Molecular analysis of Panton-Valentine Leucocidin (pvl) gene among MRSA and MSSA isolates.

The present study was conducted in order to determine the frequency of pvl gene among the pathogenic and healthy population isolates of Methicillin Resistant Staphylococcus aureus (MRSA) and Methicillin Sensitive Staphylococcus aureus (MSSA). For this purpose, nasal swab samples were collected from the healthy individuals (to be used as controls, all the samples were collected irrespective of the sex and age factors), the pathogenic samples were collected from different patients suffering from skin &soft tissue infections caused by S. aureus (to be used as test samples).Both of these population samples were analyzed for the presence of pvl gene. S.aureus were identified through conventional microbiological identification procedures. In the case of normal samples, 70 nasal swabs were collected and only 33 (47%) proved to be S. aureus while 20 pathogenic samples were collected and all (100%) were cleared as S. aureus. For further distribution of samples into MRSA and MSSA, antibiotic susceptibility pattern was checked by using the standard protocols of Kirby-Bauer disc diffusion method. Two antibiotic discs Oxacillin (OX: 1ug) and cefoxitin (FOX: 30ug) were used. Among healthy population, MRSA was found to be 79% (n=26) and MSSA were present as 21% (n= 7). Among pathogenic strains 100% MRSA was detected where n= 20. Detection of pvl gene among the MRSA and MSSA isolates was done by using the uniplex PCR followed by gel electrophoresis. MRSA and MSSA of normal healthy population carried 49% and 7% pvl gene respectively. While, pathogenic MRSA samples carried 46% pvl gene among them. Potentially alarming percentage of pvl gene is present among the normal healthy individuals which indicates a future threat and a major health concern.

An ethno-botanical study of indigenous medicinal plants and their usage in rural valleys of Swabi and Hazara region of Pakistan / Um estudo etnobotânico de plantas medicinais indígenas e seu uso nos vales rurais da região de Swabi e Hazara, no Paquistão

An ethnobotanical study was conducted to document indigenous medicinal plants and their usage from knowledgeable and elderly persons in Razzar and Gadoon valley of Swabi and Allai and Tanawal valley of Hazara region of Pakistan during 2016-2019. Several systematic field visits and questionnaire surveys were carried out in selected sites of the study area to gather relevant information from the local community. Rapid assessment method was adopted for data collection by interviewing the local people having enough knowledge of medicinal plants use for treatment of different ailments. UV (UV) formula was applied to calculate the relative importance of medicinal plant species in each site of the study area. In the present study, 221 medicinal plants belonging to 105 families have been reported through 580 respondents (385 males, 138 females and 57 local health healer) from the Swabi and Hazara region. The main sources of herbal medicines were leaves (21%) followed by fruits (21%), seeds (17%), whole plants (14%), roots (9%), bark (9%), flowers (7%) and gum (2%). Mentha spicata L. and Berberis lycium Royle were reported with highest UV (UV) i.e. 0.92 and 0.68 in Razzar tehsil and Gadoon valley of Swabi, whereas Mentha longifolia L and Geranium wallichianum D were reported with highest UV i.e. (0.65) and (0.88) in Allai and Tanawal valley of Hazara region, respectively. It was concluded that Swabi and Hazara region is rich in medicinal plants species and associated traditional knowledge. Moreover, ethno-medicines have played significant role in the indigenous healthcare system of the study area. However, uprooting the entire plant for ethno-medicine is a big threat to conservation of medicinal plants diversity in the study area.

Um estudo etnobotânico foi realizado para documentar as plantas medicinais indígenas e seu uso por pessoas experientes e idosas em Razzar e Gadoon, no vale de Swabi e no vale Allai e Tanawal da região de Hazara, no Paquistão, durante 2016 a 2019. Várias visitas sistemáticas de campo e pesquisas por questionário foram realizadas em locais selecionados da área de estudo para coletar informações relevantes da comunidade local. O método de avaliação rápida foi adotado para a coleta de dados por meio de entrevistas com a população local, com conhecimento suficiente do uso de plantas medicinais para o tratamento de diferentes enfermidades. A fórmula UV (UV) foi aplicada para calcular a importância relativa das espécies de plantas medicinais em cada local da área de estudo. No presente estudo, 221 plantas medicinais pertencentes a 105 famílias foram relatadas por 580 entrevistados (385 homens, 138 mulheres e 57 curandeiros locais) da região de Swabi e Hazara. As principais fontes de medicamentos fitoterápicos foram folhas (21%), seguidas de frutas (21%), sementes (17%), plantas inteiras (14%), raízes (9%), cascas (9%), flores (7%) e goma (2%). Mentha spicata L. e Berberis lycium Royle foram relatados com maior UV (UV), ou seja, 0,92 e 0,68 em Razzar tehsil e vale Gadoon de Swabi, enquanto Mentha longifolia L. e Geranium wallichianum D. foram relatados com maior UV, isto é, 0,65 e 0,88 no vale Allai e Tanawal da região de Hazara, respectivamente. Concluiu-se que a região de Swabi e Hazara é rica em espécies de plantas medicinais e conhecimentos tradicionais associados. Além disso, etnomedicamentos têm desempenhado um papel significativo no sistema de saúde indígena da área de estudo. No entanto, arrancar a planta inteira para etnomedicina é uma grande ameaça à conservação da diversidade de plantas medicinais na área de estudo.

