Unraveling Azospirillum's colonization ability through microbiological and molecular evidence.

It is known that members of the bacterial genus Azospirillum can promote the growth of a great variety of plants, an ability harnessed by the industry to create bioproducts aimed to enhance the yield of economically relevant crops. Its versatile metabolism allows this bacterium to adapt to numerous environments, from optimal to extreme or highly polluted. The fact of having been isolated from soil and rhizosphere samples collected worldwide and many other habitats proves its remarkable ubiquity. Azospirillum rhizospheric and endophytic lifestyles are governed by several mechanisms, leading to efficient niche colonization. These mechanisms include cell aggregation and biofilm formation, motility, chemotaxis, phytohormone and other signaling molecules production, and cell-to-cell communication, in turn, involved in regulating Azospirillum interactions with the surrounding microbial community. Despite being infrequently mentioned in metagenomics studies after its introduction as an inoculant, an increasing number of studies detected Azospirillum through molecular tools (mostly 16S rRNA sequencing) as part of diverse, even unexpected, microbiomes. This review focuses on Azospirillum traceability and the performance of the available methods, both classical and molecular. An overview of Azospirillum occurrence in diverse microbiomes and the less-known features explaining its notorious ability to colonize niches and prevail in multiple environments is provided.

Metasurface-enhanced mid-infrared spectrochemical imaging of tissues.

Label-free and nondestructive mid-infrared vibrational hyperspectral imaging is emerging as an important ex-vivo tissue analysis tool, providing spatially resolved biochemical information critical to understanding physiological and pathological processes. However, the chemically complex and spatially heterogeneous composition of tissue specimens and the inherently weak interaction of infrared light with biomolecules limit the analytical performance of infrared absorption spectroscopy. Here, we introduce an advanced mid-infrared spectrochemical tissue imaging modality using metasurfaces that support strong surface-localized electromagnetic fields to capture quantitative molecular maps of large-area murine brain-tissue sections. Our approach leverages polarization-multiplexed multi-resonance plasmonic metasurfaces to simultaneously detect many different functional biomolecules. The resulting surface-enhanced mid-infrared spectral imaging (SE-MIRSI) method eliminates the non-specific effects of bulk tissue morphology on the quantitative analysis of fingerprint spectra and improves the chemical selectivity. We show that the metasurface enhancement increases the retrieval of amide I and II absorption bands associated with secondary structures of proteins. Moreover, we demonstrate that plasmonic metasurfaces enhance the chemical contrast in infrared images and enable the detection of ultrathin tissue regions that are not otherwise visible to conventional mid-infrared spectral imaging. While we tested our approach on murine brain tissue sections, this chemical imaging method is well-suited for any tissue type, which significantly broadens the potential impacts of our method for both translational research and clinical histopathology.

Day and blue light modify growth, cell physiology and indole-3-acetic acid production of Azospirillum brasilense Az39 under planktonic growth conditions.

AIM: In this work, we evaluated the effects of light on growth, cell physiology and stress response of Azospirillum brasilense Az39, a non-photosynthetic rhizobacteria, under planktonic growth conditions. METHODS AND RESULTS: Exponential cultures of Az39 were exposed to blue (BL), red (RL) and daylight (DL) or maintained in darkness for 24, 48 and 72 h. The biomass production and indole 3-acetic acid (IAA) biosynthesis increased by exposition to DL. Conversely, BL decreased IAA concentration through a direct effect on the molecule. The DL increased superoxide dismutase activity, hydrogen peroxide and thiobarbituric acid reactive substances levels, but the last one was also increased by BL. Both DL and BL increased cell aggregation but only BL increased biofilm formation. CONCLUSIONS: We demonstrated that both BL and DL are stress effectors for A. brasilense Az39 under planktonic growth conditions. The DL increased biomass production, IAA biosynthesis and bacterial response to stress, whereas BL induced cell aggregation and biofilms formation, but decreased the IAA concentration by photooxidation. SIGNIFICANCE AND IMPACT OF THE STUDY: Blue light and DL changes growth capacity, cell physiology and plant growth promotion ability of A. brasilense Az39 and these changes could be considered to improve the production and functionality of biofertilizers.

