生物工程学报

Iron is an essential element for living organisms that plays critical roles in the process of bacterial growth and metabolism. However, it remains to be elucidated whether piuB encoding iron-uptake factor is involved in iron uptake and pathogenicity of Xanthomonas axonopodis pv. glycines (Xag). To investigate the function of piuB, we firstly generated a piuB deletion mutant (ΔpiuB) by homologous recombination. Compared with the wild-type, the piuB mutant exhibited significantly reduced growth and virulence in host soybean. The mutant displayed markedly increased siderophore secretory volume, and its sensitivity to Fe3+, Cu2+, Zn2+ and Mn2+ was significantly enhanced. Additionally, the H2O2 resistance, exopolysaccharide yield, biofilm formation, and cell mobility of ΔpiuB were significantly diminished compared to that of the wild-type. The addition of exogenous Fe3+ cannot effectively restore the above characteristics of ΔpiuB. However, expressing piuB in trans rescued the properties lost by ΔpiuB to the levels in the wild-type. Taken together, our results demonstrated that PiuB is a potential factor for Xag to assimilate Fe3+, and is necessary for Xag to be pathogenic in host soybean.

Identification and Characterization of the Pathogen Causing Canker Disease in Acid Lime (Citrus aurantiifolia) cv. Petlur Acid Lime

In this study, the pathogen responsible for canker disease in Acid Lime Cv. Petlur Acid Lime was identified through a comprehensive analysis involving both morphological and molecular characterization techniques. Cultures of the Xanthomonas campestris pv. citri (Xcc) bacterial isolate, grown on artificial media, exhibited distinct characteristics: small, round, and mucoid colonies with a gram-negative nature, as revealed by Gram staining. Molecular identification was performed using universal primers 27F and 1492R, resulting in a specific DNA band of 424 bp, confirming the presence of the pathogen in the samples.

Photovoltaics: Panacea to Africa Sustainable Development

African countries are falling behind in attaining the sustainable development goals. They are in dire need of electricity, which had been declared the essential tool to achieve most goals, to meet their population’s utilizable energy requirement. With renewables being globally adopted energy options to save the changing climate and preserve the environment and biodiversity, photovoltaic (PV) is advocated to remedy the energy deficiency for sustainable development. All over Africa indicates viability in PV energy owing to the incident global solar radiation (Gh). Nigeria being a case study shows awareness of PV energy, which has manifested in the attitude towards its penetration and the expected benefits. With the anticipated threat of climate change on PV performances, the projected incident Gh across Nigeria has shown two epochs which are yet viable for PV energy by the year 2100. Foreseeable challenges in the PV penetration and investments have been presented as wrongdoings and whistleblowing is advocated as the policy that will ensure the PV penetration for sustainable development.

Biological Management of Bacterial Leaf Blight of Rice (Xanthomonas oryzae pv. oryzae) through Rice Rhizosphere Antagonist Actinobacteria

Rice (Oryza sativa L.) is a staple food crop for more than two billion people around the world. Bacterial leaf blight (BLB) caused by Xanthomonas oryzae pv. oryzae (Xoo) (Ishimiya) Swings et al. is one of the most destructive diseases of rice in all the major rice-growing. Use of antibiotics is the common practice for the management of BLB, but it has detrimental effect on human health and environment. Hence other eco-friendly strategies like biological control and host plant resistance (HPR) should be effectively exploited for BLB management. An attempt was made to manage the disease using actinobacteria isolated from rice rhizosphere. Among sixteen actinobacterial isolates, six were selected for secondary screening based on disc diffusion assay. Out of six isolates AUDT-502 produced maximum zone of inhibition (1.81 cm) followed by AUDT-508 (1.62 cm) and were evaluated under glasshouse condition. AUDT-502 (Streptomyces rimosus) appeared as the most promising bio agent which was able to inhibit the pathogen (Xoo) both in in vitro and in vivo condition. AUDT-502 increased the plant growth parameters like number of tillers (5.73), plant height (56.85 cm), root length (34.33cm) and dry root weight (3.90g) and also reduced the leaf blight lesion length and per cent disease index (4.43 per cent) significantly when compared to other treatments. The current study thus showed that there was a good correlation between in vitro inhibition of the pathogen and the in vivo control of the disease and AUDT-502 (Streptomyces rimosus) appears to be a hopeful bioagent for the management of bacterial leaf blight of rice.

