Presence of Indicator Bacteria and Occurrence of Potentially Pathogenic Nontuberculous Mycobacteria Species in Packaged Ice Cubes in Central Mexico.

This study aimed to determine the bacteriological quality and presence of diarrheagenic Escherichia coli pathotypes (DEP) and nontuberculous mycobacteria (NTM) species in 85 packaged ice samples from 12 different states of central Mexico. Three samples had a pH of 9.8 and therefore fell outside of the acceptable range for pH. All samples were positive for aerobic-mesophilic bacteria, with limits ranging from 1 to 3.47 log CFU/mL. In total, 35, 11, and 3 ice samples were positive for total coliforms (TC), fecal coliforms (FC), and E. coli, respectively. In the samples, the TC concentration ranged from <1.1 to >23 MPN/100 mL and from <1.1 to 23 MPN/100 mL for FC and E. coli. In total, 38 (44.7%) ice samples were outside of Mexico's official guidelines. None of the 12 E. coli strains isolated from the three ice samples belonged to DEP. NTM were recovered from 20 ice samples and included M. neoaurum (n = 7), M. porcinum (n = 2), M. flavescens (n = 2), M. fortuitum (n = 1), M. abscessus (n = 1), M. senegalense (n = 1), M. conceptionense (n = 1), and M. sp. (n = 1). In the remaining four samples, two NTM were isolated simultaneously. Thus, we recommend that producers should evaluate the microbiological quality of purified water used as a raw material as well as that of the final product, the ice should be packed in thick bags to avoid stretching and tearing during transportation or storage to prevent environmental contamination of ice, personnel involved in the production, and handling of ice should be trained in relative hygiene matters and how ice-machines should be cleaned and disinfected and the implementation of hazard analysis and critical control points must be applied throughout the chain of production. Finally, regular inspection by the authorities is also of great importance. These recommendations can be applied in different countries with low microbiological quality packaged ice.

Microbial-Based Biofungicides Mitigate the Damage Caused by Fusarium oxysporum f. sp. cubense Race 1 and Improve the Physiological Performance in Banana.

Fusarium wilt of banana (FWB) is the most limiting disease in this crop. The phytosanitary emergency caused by FWB since 2019 in Colombia has required the development of ecofriendly control methods. The aim of this study was to test the effectiveness of microbial-based biofungicides against FWB caused by Fusarium oxysporum f. sp. cubense race 1 (Foc R1) and correlate such effect with plant physiological parameters. Five Trichoderma (T1 to T4 and T9) and four Bacillus (T5 to T8)-based biofungicides were evaluated in pot experiments. In vitro, dual confrontation tests were also carried out to test whether the in vitro effects on Foc growth were consistent with the in vivo effects. While Trichoderma-based T3, T4, and T9, and Bacillus-based T8, significantly reduced the growth of Foc R1 in vitro, Trichoderma-based T1, T3, T4, and T9 temporarily reduced the Foc population in the soil. However, the incidence progress of FWB was significantly reduced by Bacterial-based T7 (74% efficacy) and Trichoderma-based T2 (50% efficacy). The molecular analysis showed that T7 prevented the inner tissue colonization by Foc R1 in 80% of inoculated plants. The T2, T4, T7, and T9 treatments mitigated the negative effects caused by Foc R1 on plant physiology and growth. Our data allowed us to identify three promising treatments to control FWB, reducing the progress of the disease, delaying the colonization of inner tissue, and mitigating physiological damages. Further studies should be addressed to determine the modes of action of the biocontrol agents against Foc and validate the utilization in the field.

Genetic Diversity of Mycobacterium tuberculosis Strains Isolated from HIV-Infected Patients in Mexico.

There has been very limited investigation regarding the genetic diversity of Mycobacterium tuberculosis (MTb) strains isolated from human immunodeficiency virus (HIV)-infected patients in Mexico. In this study, we isolated 93 MTb strains from pulmonary and extrapulmonary samples of HIV-infected patients treated in a public hospital in Mexico City to evaluate the genetic diversity using spoligotyping and mycobacterial interspersed repetitive unit-variable-number tandem-repeat (MIRU-VNTR) typing (based on 24 loci). The cohort comprised 80 male and 13 female individuals. There was a positive correlation between a high HIV viral load (>100,000 copies) and extrapulmonary tuberculosis (TB) (r = 0.306, p = 0.008). Lineage 4 was the most frequent lineage (79 strains). In this lineage, we found the H clade (n = 24), including the Haarlem, H3, and H1 families; the T clade (n = 22), including T1 and T2; the X clade (n = 15), including X1 and X3; the LAM clade (n = 14), including LAM1, LAM2, LAM3, LAM6, and LAM9; the S clade (n = 2); Uganda (n = 1); and Ghana (n = 1). We also found 12 strains in the EAI clade belonging to lineage 1, including the EAI2-Manila and EAI5 families. Interestingly, we identified one strain belonging to the Beijing family, which is part of lineage 2. One strain could not be identified. This study reports high genetic diversity among MTb strains, highlighting the need for a molecular epidemiological surveillance system that can help to monitor the spread of these strains, leading to more appropriate measures for TB control in HIV-infected patients.