New insights into indole-3-acetic acid metabolism in Azospirillum brasilense.

AIMS: The aim of this research was to analyse the global indole-3-acetic acid (IAA) metabolism in three commercially used strains of Azospirillum brasilense. METHODS AND RESULTS: Azospirillum brasilense Sp245, Az39 and Cd, containing a plasmid with the ipdC-gusA fusion (pFAJ64), were cultured in minimal medium MMAB with or without 10 mg l-1 of l-trp till exponential or stationary growth phase. The cultures were then split into 10 ml tubes and individually treated with 10 mg ml-1 IAA, IBA or NAA (auxin catabolism and homeostasis); IAPhe, IALeu, IAA-ala, IAA-glucose (IAA conjugate hydrolysis); or l-lys, l-leu, l-ileu, l-phe, l-ala, l-val, l-arg, l-glu, l-his, l-met, l-asp, l-cys, l-ser, l-pro, l-thr and l-trp (regulation of IAA biosynthesis and IAA conjugation). Bacterial growth, IAA production and ipdC expression were evaluated. None of the A. brasilense strains were able to hydrolyse IAA conjugates, catabolize auxins, or conjugate IAA with amino acids or glucose. l-amino acids l-met, l-val, l-cys and l-ser inhibited bacterial growth and decreased IAA biosynthesis. The expression of ipdC and IAA biosynthesis but not bacterial growth was affected by l-leu, l-phe, l-ala, l-ile, l-pro. l-arg, l-glu, l-his, l-lys, l-asp and l-thr did not affect any of the measured parameters. CONCLUSIONS: In this paper, we confirmed that A. brasilense produces IAA only in presence of l-trp is not able to degrade auxins, conjugate IAA with sugars and/or l-amino acids, or hydrolyse such conjugates to release free IAA. Finally, we found that bacterial growth and/or IAA biosynthesis were inhibited by the presence of several l-amino acids probably by diversion of the cellular metabolism. SIGNIFICANCE AND IMPACT OF THE STUDY: We propose a renewed model to explain IAA metabolism in A. brasilense, one of the most studied phytostimulatory bacteria.

Impact of maternal risky behaviors on the behaviors of children born to adolescent and young mothers / 中华儿科杂志

Objective@#To examine the impact of maternal risky behaviors on the behaviors of children born to adolescent and young mothers.@*Methods@#Adolescents and young Chinese mothers were recruited from an integrated young mother supportive program in Hong Kong between January and June 2015. Eligible mothers were asked to complete a questionnaire on their sociodemographic characteristics and history of risky behavior as well as their children's behaviors. Multiple regression analyses were conducted to explore the association between maternal risky behaviors and their children's behaviors.@*Results@#Among 201 respondents, there were 187 (93.0%) ex-drinkers, 136 (67.7%) ex-smokers, and 83 (41.3%) ex-addicts. Compared to the reference group, children of mothers with drug use behaviors were more likely to have abnormal SDQ total difficulties scores (odds ratio 2.60, P=0.01), those of ex-drinking mothers had more behavioral difficulties and more conduct problems (B=3.82 and 1.37, P both=0.01) and those of ex-smoking mothers had more conduct problems (B=0.74, P=0.01) after adjustment for confounders. Children of active drug-taking mothers also had more emotional symptoms (B=1.77, P=0.04) and hyperactivity/inattention problems (B=2.14, P=0.03).@*Conclusion@#The history of mother's risky behavior was significantly associated with the behavioral problems of the children.

Desarrollo tardío de múltiples dientes supernumerarios en una paciente con miastenia gravis juvenil tratada con azatioprina / Delayed development of multiple supernumerary teeth in a patient with juvenile myasthenia gravis treated with azathioprine

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Análisis del tipo de publicación y áreas del conocimiento de Enfermería en la evaluación interna de la Revista Enfermería Universitaria 2007-2011 / Publication type and nursing knowledge areas analysis as part of the internal assessment of Enfermería Universitaria Journal, 2007-2011

