Depression as a prognostic factor for lumbar spinal stenosis outcomes: a systematic review.

PURPOSE: Lumbar spinal stenosis (LSS) is associated with increased levels of psychological distress, including depression; however, the prognostic value of depression remains unclear. The purpose of this systematic review was to synthesize the evidence on the prognostic value of depression for a range of outcomes in patients with LSS. METHODS: Inclusion criteria were prospective cohort studies that investigated depression in patients diagnosed with LSS. Searches were conducted in 7 databases. Critical appraisal, data extraction, and judgement of cumulative evidence were conducted independently by two reviewers. A meta-analysis was not conducted due to a lack of unique cohorts for each outcome, varying follow-up times, and differences in measurements for both prognostic factors and outcomes. RESULTS: Twenty-three articles were included. There was evidence for an association between preoperative depression and postoperative disability and symptom severity outcomes for patients with LSS. Odds ratios ranged from 1.15 to 2.94 for postoperative disability and 1.16-1.20 for symptom severity at various follow-up times. Using GRADE, evidence supporting depression as a prognostic factor for these LSS outcomes was deemed to be of moderate quality. Similarly strong evidence suggested depressive symptoms are of no prognostic value for postoperative walking capacity. CONCLUSION: Depression appears to have small to moderate prognostic value for LSS outcomes, with the strongest evidence for postoperative disability and symptom severity. The prognostic value of depression for LSS outcomes should be further explored using standardized measures in additional cohorts, including patients managing their condition conservatively, who have been neglected in related research.

Spinal Cord Stimulation: Comparing Traditional Low-frequency Tonic Waveforms to Novel High Frequency and Burst Stimulation for the Treatment of Chronic Low Back Pain.

PURPOSE OF REVIEW: The purpose of the present investigation is to summarize supporting evidence for novel sub-perception spinal cord stimulation (SCS) therapy over traditional paresthesia inducing low-frequency waveforms for the treatment of chronic pain. The focus of this review is to summarize key studies comparing traditional low-frequency tonic waveforms to modern high frequency and burst stimulation for the treatment of patients with chronic intractable low back pain and/or leg pain. RECENT FINDINGS: Several recent studies have demonstrated the benefit of novel SCS therapies over traditional low-frequency SCS for the treatment of patients with chronic low back and/or leg pain. SENZA-RTC showed that paresthesia-free high-frequency SCS was superior to low-frequency stimulation for treatment of chronic low back pain with leg pain. The SUNBURST crossover trial recently found that high-frequency burst stimulation was preferred over low-frequency tonic SCS with patients citing better pain relief and a preference for paresthesia-free SCS. The new ongoing EVOLVE workflow retrospective multicenter study uses technology that can deliver both low-dose and high-dose SCS. Further, the wavewriter technology addresses patient variability with its ability to layer sub-perception waveforms and paresthesia inducing low-frequency stimulation tailored to patient needs via an interactive feedback feature. Neuromodulation for the treatment of chronic pain is rapidly evolving with technology at its forefront. Modern SCS systems use novel waveforms, frequencies, and stimulation modes to deliver paresthesia-free pain relief to patients suffering from chronic low back pain and/or leg pain with better results than traditional tonic low-frequency SCS. As the field advances, new studies are needed comparing new waveform and delivery systems to optimize patient selection and treatment response.

Take -home naloxone rescue kits following heroin overdose in the emergency department to prevent opioid overdose related repeat emergency department visits, hospitalization and death- a pilot study.

