Effect of plasma-activated water on the supramolecular structure and techno-functional properties of starch: A review.

This review discusses the changes in the multi-scale structure and functionality of starch after its hydrothermal modification using plasma-activated water (PAW). PAW contains reactive species that decrease the pH of the water and increase the oxidation-reduction potential, which promotes the oxidation and degradation of the surface of the starch granules to varying extents, depending on the botanical source and treatment conditions. In this article, we compile the information published so far on the effects of using PAW during heat-moisture and annealing treatments and discuss the results of the substitution of water with PAW on the long and short-range crystallinity, helical order, thermal behavior, functional properties, and digestibility. Additionally, we highlighted the possible application of PAW-modified starches. Finally, we provided an overview of future challenges, suggesting several potential directions to understand the mechanisms behind PAW use for developing sustainable modified starches for the food industry.

Benign Paroxysmal Torticollis.

BACKGROUND: The purpose of this review is to clarify the natural course of benign paroxysmal torticollis (BPT) and update the information on the relationship of this disorder with migraine. BPT belongs to a group of "episodic syndromes that may be associated with migraine" and is diagnosed according to diagnostic criteria of the International Classification of Headache Disorders, 3rd edition. BPT affects infants and young children and is often an underdiagnosed manifestation since it is not recognized in cases with a benign evolution, requiring a careful differential diagnosis. It was first described by Snyder in 1969 as a movement disorder, a cervical dystonia consequent to labyrinthic disorder. MATERIALS AND METHODS: The PubMed and Web of Science databases were consulted from 1968 to 2024, according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) 2020 guidelines. RESULTS: In total, 113 articles were identified, 86 selected, and 25 considered for the purpose of this review. Clinical studies were considered in relation to evolution, cognitive, and motor development; genetic and not genetic etiology; the relationship with migraine with and without aura; vestibular migraine; hemiplegic migraine; and paroxysmal vertigo.

Sodium Tungstate (NaW) Decreases Inflammation and Renal Fibrosis in Diabetic Nephropathy.

BACKGROUND: Diabetic Nephropathy is one of the most severe complications of Diabetes Mellitus and the main cause of end-stage kidney disease worldwide. Despite the therapies available to control blood glucose and blood pressure, many patients continue to suffer from progressive kidney damage. Chronic hyperglycemia is the main driver of changes observed in diabetes; however, it was recently discovered that inflammation and oxidative stress contribute to the development and progression of kidney damage. Therefore, it is important to search for new pharmacological therapies that stop the progression of DN. Sodium tungstate (NaW) is an effective short and long-term antidiabetic agent in both type 1 and type 2 diabetes models. METHODS: In this study, the effect of NaW on proinflammatory signalling pathways, proinflammatory proteins and fibrosis in the streptozotocin (STZ)-induced type 1 diabetic rat model was analysed using histological analysis, western blotting and immunohistochemistry. RESULTS: NaW treatment in diabetic rats normalize parameters such as glycemia, glucosuria, albuminuria/creatinuria, glomerular damage, and tubulointerstitial damage. NaW decreased the proinflammatory signaling pathway NF-κB, inflammatory markers (ICAM-1, MCP-1 and OPN), profibrotic pathways (TGFß1/Smad2/3), reduced epithelial-mesenchymal transition (α -SMA), and decreased renal fibrosis (type IV collagen). CONCLUSION: NaW could be an effective drug therapy for treating human diabetic nephropathy.

The Impact of A3AR Antagonism on the Differential Expression of Chemoresistance-Related Genes in Glioblastoma Stem-like Cells.

Glioblastoma (GB) is the most aggressive and common primary malignant tumor of the brain and central nervous system. Without treatment, the average patient survival time is about six months, which can be extended to fifteen months with multimodal therapies. The chemoresistance observed in GB is, in part, attributed to the presence of a subpopulation of glioblastoma-like stem cells (GSCs) that are characterized by heightened tumorigenic capacity and chemoresistance. GSCs are situated in hypoxic tumor niches, where they sustain and promote the stem-like phenotype and have also been correlated with high chemoresistance. GSCs have the particularity of generating high levels of extracellular adenosine (ADO), which causes the activation of the A3 adenosine receptor (A3AR) with a consequent increase in the expression and activity of genes related to chemoresistance. Therefore, targeting its components is a promising alternative for treating GB. This analysis determined genes that were up- and downregulated due to A3AR blockades under both normoxic and hypoxic conditions. In addition, possible candidates associated with chemoresistance that were positively regulated by hypoxia and negatively regulated by A3AR blockades in the same condition were analyzed. We detected three potential candidate genes that were regulated by the A3AR antagonist MRS1220 under hypoxic conditions: LIMD1, TRIB2, and TGFB1. Finally, the selected markers were correlated with hypoxia-inducible genes and with the expression of adenosine-producing ectonucleotidases. In conclusion, we detected that hypoxic conditions generate extensive differential gene expression in GSCs, increasing the expression of genes associated with chemoresistance. Furthermore, we observed that MRS1220 could regulate the expression of LIMD1, TRIB2, and TGFB1, which are involved in chemoresistance and correlate with a poor prognosis, hypoxia, and purinergic signaling.