An ethno-botanical study of indigenous medicinal plants and their usage in rural valleys of Swabi and Hazara region of Pakistan / Um estudo etnobotânico de plantas medicinais indígenas e seu uso nos vales rurais da região de Swabi e Hazara, no Paquistão

An ethnobotanical study was conducted to document indigenous medicinal plants and their usage from knowledgeable and elderly persons in Razzar and Gadoon valley of Swabi and Allai and Tanawal valley of Hazara region of Pakistan during 2016-2019. Several systematic field visits and questionnaire surveys were carried out in selected sites of the study area to gather relevant information from the local community. Rapid assessment method was adopted for data collection by interviewing the local people having enough knowledge of medicinal plants use for treatment of different ailments. UV (UV) formula was applied to calculate the relative importance of medicinal plant species in each site of the study area. In the present study, 221 medicinal plants belonging to 105 families have been reported through 580 respondents (385 males, 138 females and 57 local health healer) from the Swabi and Hazara region. The main sources of herbal medicines were leaves (21%) followed by fruits (21%), seeds (17%), whole plants (14%), roots (9%), bark (9%), flowers (7%) and gum (2%). Mentha spicata L. and Berberis lycium Royle were reported with highest UV (UV) i.e. 0.92 and 0.68 in Razzar tehsil and Gadoon valley of Swabi, whereas Mentha longifolia L and Geranium wallichianum D were reported with highest UV i.e. (0.65) and (0.88) in Allai and Tanawal valley of Hazara region, respectively. It was concluded that Swabi and Hazara region is rich in medicinal plants species and associated traditional knowledge. Moreover, ethno-medicines have played significant role in the indigenous healthcare system of the study area. However, uprooting the entire plant for ethno-medicine is a big threat to conservation of medicinal plants diversity in the study area.

Um estudo etnobotânico foi realizado para documentar as plantas medicinais indígenas e seu uso por pessoas experientes e idosas em Razzar e Gadoon, no vale de Swabi e no vale Allai e Tanawal da região de Hazara, no Paquistão, durante 2016 a 2019. Várias visitas sistemáticas de campo e pesquisas por questionário foram realizadas em locais selecionados da área de estudo para coletar informações relevantes da comunidade local. O método de avaliação rápida foi adotado para a coleta de dados por meio de entrevistas com a população local, com conhecimento suficiente do uso de plantas medicinais para o tratamento de diferentes enfermidades. A fórmula UV (UV) foi aplicada para calcular a importância relativa das espécies de plantas medicinais em cada local da área de estudo. No presente estudo, 221 plantas medicinais pertencentes a 105 famílias foram relatadas por 580 entrevistados (385 homens, 138 mulheres e 57 curandeiros locais) da região de Swabi e Hazara. As principais fontes de medicamentos fitoterápicos foram folhas (21%), seguidas de frutas (21%), sementes (17%), plantas inteiras (14%), raízes (9%), cascas (9%), flores (7%) e goma (2%). Mentha spicata L. e Berberis lycium Royle foram relatados com maior UV (UV), ou seja, 0,92 e 0,68 em Razzar tehsil e vale Gadoon de Swabi, enquanto Mentha longifolia L. e Geranium wallichianum D. foram relatados com maior UV, isto é, 0,65 e 0,88 no vale Allai e Tanawal da região de Hazara, respectivamente. Concluiu-se que a região de Swabi e Hazara é rica em espécies de plantas medicinais e conhecimentos tradicionais associados. Além disso, etnomedicamentos têm desempenhado um papel significativo no sistema de saúde indígena da área de estudo. No entanto, arrancar a planta inteira para etnomedicina é uma grande ameaça à conservação da diversidade de plantas medicinais na área de estudo.