BACKGROUND: Opioid overdoses are at an epidemic in the United States causing the deaths of thousands each year. Project DAWN (Deaths Avoided with Naloxone) is an opioid overdose education and naloxone distribution program in Ohio that distributes naloxone rescue kits at clinics and in the emergency departments of a single hospital system. METHODS: We performed a retrospective analytic cohort study comparing heroin overdose survivors who presented to the emergency department and were subsequently discharged. We compared those who received a naloxone rescue kit at discharge with those who did not. Our composite outcome was repeat opioid overdose related emergency department visit(s), hospitalization and death at 0-3 months and at 3-6 months following emergency department overdose. Heroin overdose encounters were identified by ICD- 9 or 10 codes and data was abstracted from the electronic medical record for emergency department patients who presented for heroin overdose and were discharged over a 31- month period between 2013 and 2016. Patients were excluded for previous naloxone access, incarceration, suicidal ideation, admission to the hospital or death from acute overdose on initial emergency department presentation. Data was analyzed with the Chi- square statistical test. RESULTS: We identified 291emergency department heroin overdose encounters by ICD-9 or 10 codes and were analyzed. A total of 71% of heroin overdose survivors received a naloxone rescue kit at emergency department discharge. Between the patients who did not receive a naloxone rescue kit at discharge, no overdose deaths occurred and 10.8% reached the composite outcome. Of the patients who received a naloxone rescue kit, 14.4% reached the composite endpoint and 7 opioid overdose deaths occurred in this cohort. No difference in mortality at 3 or 6 months was detected, p = 0.15 and 0.36 respectively. No difference in the composite outcome was detected at 3 or 6 months either, p = 0.9 and 0.99 respectively. CONCLUSIONS: Of our emergency department patients receiving a naloxone rescue kit we did not find a benefit in the reduction of repeat emergency department visits hospitalizations, or deaths following a non-fatal heroin overdose.

A 14-20kDa protein binds to the upstream region of the phtM operon involved in the synthesis of phaseolotoxin in Pseudomonas syringae pv. phaseolicola NPS3121.

Pseudomonas syringae pv. phaseolicola is a phytopathogenic bacterium in beans that produces a phytotoxin called phaseolotoxin, in whose synthesis a group of genes that belong to the "Pht cluster" are involved. This cluster comprises 23 genes arranged in 5 transcriptional units, two monocistronic (argK, phtL) and three polycistronic (phtA, phtD, phtM) operons, whose expression is increased at 18°C, correlating with the production of phaseolotoxin by the bacterium. So far, the regulatory mechanisms involved in phaseolotoxin synthesis are poorly understood and only the requirement of low temperatures for its synthesis has been demonstrated. Therefore, in this study we searched for regulatory proteins that could be involved in the phaseolotoxin synthesis, focusing on the regulation of the phtM operon. Gel shift assays showed that the promoter region of the phtM operon contains binding sites for putative regulatory proteins, which are encoded outside the Pht cluster and are independent of the GacS-GacA two-component system. Deletion assays with the promoter region of the phtM operon show that the binding site for a putative transcription factor is located within a 58bp region. The putative transcription factor of the phtM operon has an apparent molecular mass in the 14-20kDa range. Furthermore, the results demonstrate that the transcription factor recognizes and binds the upstream phtM region as monomer o multimer of a single polypeptide. Our findings provide new insights into the regulatory mechanisms involved in phaseolotoxin production, and suggest that the Pht cluster was integrated into the global regulatory mechanism of P. syringae pv. phaseolicola.

Transcriptional profile of P. syringae pv. phaseolicola NPS3121 at low temperature: physiology of phytopathogenic bacteria.

BACKGROUND: Low temperatures play key roles in the development of most plant diseases, mainly because of their influence on the expression of various virulence factors in phytopathogenic bacteria. Thus far, studies regarding this environmental parameter have focused on specific themes and little is known about phytopathogenic bacteria physiology under these conditions. To obtain a global view regarding phytopathogenic bacteria strategies in response to physiologically relevant temperature changes, we used DNA microarray technology to compare the gene expression profile of the model bacterial pathogen P. syringae pv. phaseolicola NPS3121 grown at 18°C and 28°C. RESULTS: A total of 236 differentially regulated genes were identified, of which 133 were up-regulated and 103 were down-regulated at 18°C compared to 28°C. The majority of these genes are involved in pathogenicity and virulence processes. In general, the results of this study suggest that the expression profile obtained may be related to the fact that low temperatures induce oxidative stress in bacterial cells, which in turn influences the expression of iron metabolism genes. The expression also appears to be correlated with the profile expression obtained in genes related to motility, biofilm production, and the type III secretion system. CONCLUSIONS: From the data obtained in this study, we can begin to understand the strategies used by this phytopathogen during low temperature growth, which can occur in host interactions and disease development.

Spectro-temporal acoustic elements of music interact in an integrated way to modulate emotional responses in pigs.