Effectiveness of Laser Acupuncture for Reducing Pain and Increasing Mouth Opening Range in Individuals with Temporomandibular Disorder: A Systematic Review and Network Meta-Analysis.

PURPOSE OF REVIEW: Laser acupuncture (LA) demonstrates promising results in the treatment of musculoskeletal disorders. However, its effects on temporomandibular disorder (TMD) are not yet fully understood. Thus, the aim of this systematic review and network meta-analysis was to assess the effectiveness of LA on pain intensity and maximum mouth opening range (MMO) related to TMD. A search was carried out in 11 electronic databases and references of included studies to locate randomized clinical trials (RCTs) that evaluated LA as a primary treatment for TMD. The risk of bias was assessed using the RoB 2 tool. Network meta-analysis was conducted on the MetaInsight platform, considering the pain intensity and counseling (C) as the outcome of reference. The GRADE system was used to assess the certainty of the evidence. RECENT FINDINGS: Five studies evaluated pain intensity, four with a high risk of bias and one with a low risk. Two studies evaluated pain intensity on palpation (one with high and one with low risk of bias), and one study with high risk of bias evaluated MMO. Laser parameters were: 690-810 nm, 40-150 mW, and 7.5-112.5 J/cm2. Occlusal splint (OS) [- 2.47; CI 95% - 3.64, - 1.30] and Physiotherapy (PT) [-2.64; CI 95% - 3.94, - 1.34] reduced pain intensity compared to C. The ranking of treatments in order of effectiveness was PT > OS > LA > C > CR (craniopuncture). The certainty of the evidence was very low or low. The data do not support the indication of LA for the treatment of TMDs and new placebo-controlled RCTs must be conducted to demonstrate its effectiveness more precisely.

Morbidity trend and space-time clusters of COVID-19 occurrence in children and adolescents

Abstract Objective: To analyze the morbidity trend and space-time distribution clusters of confirmed COVID-19 cases in children and adolescents. Method: An ecological study of COVID-19 cases confirmed in the Information System from 2020 to 2022 in the age group from 0 to 19 years old, residents in Mato Grosso municipalities, Brazilian Midwest region. A trend analysis of the monthly morbidity rate of cases/100,000 inhabitants was used, following Prais-Winsten's regression. A space-time distribution of the Bayesian incidence rate per 100,000 inhabitants was performed, in addition to a space-time scan to identify high-risk clusters. Results: Of all 79,592 COVID-19 cases studied, 51.6% were in females and 44.21% in people aged 15-19 years old. The mean monthly rate was 265.87 cases per 100,000 inhabitants, with a stationary trend in the period analyzed (Monthly Percentage Variation [MPV]) = 12.15; CI95%[MPV]: -0.73;26.70). The morbidity rate due to COVID-19 was higher in the female gender (283.14/ 100,000 inhabitants) and in the age group from 15 to 19 years old (485.90/100,000 inhabitants). An increasing trend was observed with a greater monthly time variation of 14.42% (CI95%[MPV]: 1.28;29.28)] among those aged from 10 to 14 years old. The primary cluster, which was also the one with the highest Relative Risk (RR = 5,16, p-value = 0.001), included 19 municipalities located in the North health macro-region. Conclusion: The findings indicated a monthly stationary trend in the study population, an increase in the age group from 10 to 14 years old, and areas at a higher risk for the disease in the North health macro-region of the state.

Microbiota of dental caries in primary teeth of a Costa Rican Child population / Microbiota de la caries dental en piezas primarias de una población infantil costarricense

Abstract The objective of this research was to identify bacteria present in the microbiota of dentinal carious lesions in primary molars of some Costa Rican pediatric patients. Data were collected from 15 children aged between 4 and 8 years old who attended the Pediatric Dentistry Clinic at the Faculty of Dentistry from the University of Costa Rica (UCR). The inclusion criteria were: infants between 4 and 8 years old who presented cavitated carious lesions in primary teeth, who were actively attended by students at the Faculty of Dentistry from the UCR, and whose parents or legal guardians signed the informed consent to participate in this research. Samples were taken using a sterile spoon, placed in storage vials, and subjected to various conventional and molecular microbial identification techniques, such as Gram stain identification, catalase tests, oxidase, TSI, API 20E, API STAPH, and VITEK 2. Of the 60 bacterial strains subjected to Gram staining, the following was obtained: 28 Gram-positive bacteria and 32 Gram-negative bacteria. The main isolated organisms were species of Staphylococcus epidermidis, Pasteurella pneumotropica/Mannheimia haemolytica, Pantoea spp, and Streptococcus mutans.

