Brazilin from Caesalpinia sappan L. as a Proprotein Convertase Subtilisin/Kexin Type 9 (PCSK9) Inhibitor: Pharmacophore-Based Virtual Screening, In Silico Molecular Docking, and In Vitro Studies.

Background: Proprotein convertase subtilisin/kexin type 9 (PCSK9) is a crucial regulator of low-density lipoprotein cholesterol (LDL-c) levels, as it binds to and degrades the LDL receptor (LDLR) in the lysosome of hepatocytes. Elevated levels of PCSK9 have been linked to an increased LDL-c plasma levels, thereby increasing the risk of cardiovascular disease (CVD), making it an attractive target for therapeutic interventions. As a way to inhibit PCSK9 action, we searched for naturally derived small molecules which can block the binding of PCSK9 to the LDLR. Methods: In this study, we carried out in silico studies which consist of virtual screening using an optimized pharmacophore model and molecular docking studies using Pyrx 0.98. Effects of the candidate compounds were evaluated using in vitro PCSK9-LDLR binding assays kit. Results: Eleven natural compounds that bind to PCSK9 were virtually screened form HerbalDB database, including brazilin. Next, molecular docking studies using Pyrx 0.98 showed that brazilin had the highest binding affinity with PCSK9 at -9.0 (Kcal/mol), which was higher than that of the other ten compounds. Subsequent in vitro PCSK9-LDLR binding assays established that brazilin decreased the binding of PCSK9 to the EGF-A fragment of the LDLR in a dose-dependent manner, with an IC50 value of 2.19 µM. Conclusion: We have identified brazilin, which is derived from the Caesalpinia sappan herb, which can act as a small molecule inhibitor of PCSK9. Our findings suggest that screening for small molecules that can block the interaction between PCSK9 and the LDLR in silico and in vitro may be a promising approach for developing novel lipid-lowering therapy.

Hypofractionated (2.75 Gy per fraction) versus Conventionally Fractionated Primary Radiotherapy for T2N0M0 Carcinoma of the Glottis

Abstract Introduction Radiotherapy provides excellent outcome in early stage glottic cancer; however, the optimal radiotherapy dose fractionation remains unknown. Objective To investigate the outcome of patients with T2N0M0 treated with either hypofractionated (HypoFxn) or conventionally fractionated radical (ConFxn) radiotherapy. Methods According to our institutional protocol, patients with T2N0M0 glottic cancer can be treated either with ConfFxn or HypoFxn radiotherapy, as per clinician's and patient's choice, following shared decision making discussing the advantages and disadvantages of both modalities. A total of 77 patients with T2N0M0 squamous cell carcinoma of glottis treated with either HypoFxn 55Gy in 20 fractions (n = 19) or ConFxn 63 to 65Gy in 30 fractions (n = 58) were included. Results With median follow-up of 3.4 years, there was no significant difference in disease-free survival (median: HypoFxn = 65.2 months, and ConFxn = 75.3 months; p = 0.874), local recurrence free survival rates (median: HypoFxn = 78.8 months vs. ConFxn = 81.2 months; p = 0.274), and overall survival (median: HypoFxn = 65.9 months vs. ConFxn = 67.7 months; p = 0.532). Elective neck irradiation was given to 43 patients, all in the ConFxn group, and this was associated with poorer local control (p = 0.027). The use of radiotherapy modality, three-dimensional conformal radiotherapy (3DRT) versus intensity modulated radiotherapy (IMRT), was not a prognostic factor (p = 0.36). In the HypoFxn group, grade III acute dysphagia requiring nasogastric tube was 16%, compared with 25% in the ConFxn group (p = 0.446). Conclusion HypoFxn radiotherapy provides a comparable treatment outcome with acceptable toxicity. The addition of prophylactic irradiation of the neck lymph nodes has no impact on regional control.

Hypofractionated (2.75 Gy per fraction) versus Conventionally Fractionated Primary Radiotherapy for T2N0M0 Carcinoma of the Glottis.