Music is a complex stimulus, with various spectro-temporal acoustic elements determining one of the most important attributes of music, the ability to elicit emotions. Effects of various musical acoustic elements on emotions in non-human animals have not been studied with an integrated approach. However, this knowledge is important to design music to provide environmental enrichment for non-human species. Thirty-nine instrumental musical pieces were composed and used to determine effects of various acoustic parameters on emotional responses in farm pigs. Video recordings (n = 50) of pigs in the nursery phase (7-9 week old) were gathered and emotional responses induced by stimuli were evaluated with Qualitative Behavioral Assessment (QBA). Non-parametric statistical models (Generalized Additive Models, Decision Trees, Random Forests, and XGBoost) were applied and compared to evaluate relationships between acoustic parameters and pigs' observed emotional responses. We concluded that musical structure affected emotional responses of pigs. The valence of modulated emotions depended on integrated and simultaneous interactions of various spectral and temporal structural components of music that can be readily modified. This new knowledge supports design of musical stimuli to be used as environmental enrichment for non-human animals.

Muscle cyclo-oxygenase-2 pathway contributes to the exaggerated muscle mechanoreflex in rats with congestive heart failure.

Cyclo-oxygenase (COX) enzymes are responsible for the formation from arachidonic acid of prostaglandins, among other metabolites. Prior studies have suggested that inhibition of the COX pathway attenuates the responses of sympathetic nerve activity and blood pressure during static muscle contraction. Static muscle contraction activates the exercise pressor reflex, which in turn increases sympathetic nerve activity and blood pressure. Also, COX products contribute to exaggeration of the exercise pressor reflex in heart failure (HF). This dysfunction of the exercise pressor reflex has previously been shown to be mediated primarily by muscle mechanoreflex overactivity. It is well known that COX-1 and COX-2 are two isoforms of the enzyme that lead to formation of these important biological mediators involved in the muscle reflex. Thus, in the present study, we determined whether the COX-1 and/or COX-2 pathway contribute(s) to the augmented mechanoreflex activity in HF. First, Western blot analysis was employed to examine protein expression of COX-1 and COX-2 in skeletal muscle tissue of control rats and rats with HF induced by myocardial infarction. Our data show that there is no significant difference in COX-1 expression in both experimental groups. However, COX-2 displays significant overexpression in rats with HF compared with control rats (optical density 1.06 ± 0.05 in control and 1.6 ± 0.05 in HF, P < 0.05 versus control). Second, the mechanoreflex was evoked by passive tendon stretch, and the reflex sympathetic and pressor responses to muscle stretch were examined after COX-1 and COX-2 inhibitors (FR-122047 and SC-236) were individually injected into the arterial blood supply of the hindlimb muscles. The results demonstrate that the stretch-evoked reflex responses in rats with HF were significantly attenuated by administration of SC-236, but not by FR-122047, i.e. renal sympathetic nerve activity and mean arterial pressure responses evoked by 0.5 kg of muscle tension were 52.3 ± 8.9% and 19 ± 1.4 mmHg, respectively, in control conditions and 26.4 ± 5.6% and 5.7 ± 1.6 mmHg (P < 0.05 versus control group) after 0.25 mg kg(-1) of SC-236. Muscle stretch-evoked renal sympathetic nerve activity and mean arterial pressure responses were 51.8 ± 8.2% and 18.7 ± 1.2 mmHg, respectively, in control conditions and 48.3 ± 5.3% and 17.5 ± 1.9 mmHg (P > 0.05 versus control group) after 1.0 mg kg(-1) of FR-122047. Accordingly, the results obtained from this study support our hypothesis that heightened COX-2 expression within the hindlimb muscles contributes to the exaggerated muscle mechanoreflex in congestive HF.

Music modulates emotional responses in growing pigs.