An ethno-botanical study of indigenous medicinal plants and their usage in rural valleys of Swabi and Hazara region of Pakistan

Abstract An ethnobotanical study was conducted to document indigenous medicinal plants and their usage from knowledgeable and elderly persons in Razzar and Gadoon valley of Swabi and Allai and Tanawal valley of Hazara region of Pakistan during 2016-2019. Several systematic field visits and questionnaire surveys were carried out in selected sites of the study area to gather relevant information from the local community. Rapid assessment method was adopted for data collection by interviewing the local people having enough knowledge of medicinal plants use for treatment of different ailments. UV (UV) formula was applied to calculate the relative importance of medicinal plant species in each site of the study area. In the present study, 221 medicinal plants belonging to 105 families have been reported through 580 respondents (385 males, 138 females and 57 local health healer) from the Swabi and Hazara region. The main sources of herbal medicines were leaves (21%) followed by fruits (21%), seeds (17%), whole plants (14%), roots (9%), bark (9%), flowers (7%) and gum (2%). Mentha spicata L. and Berberis lycium Royle were reported with highest UV (UV) i.e. 0.92 and 0.68 in Razzar tehsil and Gadoon valley of Swabi, whereas Mentha longifolia L and Geranium wallichianum D were reported with highest UV i.e. (0.65) and (0.88) in Allai and Tanawal valley of Hazara region, respectively. It was concluded that Swabi and Hazara region is rich in medicinal plants species and associated traditional knowledge. Moreover, ethno-medicines have played significant role in the indigenous healthcare system of the study area. However, uprooting the entire plant for ethno-medicine is a big threat to conservation of medicinal plants diversity in the study area.

Resumo Um estudo etnobotânico foi realizado para documentar as plantas medicinais indígenas e seu uso por pessoas experientes e idosas em Razzar e Gadoon, no vale de Swabi e no vale Allai e Tanawal da região de Hazara, no Paquistão, durante 2016 a 2019. Várias visitas sistemáticas de campo e pesquisas por questionário foram realizadas em locais selecionados da área de estudo para coletar informações relevantes da comunidade local. O método de avaliação rápida foi adotado para a coleta de dados por meio de entrevistas com a população local, com conhecimento suficiente do uso de plantas medicinais para o tratamento de diferentes enfermidades. A fórmula UV (UV) foi aplicada para calcular a importância relativa das espécies de plantas medicinais em cada local da área de estudo. No presente estudo, 221 plantas medicinais pertencentes a 105 famílias foram relatadas por 580 entrevistados (385 homens, 138 mulheres e 57 curandeiros locais) da região de Swabi e Hazara. As principais fontes de medicamentos fitoterápicos foram folhas (21%), seguidas de frutas (21%), sementes (17%), plantas inteiras (14%), raízes (9%), cascas (9%), flores (7%) e goma (2%). Mentha spicata L. e Berberis lycium Royle foram relatados com maior UV (UV), ou seja, 0,92 e 0,68 em Razzar tehsil e vale Gadoon de Swabi, enquanto Mentha longifolia L. e Geranium wallichianum D. foram relatados com maior UV, isto é, 0,65 e 0,88 no vale Allai e Tanawal da região de Hazara, respectivamente. Concluiu-se que a região de Swabi e Hazara é rica em espécies de plantas medicinais e conhecimentos tradicionais associados. Além disso, etnomedicamentos têm desempenhado um papel significativo no sistema de saúde indígena da área de estudo. No entanto, arrancar a planta inteira para etnomedicina é uma grande ameaça à conservação da diversidade de plantas medicinais na área de estudo.

An ethno-botanical study of indigenous medicinal plants and their usage in rural valleys of Swabi and Hazara region of Pakistan.