Performance evaluation of nucleic acid testing system for blood screening in high-altitude areas / 中国输血杂志

【Objective】 To validate the performance of a nucleic acid testing(NAT) system for blood screening in the high-altitude Nagqu region of Tibet, in order to assess the capability of NAT in high-altitude areas and further enhance blood safety. 【Methods】 Various methods were employed to evaluate the analytical sensitivity, reproducibility, ability to prevent cross-contamination, and comparison between different NAT systems. 【Results】 The NAT system in the Nagqu region of Tibet achieved a 100% detection rate for high-concentration HBV DNA and HIV-1 RNA samples, and over 90% for medium-concentration samples. PROBIT analysis revealed the lower limits of detection (LOD) for HBV DNA and HIV-1 RNA to be 8.29 IU/mL (95% CI, 5.88~20.55 IU/mL) and 40.52 IU/mL (95% CI, 30.26~85.92 IU/mL), respectively. For HCV RNA genotype 2a, the LOD was 97.14 IU/mL (95% CI, 71.00~182.67 IU/mL), all of which were lower than the declared minimum detectable concentrations in the instructions. Reproducibility analysis demonstrated a 100% level of consistency within the system. Cross-contamination performance verification showed a strong ability to resist cross-contamination. Comparative analysis of repeated testing of low-concentration HBV DNA samples and multi-system testing in plain areas revealed consistency rates of 77.78%(14/18) and 77.27%(17/22), respectively, indicating certain differences between the NAT system in Nagqu region and other systems. 【Conclusion】 The NAT system exhibited excellent performance in blood screening at high altitudes. The results of performance validation in high-altitude blood screening NAT systems were largely consistent with those in plain areas, providing a reliable basis for enhancing blood safety in high-altitude regions.

Establishment and internal validation of a risk prediction model for urinary incontinence after transurethral holmium laser enucleation of the prostate / 现代泌尿外科杂志

【Objective】 To establish a model for predicting the risk of urinary incontinence after holmium laser enucleation of the prostate (HoLEP). 【Methods】 The clinical data of 258 patients with benign prostatic hyperplasia (BPH) who underwent HoLEP in our hospital during Jan.2019 and Feb.2022 were retrospectively analyzed. According to the occurrence of urinary incontinence after surgery, they were divided into the urinary incontinence group (n=84) and non-urinary incontinence group (n=174). Lasso regression was used to screen the predictors of urinary incontinence after HoLEP. Logistic regression was used to establish a suitable model, and a nomogram of urinary incontinence after HoLEP was drawn. Bootstrap was used to verify and draw the calibration curve of the model, calculate the C index, and draw the clinical decision curve to further verify the accuracy and identification ability of the model. 【Results】 Predictors including International Prostate Symptom Score (IPSS), Quality of Life Score (QoL), body mass index (BMI), diabetes, prostate volume (PV), and prostate-specific antigen (PSA) were selected, based on which a prediction model was constructed. The area under the receiver operating characteristic (ROC) curve of the prediction model was 0.766 0, and the 95% confidence interval was 0.704-0.828. Bootstrap internal validation showed a C-index of 0.766 2, and the calibration model curve coincided well with the actual model curve. The clinical decision curve analysis showed that the model had high accuracy, and net benefit in the probability of urinary incontinence was within 10% to 82%. 【Conclusion】 IPSS, QoL, diabetes, prostate volume, and PSA are predictors that can affect the occurrence of urinary incontinence after HoLEP. The model has high accuracy, identification ability and net benefit.