There is a lack of clarity on whether pigs can emotionally respond to musical stimulation and whether that response is related to music structure. Qualitative Behavioral Assessment (QBA) was used to evaluate effects of 16 distinct musical pieces (in terms of harmonic structure) on emotional responses in nursery pigs (n = 30) during four periods: "habituation", "treatments", "breaks" and "final". Data were evaluated using Principal component analysis (PCA). Two principal components (PC) were considered in the analysis: PC1, characterized as a positive emotions index, included the emotional responses content, playful, sociable, and happy, whereas PC2, characterized as a negative emotions index, included fearful, inquisitive, and uneasy with positive loadings, and relaxed and calm with negative loadings. Musical stimulation (treatment) increased (P < 0.01) both emotional indices, compared to other periods and this response was influenced by harmonic characteristics of the music. We concluded that pigs have a wide variety of emotional responses, with different affective states related to the music structure used, providing evidence of its potential use as environmental enrichment for this species.

Integration Host Factor (IHF) binds to the promoter region of the phtD operon involved in phaseolotoxin synthesis in P. syringae pv. phaseolicola NPS3121.

BACKGROUND: Pseudomonas syringae pv. phaseolicola, the causal agent of halo blight disease in beans, produces a toxin known as phaseolotoxin, in whose synthesis participate a group of genes organized within the genome in a region known as the "Pht cluster". This region, which is thought to have been acquired by horizontal gene transfer, includes 5 transcriptional units, two monocistronic (argK, phtL) and three polycistronic (phtA, phtD, phtM), whose expression is temperature dependent. So far, the regulatory mechanisms involved in phaseolotoxin synthesis have not been elucidated and the only well-established fact is the requirement of low temperatures for its synthesis. In this work, we searched for regulatory proteins that could be involved in phaseolotoxin synthesis, focusing on the regulation of the phtD operon. RESULTS: In this study we identified the global regulator IHF (Integration Host Factor), which binds to the promoter region of the phtD operon, exerting a negative effect on the expression of this operon. This is the first regulatory protein identified as part of the phaseolotoxin synthesis system. Our findings suggest that the Pht cluster was similarly regulated in the ancestral cluster by IHF or similar protein, and integrated into the global regulatory mechanism of P. syringae pv. phaseolicola, after the horizontal gene transfer event by using the host IHF protein. CONCLUSION: This study identifies the IHF protein as one element involved in the regulation of phaseolotoxin synthesis in P. syringae pv. phaseolicola NPS3121 and provides new insights into the regulatory mechanisms involved in phaseolotoxin production.

Exogenous Application of Polycationic Nanobactericide on Tomato Plants Reduces the Candidatus Liberibacter Solanacearum Infection.

Candidatus Liberibacter solanacearum (CaLso) is associated with diseases in tomato crops and transmitted by the tomato psyllid Bactericera cockerelli. A polymeric water-dispersible nanobactericide (PNB) was evaluated against CaLso as a different alternative. PNB is a well-defined polycationic diblock copolymer designed to permeate into the vascular system of plants. Its assessment under greenhouse conditions was carried out with tomato plants previously infected with CaLso. Using a concentration as low as 1.0 mg L-1, a small but significant reduction in the bacterial load was observed by real-time qPCR. Thus, to achieve an ecologically friendly dosage and set an optimum treatment protocol, we performed experiments to determine the effective concentration of PNB to reduce ~65% of the initial bacterial load. In a first bioassay, a 40- or 70-fold increase was used to reach that objective. At this concentration level, other bioassays were explored to determine the effect as a function of time. Surprisingly, a real reduction in the symptoms was observed after three weeks, and there was a significant decrease in the bacterial load level (~98%) compared to the untreated control plants. During this period, flowering and formation of tomato fruits were observed in plants treated with PNB.

Transcriptional profile of Pseudomonas syringae pv. phaseolicola NPS3121 in response to tissue extracts from a susceptible Phaseolus vulgaris L. cultivar.