Infection of Fetal Membranes with Mycobacterium tuberculosis and Tissue Processing to Isolate RNA for Expression Analysis.

This chapter outlines the methodology employed to infect the chorionic and amniotic membranes with Mycobacterium tuberculosis during pregnancy. Particularly, congenital tuberculosis, a rare and serious condition associated with cases in neonates and reactivation of latent tuberculosis in pregnant mothers, is interesting to study. Understanding the mechanisms of infection and the response of fetal membranes is crucial for developing effective treatments in these cases, which will promote better neonatal and maternal health in situations of tuberculosis during pregnancy. Establishing a standardized infection model in the chorioamniotic membranes is imperative, followed by a treatment protocol for isolating both cellular and mycobacterial RNA. This will enable the expression analysis during the maternal-fetal interface interaction with M. tuberculosis. The proposed methodology might be invaluable for qRT-PCR, microarrays, and sequencing research.

Seguimiento farmacoterapéutico a pacientes polimedicados / Pharmaceutical care to polimedicated patients

Los servicios profesionales farmacéuticos asistenciales contribuyen a un mejor control de los problemas de salud, sobre todo en pacientes polimedicados, y a una optimización de los recursos sanitarios, lo cual conlleva una menor prescripción de medicamentos y un menor número de visitas a los centros sanitarios. Formaron parte de este estudio 78 pacientes, siendo el sexo femenino el más prevalente, con una edad media 72,49 ± 13,92 años; de ellos, 60 pacientes eran mayores de 65 años y, por tanto, considerados pacientes geriátricos. Fueron incluidos en un servicio de sistemas personalizados de dosificación y, posteriormente, se realizó un seguimiento farmacoterapéutico. Se detectaron 450 problemas relacionados con los medicamentos siendo los más frecuentes los asociados al paciente: falta de conocimiento de uso, mala adherencia terapéutica, errores en la administración…, siendo la probabilidad de presentarlos mayor en el sexo masculino. Asociados a estos problemas con los medicamentos se detectaron 160 resultados negativos asociados a la medicación; solo 5 de ellos no fueron resueltos durante la fase estudio, los más frecuentes fueron los de inefectividad no cuantitativa, es decir, aquellos que no se resuelven con una modificación de la dosis farmacológica sino incluyendo o eliminando otros principios activos. En conclusión, el servicio de sistema personalizado de dosificación precisa de una revisión de la medicación como, por ejemplo, el seguimiento farmacoterapéutico, ya que ambos permiten optimizar el tratamiento de los pacientes polimedicados, así como, un aumento del control de los problemas de salud.(AU)

Professional pharmaceutical care services contribute to better health problem control, especially in polymedicated patients, and to the optimization of health resources, which leads to less medication prescription and fewer visits to healthcare centers. This study included 78 patients, with the female sex being the most prevalent, with an average age of 72.49 ± 13.92 years; of these, 60 patients were over 65 years of age and, herefore, considered geriatric patients. They were included in a personalized medication dispensing service and, subsequently, a pharmacotherapeutic follow-up was carried out. 450 medication-related problems were detected, the most frequent of which were associated with the patient: lack of knowledge of use, poor therapeutic adherence, administration errors, etc., with the probability of presenting them being higher in the male sex. Associated with these medication problems, 160 negative medication-related results were detected; only 5 of them were not resolved during the study phase, the most frequent being those of non-quantitative ineffectiveness, that is, those that are not resolved with a modification of the pharmacological dose but by including or eliminating other active ingredients. In conclusion, the personalized medication dispensing service requires a medication review, such as pharmacotherapeutic follow-up, as both allow the optimization of the treatment of polymedicated patients, as well as an increase in the control of health problems.(AU)

Phenotypic and Genotypic Drug Resistance of Mycobacterium tuberculosis Strains Isolated from HIV-Infected Patients from a Third-Level Public Hospital in Mexico.