Introduction Radiotherapy provides excellent outcome in early stage glottic cancer; however, the optimal radiotherapy dose fractionation remains unknown. Objective To investigate the outcome of patients with T2N0M0 treated with either hypofractionated (HypoFxn) or conventionally fractionated radical (ConFxn) radiotherapy. Methods According to our institutional protocol, patients with T2N0M0 glottic cancer can be treated either with ConfFxn or HypoFxn radiotherapy, as per clinician's and patient's choice, following shared decision making discussing the advantages and disadvantages of both modalities. A total of 77 patients with T2N0M0 squamous cell carcinoma of glottis treated with either HypoFxn 55Gy in 20 fractions ( n = 19) or ConFxn 63 to 65Gy in 30 fractions ( n = 58) were included. Results With median follow-up of 3.4 years, there was no significant difference in disease-free survival (median: HypoFxn = 65.2 months, and ConFxn = 75.3 months; p = 0.874), local recurrence free survival rates (median: HypoFxn = 78.8 months vs. ConFxn = 81.2 months; p = 0.274), and overall survival (median: HypoFxn = 65.9 months vs. ConFxn = 67.7 months; p = 0.532). Elective neck irradiation was given to 43 patients, all in the ConFxn group, and this was associated with poorer local control ( p = 0.027). The use of radiotherapy modality, three-dimensional conformal radiotherapy (3DRT) versus intensity modulated radiotherapy (IMRT), was not a prognostic factor ( p = 0.36). In the HypoFxn group, grade III acute dysphagia requiring nasogastric tube was 16%, compared with 25% in the ConFxn group ( p = 0.446). Conclusion HypoFxn radiotherapy provides a comparable treatment outcome with acceptable toxicity. The addition of prophylactic irradiation of the neck lymph nodes has no impact on regional control.

Cytokinin Production by Azospirillum brasilense Contributes to Increase in Growth, Yield, Antioxidant, and Physiological Systems of Wheat (Triticum aestivum L.).

Plant growth-promoting rhizobacteria are known to associate with several cereal crops. The rhizobacterium exerts its function by synthesizing diverse arrays of phytohormones, such as cytokinin (Ck). However, it is difficult to determine the plant growth promotion when a bacterium produces many different kinds of phytohormones. Therefore, to assess the involvement of Ck in growth promotion and activation of antioxidant and physiological systems, we set up this experiment. Wheat seeds (Triticum aestivum L.) were inoculated with Azospirillum brasilense RA-17 (which produces zeatin type Ck) and RA-18 (which failed to produce Ck). Results showed that seed inoculation with RA-17 significantly improved growth and yield-related parameters compared with RA-18. The activity of enzymes, proline contents, and endogenous hormonal levels in wheat kernels were improved considerably with RA-17 than with RA-18. Strain RA-17 enhanced grain assimilation more than strain RA-18 resulting in a higher crop yield. These results suggest that microbial Ck production may be necessary for stimulating plant growth promotion and activating antioxidant and physiological systems in wheat.

Seasonal Fluctuations of N, P and K in Leaves Influenced Nutrient Requirement During Fruit Development Stages in Different Olive Genotypes

Abstract: Olive is grown in semi-arid climatic conditions; however, little is known about mineral changes in olive plant and nutrient requirements during the production period. Hence, the current study was conducted under Pothwar agro-climatic conditions in order to select appropriate stage of macronutrients (N, P, K) application in relation to soil and leaf nutritional status during 2017 and 2018 growing seasons. Soil and leaf analysis were performed at four different phenological stages (i.e. flowering, fruit setting, fruit enlargement and fruit maturity stages). The results revealed that the assessed macronutrient in leaf and soil varied significantly among varieties, phenological stages and growing year. The results revealed also that nitrogen level was found to decrease from fruit set (1.56%) to fruit enlargement stage (1.47%). Leaf and soil N, P and K contents were found higher before the flowering (stage 1) and depleted after fruit harvesting (stage 4), regardless of olive varieties. However, high yielding varieties showed lower nutrients after fruit harvesting (stage 4). Therefore, N content in leaf and soil gradually decreased during fruit growth and development. Whereas, K content in leaf and soil sharply declined from fruit maturity to fruit ripening stage. Overall, the trend of nutrient depletion showed that plants need phosphorus for fruit setting, nitrogen before and after fruit setting, and potash after pit hardening or at oil accumulation stages.