BACKGROUND: Pseudomonas syringae pv. phaseolicola is a Gram-negative plant-pathogenic bacterium that causes "halo blight" disease of beans (Phaseolus vulgaris L.). This disease affects both foliage and pods, and is a major problem in temperate areas of the world. Although several bacterial genes have been determined as participants in pathogenesis, the overall process still remains poorly understood, mainly because the identity and function of many of the genes are largely unknown. In this work, a genomic library of P. syringae pv. phaseolicola NPS3121 was constructed and PCR amplification of individual fragments was carried out in order to print a DNA microarray. This microarray was used to identify genes that are differentially expressed when bean leaf extracts, pod extracts or apoplastic fluid were added to the growth medium. RESULTS: Transcription profiles show that 224 genes were differentially expressed, the majority under the effect of bean leaf extract and apoplastic fluid. Some of the induced genes were previously known to be involved in the first stages of the bacterial-plant interaction and virulence. These include genes encoding type III secretion system proteins and genes involved in cell-wall degradation, phaseolotoxin synthesis and aerobic metabolism. On the other hand, most repressed genes were found to be involved in the uptake and metabolism of iron. CONCLUSION: This study furthers the understanding of the mechanisms involved, responses and the metabolic adaptation that occurs during the interaction of P. syringae pv. phaseolicola with a susceptible host plant.

Endogenously generated hydrogen peroxide induces apoptosis via mitochondrial damage independent of NF-kappaB and p53 activation in bovine embryos.

Hydrogen peroxide (H(2)O(2)) has been implicated as a key molecule in arresting embryonic development; however, its mechanism of action is not fully established. The aim of the present study was to determine the chronological generation of H(2)O(2) from oocyte to morula, and to examine the relationship of H(2)O(2) with loss of mitochondrial membrane potential, nuclear factor kappa-B (NF-kappaB), p53, caspase-3 activation, and cell death in bovine embryos in vitro. Accordingly, superoxide anion radicals were detected between 32 and 120 h after in vitro fertilization, but higher percentages of oxygen radicals were found in non-competent embryos (n=73, 22 to 34%) than in competent embryos (n=73, 0 to 1%; P<0.005). Similarly, H(2)O(2) levels were higher in non-competent embryos (n=249, 39 to 71%) than in competent embryos (n=278, 0 to 3.4%) at all developmental stages tested (P<0.005). The percentage of cells with apoptotic morphology were higher in non-competent embryos (n=411, 3 to 54%) than in competent embryos (n=306, 0 to 0.6%; P<0.005). Based on assessment of mitochondrial membrane potential, competent embryos (n=305) had the highest percentages of JC-1 staining (31 to 50%) when compared with non-competent embryos (n=411; 1 to 15%, P<0.005). The percentage of activation of general caspases was different in non-competent embryos (n=291, 15 to 57%) when compared to competent embryos (n=304, 0 to 0.5%; P<0.005). Pharmacological inhibition of caspase-3, NF-kappaB and p53 triggered aberrant embryo cytoplasmic fragmentation with and without nuclei. We concluded that the sequential mechanism of O(2)(-) and H(2)O(2) generation, mitochondrial damage, caspase activation, and apoptotic morphology might be responsible for the developmental arrest of preimplantation embryos.

Temperature-mediated biosynthesis of the phytotoxin phaseolotoxin by Pseudomonas syringae pv. phaseolicola depends on the autoregulated expression of the phtABC genes.

Pseudomonas syringae pv. phaseolicola produces phaseolotoxin in a temperature dependent manner, being optimally synthesized between 18°C and 20°C, while no detectable amounts are present above 28°C. The Pht cluster, involved in the biosynthesis of phaseolotoxin, contains 23 genes that are organized in five transcriptional units. The function of most of the genes from the Pht cluster is still unknown and little information about the regulatory circuitry leading to expression of these genes has been reported. The purpose of the present study was to investigate the participation of pht genes in the regulation of the operons coded into the Pht cluster. We conducted Northern blot, uidA fusions and reverse transcription-PCR assays of pht genes in several mutants unable to produce phaseolotoxin. This allowed us to determine that, in P. syringae pv. phaseolicola NPS3121, genes phtABC are essential to prevent their own expression at 28°C, a temperature at which no detectable amounts of the toxin are present. We obtained evidence that the phtABC genes also participate in the regulation of the phtD, phtM and phtL operons. According to our results, we propose that PhtABC and other Pht product activities could be involved in the synthesis of the sulfodiaminophosphinyl moiety of phaseolotoxin, which indirectly could be involved in the transcriptional regulation of the phtA operon.

Physical map of the chromosome of the phytopathogenic bacterium Pseudomonas syringae pv. phaseolicola.