BACKGROUND: Drug-resistant tuberculosis (TB) is associated with higher mortality rates in patients with human immunodeficiency virus (HIV). In Mexico, the number of deaths due to TB among the HIV-positive population has tripled in recent years. METHODS: Ninety-three Mycobacterium tuberculosis strains isolated from the same number of HIV-infected patients treated in a public hospital in Mexico City were studied to determine the drug resistance to first- and second-line anti-TB drugs and to identify the mutations associated with the resistance. RESULTS: Of the 93 patients, 82.7% were new TB cases, 86% were male, and 73% had extrapulmonary TB. Most patients (94%) with a CD4 T-lymphocyte count <350 cells/mm3 were associated with extrapulmonary TB (p <0.0001), whilst most patients (78%) with a CD4 T-lymphocyte count >350 cells/mm3 were associated with pulmonary TB (p = 0.0011). Eighty-two strains were pan-susceptible, four mono-resistant, four poly-resistant, two multidrug-resistant, and one was extensively drug-resistant. In the rifampicin-resistant strains, rpoB S531L was the mutation most frequently identified, whereas the inhA C15T and katG S315T1 mutations were present in isoniazid-resistant strains. The extensively drug-resistant strain also contained the mutation gyrA D94A. CONCLUSIONS: These data highlight the need to promptly diagnose the drug resistance of M. tuberculosis among all HIV-infected patients by systematically offering access to first- and second-line drug susceptibility testing and to tailor the treatment regimen based on the resistance patterns to reduce the number of deaths in HIV-infected patients.

Influence of feeding practices in the composition and functionality of infant gut microbiota and its relationship with health.

Establishing the infant's gut microbiota has long-term implications on health and immunity. Breastfeeding is recognized as the best practice of infant nutrition in comparison with formula feeding. We evaluated the effects of the primary feeding practices by analyzing the infant growth and the potential association with gut diseases. A cross-sectional and observational study was designed. This study included 55 mothers with infants, who were divided according to their feeding practices in breastfeeding (BF), formula feeding (FF), and combined breast and formula feeding (CF). Anthropometric measurements of the participants were recorded. Additionally, non-invasive fecal samples from the infants were collected to analyze the microbiota by sequencing, immunoglobulin A (IgA) concentration (ELISA), and volatile organic compounds (gas chromatography with an electronic nose). Results showed that the microbiota diversity in the BF group was the highest compared to the other two groups. The IgA levels in the BF group were twice as high as those in the FF group. Moreover, the child´s growth in the BF group showed the best infant development when the data were compared at birth to the recollection time, as noted by the correlation with a decreased concentration of toxic volatile organic compounds. Interestingly, the CF group showed a significant difference in health status when the data were compared with the FF group. We conclude that early health practices influence children's growth, which is relevant to further research about how those infants' health evolved.

Isoreticular Chemistry and Applications of Supramolecularly Assembled Copper-Adenine Porous Materials.

The useful concepts of reticular chemistry, rigid and predictable metal nodes together with strong and manageable covalent interactions between metal centers and organic linkers, have made the so-called metal-organic frameworks (MOFs) a flourishing area of enormous applicability. In this work, the extension of similar strategies to supramolecularly assembled metal-organic materials has allowed us to obtain a family of isoreticular compounds of the general formula [Cu7(µ-adeninato-κN3:κN9)6(µ3-OH)6(µ-OH2)6](OOC-R-COO)·nH2O (R: ethylene-, acetylene-, naphthalene-, or biphenyl-group) in which the rigid copper-adeninato entities and the organic dicarboxylate anions are held together not by covalent interactions but by a robust and flexible network of synergic hydrogen bonds and π-π stacking interactions based on well-known supramolecular synthons (SMOFs). All compounds are isoreticular, highly insoluble, and water-stable and show a porous crystalline structure with a pcu topology containing a two-dimensional (2D) network of channels, whose dimensions and degree of porosity of the supramolecular network are tailored by the length of the dicarboxylate anion. The partial loss of the crystallization water molecules upon removal from the mother liquor produces a shrinkage of the unit cell and porosity, which leads to a color change of the compounds (from blue to olive green) if complete dehydration is achieved by means of gentle heating or vacuuming. However, the supramolecular network of noncovalent interactions is robust and flexible enough to reverse to the expanded unit cell and color after exposure to a humid atmosphere. This humidity-driven breathing behavior has been used to design a sensor in which the electrical resistance varies reversibly with the degree of humidity, very similar to the water vapor adsorption isotherm of the SMOF. The in-solution adsorption properties were explored for the uptake and release of the widely employed 5-fluorouracil, 4-aminosalycilic acid, 5-aminosalycilic acid, and allopurinol drugs. In addition, cytotoxicity activity assays were completed for the pristine and 5-fluorouracil-loaded samples.