Copper formulations in bacterial blight control and toxic effects on coffee seedlings

Bacterial blight of coffee (Pseudomonas syringae pv. garcae) is an important coffee disease and can be controlled using antibiotics and copper-based compounds. However, copper-based compounds raise doubts among coffee growers regarding bacterial blight control efficiency and phytotoxic potential. In this work, coffee plants were sprayed with different copper molecules in order to study their efficiency on bacterial blight control and the phytotoxic potential. Seven copper formulations, cuprous oxide, copper oxychloride, copper nitrate, copper hydroxide 1 (water-dispersible granules) and 2 (concentrated suspension), copper sulfate 1 (complexed with gluconic acid) and 2 (Bordeaux mixture) were studied. The copper formulations efficiency was compared with the antibiotic kasugamycin, saline solution, and control. In controlled environmental conditions of temperature, relative humidity, and photoperiod, coffee seedlings were sprayed with the treatments and after 24 hours they were inoculated with Pseudomonas syringae pv. garcae suspension. Disease incidence and severity assessments were performed in a 2-day interval during a 16-day period. Phytotoxicity incidence and severity, mapping, and quantification of copper on the leaf tissue surface, dried leaves weight, and total copper leaf content were assessed 16 days after pathogen inoculation. Data were submitted to the Scott-Knott test (p < 0.05). Cuprous oxide and copper sulfate 2 proved most efficient to bacterial blight control, causing lower phytotoxicity effect, best covering, and persistence on leaf tissues. Copper nitrate and copper sulfate complexed with gluconic acid were more phytotoxicity compared to other copper formulations.

Suppression of Xanthomonas oryzae pv. oryzae biofilm formation by Acacia mangium methanol leaf extract / Supressão de Xanthomonas oryzae pv. oryzae: formação do biofilme oryzae pelo extrato de folhas de metanol de Acacia mangium

Abstract Xanthomonas oryzae pv. oryzae (Xoo), a pathogen responsible for rice bacterial leaf blight, produces biofilm to protect viable Xoo cells from antimicrobial agents. A study was conducted to determine the potency of Acacia mangium methanol (AMMH) leaf extract as a Xoo biofilm inhibitor. Four concentrations (3.13, 6.25, 9.38, and 12.5 mg/mL) of AMMH leaf extract were tested for their ability to inhibit Xoo biofilm formation on a 96-well microtiter plate. The results showed that the negative controls had the highest O.D. values from other treatments, indicating the intense formation of biofilm. This was followed by the positive control (Streptomycin sulfate, 0.2 mg/mL) and AMMH leaf extract at concentration 3.13 mg/mL, which showed no significant differences in their O.D. values (1.96 and 1.57, respectively). All other treatments at concentrations of 6.25, 9.38, and 12.5 mg/mL showed no significant differences in their O.D. values (0.91, 0.79, and 0.53, respectively). For inhibition percentages, treatment with concentration 12.5 mg/mL gave the highest result (81.25%) followed by treatment at concentrations 6.25 and 9.38 mg/mL that showed no significant differences in their inhibition percentage (67.75% and 72.23%, respectively). Concentration 3.13 mg/mL resulted in 44.49% of biofilm inhibition and the positive control resulted in 30.75% of biofilm inhibition. Confocal laser scanning microscopy (CLSM) analysis of Xoo biofilm inhibition and breakdown showed the presence of non-viable Xoo cells and changes in aggregation size due to increase in AMMH leaf extract concentration. Control slides showed the absence of Xoo dead cells.