Pseudomonas syringae pv. phaseolicola (P.s. phaseolicola) is one of about 45 recognized pathovars within the P. syringae group and is the causal agent of halo-blight disease of beans. DNA from this bacterium digested to completion with two different restriction enzymes, PacI and PmeI, yielded 15 and 16 fragments, respectively. These were separated using PFGE and sized by comparison to known molecular mass markers. The P.s. phaseolicola chromosome was determined to be approximately 5.64 Mb in size. To link the different fragments obtained into a circular chromosome map for both enzymes, 150 random Tn5 mutants of P.s. phaseolicola were used as a source of DNA and the identification of the band carrying the transposon 'tag' in each mutant was done after PFGE and Southern hybridization of a complete chromosomal digestion using a Tn5 probe. Partial digestions of DNA from different Tn5 mutants 'tagging' specific bands were then generated and the complete and partial products of the digestion separated by PFGE and identified with a Tn5 probe. By calculating the size of the partial products, it was then possible to link different bands into a physical map. This is the first report on the construction of a physical map of a member of the P. syringae group and should be invaluable for molecular genetic analysis in this species and in evolutionary or taxonomic studies when compared to similar data obtained for any of the other recognized pathovars.

[Non-bacterial thrombotic endocarditis as paraneoplastic manifestation of pulmonary adenocarcinoma]. / Endocarditis trombótica no bacteriana como manifestación paraneoplásica de adenocarcinoma pulmonar.

Non bacterial thrombotic endocarditis is characterized by the presence of non infected vegetation in aortic or mitral valves associated with systemic arterial emboli. Non-bacterial thrombotic endocarditis is a common complication of neoplastic diseases: adenocarcinoma of the lung, another adenocarcinomas, myeloma, lymphoma, leukemia, carcinoma of the pancreas, breast, cervix, colon and stomach. We report a case of non-bacterial thrombotic endocarditis localized in the aortic and mitral valves and systemic emboli as the first manifestation of adenocarcinoma of the lung.

The PhtL protein of Pseudomonas syringae pv. phaseolicola NPS3121 affects the expression of both phaseolotoxin cluster (Pht) and Non-Pht encoded genes.

Pseudomonas syringae pv. phaseolicola, the causal agent of halo blight disease in bean, produces a toxin known as phaseolotoxin, whose synthesis involves the products of some of the genes found within the Pht region. This region, considered a pathogenicity island, comprises 23 genes arranged in five transcriptional units: two single-gene units (argK, phtL) and three arranged as operons (phtA, phtD, phtM), most with unknown function. In P. syringae pv. phaseolicola, maximal expression of most of the genes encoded in the Pht region and the synthesis of phaseolotoxin require the product of the phtL gene, of unknown function but that has been proposed to have a regulatory role. In order to evaluate the role of phtL gene in P. syringae pv. phaseolicola, we performed a comparative transcriptional analysis with the wild type and a phtL(-) mutant strains using microarrays. The microarray data analysis showed that PhtL regulates the expression not only of genes within the Pht region, but also alters the expression of genomic genes outside it, indicating that this gene has been integrated into the regulatory machinery of the bacterium. The expression changes of many of those genes were confirmed by RT-PCR. This study also demonstrated the importance of the PhtL protein in the process of iron response, and suggests that the effect of PhtL on the expression of pathogenicity related, respiration and oxidative stress genes, observed in this study, appears to be indirect through its influence on the Fur protein expression.

Genetic diversity of four broodstocks of tilapia (Oreochromis sp.) from Antioquia, Colombia / Diversidad genética de cuatro grupos de reproductores de tilapia (Oreochromis sp.) de Antioquia, Colombia / Diversidade genética de quatro estoques de reprodutores de tilápia (Oreochromis sp.) de Antioquia, Colombia