Thermochemical CO2 Reduction Catalyzed by Homometallic and Heterometallic Nanoparticles Generated from the Thermolysis of Supramolecularly Assembled Porous Metal-Adenine Precursors.

A family of unprecedented supramolecularly assembled porous metal-organic compounds (SMOFs), based on [Cu6M(µ-adeninato)6(µ3-OH)6(µ-H2O)6]2+ cations (MII: Cu, Co, Ni, and Zn) and different dicarboxylate anions (fumarate, benzoate, and naphthalene-2,6-dicarboxylate), have been employed as precursors of catalysts for the thermocatalytic reduction of CO2. The selected metal-organic cation allows us to tune the composition of the SMOFs and, therefore, the features and performance of the final homometallic and bimetallic catalysts. These catalysts were obtained by thermolysis at 600 °C under a N2 atmosphere and consist of big metal particles (10-20 µm) placed on the surface of the carbonaceous matrix and very tiny metal aggregates (<10 nm) within this carbonaceous matrix. The latter are the most active catalytic sites for the CO2 thermocatalytic reduction. The amount of this carbonaceous matrix correlates with the organic content present in the metal-organic precursor. In this sense, CO2 thermocatalytic reduction experiments performed over the homometallic, copper only, catalysts with different carbon contents indicate that above a certain value, the increase of the carbonaceous matrix reduces the overall performance by encapsulating the nanoparticles within this matrix and isolating them from interacting with CO2. In fact, the best performing homometallic catalyst is that obtained from the precursor containing a small fumarate counterion. On the other hand, the structural features of these precursors also provide a facile route to work with a solid solution of nanoparticles as many of these metal-organic compounds can replace up to 1/7 of the copper atoms by zinc, cobalt, or nickel. Among these heterometallic catalysts, the best performing one is that of copper and zinc, which provides the higher conversion and selectivity toward CO. XPS spectroscopy and EDX mappings of the latter catalyst clearly indicate the presence of Cu1-xZnx nanoparticles covered by small ZnO aggregates that provide a better CO2 adsorption and easier CO release sites.

Targeted Expression to Liver of an antimiR-33 Sponge as a Gene Therapy Strategy against Hypercholesterolemia: In Vitro Study.

Atherosclerosis is the leading cause of cardiovascular diseases in Mexico and worldwide. The membrane transporters ABCA1 and ABCG1 are involved in the reverse transport of cholesterol and stimulate the HDL synthesis in hepatocytes, therefore the deficiency of these transporters promotes the acceleration of atherosclerosis. MicroRNA-33 (miR-33) plays an important role in lipid metabolism and exerts a negative regulation on the transporters ABCA1 and ABCG1. It is known that by inhibiting the function of miR-33 with antisense RNA, HDL levels increase and atherogenic risk decreases. Therefore, in this work, a genetic construct, pPEPCK-antimiR-33-IRES2-EGFP, containing a specific antimiR-33 sponge with two binding sites for miR-33 governed under the PEPCK promoter was designed, constructed, and characterized, the identity of which was confirmed by enzymatic restriction, PCR, and sequencing. Hep G2 and Hek 293 FT cell lines, as well as a mouse hepatocyte primary cell culture were transfected with this plasmid construction showing expression specificity of the PEPCK promoter in hepatic cells. An analysis of the relative expression of miR-33 target messengers showed that the antimiR-33 sponge indirectly induces the expression of its target messengers (ABCA1 and ABCG1). This strategy could open new specific therapeutic options for hypercholesterolemia and atherosclerosis, by blocking the miR-33 specifically in hepatocytes.

Organization and Characterization of the Promoter Elements of the rRNA Operons in the Slow-Growing Pathogen Mycobacterium kumamotonense.