Soybean herbage yield, nutritional value and profitability under integrated manures management.

Organic manures are more preferred and environmentally friendly than chemical fertilizers for minimally contaminating soil, water and environmental resources, but the determination of right source of organic manures continues to remain an unexplored aspect. Considering the important issue, a multi-year field trial was carried out to determine the response of forage soybean to four sources of nutrients such as chemical fertilizers (IF), poultry litter (PL), bovine's farm yard slurry (BFYS) and sewage sludge (SS) and their seven binary combinations (PL+BFYS, PL+SS, PL+IF, BFYS+SS, BFYS+IF, SS+IF and PL+BFYS+SS). Supplementation of organic manures with mineral fertilizers remained superior to their sole application, particularly BFYS + IF was found significantly (p≤0.05) superior for yielding the highest fresh biomass (23.9, 26.4 and 25.7 t ha-1) with improved nutritional quality. The same combination of integrated fertilizer management also recorded higher sustainability as per sustainable forage yield index along with the highest net income and the benefit-cost ratio. PL and SS applied in conjunction with IF performed better than sole or binary application of organic manures. Therefore, BFYS + IF may be recommended for adoption to produce comparable forage yield and nutritional quality of soybean along with reducing dependency on chemical fertilizers.

Relative efficacy of organic substrates on maize root proliferation under water stress / Eficácia relativa de substratos orgânicos na proliferação de raízes de milho sob estresse hídrico

The aggravating threat for today's agriculture is provision of food security to ever-escalating population utilizing scarce resources. Water scarcity is restraining humans to produce more from drops of water in place of gallons. Root is present at soil-plant interface and is main water extractor for plant. Its growth pattern varies as soil moisture conditions fluctuates. Present pot study consisting of two factors i.e. organic substrates (Farm manure, Poultry Manure and Molasses) and different water stress levels {50, 75, 100 and 125% of available water contents (AWCs)} using maize as test crop to assess their impact on different growth parameters (especially root growth). The experiment was conducted using completely randomized design CRD under factorial arrangement. Root length (44.5 cm), root fresh & dry biomass (71.1 g and 24.3 g, respectively), root diameter (1.73 mm), root volume (0.24 cm3) and root length density (7.4 x 10-3 cm cm-3) were observed in farm manure treated pots at 75% AWC that was statistically indistinguishable from all other treatments at same water level and 100% water availability but eloquently greater than plants of all treatments at 50% and 125% available water contents. Shoot length, dry and fresh weights were observed greater in plants having 100% available moistures. They were statistically at par with 75% water treated plants. Comparing treatments for all the parameters in multivariate cluster analysis it was concluded that 75% available water contents produce almost similar to 100% along with the benefit of water security.

A ameaça agravante para a agricultura atual é a provisão de segurança alimentar para populações cada vez maiores, utilizando recursos escassos. A escassez de água está restringindo os seres humanos a produzir mais a partir de gotas de água no lugar de galões. A raiz está presente na interface solo-planta e é o principal extrator de água para a planta. Seu padrão de crescimento varia conforme as condições de umidade do solo flutuam. O presente estudo em vaso é constituído por dois fatores, ou seja, substratos orgânicos (esterco bovino, esterco de aves e melaço) e diferentes níveis de stress hídrico {50, 75, 100 e 125% do teor de água disponível (AWCs)} utilizando milho como cultura de teste para avaliar o seu impacto em diferentes parâmetros de crescimento (especialmente crescimento de raiz). O experimento foi conduzido utilizando delineamento experimental inteiramente casualizado, em arranjo fatorial. Raiz radicular (44,5 cm), biomassa seca e fresca da raiz (71,1 ge 24,3 g, respectivamente), diâmetro radicular (1,73 mm), volume radicular (0,24 cm3) e densidade radicular (7,4 x 10-3 cm cm-3) foram observados em vasos tratados com esterco de fazenda a 75% AWC que foi estatisticamente indistinguível de todos os outros tratamentos no mesmo nível de água e 100% de disponibilidade de água, mas eloqüentemente maior que plantas de todos os tratamentos a 50% e 125% de conteúdo de água disponível. O comprimento do rebento, peso seco e fresco foram maiores nas plantas com 100% de umidade disponível. Eles foram estatisticamente equivalentes a 75% de plantas tratadas com água. Comparando os tratamentos para todos os parâmetros na análise multivariada de clusters, concluiu-se que 75% do conteúdo de água disponível produz quase o mesmo que 100%, juntamente com o benefício da segurança hídrica.