Resumo Xanthomonas oryzae pv. oryzae (Xoo), um patogênico responsável pela influência bacteriana na folha do arroz, produz biofilme para proteger células Xoo viáveis de agentes antimicrobianos. Foi conduzido um estudo para determinar a potência do extrato de folha de Acacia mangium methanol (AMMH) como um inibidor de biofilme Xoo. Quatro concentrações (3,13, 6,25, 9,38 e 12,5 mg/mL) de extrato de folha de AMMH foram testadas quanto à sua capacidade de inibir a formação de biofilme Xoo em uma placa de microtitulação de 96 poços. Os resultados mostraram que os controles negativos tiveram o maior valor de OD do que os outros tratamentos, indicando a intensa formação de biofilme. Isso foi seguido do controle positivo (sulfato de estreptomicina, com concentração de 0,2 mg/mL, e extrato de folha de AMMH, com concentração de 3,13 mg/mL), que não apresentou diferenças significativas nos seus valores OD (1,96 e 1,57, respectivamente). Todos os outros tratamentos com concentrações de 6,25, 9,38, e 12,5 mg/mL não tiveram diferenças significativas nos seus valores OD (0,91, 0,79, e 0,53, respectivamente). Para percentagens de inibição, o tratamento com concentração 12,5 mg/mL apresentou o maior resultado (81,25%), seguido do tratamento em concentrações de 6,25 e 9,38 mg/mL, que não mostraram diferenças significativas na sua percentagem de inibição (67,75 e 72,23%, respectivamente). Concentração 3,13 mg/mL resultou em 44,49% de inibição do biofilme, e o controle positivo resultou em 30,75% de inibição do biofilme. Análise por microscopia confocal de leitura a laser de inibição e separação de biofilme Xoo revelou a presença de células Xoo não viáveis e alterações no tamanho da agregação por causa do aumento na concentração de extrato de folha de AMMH. Slides de controle mostraram a ausência de células Xoo mortas.

Assessment of resistance of hybrids and lineages of passion fruit to bacteriosis under protected cultivation / Avaliação da resistência de híbridos e linhagens de maracujazeiro à bacteriose sob cultivo protegido

Yield and longevity of yellow passion fruit have been reduced by diseases such as the bacterial spot caused by Xanthomonas axonopodis pv. passiflorae. Genetic resistance has been confirmed as the most efficient and economical correct option to minimize this disease problem. Aiming at it, the objective of this research was to evaluate the incidence, severity and progression of the disease in 12 genotypes of sour passion fruit, in seedling stage in nursery greenhouse after inoculation of Xanthomonas axonopodis pv. passiflorae. The inoculation was performed with an isolate collected in the Pipiripau Rural Nucleus, Brasilia-DF, named UnB-1397 (1x106 CFU/mL), through induction of injuries. There were performed four assessments, with interval of 7 days except the first which was performed 11 days after inoculation. The incidence was estimated by the percentage of plants affected. To evaluate the severity, it was used the diagrammatic scale validated by Costa et al. (2018), with adaptations, using the measurement of the affected area by necrotic lesions on the leaf. All genotypes were susceptible to bacteriosis, 5 being considered moderately susceptible: F1BRS Pérola do Cerrado x Rosa Intenso, Mar20#21, Mar20#15b, Mar20#24xMar20#40 and FB200PL4R2 x Mar20#2005, with a mean of severity ranging from 11 to 25% of injured area in leaves.

A produtividade e a longevidade dos pomares de maracujazeiro-azedo têm sido comprometidas em razão de doenças como a bacteriose, causada por Xanthomonas axonopodis pv. passiflorae. A resistência genética tem se confirmado como a opção mais eficiente e econômica para minimizar tal problema. Dessa forma, o objetivo deste trabalho foi avaliar a reação de 12 genótipos de maracujazeiro, em fase de mudas, sob cultivo protegido, à Xanthomonas axonopodis pv. passiflorae. A inoculação com isolado denominado UnB-1397 (1x106 CFU/mL),coletado no Núcleo Rural de Pipiripau, Brasília-DF, se deu pela indução de ferimentos. Foram realizadas 4 avaliações, com intervalo de 7 dias, sendo a primeira avaliação realizada 11 dias após a inoculação. A incidência foi estimada pela porcentagem de plantas afetadas. Para avaliação da severidade, foi utilizada escala diagramática validada por Costa et al. (2018), com adaptações, utilizando-se a mensuração da área foliar atingida por lesões nas folhas. Todos os genótipos se mostraram suscetíveis à bacteriose, sendo 5 considerados moderadamente suscetíveis: F1 BRS Pérola do Cerrado x Rosa Intenso, Mar20#21, Mar20#15b, Mar20#24 x Mar20#40 e FB200PL4R2 x Mar20#2005, apresentando uma média de severidade que variou de 11 a 25% de área ou tecidos foliares lesionados.