Resumen El objetivo de esta investigación fue identificar las bacterias presentes en la microbiota de lesiones cariosas dentinales en molares primarias de pacientes pediátricos costarricenses. Las muestras fueron recolectadas de 15 niños entre los 4 y 8 años que fueron atendidos en la Clínica de Odontopediatría en la Facultad de Odontología de la Universidad de Costa Rica (UCR). Los criterios de inclusión fueron: pacientes entre los 4 y los 8 años de edad que presentaran lesiones cariosas cavitadas en dientes primarios, que se encuentraran activos para su atención por estudiantes de la Facultad de Odontología de la UCR, y que los padres o encargados legales firmaran el consentimiento informado para participar en esta investigación. Las muestras se tomaron utilizando una cuchareta estéril, colocándolas en viales de almacenamiento y fueron sometidas a diversas técnicas de identificación microbiana convencionales y moleculares tales como: identificación por Tinción de Gram, pruebas catalasa, oxidasa, TSI, API 20E, API STAPH y VITEK. De las 60 cepas bacterianas sometidas a tinción de Gram se obtuvo: 28 bacterias Gram Positivas y 32 bacterias Gram Negativas. Los principales organismos aislados fueron: especies de Staphylococcus epidermidis, Pasteurella pneumotropica/ Mannheimia haemolytica, Pantoea spp y Streptococcus mutans.

Morbidity trend and space-time clusters of COVID-19 occurrence in children and adolescents.

OBJECTIVE: To analyze the morbidity trend and space-time distribution clusters of confirmed COVID-19 cases in children and adolescents. METHOD: An ecological study of COVID-19 cases confirmed in the Information System from 2020 to 2022 in the age group from 0 to 19 years old, residents in Mato Grosso municipalities, Brazilian Midwest region. A trend analysis of the monthly morbidity rate of cases/100,000 inhabitants was used, following Prais-Winsten's regression. A space-time distribution of the Bayesian incidence rate per 100,000 inhabitants was performed, in addition to a space-time scan to identify high-risk clusters. RESULTS: Of all 79,592 COVID-19 cases studied, 51.6% were in females and 44.21% in people aged 15-19 years old. The mean monthly rate was 265.87 cases per 100,000 inhabitants, with a stationary trend in the period analyzed (Monthly Percentage Variation [MPV]) = 12.15; CI95%[MPV]: -0.73;26.70). The morbidity rate due to COVID-19 was higher in the female gender (283.14/100,000 inhabitants) and in the age group from 15 to 19 years old (485.90/100,000 inhabitants). An increasing trend was observed with a greater monthly time variation of 14.42% (CI95%[MPV]: 1.28;29.28)] among those aged from 10 to 14 years old. The primary cluster, which was also the one with the highest Relative Risk (RR = 5,16, p-value = 0.001), included 19 municipalities located in the North health macro-region. CONCLUSION: The findings indicated a monthly stationary trend in the study population, an increase in the age group from 10 to 14 years old, and areas at a higher risk for the disease in the North health macro-region of the state.

Avaliação da puericultura na Estratégia Saúde da Família em município-sede de macrorregião de saúde / Evaluation of childcare in the Family Health Strategy in a municipality in the health macro-region

Resumo Avaliou-se a puericultura na ESF em município-sede de macrorregião de saúde do Nordeste brasileiro. Realizou-se estudo avaliativo com dois níveis de análise: a gestão municipal e local. Aprofundou-se a avaliação local em quatro EqSF mediante entrevistas com gestores, profissionais e puérperas, análise documental e observação não-participante. As fontes de dados foram trianguladas e processadas no software Nvivo, e o material empírico cotejado com uma situação-objetivo segundo os níveis de análise da pesquisa e elaborada pelos pesquisadores. No nível municipal, a definição clara de atribuições para os pontos de atenção em puericultura e fluxos assistenciais foi um dos critérios mais críticos e, no âmbito local, a integração de ações com outros serviços primários, a contrarreferência e ações educativas coletivas no território. A implementação das práticas de puericultura nas EqSF é fortemente dependente dos determinantes político-organizacionais entre o âmbito municipal e local, e os resultados aqui evidenciados podem se aplicar a outros sistemas municipais de saúde com configurações semelhantes. Este estudo contribuiu para a avaliação da puericultura sustentada na proposição de critérios que consideraram a interdependência municipal e local na organização das ações de avaliação do crescimento e desenvolvimento infantil.