Revamping soil quality and correlation studies for yield and yield attributes in sorghum-legumes intercropping systems / Melhoria da qualidade do solo e estudos de correlação para a produtividade e os atributos de produtividade em sistemas de plantio consorciado de sorgo e legumes

Restoration, preservation and sustainability of agricultural resources including soil demands biologically and economically viable farming practices. For boosting physico-chemical properties of soil from 2012 to 2015, three legumes including cowpea, cluster bean and soybean were intercropped with sorghum in 40-60, 75-25 and 100-100 seed blending ratios, while pure stand of sorghum was kept for comparison. Correlation studies were also conducted to determine linear or inverse relationship between physiological and agronomic attributes with green forage yield of sorghum. Results revealed that intercropping systems were not effective in significantly improving the physical properties of soil, however soybean and sorghum mixed seeded crop in 100-100 seed ratio resulted in the highest total and available nitrogen especially during last two years of the study and it was followed by mixed seeded crop of cluster bean and sorghum (100-100 seed ratio), while pure stand of sorghum appeared to be the most exhaustive farming system. Correlation analysis depicted a linear relationship between all agronomic attributes (seedling count, plant population at harvest, plant height, stem diameter, number of leaves and leaf area per plant, fresh and dry weights per plant) and physiological growth parameters (leaf area index and leaf area duration, crop growth rate and net assimilation rate) with forage yield of sorghum. Thus, legumes inclusion was found to be effective in restoring soil fertility, while long term legumes inclusion by following rotation may be suggested.

A restauração, a preservação e a sustentabilidade dos recursos agrícolas, incluindo o solo, exigem práticas agrícolas biologicamente e economicamente viáveis. Para aumentar as propriedades físico-químicas do solo de 2012 a 2015, três leguminosas, incluindo feijão-fradinho, guar e soja, foram consorciadas com sorgo nas proporções de mistura de 40-60, 75-25 e 100-100, enquanto que o sorgo puro foi mantido por comparação. Estudos de correlação também foram conduzidos para determinar a relação linear ou inversa entre atributos fisiológicos e agronômicos com a produtividade de forragem verde do sorgo. Os resultados revelaram que os sistemas consorciados não foram eficazes em afetar significativamente as propriedades físicas do solo, no entanto, soja e sorgo misturados na proporção de sementes 100-100 resultou no maior nitrogênio total e nitrogênio disponível, especialmente durante os últimos dois anos do estudo e foi seguido do plantio de sementes mistas de guar e sorgo (100-100 rácio de semente), enquanto a monocultura de sorgo parecia ser o sistema de agricultura mais exaustivo. A análise de regressão mostrou uma correlação linear entre todos os atributos agronômicos (contagem de plântulas, população de plantas na colheita, altura de planta, diâmetro de caule, número de folhas e área foliar por planta, peso fresco e seco por planta) e parâmetros de crescimento fisiológico (índice de área foliar e duração da área foliar, taxa de crescimento da cultura e taxa de assimilação líquida) com o rendimento de forragem do sorgo. Assim, a inclusão de leguminosas foi eficaz no aumento da fertilidade do solo, enquanto a inclusão de leguminosas a longo prazo por rotação de diferentes leguminosas é sugerida.