Introducción: Analizar las publicaciones de una disciplina determinada permite evaluar su avance o consolidación en la construcción del conocimiento, identificar las áreas de interés de los propios investigadores y aquellas que requieren de fomento. Objetivo: Analizar el tipo de artículo y áreas del conocimiento en los artículos publicados en la revista enfermería universitaria en el periodo 2007-2011. Material y métodos: el diseño metodológico fue de corte cuantitativo, transversal, descriptivo. Se evaluó el tipo de artículos publicados y las áreas del conocimiento de Enfermería. La muestra de estudio fue el total de artículos publicados durante los años 2007 a 2011. Resultados: Respecto al tipo de artículos producidos: el 45.9% fueron de investigación, 21.8% de innovación para la práctica, 8.3% de revisión y 7.5% de docencia. En cuanto a las áreas de conocimiento se publicaron en mayor medida artículos de aspectos metodológicos, relacionados con la enseñanza-aprendizaje y con la administración-gestión. Conclusiones: el predominio, en cuanto a tipo, de artículos de investigación pudiera considerarse un avance en esta área, aunque es preciso continuar mejorando este indicador. Además se requiere apoyar a profesionales de Enfermería del área clínico-asistencial para que incursionen en la investigación y publicación de aspectos vinculados a su práctica cotidiana.

Introduction: Analyzing a determined discipline's publications allows an assessment of its development or consolidation towards knowledge construction, and an identification of the researchers' areas of interest, as well as those which require further impulse. Objective: to analyze the study type and knowledge areas of the published articles of Enfermería Universitaria Journal during the period 2007-2011. Material and methods: the study was quantitative, transversal, and descriptive. The article type and nursing knowledge areas of the journal publications from 2007 to 2011 were assessed. Results: Regarding the study types, 45.9% were identified as research, 21.8% as practice innovation, 8.3% as revision, and 7.5% as academic. Regarding the areas of knowledge, most published articles were related to methodological, teaching-learning, and managerial aspects. Conclusions: An interesting finding was the proportion of research articles published. Moreover, supporting nursing professionals within the clinical-assistance areas to conduct and publish re-search studies related to their practices is also suggested.

Determinants of coronary artery calcification progression in renal transplant recipients.

Coronary artery calcification (CAC) is associated with increased atherosclerotic burden and cardiovascular events. The objective of this study was to determine the natural history and risk factors associated with CAC progression in a cohort of incident asymptomatic renal transplant recipients with no history of coronary revascularization. Electron-beam computed tomography was performed in 82 subjects at time of transplantation and at least 1 year later. Mean (SD) and median CAC score increased for all subjects from 392.4 (747.9) and 75.8 at time of transplant to 475.3 (873.5), (p = 0.002[log]) and 98.9 (p < 0.001), respectively. Most subjects (89%) with no calcifications remained without calcification. Mean annualized rate (SD) of CAC score change was 52.5 (150) with a median of 0.5. Average yearly percent change was 67.3 (409.6) with a median of 1.4. In multivariate analysis, diastolic blood pressure at 3 months post-transplant, Caucasian race, glomerular filtration rate at 3.0, months post-transplant, body mass index and baseline CAC score were independent predictors of annualized rate of CAC change. There is significant progression of CAC post-renal transplantation in most subjects. Progression is most likely to occur in white patients and is associated with clinical factors such as blood pressure, body mass index, renal function and baseline CAC score.

Assessment of differences in HLA-A, -B, and -DRB1 allele mismatches among African-American and non-African-American recipients of deceased kidney transplants.

Among recipients of deceased donor kidney transplants, African-Americans experience a more rapid rate of kidney allograft loss than non-African-Americans. The purpose of this study was to characterize and quantify the HLA-A, -B, and -DRB1 allele mismatches and amino acid substitutions at antigen recognition sites among African-American and non-African-American recipients of deceased donor kidney transplants matched at the antigen level. In recipients with zero HLA antigen mismatches, the degree of one or two HLA allele mismatches for both racial groups combined was 47%, 29%, and 11% at HLA-DRB1, HLA-B, and HLA-A, respectively. There was a greater number of allele mismatches in African-Americans than non-African-Americans at HLA-A (P < .0001), -B (P = .096), and -DRB1 loci (P < .0001). For both racial groups, the HLA allele mismatches were predominantly at A2 for HLA-A; B35 and B44 for HLA-B; but multiple specificities for HLA-DRB1. The observed amino acid mismatches were concentrated at a few functional positions in the antigen binding site of HLA-A and -B and -DRB1 molecules. Future studies are ongoing to assess the impact of these HLA mismatches on kidney allograft loss.