Abstract Background: Tilapia is the most farmed fish in Colombia. However, the genetic diversity and structure of broodstocks in the hatcheries of Antioquia province remains unknown. Objective: To analyze the genetic diversity and structure of one Nile and three red tilapia broodstocks in Antioquia, Colombia. Methods: Fish were genotyped using 24 microsatellite markers of 13 linkage groups in five multiple reactions. Genetic diversity metrics were estimated and null alleles were detected. Analysis of Molecular Variance and analysis of number of clusters were used to describe the relationship between broodstocks. Results: Two microsatellites could not be amplified, and 22 were polymorphic. Average number of alleles per locus ranged 5.77 to 7.91. Locus UNH211 had the most alleles (17), whereas OMO032 had the fewest (4). Except for GM234 and OMO032, the analyzed loci had at least one private allele per population. Average effective number of alleles (3.37-4.03) was always less than the number of observed alleles. Significant deviations from Hardy-Weinberg equilibrium with heterozygote deficiencies were registered. Nine markers showed evidence of null alleles. The expected heterozygosity (0.65 to 0.67 per broodstock) was significantly higher than the observed heterozygosity (0.601 to 0.649) in the four populations. The fixation index for all broodstocks (excluding null alleles) was 0.0766 (95% confidence interval, 0.05092 to 0.10289). According to the molecular variance analysis, the greatest variation was between individuals rather than between groups of broodstocks or individuals within broodstocks. The genetic distance between the Nile and red broodstocks ranged from 0.43 to 0.54. Conclusions: Overall, these findings provide baseline information about the genetic diversity and structure of tilapia broodstocks in Antioquia, Colombia, useful for the management of hatcheries.

Resumen Antecedentes: La tilapia es el pez más cultivado en Colombia; sin embargo, hay gran desconocimiento sobre la estructura genetica actual de los reproductores. Objetivo: Analizar la diversidad y estructura genética de los reproductores de tres granjas de tilapia roja y una de tilapia Nilótica en Antioquia, Colombia. Métodos: Se utilizaron 24 microsatélites de 13 grupos de ligamiento amplificados en cinco reacciones múltiples. Se calcularon diferentes medidas de diversidad y se detectaron alelos nulos. Se utilizó un análisis de varianza molecular y uno de número de grupos para describir las relaciones entre las granjas de reproductores. Resultados: Dos marcadores no fueron amplificados y los 22 restantes fueron polimórficos. El promedio de alelos por locus varió entre 5,77 y 7,91. El mayor número de alelos (17) se encontró en el locus UNH 211, mientras que el menor se observó en OMO032 (cuatro). Veinte loci presentaron por lo menos un alelo privado. El número de alelos efectivos promedio fue menor al número de alelos observado y estuvo entre 3,37 y 4,03. Se registraron desviaciones significativas en el equilibrio Hardy-Weinberg, en su mayoría con deficiencias de heterocigotos. Se encontraron evidencias de alelos nulos en nueve marcadores. La heterocigosidad observada estuvo entre 0,601 y 0,649. El índice de fijación fue de 0.0766 (intervalo de confianza de 95%, entre 0,05092 y 0,10289). Según el análisis de varianza molecular, la mayor fuente de variación se encontró entre individuos. El valor de la distancia de Nei entre los reproductores Nilóticos y rojos estuvo entre 0,43 y 0,54. Conclusión: Los resultados de la presente investigación proveen una línea base acerca de la diversidad y estructura genética de los reproductores de tilapia en Antioquia, Colombia, y son útiles para el manejo de granjas dedicadas a la reproducción de tilapia.

Resumo Antecedentes: A tilápia é o peixe mais cultivado na Colômbia. É importante examinar a diversidade genética de peixes reprodutores. Objetivo: Avaliar a diversidade e estrutura genética de três estoques de reprodutores de tilápias vermelhas e um de tilápia Nilótica em Antioquia, Colômbia. Métodos: Utilizaram-se 24 microssatélites de 13 grupos de ligação em cinco reações múltiplas. Métricas de diversidade genética foram estimadas e alelos nulos foram detectados. Análise da Variância Molecular e análise do número de clusters foram utilizados para descrever a relação entre os estoques. Resultados: Dois marcadores não foram amplificados e vinte e dois microssatélites analisados mostraram-se polimórficos. O número médio de alelos por locus variou entre 5,77 e 7,91. O Locus UNH211 apresentou o maior número de alelos (17), enquanto o OMO032 apresentou o menor número (4). Exceto GM234 e OMO032, os loci analisados mostrou um pelo menos um alelo privado por população. O número efetivo médio de alelos (3,37-4,03) foi sempre menor do que o número de alelos observados. Foram observados desvios significativos do equilíbrio Hardy-Weinberg e deficiência de heterozigotos. Nove loci mostraram evidências de alelos nulos. A heterozigosidade esperada (0,6504-0,6748 por população) foi significativamente maior do que a heterozigosidade observada (0,601-0,649). O índice de fixação foi de 0,0766 (intervalo de confiança de 95%, 0,05092-0,10289). De acordo com a análise da variância molecular, a maior variação foi entre indivíduos. Adistância genética entre o Nilo e os reprodutores vermelhos variou de 0,43 a 0,54. Conclusão: No geral, esses resultados fornecem informação básica sobre sobre diversidade e estrutura genética de reprodutores de tilápia em Antioquia, Colômbia, e são significativos para o manejo de plantéis de reprodutores.