Cotton productivity enhanced through transplanting and early sowing

Due to high temperatures in arid regions, cotton crop emergence and early establishment of seedlings are adversely affected. Field studies were undertaken to quantify the effects of transplantation of cotton (Gossypium hirsutum L.) seedlings during the early part of the growing season (March) and crop season (May) for potential realization of cotton productivity under the harsh weather of the southern part of Punjab province, Pakistan. Treatments, consisting of (a) two planting dates (March and May), and (b) two sowing methods (transplanting of seedlings and direct seeding), were arranged in a randomized complete-block design with four replications. Results showed that transplanting seedlings improved the productivity of cotton by 14.2% over direct seeding. Productivity was also increased substantially (34.8%) by planting during March over May sowing. The practice of planting cotton by transplanting seedlings and early sowing could be successfully adapted in areas where high temperatures coincide with the May planting and peak blooming periods in different cotton growing areas.

Cotton productivity enhanced through transplanting and early sowing / Aumento da produtividade do algodão por meio de transplante de mudas e semeadura precoce

Due to high temperatures in arid regions, cotton crop emergence and early establishment ofseedlings are adversely affected. Field studies were undertaken to quantify the effects of transplantation ofcotton (Gossypium hirsutum L.) seedlings during the early part of the growing season (March) and cropseason (May) for potential realization of cotton productivity under the harsh weather of the southern partof Punjab province, Pakistan. Treatments, consisting of (a) two planting dates (March and May), and (b)two sowing methods (transplanting of seedlings and direct seeding), were arranged in a randomizedcomplete-block design with four replications. Results showed that transplanting seedlings improved theproductivity of cotton by 14.2% over direct seeding. Productivity was also increased substantially (34.8%)by planting during March over May sowing. The practice of planting cotton by transplanting seedlings andearly sowing could be successfully adapted in areas where high temperatures coincide with the Mayplanting and peak blooming periods in different cotton growing areas.

Altas temperaturas, em torno de 48ºC, prevalecem durante os meses de maio na parte do sul da província de Punjab, no Paquistão. A esta temperatura a emergência da colheita de algodão e o estabelecimento precoce da plântula são adversamente afetados. Estudos de campo foram realizados paraquantificar os efeitos do transplante de mudas de algodão (Gossypium hirsutum L.) durante a primeira parteda estação de crescimento (março) e da safra (maio) para a realização potencial da produtividade do algodãosob condições adversas da parte sul do Punjab Província-Paquistão. Os tratamentos, consistindo em(a) duas datas de plantio (março e maio), e (b) dois métodos de semeadura (transplante de mudas e semeadura direta), foram dispostos em delineamento em blocos ao acaso com quatro repetições. Os resultados mostraram que o transplante de mudas melhorou a produtividade do algodão em 14,2% em relação à semeadura direta. A produtividade também aumentou substancialmente (34,8%) com o plantioem março sobre a semeadura de maio. A prática de plantio de algodão por transplante de mudas e sua semeadura precoce poderia ser adaptada com sucesso, onde a alta temperatura coincide com o tempo de plantio durante o mês de maio e períodos de florescimento máximo em diferentes áreas de cultivo de algodão.

Cotton productivity enhanced through transplanting and early sowing

Due to high temperatures in arid regions, cotton crop emergence and early establishment of seedlings are adversely affected. Field studies were undertaken to quantify the effects of transplantation of cotton (Gossypium hirsutum L.) seedlings during the early part of the growing season (March) and crop season (May) for potential realization of cotton productivity under the harsh weather of the southern part of Punjab province, Pakistan. Treatments, consisting of (a) two planting dates (March and May), and (b) two sowing methods (transplanting of seedlings and direct seeding), were arranged in a randomized complete-block design with four replications. Results showed that transplanting seedlings improved the productivity of cotton by 14.2% over direct seeding. Productivity was also increased substantially (34.8%) by planting during March over May sowing. The practice of planting cotton by transplanting seedlings and early sowing could be successfully adapted in areas where high temperatures coincide with the May planting and peak blooming periods in different cotton growing areas.