Optimization of RNA yield, purity and mRNA copy number by treatment of urine cell pellets with RNAlater.

BACKGROUND: We have shown that measurement of mRNA for cytotoxic attack proteins perforin and granzyme B in urinary cells is a noninvasive means of diagnosing acute rejection of human renal allografts. Urinary cell mRNA studies have yielded useful information in other patient populations such as patients with cancer. The isolation of sufficient and high quality ribonucleic acid (RNA) from urinary cells however is problematic. RNAlater, an RNA stabilization solution, has been reported to optimize RNA isolation from tumor tissues stored at room temperature and from pigment-rich ocular tissues. METHODS: We explored whether the addition of RNAlater to urine cell pellets improves RNA yield, enhances purity and facilitates measurement of low abundance mRNAs. We measured, with the use of real-time quantitative polymerase chain reaction (PCR) assay, levels of expression of a constitutively expressed gene 18S rRNA and mRNA for granzyme B and transforming growth factor-beta(1) (TGF-beta(1)) in urine specimens and renal biopsies obtained from renal allograft recipients. RESULTS: RNA yield (P<0.01, Wilcoxon signed rank test) and the A260/A280 ratio (P<0.01) were both higher with urine cell pellets treated with RNAlater prior to snap freezing compared to cell pellets that were not treated with RNAlater prior to snap freezing. Levels (copy number per 1 microg of total RNA) of 18S rRNA (P<0.02), granzyme B mRNA (P=0.002) and TGF-beta(1) (P=0.02) were all higher with treated urine cell pellets compared to untreated cell pellets. Kruskall-Wallis one way analysis of variance and pair-wise comparisons with Student-Newman-Keuls test showed that the levels of mRNA for granzyme B (P<0.05) and TGF-beta(1) (P<0.05) are significantly different between renal allograft biopsies and untreated urine cell pellets but not between the biopsy specimens and RNAlater-treated urine cell pellets. CONCLUSIONS: The addition of RNAlater to urine cell pellets improves RNA isolation from urinary cells and facilitates measurement of low abundance mRNAs.

Methylation status in the promoter region of the human PGP9.5 gene in cancer and normal tissues.

PGP 9.5 is a neurospecific peptide that functions to remove ubiquitin from ubiquitinated cellular proteins, thereby preventing them from targeted degradation by the proteasome-dependent pathway or regulating their localization, activity or structure. Using the serial analysis of gene expression method (SAGE), we initially found that the PGP9.5 transcript and protein was highly expressed in more than 50% of primary lung cancers and nearly all lung cancer cell lines but was not detectable in the normal lung. This increased expression could be the result of transcriptional regulation accompanied by methylation changes at the CpG island of the promoter region. We studied the methylation status of the cytosines at the promoter region of human PGP9.5 using sodium bisulfite genomic sequencing in normal and neoplastic cells. Although no methylation of PGP9.5 promoter was observed in the normal lung, normal cervical tissue, and lung cancer cell lines, this region was densely methylated in the HeLa cell line. Exposure to HeLa cells to the demethylating agent, 5-aza-2'-deoxycytidine, led to re-expression of PGP9.5. This data suggested that while other mechanisms may be involved in the frequent overexpression of PGP9.5 gene in lung tumors and lung cancer cell lines, promoter methylation may play a role in the transcriptional suppression of PGP9.5 gene expression in the cervical tissue-derived HeLa cell line.

Prevalence and determinants of erectile dysfunction in hemodialysis patients.