The slow-growing, nontuberculous mycobacterium Mycobacterium kumamotonense possesses two rRNA operons, rrnA and rrnB, located downstream from the murA and tyrS genes, respectively. Here, we report the sequence and organization of the promoter regions of these two rrn operons. In the rrnA operon, transcription can be initiated from the two promoters, named P1 rrnA and PCL1, while in rrnB, transcription can only start from one, called P1 rrnB. Both rrn operons show a similar organization to the one described in Mycobacterium celatum and Mycobacterium smegmatis. Furthermore, by qRT-PCR analyses of the products generated from each promoter, we report that stress conditions such as starvation, hypoxia, and cellular infection affect the contribution of each operon to the synthesis of pre-rRNA. It was found that the products from the PCL1 promoter of rrnA play a pivotal role in rRNA synthesis during all stress conditions. Interestingly, the main participation of the products of transcription from the P1 promoter of rrnB was found during hypoxic conditions at the NRP1 phase. These results provide novel insights into pre-rRNA synthesis in mycobacteria, as well as the potential ability of M. kumamotonense to produce latent infections.

The CpxRA two-component system represses gene expression of the heat-labile toxin of enterotoxigenic Escherichia coli.

Enterotoxigenic Escherichia coli (ETEC) strains produce at least one of two types of enterotoxins: the heat-labile (LT) and heat-stable (ST) toxins, which are responsible for the watery secretory diarrhoea that is a hallmark of the human ETEC infection. One regulatory system that controls the transcription of virulence genes in pathogenic bacteria is the CpxRA two-component system (TCS). We reported that the eltAB bicistronic operon, which encodes for the A and B subunits of LT, was repressed for the CpxRA TCS by direct binding of CpxR-P from -12 to +6 bp with respect to the transcription start site of eltAB. Moreover, the Cpx-response activation down-regulated the transcription of eltAB genes, and this negative effect was CpxRA-dependent. Our data show that CpxRA TCS is a negative regulator of the LT, one of the main virulence determinants of ETEC.

Quality Care in Outpatient Chemotherapy. Design and Validation of the Novel Care Quality Ambulatory Instrument / Cuidado con calidad en quimioterapia ambulatoria. Diseño y validación del nuevo instrumento Care Quality Ambulatory / Cuidado com qualidade em quimioterapia ambulatória. Desenho e validação do novo instrumento Care Quality Ambulatory

Introduction: Neither validated scales nor scales translated into Spanish currently exist to evaluate the perception of quality care in people receiving outpatient chemotherapy, despite their usefulness in evaluating the care provided by nurses and in promoting changes in the delivery of these services. Objective: To design and validate the novel Care Quality Ambulatory-I (CQAMB-I) instrument in outpatient chemotherapy services. Materials and methods: This is an instrument validation study carried out in three stages: literature review, design, and validation of the instrument. Content validation was performed with 14 specialists with at least a master's degree and experience in chemotherapy or quality services; face validation was carried out through a pilot test with 31 participants diagnosed with cancer who received curative chemotherapy, and construct validation was carried out through an exploratory factor analysis with 436 users who received outpatient chemotherapy with curative intent. Results: A total of 15 items were removed in the content validation stage; none were removed in face validation. In construct validation, participants had a median age of 56 years, a median treatment time of 5 months, and breast cancer was the most commonly diagnosed form of cancer. Eighteen models were run, 30 items were removed, 6 factors emerged, and a KMO of 0.80 and a Cronbach's alpha of 0.82 were obtained. Conclusion: The CQAMB-I proved to be valid and reliable for the evaluation of quality care in outpatient chemotherapy services. Additionally, it enabled expanding the conception of such care beyond the attributes of structure, process, and outcome.

Introducción: no existen escalas validadas, ni traducidas al español para evaluar la percepción del cuidado con calidad en personas que reciben quimioterapia ambulatoria, a pesar de su utilidad para evaluar la prestación de servicios enfermeros y generar cambios en la entrega de estos. Objetivo: diseñar y validar el nuevo instrumento Care Quality Ambulatory-I (CQAMB-I) en servicios de quimioterapia ambulatoria. Materiales y métodos: estudio de validación de instrumentos en tres fases: revisión de la literatura, diseño y validación del instrumento. Se realizó validación de contenido con 14 expertos con formación mínima de maestría y experiencia en servicios de quimioterapia o calidad; validación facial a través de prueba piloto con 31 participantes con diagnóstico de cáncer que recibían quimioterapia curativa; y validación de constructo por medio de un análisis factorial exploratorio con 436 usuarios que recibían quimioterapia ambulatoria con intención curativa. Resultados: en la validación de contenido se eliminaron 15 ítems; en la validación facial no se eliminó ninguno; y en la validación de constructo los participantes tuvieron una mediana de edad de 56 años, mediana de tiempo de tratamiento de 5 meses y cáncer de mama como el más diagnosticado. Se ejecutaron 18 modelos, se eliminaron 30 ítems, emergieron 6 factores y se obtuvo un KMO de 0.80 y un alpha de Cronbach de 0.82. Conclusión: el instrumento CQAMB-I demostró ser válido y confiable para evaluar el cuidado con calidad en servicios de quimioterapia ambulatoria y, adicionalmente, permitió ampliar la concepción de este más allá de los atributos de estructura, proceso y resultado.