An ethnobotanical study was conducted to document indigenous medicinal plants and their usage from knowledgeable and elderly persons in Razzar and Gadoon valley of Swabi and Allai and Tanawal valley of Hazara region of Pakistan during 2016-2019. Several systematic field visits and questionnaire surveys were carried out in selected sites of the study area to gather relevant information from the local community. Rapid assessment method was adopted for data collection by interviewing the local people having enough knowledge of medicinal plants use for treatment of different ailments. UV (UV) formula was applied to calculate the relative importance of medicinal plant species in each site of the study area. In the present study, 221 medicinal plants belonging to 105 families have been reported through 580 respondents (385 males, 138 females and 57 local health healer) from the Swabi and Hazara region. The main sources of herbal medicines were leaves (21%) followed by fruits (21%), seeds (17%), whole plants (14%), roots (9%), bark (9%), flowers (7%) and gum (2%). Mentha spicata L. and Berberis lycium Royle were reported with highest UV (UV) i.e. 0.92 and 0.68 in Razzar tehsil and Gadoon valley of Swabi, whereas Mentha longifolia L and Geranium wallichianum D were reported with highest UV i.e. (0.65) and (0.88) in Allai and Tanawal valley of Hazara region, respectively. It was concluded that Swabi and Hazara region is rich in medicinal plants species and associated traditional knowledge. Moreover, ethno-medicines have played significant role in the indigenous healthcare system of the study area. However, uprooting the entire plant for ethno-medicine is a big threat to conservation of medicinal plants diversity in the study area.

Characterisation of plant growth-promoting rhizobacteria from rhizosphere soil of heat-stressed and unstressed wheat and their use as bio-inoculant.

High temperature induces several proteins in plants that enhance tolerance to high temperature shock. The fate of proteins synthesised in microbial cells or secreted into culture media by interacting microbes has not been fully elucidated. The present investigation aimed to characterise plant growth-promoting rhizobacteria (PGPR) isolated from the rhizosphere of wheat genotypes (differing in tolerance to high temperature stress) and evaluate their performance as bioinoculant for use in wheat. Four bacterial strains, viz. Pseudomonas brassicacearum, Bacillus thuringiensis, Bacillus cereus strain W6 and Bacillus subtilis, were isolated from the rhizosphere of heat-stressed and unstressed wheat genotypes. The wheat genotypes were exposed to high temperature stress at 45 °C for 10 days (3 h daily) at pre-anthesis phase. Isolates were identified on the basis of morphology and biochemical characteristics, 16S rRNA gene sequencing and whole cell protein profiles. Results were further complemented by size exclusion chromatography (SEC) with fast protein liquid chromatography (FPLC) and SDS PAGE of 80% ammonium sulphate precipitates of the cell-free supernatants. Isolates were positive for catalase, oxidases and antimicrobial activity. P. brassicacearum from the rhizosphere of the heat-tolerant genotype was more efficient in phosphate solubilisation, bacteriocin production, antifungal and antibacterial activity against Helminthosporium sativum, Fusarium moniliforme and Klebsiella pneumonia, respectively. The inoculated seedlings had significantly higher root and shoot fresh weight, enhanced activity of antioxidant enzymes, proline and protein content. Total profiling of the culture with SDS-PAGE indicated expression of new protein bands in 95 kDa in P. brassicacearum. Temperature-induced changes in PGPR isolates are similar to those in the host plant. P. brassicacearum may be a good candidate for use in biofertiliser production for plants exposed to high temperature stress.

Impact of phosphate solubilizing bacteria on wheat (Triticum aestivum ) in the presence of pesticides.

Three phosphate solubilizing bacteria were isolated and identified by 16S rRNA sequencing as Pseudomonas putida, Pseudomonas sp and Pseudomonas fulva . The strains were subjected to plant biochemical testing and all the PGPR attributes were checked in the presence of pesticides (chlorpyrifos and pyriproxyfen). The phosphate solubilizing index of strain Ros2 was highest in NBRIP medium i.e 2.23 mm. All the strains showed acidic pH (ranges from 2.5-5) on both medium i.e PVK and NBRIP. Strain Ros2 was highly positive for ammonia production as well as siderophore production while strain Rad2 was positive for HCN production. The results obtained by the strains Rad1, Rad2 and Ros2 for auxin production were 33.1, 30.67 and 15.38 µg ml-1, respectively. Strain Rad1 showed 16% increase in percentage germination in comparison to control in the presence of pesticide stress. Most promising results for chlorophyll content estimation were obtained in the presence of carotenoids upto 6 mgg-1 without stress by both strains Rad1 and Rad2. Study suggests that especially strain Ros2 can enhance plant growth parameters in the pesticide stress.