BACKGROUND: The prevalence of erectile dysfunction (ED) among patients with end-stage renal disease (ESRD) is not known. METHODS: A cross-sectional study was conducted to determine the prevalence of ED among a community-based hemodialysis (HD) population using a two-stage cluster random sampling design. The presence and severity of ED were assessed among 302 ESRD patients using the self-administered International Index of Erectile Function-5 (IIEF-5). Logistic regression was used to examine and test associations between ED and other medical conditions. RESULTS: The prevalence of any level of ED was 82% (95% CI, 76 to 87%) for all HD subjects. The prevalence of severe ED was 45% (CI, 36 to 55%). Subjects younger than 50 years had a prevalence of ED of 63% (CI, 53 to 71%), while in subjects 50 years or older, it was 90% (CI, 84 to 94%). A multivariable analysis demonstrated increasing age (50 to 59, OR = 2.04, 95% CI, 1.3 to 3.1; 60 to 69, OR = 5.5, 95% CI, 1.9 to 15.6) and diabetes (OR = 2.0, 95% CI, 1.2 to 3.3) to be independently associated with the presence of any level of ED. However, neither the subjects' age nor history of diabetes predicted the severity of ED among subjects with ED. The use of angiotensin-converting enzyme inhibitors (ACEIs) was inversely associated with ED (OR = 0.41, 95% CI, 0.17 to 0.98). Poor functional status (Karnofsky score or the Index of Physical Impairment) was not associated with ED. CONCLUSIONS: ED is extremely prevalent among HD patients. Increasing age, diabetes, and nonuse of ACEIs were associated with higher prevalence of ED. The high prevalence of ED was seen even among patients with good functional status.

Promoter hypermethylation patterns of p16, O6-methylguanine-DNA-methyltransferase, and death-associated protein kinase in tumors and saliva of head and neck cancer patients.

Aberrant promoter hypermethylation is common in head and neck cancer and may be useful as a marker for cancer cells. We examined whether cells with tumor-specific aberrant DNA-methylation might be found in the saliva of affected patients. We tested 30 patients with primary head and neck tumors using methylation-specific PCR searching for promoter hypermethylation of the tumor suppressor gene p16 (CDKN2A), the DNA repair gene O6-methylguanine-DNA-methyltransferase (MGMT) and the putative metastasis suppressor gene death-associated protein kinase (DAP-K). Aberrant methylation of at least one of these genes was detected in 17 (56%) of 30 head and neck primary tumors; 14 (47%) of 30 at p16, 10 (33%) of 30 at Dap-K and 7 (23%) of 30 at MGMT. In 11 (65%) of 17 methylated primary tumors abnormal methylated DNA was detected in the matched saliva samples. Abnormal promoter methylation in saliva DNA was found in all tumor stages and more frequently in tumors located in the oral cavity. Moreover, none of the saliva from patients with methylation-negative tumors displayed methylation of any marker. Of 30 saliva samples from healthy control subjects (15 smokers and 15 nonsmokers), only one sample from a smoking patient was positive for DNA methylation at two target genes. Detection of aberrant promoter hypermethylation patterns of cancer-related genes in saliva of head and cancer patients is feasible and may be potentially useful for detecting and monitoring disease recurrence. Long-term longitudinal studies are needed to evaluate this approach for early detection of head and neck cancer in at-risk populations.

Preliminary observations of sildenafil treatment for erectile dysfunction in dialysis patients.

Erectile dysfunction is common in dialysis patients. We report our experience with sildenafil citrate in patients undergoing dialysis therapy. Male subjects attending the Outpatient Dialysis Unit at the University of Pennsylvania (Philadelphia, PA) who were prescribed sildenafil by their primary physician or nephrologist were asked to complete the International Index of Erectile Function before their first dose of sildenafil and after at least 4 weeks of therapy. Subjects' mean age was 50.3 +/- 14.63 (SD) years. Ninety-three percent of the subjects were black. Based on a global efficacy question, 66.7% of the subjects believed that treatment had improved their erections. Subjects reported no increase in the sexual desire domain despite experiencing a significant increase in erectile function, orgasmic function, and satisfaction with intercourse. Sildenafil was well tolerated in a selected group of patients who reported improved sexual function with no major adverse effects.

Management of selected lipid abnormalities. Hypertriglyceridemia, low HDL cholesterol, lipoprotein(a), in thyroid and renal diseases, and post-transplantation.