Cotton productivity enhanced through transplanting and early sowing / Aumento da produtividade do algodão por meio de transplante de mudas e semeadura precoce

Due to high temperatures in arid regions, cotton crop emergence and early establishment ofseedlings are adversely affected. Field studies were undertaken to quantify the effects of transplantation ofcotton (Gossypium hirsutum L.) seedlings during the early part of the growing season (March) and cropseason (May) for potential realization of cotton productivity under the harsh weather of the southern partof Punjab province, Pakistan. Treatments, consisting of (a) two planting dates (March and May), and (b)two sowing methods (transplanting of seedlings and direct seeding), were arranged in a randomizedcomplete-block design with four replications. Results showed that transplanting seedlings improved theproductivity of cotton by 14.2% over direct seeding. Productivity was also increased substantially (34.8%)by planting during March over May sowing. The practice of planting cotton by transplanting seedlings andearly sowing could be successfully adapted in areas where high temperatures coincide with the Mayplanting and peak blooming periods in different cotton growing areas.(AU)

Altas temperaturas, em torno de 48ºC, prevalecem durante os meses de maio na parte do sul da província de Punjab, no Paquistão. A esta temperatura a emergência da colheita de algodão e o estabelecimento precoce da plântula são adversamente afetados. Estudos de campo foram realizados paraquantificar os efeitos do transplante de mudas de algodão (Gossypium hirsutum L.) durante a primeira parteda estação de crescimento (março) e da safra (maio) para a realização potencial da produtividade do algodãosob condições adversas da parte sul do Punjab Província-Paquistão. Os tratamentos, consistindo em(a) duas datas de plantio (março e maio), e (b) dois métodos de semeadura (transplante de mudas e semeadura direta), foram dispostos em delineamento em blocos ao acaso com quatro repetições. Os resultados mostraram que o transplante de mudas melhorou a produtividade do algodão em 14,2% em relação à semeadura direta. A produtividade também aumentou substancialmente (34,8%) com o plantioem março sobre a semeadura de maio. A prática de plantio de algodão por transplante de mudas e sua semeadura precoce poderia ser adaptada com sucesso, onde a alta temperatura coincide com o tempo de plantio durante o mês de maio e períodos de florescimento máximo em diferentes áreas de cultivo de algodão.(AU)

Primary Concurrent Chemoradiation in Head and Neck Cancers withWeekly Cisplatin Chemotherapy: Analysis of Compliance, Toxicity and Survival

Abstract Introduction Concurrent chemoradiation is the standard of care in inoperable locally advanced squamous cell head and neck cancers. The most widely accepted schedule of concomitant cisplatin is 100mg/m2 given on a 3 weekly basis but the optimal regime is unknown. Objective The objective of this study is to assess the tolerability, compliance, and clinical outcomes of weekly cisplatin (40mg/m2). Methods During the period of January 2007-December 2009, we analyzed retrospectively 122 patients with histologically proven squamous cell carcinoma of head and neck (nasopharynx, oropharynx, larynx, hypopharynx, and oral cavity) treated with definitive chemoradiation. All patients received 63 Gy in 30 daily fractions with concomitant weekly cisplatin 40mg/m2. We assessed treatment toxicities and patient compliance. We estimated overall survival using the Kaplan-Meier method. Results Sixty-eight percent of patients managed to complete all six cycles of chemotherapy while 87% of patients completed at least 5 cycles of weekly cisplatin. Incidence of grade 3/4 toxicity was as follows: mucositis 33%, dermatitis 41%, dysphagia 15%, mouth/neck pain 17%, neutropenia 2%, and renal impairment 3%. 53% patients required at least one hospital admission for symptom control. The 5-year overall survival rate was 60%. Conclusion Concurrent chemoradiotherapy using weekly cisplatin at 40mg/m2 per week is an effective, well tolerated regimen allowing most patients to receive at least 5 cycles of chemotherapy. However, a phase III randomized control trial comparing the standard dose of 100mg/m2 cisplatin tri-weekly with a weekly regimen is needed to establish the long term clinical outcome.

Primary Concurrent Chemoradiation in Head and Neck Cancers with Weekly Cisplatin Chemotherapy: Analysis of Compliance, Toxicity and Survival.