Introdução: não existem escalas validadas nem traduzidas ao espanhol para avaliar a percepção do cuidado com qualidade em pessoas que recebem quimioterapia ambulatória, apesar de sua utilidade para avaliar a prestação de serviços de enfermagem e gerar mudanças na entrega destes. Objetivo: desenhar e validar o novo instrumento Care Quality Ambulatory-I (CQAMB-I) em serviços de quimioterapia ambulatória. Materiais e método: estudo de validação de instrumentos em três fases: revisão da literatura, desenho e validação do instrumento. Foi realizada validação de conteúdo com 14 especialistas com formação mínima de mestrado e experiência em serviços de quimioterapia ou qualidade; validação facial por meio de teste-piloto com 31 participantes com diagnóstico de câncer que recebiam quimioterapia curativa e validação de constructo por meio de uma análise fatorial exploratória com 436 usuários que recebiam quimioterapia ambulatória com intenção curativa. Resultados: na validação de conteúdo, foram eliminados 15 itens; na validação facial, não foi eliminado nenhum; na validação de constructo, os participantes tiveram uma média de idade de 56 anos, média de tempo de tratamento de 5 meses e câncer de mama como o mais diagnosticado. Foram executados 18 modelos, foram eliminados 30 itens, emergidos 6 fatores e obtido um KMO de 0,80 e um alpha de Cronbach de 0,82. Conclusões: o instrumento CQAMB-I demonstrou ser válido e confiável para avaliar o cuidado com qualidade em serviços de quimioterapia ambulatória e, além disso, permitiu ampliar a concepção deste mais além dos atributos de estrutura, processo e resultado.

Identification of a virulent phage infecting species of Nitrosomonas.

In the first and limiting step of nitrification, ammonia (NH3) is oxidised to nitrite (NO2-) by the action of some prokaryotes, including bacteria of the Nitrosomonas genus. A potential approach to nitrification inhibition would be through the application of phages, but until now this method has been unexplored and no virulent phages that infect nitrifying bacteria have been described. In this study, we report the isolation of the first phage infecting some Nitrosomonas species. This polyvalent virulent phage (named ΦNF-1) infected Nitrosomonas europaea, Nitrosomonas communis, and Nitrosomonas nitrosa. Phage ΦNF-1 has the morphology of the Podoviridae family, a dsDNA genome of 41,596 bp and a 45.1 % GC content, with 50 predicted open reading frames. Phage ΦNF-1 was found to inhibit bacterial growth and reduce NH4+ consumption in the phage-treated cultures. The application of phages as biocontrol agents could be a useful strategy for nitrification inhibition without the restrictions associated with chemical inhibitors.

The Two-Component System CpxRA Represses Salmonella Pathogenicity Island 2 by Directly Acting on the ssrAB Regulatory Operon.