Although the focus in treating lipid disorders is on reducing LDL cholesterol levels, triglycerides, HDL cholesterol, and Lp(a) are all independent risk factors that can be used clinically to assess cardiovascular risk. Decisions to initiate drug therapy for LDL cholesterol reduction may be influenced by levels of these other lipoprotein fractions. Data supporting intervention to modify these factors is less abundant than for LDL cholesterol reduction, but in certain circumstances drug therapy targeted at triglycerides or HDL cholesterol may be appropriate. Patients with nephrotic syndrome and end-stage renal disease are at particularly high risk for the development of cardiovascular disease and should be treated aggressively for lipid disorders. Finally, solid organ transplant recipients are almost always hyperlipidemic, and appropriate therapy could reduce cardiovascular events.

[Posterior genital prolapse following the Burch method of colposuspension]. / Prolapso genital posterior a colposuspensión de Burch.

The aim of this paper was to know the prevalence of genital prolapse after the Burch procedure. Also to intend knowing if the associate surgery for vaginal relaxation used in our Urogynecologic Service is working well. Seventy four patients with more than six months after the Burch procedure were addressed from October 1995 to January 1996. Special attention was put to detect pelvis floor defects in specific vaginal sites (urethra, bladder, vaginal ápex, cul de sac and distal posterior wall). The prevalence of urethrocele was 2.7%, cystocele 50.6%, almost of all were mild. None severe cystocele was detected. Cervical prolapse was present in 21.3%, vault prolapse in 9.3% and enterocele 41.9%. The posterior colporraphy seems to be useful to prevent the vault prolapse and the enterocele. On the contrary the Moschowitz procedure seems not to be useful in this aspect. In order to diminish the prevalence of genital prolapse posterior to Burch procedure it would be better to improve the preoperative vaginal examination in order to do segmentary prolapse diagnosis and also to stablish an individualized surgical treatment of them.

Membranoproliferative glomerulonephritis type I, mixed cryoglobulinemia and lymphoma in the absence of hepatitis C infection.

Chronic hepatitis C virus infection has been linked to cryoglobulinemia, membranoproliferative glomerulonephritis, and malignant B-cell lymphoproliferation, suggesting a possible pathogenetic link between these disorders. We report a patient with the latter clinical triad in the absence of hepatitis C infection. We postulate that the persistent and dysregulated immunologic activity associated with chronic antigen stimulation, inflammation and/or B-cell malignancy induces nephritogenic autoantibodies, including cryoglobulins, that produce a similar clinical syndrome in genetically susceptible individuals.

Prolapso genital posterior a colposuspensión de Burch / Genital prolapse after the Burch procedure

Los objetivos del estudio fueron: conocer la prevalencia de prolapso genital posterior al procedimiento de Burch. Evaluar resultados de la cirugía efectuada en asociación con procedimiento de colposuspensión con el propósito de corregir el prolapso existente o prevenir el prolapso postoperatorio. Estudio clínico retrospectivo, longitudinal. Se exploraron 74 pacientes que con diagnóstico de incontinencia genuina de esfuerzo habían sido tratadas quirúrgicamente con el procedimiento de Burch en el Servicio de Urología Ginecológica del Hospital de Gineco-Obstetricia del C.M.N.O. del IMSS en Guadalajara. Control postoperatorio mínimo de seis meses. Método estadístico utilizado: frecuencias simples, porcentajes y prueba de Chi-cuadrada. La incidencia de prolapso genital postoperatorio en pared anterior de vagina, ápex y pared posterior, fue: Uretrocele 2.7 por ciento, cistocele 51.3 por ciento, ápex 17.9 por ciento en pacientes histerectomizadas; rectocele 45.9 por ciento. La mayoría de los casos fueron de grado leve, sin embargo siete pacientes (9.5 por ciento) requirieron de cirugía complementaria por el prolapso mismo. La colporrafia posterior parece ser útil para evitar el enterocele; no así el cierre de Douglas con la técnica de Moschowitz. Al evaluar con criterio estricto cada uno de los segmentos de vagina, se pudo apreciar una alta incidencia de prolapso genital posterior al procedimiento de Burch. Si bien la gran mayoría de los casos fueron prolapsos leves y asintomáticos, es aconsejable estudiar preoperatoriamente a la paciente con gran cuidado, a fin de establecer una estrategia de manejo individualizado con resultados más eficientes