Development and validation of a standard area diagram set as assessment aid for estimating the severity of bacterial spot on tri-lobed leaves of yellow passion fruit / Desenvolvimento e validação de escala diagramática como auxílio na avaliação da severidade da bacteriose em folhas trilobadas do maracujazeiro azedo

Bacterial spot (Xanthomonas axonopodis pv. passiflorae) significantly reduces yellow passion fruit (Passiflora edulis Sims) yield and longevity. A standard area diagram set (SADs) for severity assessment of bacterial spot on tri-lobed leaves of yellow passion was developed and validated in this study. The SADs consisted of eight severity levels (2; 4; 9; 18; 35; 58; 80; and 94%). For its validation, 20 raters, who initially estimated the disease severity without the aid of the SADs, were divided into four groups (G1 and G3, inexperienced; G2 and G4, experienced). Subsequently, G1 and G2 performed the second evaluation without the SADs, and G3 and G4 completed the second evaluation with the proposed SADs. The accuracy and precision of the assessments were determined by simple linear regression and by the Lin's concordance correlation coefficient (LCCC). The proposed SADs allowed accurate and precise quantification of bacterial spot severity, increasing the agreement between estimated and actual values. Inexperienced raters benefited the most from the use of the SADs. The increase in accuracy and precision in the non-aided groups, when present, was less pronounced than those increments observed in the SADs-aided groups. The LCCC confirmed the increases in accuracy and precision detected by the linear regression analysis.

A bacteriose (Xanthomonas axonopodis pv. passiflorae) reduz significativamente a produção e longevidade do maracujazeiro azedo (Passiflora edulis Sims). Uma escala diagramática para a avaliação da severidade da bacteriose em folhas trilobadas do maracujazeiro azedo foi desenvolvida e validada neste estudo. A escala diagramática apresentou oito níveis de severidade (2; 4; 9; 18; 35; 58; 80 e 94%). Para a sua validação, os 20 avaliadores foram divididos em quatro grupos (G1 e G3, sem experiência; G2 e G4, com experiência), que inicialmente estimaram a severidade da doença sem auxílio da escala. Posteriormente, G1 e G2 fizeram outra avaliação sem escala, e G3 e G4 realizaram a avaliação com a escala proposta. A acurácia e a precisão das estimativas foram determinadas por regressão linear simples e pelo coeficiente de correlação de concordância de Lin (LCCC). A escala diagramática proposta permitiu quantificar a severidade da bacteriose de forma acurada e precisa, aumentando a concordância entre os valores estimados e os reais. Os avaliadores inexperientes foram os mais beneficiados pelo uso da escala. O aumento da acurácia e precisão nos grupos que realizaram dupla avaliação sem escala, quando ocorreu, foi mais discreto que os incrementos observados nos grupos que utilizaram a escala. O LCCC confirmou os incrementos da acurácia e precisão detectados pela análise de regressão linear.

Characterization of Xanthomonas campestris pv. viticola causing bacterial leaf spot of grapes in Maharashtra, India

Aim: The aim of the present study was to isolate, characterize and identify pathogenic bacteria from bacterial leaf spot infected grape vineyards of Maharashtra. Methodology: Collected diseased samples were subjected to isolation. All isolates were further subjected to different morphological and biochemical characterization. These isolates were also used to standardize inoculation methods to study host range and test pathogenicity to confirm infectivity. Genomic DNA was isolated from bacteria for phylogeneitc analysis using 16 rDNA. Results: The disease is characterized by water soaked, angular leaf spot which later turns to irregular, dark brown to black necrotic region on the leaf surface. Microbial studies confirmed that it is a Gram negative, rod shaped bacterium with white, mucoid, glistening and convex circular colonies on artificial medium. Pathogenicity test was performed to confirm virulence. Phylogenetic analysis of 16S rDNA gene exhibited more than 99% similarity with other Xanthomonas campestris pv. viticola. The bacterium was found resistant to vancomycin, peniciliin and oxacillin antibiotics. The bacterium is also capable of infecting Mangifera indica and Citrus limon along with eight weed species.