Impact of phosphate solubilizing bacteria on wheat (Triticum aestivum ) in the presence of pesticides / Impacto das bactérias solubilizantes de fosfato no trigo (Triticum aestivum ) na presença de pesticidas

Abstract Three phosphate solubilizing bacteria were isolated and identified by 16S rRNA sequencing as Pseudomonas putida, Pseudomonas sp and Pseudomonas fulva . The strains were subjected to plant biochemical testing and all the PGPR attributes were checked in the presence of pesticides (chlorpyrifos and pyriproxyfen). The phosphate solubilizing index of strain Ros2 was highest in NBRIP medium i.e 2.23 mm. All the strains showed acidic pH (ranges from 2.5-5) on both medium i.e PVK and NBRIP. Strain Ros2 was highly positive for ammonia production as well as siderophore production while strain Rad2 was positive for HCN production. The results obtained by the strains Rad1, Rad2 and Ros2 for auxin production were 33.1, 30.67 and 15.38 µg ml-1, respectively. Strain Rad1 showed 16% increase in percentage germination in comparison to control in the presence of pesticide stress. Most promising results for chlorophyll content estimation were obtained in the presence of carotenoids upto 6 mgg-1 without stress by both strains Rad1 and Rad2. Study suggests that especially strain Ros2 can enhance plant growth parameters in the pesticide stress.

Resumo Três bactérias solubilizantes de fosfato foram isoladas e identificadas por seqüenciamento de rRNA 16S como Pseudomonas putida, Pseudomonas sp e Pseudomonas fulva. As estirpes foram submetidas a testes bioquímicos de plantas e todos os atributos PGPR foram verificados na presença de pesticidas (clorpirifos e piriproxifeno). O índice de solubilização de fosfato da estirpe Ros2 foi mais elevado no meio NBRIP, isto é, 2,23 mm. Todas as estirpes apresentaram um pH ácido (varia de 2,5-5) em ambos os meios, isto é PVK e NBRIP. A estirpe Ros2 foi altamente positiva para a produção de amoníaco, bem como a produção de sideróforos enquanto a estirpe Rad2 foi positiva para a produção de HCN. Os resultados obtidos pelas estirpes Rad1, Rad2 e Ros2 para a produção de auxina foram 33,1, 30,67 e 15,38 μg ml-1 , respectivamente. A deformação Rad1 mostrou aumento de 16% na germinação percentual em comparação com o controlo na presença de stress de pesticida. Os resultados mais promissores para a estimativa do teor de clorofila foram obtidos na presença de carotenóides até 6 mgg-1 sem estresse por ambas as cepas Rad1 e Rad2. Estudo sugere que especialmente a estirpe Ros2 pode melhorar parâmetros de crescimento de plantas no estresse de pesticidas.

Assessment of microbial diversity in the rhizosphere of Pinus roxburghii (Sarg.) and bio-inoculant potential of selected pine bacterial isolates for wheat varieties based on cultureindependent and culture-dependent techniques.

Evidence is lacking regarding compatibility of pine bacteria as bio-inoculants for crops. The diversity and abundance of rhizosphere bacteria of Pinus roxburghii has never been investigated with simultaneous application of culture-dependent and culture-independent techniques. The present study was aimed to isolate, characterise, check the bio-inoculant potential of pine bacteria and assess rhizosphere bacterial diversity using culture-independent advanced approaches. Forty bacteria isolated from the rhizoplane of P. roxburghii growing in a cold climate at high altitude in Murree, were morphologically characterised; nine were identified by 16S rRNA sequence analyses and used in experiments. Diversity and abundance of the 16S rRNA gene and nifH gene in the rhizosphere was assessed by cloning, RFLP analysis, 454-amplicon pyrosequencing and qPCR. The bacterial isolates significantly improved dry weight of shoot, root, root area, IAA and GA3 content, number of grains plant-1 , weight of grains plant-1 in wheat varieties Chakwal-50 and Fareed-06 under axenic and field conditions. The number of 16S rRNA sequences (2979) identified by pyrosequencing shared similarity with 13 phyla of bacteria and archaea. The results confirm the existence of diverse bacteria of agricultural and industrial importance in the rhizosphere and compatibility of rhizoplane bacteria as bio-inoculants for wheat varieties.

Plant defence induced by PGPR against Spodoptera litura in tomato (Solanum lycopersicum L.).