A 14-20 kDa protein binds to the upstream region of the phtM operon involved in the synthesis of phaseolotoxin in Pseudomonas syringae pv. phaseolicola NPS3121 / Una proteína de 14-20 kDa se une a la región río arriba del operón phtM involucrado en la síntesis de faseolotoxina en Pseudomonas syringae pv. phaseolicola NPS3121

Pseudomonas syringae pv. phaseolicola is a phytopathogenic bacterium in beans that produces a phytotoxin called phaseolotoxin, in whose synthesis a group of genes that belong to the "Pht cluster" are involved. This cluster comprises 23 genes arranged in 5 transcriptional units, two monocistronic (argK, phtL) and three polycistronic (phtA, phtD, phtM) operons, whose expression is increased at 18°C, correlating with the production of phaseolotoxin by the bacterium. So far, the regulatory mechanisms involved in phaseolotoxin synthesis are poorly understood and only the requirement of low temperatures for its synthesis has been demon strated. Therefore, in this study we searched for regulatory proteins that could be involved in the phaseolotoxin synthesis, focusing on the regulation of the phtM operon. Gel shift assays showed that the promoter region of the phtM operon contains binding sites for putative regulatory proteins, which are encoded outside the Pht cluster and are independent of the GacS-GacA two-component system. Deletion assays with the promoter region of the phtM operon show that the binding site for a putative transcription factor is located within a 58 bp region. The putative transcription factor of the phtM operon has an apparent molecular mass in the 14-20 kDa range. Furthermore, the results demonstrate that the transcription factor recognizes and binds the upstream phtM region as monomer o multimer of a single polypeptide. Our findings provide new insights into the regulatory mechanisms involved in phaseolotoxin production, and suggest that the Pht cluster was integrated into the global regulatory mechanism of P. syringae pv. phaseolicola.

Expression of the gene for resistance to phaseolotoxin (argK) depends on the activity of genes phtABC in Pseudomonas syringae pv. phaseolicola.

The bacterium Pseudomonas syringae pv. phaseolicola produces phaseolotoxin in a temperature dependent manner, being optimally produced between 18°C and 20°C, while no detectable amounts are present above 28°C. Phaseolotoxin is an effective inhibitor of ornithine carbamoyltransferase (OCTase) activity from plant, mammalian and bacterial sources and causes a phenotypic requirement for arginine. To protect the cell from its own toxin, P. syringae pv. phaseolicola synthesizes a phaseolotoxin-resistant OCTase (ROCT). The ROCT is the product of the argK gene and is synthesized only under conditions leading to phaseolotoxin synthesis. The argK gene is included in a chromosomal fragment named Pht cluster, which contains genes involved in the synthesis of phaseolotoxin. The aim of the present work was to investigate the possible involvement of other genes included in the Pht cluster in the regulation of gene argK. We conducted transcriptional analyses of argK in several mutants unable to produce phaseolotoxin, transcriptional fusions and electrophoretic mobility shift assays, which allowed us to determine that genes phtABC, located within the Pht cluster, participate in the transcriptional repression of gene argK at temperatures not permissive for phaseolotoxin biosynthesis. This repression is mediated by a protein present in both toxigenic and nontoxigenic strains of P. syringae and in E. coli, and requires the coordinated participation of phtA, phtB and phtC products in order to carry out an efficient argK repression.