In vitro Analysis of Antimicrobial Susceptibility of Xanthomonas axonopodis pv. punicae and Comparative Study of Protein Profiling under Biotic Stress

Bacterial blight of pomegranate caused by X. axonopodis pv. Punicae (XAP) assumed epidemic form and resulted in economic burden on farmers. In the current study the pathogen infected samples were collected and the isolated XAP was identity and confirmed through the morphological, biochemical characterization and Pathogenicity test. Bacterium was reisolated from infected plant to prove Koch’s postulates. Efficacy of different chemicals and oils were tested by disc diffusion assay and turbidometrically. Bronopol 3000 ppm (25.6±1.6 mm) and Clove oil (18.0±0.7 mm) formed highest zone of inhibition Turbidometri showed the highest O.D. (0.908 nm) by Copper oxy chloride and Neem oil showed maximum inhibition of growth with O.D. (0.842 nm). Biotic stress (pathogen) induced protein response was studies by using SDS-PAGE method after protein extraction from XAP, healthy P. granatum L. and infected P. granatum L. The protein band pattern showed the unique band no. 2 (Mol.Wt.66000 Da) in infected P. granatum L. as compared to the banding pattern of XAP and healthy P. granatum L. The over expressed protein due to biotic stress could be useful as a marker for detection of the disease at the early stage and for control of the diseases after knowing the biochemical significance of the protein.

A Study on Demographic Profile and Risk Factor Association in Pityriasis Versicolor Cases Attending at a Tertiary Care Hospital in Kolkata

Pityriasis versicolor (PV), also known as tinea versicolor, is a chronic, mild, superficial and recurrent infection of the stratum corneum, caused by different Malassezia spp and seen predominantly in young age group and primarily in hot and humid climates. The aim of this study was to analyze epidemiological parameters and risk factors association in clinically diagnosed PV cases and also the mycological evaluation of those PV cases. Methods: A total of 116 patients attending the OPD of Dermatology were included and analysed for detailed history, clinical examination, epidemiological parameters, risk factors and investigations. Skin scrapings collected were processed by direct microscopy with 10% KOH and culture in modified Dixon agar (mDA). Isolates were identified by colony morphology, gram staining, biochemical characteristics & tween assimilation test. Results: Females were more affected (56.03%) than the males (43.97%) with F: M ratio 1.27:1. PV affected most commonly (36.21%) in 11-20 years of age group. Students (32.29%) were affected in maximum. Majority of affected patients (65.52%) used oily body creams, whereas 34.48% cases shared their body towels with others. 10.34% cases were associated with seborrheic dermatitis. Seasonal occurrence mostly seen in May - August. Patients with type III (Medium) complexion (56.03%) with normal skin texture (49.14%) were mostly affected. Maximum patients (74.14%) were associated with excessive sweating. 18.96% patients were associated with Type II DM. Most of the cases presented with macular, scaly hypopigmented, bilaterally asymmetrically distributed and having well defined margin. Neck was the most affected site (28.45%) followed by back (20.69%). Conclusion: M.furfur was the most common isolate (47.06%) followed by M. globosa (24.71%) and M. sympodialis (15.29%).