The research conducted including its rationale: Spodoptera litura is the major pest of tomato causing significant reduction in tomato yield. Application of Plant growth promoting rhizobacteria(PGPR) prevent use of chemical fertilizer and synthetic pesticides through enhancement of plant growth and yield and induction of systemic resistance. Present investigation is an attempt to evaluate the role of PGPR, Pseudomonas putida and Rothia sp. on the physiology and yield of tomato fruit infested with the S. litura. Central methods applied: The surface sterilized seeds of tomato were inoculated with 48 h culture of P. putida and Rothia sp. At 6-7 branching stage of the plant, the larvae of S. litura at 2nd in star was used to infect the tomato plant leaves. KEY RESULTS: The S. litura infestation decreased dry weight of shoots and roots by 46% and 22%, and significant reduction was recorded in tomato fruit yield. The P. putida and Rothia sp. inoculations alleviated the adverse effects of insect infestation and resulted in 60% increase in plant biomass and 40% increase in yield over infested plants. Main conclusions including key points of discussion: PGPR: Defense appears to be mediated via increase in proline production, enhanced activities of antioxidant enzymes, stimulation in the activities of protease and polyphenol oxidases, increased contents of phenolics, protein and chlorophyll. The formulation of biopesticide involving PGPR comprise an environment friendly and sustainable approach to overcome insect infestation.

Comparative effects of wild type Stenotrophomonas maltophilia and its indole acetic acid-deficient mutants on wheat.

The present investigation evaluated the role of Stenotrophomonas maltophilia and its IAA-deficient mutant on soil health and plant growth under salinity stress in the presence of tryptophan. In the first phase, S. maltophilia isolated from roots of the halo- phytic herb, Cenchrus ciliaris was used as bio-inoculant on wheat grown in saline sodic soil. A field experiment was conducted at Soil Salinity Research Institute during 2010-2011. Treatments included seed inoculation with S. maltophilia with or without tryptophan; uninoculated untreated plants were taken as control. An aqueous solution of tryptophan was added to rhizosphere soil at 1 µg l(_1) after seed germination. Inoculation with S. maltophilia significantly increased soil organic matter, enhanced (20-30%) availability of P, K, Ca and NO3 -N and decreased Na content and electrical conductivity of rhizosphere soil. Plant height, fresh weight, proline and phytohormone content of leaves were increased 30-40% over the control. Activities of superoxide dismutase (SOD) and peroxidase (POD) were 40-50% higher than control. Addition of tryptophan further augmented (10-15%) growth parameters, whereas NO3 -N, P, K and Ca content, proline content and SOD and POD increased 20-30%. In a second phase, indoleacetic acid (IAA)-deficient mutants of S. maltophilia were constructed and evaluated for conversion of tryptophan to IAA at the University of Calgary, Canada, during 2013-2014. About 1800 trans-conjugants were constructed that were unable to produce IAA in the presence of tryptophan. The results suggest that tryptophan assisted S. maltophilia in the amelioration of salt stress, and that IAA played positive role in induction of salt tolerance.

New insight into the stability of CaCO3 surfaces and nanoparticles via molecular simulation.

Using updated and improved atomistic models for the polymorphs of calcium carbonate and their constituent ions in solution, we revisit the question of surface energetics and nanoparticle stability. Using a simple lattice-based Monte Carlo scheme, we generate nanoparticle configurations in vacuum for all three biologically relevant polymorphs of calcium carbonate and establish that the bulk energetic ordering of polymorphs persists to the nanoscale. In aqueous environments, results based on surface enthalpy alone indicate that formation of mineral-water interfaces is marginally favorable in many cases. Including an estimate of lost entropy due to formation of structured water layers is sufficient to reverse this observation, implying a delicate balance of enthalpy and entropy at crystalline CaCO3. In contradiction to some previous studies, we find that small calcite nanoparticles with diameters in the range of 1.8-4.1 nm do not retain an ordered structure on nanosecond time scales. The consequences of these results for simulation studies of biomineralization are discussed.

Medición de la capacidad pulmonar total mediante el método radiográfico planimétrico y su variante el autocad, en pacientes con trastorno ventilatorio restrictivo / Total lung capacity measurement with the planimetric radiographic method and it's variant "the autocad'' in patients with restrictive pulmonary disease

Introducción: Según la European Respiratory Society faltan estudios para pronunciarse sobre la validez de los métodos radiográficos, como métodos de medición de la capacidad pulmonar total, en pacientes con trastorno ventilatorio restrictivo. Objetivo: Valorar la validez del método radiográfico planimétrico clásico y del programa de medición de superficies planas "autocad" a través del ordenador, para la medición de la capacidad pulmonar total en pacientes con trastorno ventilatorio restrictivo. Material y Métodos: A cada uno de los 25 pacientes diagnosticados previamente de trastorno ventilatorio restrictivo, y en seguimiento periódico en la consulta de neumología, se les midió la capacidad pulmonar total, primero, por pletismografía y, después, por los métodos radigráficos planimétrico y "autocad" a partir de sus radiografías de tórax. Para la comparación de los resultados obtenidos por los distintos métodos se ha seguido el análisis estadístico de correlación lineal y cálculo del coeficiente de correlación de Pearson para una confianza del 95 por ciento (p < 0'05). Resultados: La comparación de la media de la capacidad pulmonar total por pletismografía (4675cc) con la media de la capacidad pulmonar total por el método radiográfico planimétrico (4636cc) y con el programa "autocad" (4865cc), alcanzaron un coeficiente de correlación de (r = 0'888) y (r = 0'887), respectivamente. Conclusión: Según nuestros resultados el método radiográfico planimétrico y su variante el "autocad", son válidos para la medición de la capacidad pulmonar total en pacientes con trastorno ventilatorio restrictivo (AU)