The acquisition of Salmonella pathogenicity island 2 (SPI-2) conferred on Salmonella the ability to survive and replicate within host cells. The ssrAB bicistronic operon, located in SPI-2, encodes the SsrAB two-component system (TCS), which is the central positive regulator that induces the expression of SPI-2 genes as well as other genes located outside this island. On the other hand, CpxRA is a two-component system that regulates expression of virulence genes in many bacteria in response to different stimuli that perturb the cell envelope. We previously reported that the CpxRA system represses the expression of SPI-1 and SPI-2 genes under SPI-1-inducing conditions by decreasing the stability of the SPI-1 regulator HilD. Here, we show that under SPI-2-inducing conditions, which mimic the intracellular environment, CpxRA represses the expression of SPI-2 genes by the direct action of phosphorylated CpxR (CpxR-P) on the ssrAB regulatory operon. CpxR-P recognized two sites located proximal and distal from the promoter located upstream of ssrA. Consistently, we found that CpxRA reduces the replication of Salmonella enterica serovar Typhimurium inside murine macrophages. Therefore, our results reveal CpxRA as an additional regulator involved in the intracellular lifestyle of Salmonella, which in turn adds a new layer to the intricate regulatory network controlling the expression of Salmonella virulence genes. IMPORTANCE SPI-2 encodes a type III secretion system (T3SS) that is a hallmark for the species Salmonella enterica, which is essential for the survival and replication within macrophages. Expression of SPI-2 genes is positively controlled by the two-component system SsrAB. Here, we determined a regulatory mechanism involved in controlling the overgrowth of Salmonella inside macrophages. In this mechanism, CpxRA, a two-component system that is activated by extracytoplasmic stress, directly represses expression of the ssrAB regulatory operon; as a consequence, expression of SsrAB target genes is decreased. Our findings reveal a novel mechanism involved in the intracellular lifestyle of Salmonella, which is expected to sense perturbations in the bacterial envelope that Salmonella faces inside host cells, as the synthesis of the T3SS-2 itself.

The Lack of the TetR-Like Repressor Gene BCG_2177c (Rv2160A) May Help Mycobacteria Overcome Intracellular Redox Stress and Survive Longer Inside Macrophages When Surrounded by a Lipid Environment.

Mycobacteria, like other microorganisms, survive under different environmental variations by expressing an efficient adaptive response, oriented by regulatory elements, such as transcriptional repressors of the TetR family. These repressors in mycobacteria also appear to be related to cholesterol metabolism. In this study, we have evaluated the effect of a fatty acid (oleic-palmitic-stearic)/cholesterol mixture on some phenotypic and genotypic characteristics of a tetR-mutant strain (BCG_2177c mutated gene) of M. bovis BCG, a homologous of Rv2160A of M. tuberculosis. In order to accomplish this, we have analyzed the global gene expression of this strain by RNA-seq and evaluated its neutral-lipid storage capacity and potential to infect macrophages. We have also determined the macrophage response by measuring some pro- and anti-inflammatory cytokine expressions. In comparison with wild-type microorganisms, we showed that the mutation in the BCG_2177c gene did not affect the growth of M. bovis BCG in the presence of lipids but it probably modified the structure/composition of its cell envelope. Compared to with dextrose, an overexpression of the transcriptome of the wild-type and mutant strains was observed when these mycobacteria were cultured in lipids, mainly at the exponential phase. Twelve putative intracellular redox balance maintenance genes and four others coding for putative transcriptional factors (including WhiB6 and three TetR-like) were the main elements repeatedly overexpressed when cultured in the presence of lipids. These genes belonged to the central part of what we called the "genetic lipid signature" for M. bovis BCG. We have also found that all these mycobacteria genotypic changes affected the outcome of BCG-infected macrophages, being the mutant strain most adapted to persist longer inside the host. This high persistence result was also confirmed when mutant-infected macrophages showed overexpression of the anti-inflammatory cytokine TGF-ß versus pro-inflammatory cytokines. In summary, the lack of this TetR-like repressor expression, within a lipid environment, may help mycobacteria overcome intracellular redox stress and survive longer inside their host.

Transcriptome dataset of omental and subcutaneous adipose tissues from gestational diabetes patients.

Gestational diabetes (GD) is one of the most prevalent metabolic diseases in pregnant women worldwide. GD is a risk factor for adverse pregnancy outcomes, including macrosomia and preeclampsia. Given the multifactorial etiology and the complexity of its pathogenesis, GD requires advanced omics analyses to expand our understanding of the disease. Next generation RNA sequencing (RNA-seq) was used to evaluate the transcriptomic profile of subcutaneous and omental adipose tissues (AT) collected from patients with gestational diabetes and matched controls. Samples were harvested during cesarean delivery. Results show differences based on anatomical location and provide whole-transcriptome data for further exploration of gene expression patterns unique to GD patients.