The STK domain of the cassava bacterial blight resistance protein RXam1 interacts with an E3 Ubiquitin Ligase / El dominio stk de la proteína de resistencia a la bacteriosis vascular de yuca rxam1 interactúa con una e3 ubiquitin ligasa

RESUMEN La yuca (Manihot esculenta) representa el pilar de la seguridad alimentaria para cerca de mil millones de personas, principalmente en las zonas tropicales. Uno de los factores limitantes de la producción de yuca es la bacteriosis vascular causada por la bacteria Xanthomonasaxonopodis pv. manihotis (Xam). Recientemente se identificó el gen RXaml el cual confiere resistencia parcial de yuca a cepas de Xam. RXaml codifica una proteína con un dominio LRR (Leucine Rich Repeats) extracelular y un dominio STK (Serina Treonina Kinasa) citoplasmático; estas proteínas son conocidas como RLKs (Receptor Like Kinases). En este estudio se realizó el tamizaje de una librería de ADNc de yuca mediante doble híbrido de levadura para identificar las posibles proteínas que interactúan con el dominio STK de RXam1. El tamizaje de 3x108 clones permitió identificar y confirmar cinco clones de ellos los cuales corresponden al mismo gen, el cual codifica para una proteína que presenta un dominio central de dedos de zinc CHY, seguido por un dominio C-terminal "RING finger" y un "Zinc ribbon" el cual fue denominado CRFE3-1 (Cassava RING Finger E3 ligase). La interacción entre STK y CRFE3-1 fue altamente especifica ya que se demostró también por doble híbrido que STK no interactúa con una E3 ligasa de Arabidopsis, altamente similar a CRFE3-1, así como tampoco CRFE3-1 interactúa con el dominio STK de un RLK de lechuga similar a RXam1. La identificación de CRFE3-1 sugiere que mecanismos de degradación proteica son importantes para regular la actividad de RXam1.

ABSTRACT Cassava (Manihot esculenta) represents food security support for nearly one billion people, mainly in the tropics. One of the limiting factors of cassava's production is cassava bacterial blight, caused by the bacterium Xanthomonas axonopodis pv. manihotis (Xam). Recently, the RXam1 gene was identified, which confers partial resistance to some Xam strains. RXam1 encodes a protein with an extracellular LRR (Leucine Rich Repeats) domain and a cytoplasmic STK (Serine Threonine Kinase) domain; these proteins are known as RLK (Receptor-like Kinases). In this study, a cassava cDNA library was screened using a yeast Two-hybrid assay to identify possible proteins interacting with the STK domain of RXam1. Screening of 3x108 clones allowed identifying and confirming five of them, which correspond to the same gene, and code for a protein that has a core domain of zinc fingers CHY, followed by a C-terminal "RING finger" domain and a "Zinc ribbon". This gene was called CRFE3-1 (Cassava RING Finger E3 ligase). It was also demonstrated by yeast Two-hybrid that STK does not interact with an E3 ligase of Arabidopsis that is highly like CRFE3-1. CRFE3-1 did not show interaction with the STK domain of an RLK of lettuce related to RXam1, indicating a highly specific interaction between cassava RXam1 STK and CRFE3-1. The identification of CRFE3-1 suggests that protein degradation mechanisms are important to regulate the activity of RXam1.

Development of species-specific primers for detection of Xanthomonas campestris pv. campestris causing black rot of crucifers

Aim: Black rot of crucifers caused by Xanthomonas campestris pv. campestris (Pammel) Dowson (Xcc) is a major seed-borne disease. The present study aimed to develop a rapid diagnostic protocol for the specific and sensitive detection of this pathogen. Methodology: A specific primer set was designed based on rpf gene and optimization of PCR condition was done for specific detection of Xcc. Sensitivity of PCR for primer set was then determined by diluting the Xcc DNA and cells. Results: Specific primer set was able to amplify a specific band of 304 bp in all 11 isolates of Xcc but failed to amplify other Xanthomonas species and one each of Ralstonia solanacearum, Erwinia caratovora subsp. caratovora, Bacillus subtilis, Pseudomonas fluorescens and P. aeruginosa. The primer set was highly sensitive as it was able to detect 10 pg μl-1 bacterial DNA and up to 3x103 CFU ml-1 corresponding to 12 viable cells of Xcc which were used as template for PCR reaction Interpretation: The results suggest that developed PCR primers are highly specific and sensitive and it can be used to detect the pathogen at an early stage of infection for disease management.