Isolation and 16S rRNA sequence analysis of the beneficial bacteria from the rhizosphere of rice.

The present study deals with the isolation of plant growth promoting rhizobacteria (PGPR) from rice (variety NIAB IRRI-9) and the beneficial effects of these inoculants on two Basmati rice varieties. Nitrogen-fixing activity (acetylene-reduction activity) was detected in the roots and submerged shoots of field-grown rice variety NIAB IRRI-9. Estimation of the population size of diazotrophic bacteria by ARA-based MPN (acetylene reduction assay-based most probable number) in roots and shoots indicated about 10(5)-10(6) counts/g dry weight at panicle initiation and grain filling stages. Four bacterial isolates from rice roots and shoots were obtained in pure culture which produced phytohormone indoleacetic acid (IAA) in the growth medium. Among these, three isolates S1, S4, and R3 reduced acetylene to ethylene in nitrogen-free semi-solid medium. Morphological and physiological characteristics of the isolates indicated that three nitrogen-fixing isolates S1, S4, and R3 belonged to the genus Enterobacter, while the non-fixing isolate R8 belonged to the genus Aeromonas. 16S rRNA sequence of one isolate from root (R8) and one isolate from shoot (S1) was obtained which confirmed identification of the isolates as Aeromonas veronii and Enterobacter cloacae, respectively. The 1517-nucleotide-long sequence of the isolate R8 showed 99% similarity with Aeromonas veronii (accession No. AF099023) while partial 16S rRNA sequence (two stretches of total 1271 nucleotide length) of S1 showed 97% similarity with the sequence of Enterobacter cloacae (accession No. AJ251469). The seedlings of two rice varieties Basmati 385 and Super Basmati were inoculated with the four bacterial isolates from rice and one Azospirillum brasilense strain Wb3, which was isolated from wheat. In the rice variety Basmati 385, maximum increase in root area and plant biomass was obtained in plants inoculated with Enterobacter S1 and Azospirillum Wb3, whereas in the rice variety Super Basmati, inoculation with Enterobacter R3 resulted in maximum increase of root area and plant biomass. Nitrogen fixation was quantified by using 15N isotopic dilution method. Maximum fixation was observed in Basmati 385 with the inoculants Azospirillum Wb3 and Enterobacter S1 where nearly 46% and 41% of the nitrogen was derived from atmosphere (%Ndfa), respectively. In general, higher nitrogen fixation was observed in variety Basmati 385 than in Super Basmati, and different bacterial strains were found more effective as inoculants for the rice varieties Basmati 385 and Super Basmati.

[Guidelines of the Spanish Society of Cardiology for clinical practice in exercise testing]. / Guías de práctica clínica de la Sociedad Española de Cardiología en pruebas de esfuerzo.

Most exercise testing is performed in adults with known or suspected ischemic heart disease. In the last few years cardiac imaging techniques have been applied in this field, improving the information obtained with the procedure. However, the exceptions to this rule are emerging rapidly not only in healthy people (asymptomatic individuals, athletes, handicapped people) but also in cardiac patients (advanced congestive heart failure, hypertension, rhythm disorders, congenital heart disease, etc.). All the-se issues justify the need for a multidisciplinary consensus document in Spain. This paper reviews and updates the methodological aspects of the stress test, including those related to oxygen consumption measurements. The main aim of this review was to determine the role of exercise testing in the evaluation of ischemic heart disease as well as the applications of imaging stress testing. The usefulness of this test in other non-ischemic cardiac disorders and in selected subsets of healthy people is also reviewed.

Pulmonary vagal innervation is required to establish adequate alveolar ventilation in the newborn lamb.