Introduction Concurrent chemoradiation is the standard of care in inoperable locally advanced squamous cell head and neck cancers. The most widely accepted schedule of concomitant cisplatin is 100mg/m2 given on a 3 weekly basis but the optimal regime is unknown. Objective The objective of this study is to assess the tolerability, compliance, and clinical outcomes of weekly cisplatin (40mg/m2). Methods During the period of January 2007-December 2009, we analyzed retrospectively 122 patients with histologically proven squamous cell carcinoma of head and neck (nasopharynx, oropharynx, larynx, hypopharynx, and oral cavity) treated with definitive chemoradiation. All patients received 63 Gy in 30 daily fractions with concomitant weekly cisplatin 40mg/m2. We assessed treatment toxicities and patient compliance. We estimated overall survival using the Kaplan-Meier method. Results Sixty-eight percent of patients managed to complete all six cycles of chemotherapy while 87% of patients completed at least 5 cycles of weekly cisplatin. Incidence of grade 3/4 toxicity was as follows: mucositis 33%, dermatitis 41%, dysphagia 15%, mouth/neck pain 17%, neutropenia 2%, and renal impairment 3%. 53% patients required at least one hospital admission for symptom control. The 5-year overall survival rate was 60%. Conclusion Concurrent chemoradiotherapy using weekly cisplatin at 40mg/m2 per week is an effective, well tolerated regimen allowing most patients to receive at least 5 cycles of chemotherapy. However, a phase III randomized control trial comparing the standard dose of 100mg/m2 cisplatin tri-weekly with a weekly regimen is needed to establish the long term clinical outcome.

Evaluation of different detection methods of biofilm formation in the clinical isolates.

BACKGROUND: Microorganisms growing in a biofilm are associated with chronic and recurrent human infections and are highly resistant to antimicrobial agents. There are various methods to detect biofilm production like Tissue Culture Plate (TCP), Tube method (TM), Congo Red Agar method (CRA), bioluminescent assay, piezoelectric sensors, and fluorescent microscopic examination. OBJECTIVE: This study was conducted to compare three methods for the detection of biofilms. METHOD: The study was carried out at the Department of Microbiology, Army Medical College, National University of Sciences and Technology, Pakistan, from January 2010 to June 2010. A total of 110 clinical isolates were subjected to biofilm detection methods. Isolates were identified by standard microbiological procedures. Biofilm detection was tested by TCP, TM and CRA. Antibiotic susceptibility test of biofilm producing bacteria was performed by using the Kirby-Bauer disc diffusion technique according to CLSI guidelines. RESULTS: The TCP method was considered to be superior to TM and CRA. From the total of 110 clinical isolates, TCP method detected 22.7% as high, 41% moderate and 36.3% as weak or non-biofilm producers. We have observed higher antibiotic resistance in biofilm producing bacteria than non-biofilm producers. CONCLUSION: We can conclude from our study that the TCP method is a more quantitative and reliable method for the detection of biofilm forming microorganisms as compared to TM and CRA methods, and it can be recommended as a general screening method for detection of biofilm producing bacteria in laboratories.

Evaluation of different detection methods of biofilm formation in the clinical isolates

BACKGROUND: Microorganisms growing in a biofilm are associated with chronic and recurrent human infections and are highly resistant to antimicrobial agents. There are various methods to detect biofilm production like Tissue Culture Plate (TCP), Tube method (TM), Congo Red Agar method (CRA), bioluminescent assay, piezoelectric sensors, and fluorescent microscopic examination. OBJECTIVE: This study was conducted to compare three methods for the detection of biofilms. METHOD: The study was carried out at the Department of Microbiology, Army Medical College, National University of Sciences and Technology, Pakistan, from January 2010 to June 2010. A total of 110 clinical isolates were subjected to biofilm detection methods. Isolates were identified by standard microbiological procedures. Biofilm detection was tested by TCP, TM and CRA. Antibiotic susceptibility test of biofilm producing bacteria was performed by using the Kirby-Bauer disc diffusion technique according to CLSI guidelines. RESULTS: The TCP method was considered to be superior to TM and CRA. From the total of 110 clinical isolates, TCP method detected 22.7 percent as high, 41 percent moderate and 36.3 percent as weak or non-biofilm producers. We have observed higher antibiotic resistance in biofilm producing bacteria than non-biofilm producers. CONCLUSION: We can conclude from our study that the TCP method is a more quantitative and reliable method for the detection of biofilm forming microorganisms as compared to TM and CRA methods, and it can be recommended as a general screening method for detection of biofilm producing bacteria in laboratories.