Treatment approach to a patient with catamenial epilepsy. case report

ABSTRACT Introduction: Catamenial epilepsy refers to the worsening or exacerbation of seizures due to hormonal changes during the menstrual cycle. It is thought to be secondary to the neuroactive properties of endogenous steroid hormones and the natural cyclic variation in their serum levels throughout the menstrual cycle. Case presentation: A 31-year-old female patient from Bogotá (Colombia) was admitted to the emergency department due to an episode of tonic-clonic seizure associated with the menstrual period. Since the onset of the seizures was related to menstruation (every 28 days), it was established that the patient had structural focal epilepsy with catamenial features. Advantages of medical vs. surgical treatment were discussed during a multidisciplinary medical board and it was decided to start pharmacological treatment with progestogens, which resulted in complete remission of the seizures as established during a follow-up visit. Conclusions: Catamenial epilepsy should be considered as a cause of epilepsy refractory to antiepileptic medications. Furthermore, it should be approached from a multidisciplinary perspective and its management should be focused on improving the patients' quality of life.

RESUMEN Introducción. La epilepsia catamenial se define como un empeoramiento o la exacerbación de las crisis epilépticas en relación con el cambio hormonal durante el ciclo menstrual femenino. Se cree que esta se produce por las propiedades neuroactivas de las hormonas esteroides endógenas y la variación cíclica natural en sus niveles séricos a lo largo de dicho ciclo. Presentación del caso. Mujer de 31 años de Bogotá (Colombia), quien fue llevada al servicio de urgencias por un episodio de crisis epiléptica con convulsiones tonicoclónicas asociado al período menstrual. Debido a que la aparición de las crisis epilépticas se asociaba con la menstruación (cada 28 días), se estableció que la paciente presentaba epilepsia focal estructural de características catameniales. En junta médica multidisciplinar se discutieron las ventajas del manejo médico y el manejo quirúrgico, y se decidió instaurar tratamiento farmacológico con progestágenos, el cual, tras seguimiento, evidenció supresión total de las crisis. Conclusiones. La epilepsia catamenial debe considerarse como una causa de epilepsia refractaria al tratamiento antiepiléptico. Además, su abordaje debe ser multidisciplinario y su tratamiento debe ir enfocado a mejorar la calidad de vida de los pacientes.

Loop-Mediated Isothermal Amplification for Detection of Plant Pathogens in Wheat (Triticum aestivum).

Wheat plants can be infected by a variety of pathogen species, with some of them causing similar symptoms. For example, Zymoseptoria tritici and Parastagonospora nodorum often occur together and form the Septoria leaf blotch complex. Accurate detection of wheat pathogens is essential in applying the most appropriate disease management strategy. Loop-mediated isothermal amplification (LAMP) is a recent molecular technique that was rapidly adopted for detection of plant pathogens and can be implemented easily for detection in field conditions. The specificity, sensitivity, and facility to conduct the reaction at a constant temperature are the main advantages of LAMP over immunological and alternative nucleic acid-based methods. In plant pathogen detection studies, LAMP was able to differentiate related fungal species and non-target strains of virulent species with lower detection limits than those obtained with PCR. In this review, we explain the amplification process and elements of the LAMP reaction, and the variety of techniques for visualization of the amplified products, along with their advantages and disadvantages compared with alternative isothermal approaches. Then, a compilation of analyses that show the application of LAMP for detection of fungal pathogens and viruses in wheat is presented. We also describe the modifications included in real-time and multiplex LAMP that reduce common errors from post-amplification detection in traditional LAMP assays and allow discrimination of targets in multi-sample analyses. Finally, we discuss the utility of LAMP for detection of pathogens in wheat, its limitations, and current challenges of this technique. We provide prospects for application of real-time LAMP and multiplex LAMP in the field, using portable devices that measure fluorescence and turbidity, or facilitate colorimetric detection. New technologies for detection of plant pathogen are discussed that can be integrated with LAMP to obtain elevated analytical sensitivity of detection.

A Recurrent FAM83H Mutation in an Extended Colombian Family and Variable Craniofacial Phenotypes.

Amelogenesis imperfecta (AI) is a collection of rare genetic disorders affecting the quantity and/or quality of the tooth enamel. AI can be classified into three major types according to the clinical phenotype: hypoplastic, hypocalcified, and hypomatured. Among them, the hypocalcified type shows the weakest physical properties, leaving rough and discolored enamel surfaces after tooth eruption. To date, mutations in the FAM83H gene are responsible for the autosomal-dominant hypocalcified AI. In this study, we recruited a four-generation Colombian family with hypocalcified AI and identified a recurrent nonsense mutation in the FAM83H gene (NM_198488.5:c.1289C>A, p.(Ser430 *)) by candidate gene sequencing. Cephalometric analyses revealed the anterior open bite that occurred in the proband is not correlated with the AI in this family.