To investigate the effects of bilateral intrathoracic vagotomy on the establishment of continuous breathing and effective gas exchange at birth, we studied 8 chronically instrumented, unanesthetized, sham-operated and 14 vagotomized newborn lambs after a spontaneous, unassisted vaginal delivery. Fetal lambs were instrumented in utero to record sleep states, diaphragmatic electromyogram, blood pressure, arterial pH, and blood-gas tensions. Six of eight sham-operated lambs established effective gas exchange within 10 min of birth, whereas 12 of 14 vagotomized animals developed respiratory acidosis and hypoxemia (P = 0.008). Breathing frequency in vagotomized newborns was significantly lower during the entire postnatal period compared with sham-operated newborns. Vagotomized subjects also remained hypothermic during the entire postnatal period (P < 0.05). Bronchoalveolar lavage indicated an increased minimum surface tension, whereas lung histology showed perivascular edema and partial atelectasis in the vagotomized group. We conclude that stimulation of breathing and effective gas exchange are critically dependent on intact vagal nerves during the transition from fetal to neonatal life.

Umbilical arterial blood flow and plasma prostaglandin E2 concentrations during arousal and breathing movements in fetal sheep.

To investigate the effects of lung distension and oxygenation on umbilical blood flow (UBF) and plasma prostaglandin E2 (PGE2) in relation to arousal and stimulation of breathing movements, we studied eight chronically instrumented, unanesthetized fetal sheep between 137 and 143 d of gestation. Electrocorticogram, electro-oculogram, nuchal and diaphragmatic electromyograms, arterial pH and blood gas tensions, Hb oxygen saturation, body temperature, and UBF were recorded in each fetus. Electrocorticogram, electro-oculogram, and nuchal electromyograms were used to define sleep states. No sooner than 4 d after surgery, fetal lungs were distended with 100% O2 or N2 in a randomized order via an in situ Y-endotracheal tube. PGE2 concentrations were analyzed by RIA. A significant increase in fetal arousal and stimulation of breathing during nonrapid eye movement sleep was observed during lung distension with O2 as compared with control periods and lung distension with nitrogen. In all sleep states, UBF significantly decreased during oxygenation as compared with the control values. However, no significant correlation was observed between the time of the onset of arousal and the decrease in UBF. Lung distension with N2 resulted in increased plasma PGE2 concentrations, whereas, no change was observed during oxygenation. Our data suggest that an increase in fetal partial pressure of arterial O2 leads to a decrease in UBF. However, the onset of arousal and stimulation of breathing during lung distension and oxygenation were not dependent on a decrease in plasma PGE2 concentrations.

[Small size breast cancer conservative surgical therapy and subsequent radiotherapy]. / Il cancro mammario di piccole dimensioni: terapia chirurgica conservativa e successiva radioterapia.

From february 1980 to december 1985, 115 breast carcinomas were treated with QU.A.RT. with limited surgery and following radiotherapy. Median age was 51 years (range 25 to 75). Surgical approach was quadrantectomy with axillary nodes dissection. Irradiation of residual breast was performed by an anterior high energy electron beam. 50 Gy in 25 fractions was the minimal dose to whole breast. In pN+ patients systemic adjuvant therapy was done. Local control was obtained in 111 cases. Local recurrence appeared in 4: 1 menopausal pT1 pNO and 3 premenopausal (1 pT1 pNO and 2 pT1 pN+) at respectively 31, 19, 22 months from diagnosis. Distant metastases appeared in 10 patients. 7 died for cancer and 3 are still alive. Cosmetic results were scored as good in 52, better than mastectomy in 26, unsatisfactory in 3, and not stated in 34. Our results are comparable to those of other Centers. Youth, axillary metastases and delay between surgery and radiotherapy appears to influence the risk of recurrence also in our series.

[Diagnostic aspects of aneurysm of the splenic artery (description of a case)]. / Aspetti diagnostici dell'aneurisma dell'arteria splenica (descrizione di un caso).

A case of splenic artery aneurysm with ultrasonic examination discovered is reported and analyzed. The aneurysm pas asymptomatic and founded as incidental finding during standard x-ray. Arteriography was required to confirm the diagnosis and indicate the correct surgical treatment.

[Primary carcinoma of the gallbladder: ultrasonic features (remarks on 8 cases)]. / Il carcinoma primitivo della colecisti: aspetti ecotomografici (considerazioni su otto casi).

The authors illustrate the ultrasound features of gallbladder carcinoma after reviewing literature and basing upon their own experience. The diagnosis of this neoplasia is formulated more frequently today by introduction of scanning means, but it remains very difficult on initial phase because of its precocious lymphatic and hilar infiltration.