Allelic variation at the Vrn-A1, Vrn-B1, Vrn-D1, Vrn-B3 and Ppd-D1a loci of Pakistani spring wheat cultivars

Flowering time in bread wheat (Triticum aestivum L.) is controlled by vernalization and photoperiod response, and earliness per se genes. The genetic basis of flowering time has not been investigated in Pakistani bread wheat. This study was, therefore, conducted to determine the allelic composition at Vrn-A1, Vrn-B1, Vrn-D1, Vrn-B3 and Ppd-D1a loci of 59 Pakistani spring bread wheat cultivars. These cultivars, along with 4 isogenic lines for vernalization genes were characterized with previously reported DNA markers designed for detecting allelic variation at 4 Vrn (Vernalization) and 1 Ppd (Photoperiod) loci. Spring habit Vrn-A1a allele was found in 36 percent cultivars either alone or with spring habit Vrn-B1 and Vrn-D1 alleles. Two wheat cultivars had the dominant Vrn-A1c allele, whereas none of the cultivars had Vrn-A1b. Spring habit Vrn-B1 was the most frequent allele (64 percent) present either alone or with Vrn-A1a, Vrn-A1c and Vrn-D1. Spring habit Vrn-D1 was found in 61 percent cultivars. Vrn-D1 was singly found in 25 percent cultivars and along with Vrn-B1 in 29 percent cultivars. Dominant Vrn-B3 was absent in all cultivars studied. All cultivars except Era had the photoperiod insensitive allele Ppd-D1a. We did not find any association between the flowering time and Vrn allelic composition of the studied cultivars. This indicated that the partial vernalization requirement of cultivars with Vrn-B1 and Vrn-D1 alleles is probably fulfilled during Pakistani growing season. Earliness per se and the photoperiod sensitive loci other than Ppd-D1 need to be investigated to further understand the genetic basis of flowering time in Pakistani wheat.

Assessment of genetic diversity among Pakistani wheat (Triticum aestivum L) advanced breeding lines using RAPD and SDS-PAGE

Genetic diversity was assessed among 32 advanced wheat breeding lines included in the National Uniform Wheat Yield Trials (2006-07) of Pakistan using molecular (DNA) and biochemical (SDS-PAGE) markers. Of the 72 RAPD primers used for initial screening, 15 were found polymorphic. A total of 140 bands (61.4 percent polymorphic) were generated by the 15 random decamer primers. Genetic similarity coefficients ranged from 0.81 to 0.94 for rainfed and from 0.70 to 0.93 for the normal seeding date group. Cluster analysis using the unweighted pair group method of arithmetic averages (UPGMA) clustered the 32 advanced wheat breeding lines into one major and three small groups. Maximum level of polymorphism (90 percent) was observed for the primer OPA-05. Lines N9 and N11 showed the least genetic similarities (0.70-0.82 and 0.71-0.83, respectively) with rest of the lines studied. Line RF1 had the maximum similarity (0.81-0.94) with other lines. Wheat lines included in the normal seeding date were relatively distantly related than those in the rainfed group. Seed storage protein analysis produced 19 subunits ranging from 29-120KDa. Similarity coefficients ranged from 0.53 to 1.0 for the normal seeding date and from 0.47 to 1.0 for the rainfed group. High molecular weight subunits (particularly 120KDa) showed greater polymorphism than the lower molecular weight subunits. Narrow genetic base was observed in wheat lines included in the rainfed group. DNA fingerprinting of advanced breeding lines may help to avoid release of varieties with narrow genetic base in the future.