MarR family transcription regulator HpaR and XC0449 coordinately regulate the virulence of Xanthomonas campestris pv. campestris / 生物工程学报

MarR family transcription regulators are ubiquitous among bacteria and archaea. They extensively control multiple cellular processes and elaborately regulate the expression of genes involved in virulence, stress response and antibiotics at translational level. In Xanthomonas campestris pv. campestris, insertional inactivation of MarR family transcription regulator HpaR (XC2827) resulted in significantly decrease in virulence and increase in the production of the extracellular proteases. Here, we reported that the genome of Xcc 8004 encodes nine MarR family transcription regulators. The MarR family transcription regulators, HpaR (XC2827) and XC0449, were heterologous expressed and purified. In vitro MST and Pull-down assay confirmed the physical interaction between HpaR and XC0449. Phenotypical assay determined that deletion of XC0449 resulted in substantial virulence attenuation. In vitro EMSA, in vivo qRT-PCR and GUS activity assay identified that HpaR and XC0449 coordinately act as the transcriptional activator to regulate the expression of the virulence-associated gene XC0705, and eventually control the bacterial virulence and the production of extracellular proteases.

Multiple resistance to bacterial halo blight and bacterial leaf spot in Coffea spp / Resistência múltipla à mancha aureolada e à mancha foliar bacteriana em Coffea spp

Breeding for genetic resistance is an important method of crop disease management, due to the numerous benefits and low cost of establishment. In this study, progenies of 11 Coffea species and 16 wild C. arabica accessions were tested for their response to Pseudomonas syringae pv. garcae, the causal agent of bacterial halo blight, a widespread disease in the main coffee-producing regions of Brazil and considered a limiting factor for cultivation in pathogen-favorable areas; and also to P. syringae pv. tabaci, causal agent of bacterial leaf spot, a highly aggressive disease recently detected in Brazil. Separate experiments for each disease were carried out in a greenhouse, with artificial pathogen inoculations and ideal moisture conditions for disease development. The results showed that C. canephora, C. congensis, C. eugenioides, C. stenophylla, and C. salvatrix progenies, the wild C. arabica accessions Dilla & Alghe and Palido Viridis, and cultivar IPR 102 contain satisfactory levels of simultaneous resistance against bacterial halo blight and bacterial leaf spot. These results are useful in breeding programs for durable resistance to multiple biotic agents, providing new combinations of resistance alleles by hybridization, as well as for phytopathological studies, to identify infraspecific variability of the pathogens.(AU)

O melhoramento de plantas para resistência genética é um método importante para o manejo de doenças, pelos inúmeros benefícios e baixo custo de implementação. No presente estudo, progênies de 11 espécies de Coffea e 16 acessos selvagens de C. arabica foram testados quanto à resposta a Pseudomonas syringae pv. garcae, agente causal da mancha aureolada, doença disseminada nas principais regiões produtoras de café do Brasil e considerada fator limitante para o cultivo em áreas favoráveis a patógenos; e também para P. syringae pv. tabaci, agente causal da mancha foliar bacteriana, doença altamente agressiva detectada recentemente no Brasil. Experimentos separados para cada doença foram realizados em estufa, por meio da inoculação artificial dos patógenos em condições ideais de umidade para o desenvolvimento das doenças. Os resultados mostraram que as progênies Coffea canephora, C. congensis, C. eugenioides, C. stenophylla e C. salvatrix, além dos acessos selvagens de C. arabica Dilla & Alghe e Palido Viridis e da cultivar IPR 102, possuem níveis satisfatórios de resistência simultânea contra mancha aureolada e mancha foliar bacteriana. Os resultados descritos são úteis em programas de melhoramento para resistência duradoura a múltiplos agentes bióticos, fornecendo novas combinações de alelos de resistência por hibridização, bem como para estudos fitopatológicos, para identificar a variabilidade infraespecífica dos patógenos.(AU)