Abstract Childcare was evaluated in the ESF in a municipality that is the headquarters of a health macro-region in the Brazilian Northeast. An evaluative study was carried out with two levels of analysis: municipal and local management. The local assessment was deepened in four EqSF through interviews with managers, professionals and postpartum women, document analysis and non-participant observation. The data sources were triangulated and processed in the Nvivo software, and the empirical material was compared with an objective situation according to the research analysis levels and prepared by the researchers. At the municipal level, the clear definition of assignments for childcare care points and assistance flows was one of the most critical criteria and, at the local level, the integration of actions with other primary services, counter-referral and collective educational actions in the territory. The implementation of childcare practices in EqSF is strongly dependent on political-organizational determinants between the municipal and local levels, and the results shown here may apply to other municipal health systems with similar configurations. This study contributed to the evaluation of childcare based on the proposition of criteria that considered municipal and local interdependence in the organization of actions to evaluate child growth and development.

Effect of Support Matrix and Crosslinking Agents on Nutritional Properties of Orange Juice during Enzyme Clarification: A Comparative Study.

This study investigated the impact of a support matrix and active group on the support to the nutritional properties of orange juice after juice clarification. Pectinase was immobilized on chitosan and aminated silica supports, activated with genipin or glutaraldehyde, and applied for juice clarification. The effects on various juice properties, including reducing sugars, total soluble solids, vitamin C, and phenolic compounds, juice color, and pH, were evaluated. The results revealed that the immobilization on chitosan activated using genipin resulted in the highest biocatalyst activity (1211.21 U·g-1). The juice treatments using the biocatalysts led to turbidity reduction in the juice (up to 90%), with the highest reductions observed in treatments involving immobilized enzyme on chitosan. Importantly, the enzymatic treatments preserved the natural sugar content, total soluble solids, and pH of the juice. Color differences between treated and raw juice samples were especially relevant for those treated using enzymes, with significant differences in L* and b*, showing loss of yellow vivid color. Analysis of phenolic compounds and vitamin C showed no significant alterations after the enzymatic treatment of the raw juice. According to our results, the clarification of orange juice using immobilized enzymes can be a compromise in turbidity reduction and color reduction to maintain juice quality.

"Prevalence of middle crossed syndrome in young healthy individuals from a health science university in India".

BACKGROUND: A myofascial sling is a chain of anatomically interconnected muscles. Anatomical slings work by transferring forces between upper and lower limbs while maintaining a steady core. Middle Crossed Syndrome refers to an imbalance in the transverse plane musculature through sling systems of the trunk; often associated with laterality patterns. OBJECTIVE: The primary objective of this study was to find the prevalence of Middle Crossed Syndrome(MCS) and secondary objective was to find an association of MCS with leg dominance in young healthy individuals. METHODS: This observational study, included 382 participants aged 19-26 years. The participants were screened for presence of Middle Crossed Syndrome using Supine Lateral Ball Roll Test, Active Straight Leg Raise and Gait Parameters (step length, stride length, degree of toe out, cadence, base of support). RESULTS: The results were obtained by statistical analysis using Chi-square test for prevalence of MCS in young healthy individuals with respect to age groups, gender and leg dominance. Of the total 382 screened, 25 (69.37%) were positive either unilaterally or bilaterally for MCS. CONCLUSION: The study concluded that over half of the participants clinically tested positive for MCS with no correltion of MCS with leg dominance.

Direct costs of treating men with prostate cancer with High Intensity Focused Ultrasound.

OBJECTIVE: To analyze the direct costs of materials, medicines/solutions and healthcare professionals required to treat men with prostate cancer using High Intensity Focused Ultrasound. METHOD: Quantitative, exploratory-descriptive research, single case study type. Data were collected from electronic medical records/printed documentation from the Operating Room of a public teaching and research hospital. Health professionals estimated the respective time spent on activities in the following stages: "Before anesthetic induction", "Before performing thermal ablation", "During thermal ablation" and "After performing thermal ablation". Costs were calculated by multiplying the (estimated) time spent by the unit cost of direct labor, adding to the measured cost of materials, medicines/solutions. RESULTS: The measured costs with materials corresponded to US$851.58 (SD = 2.17), with medicines/solutions to US$72.13 (SD = 25.84), and estimated personnel costs to US$196.03, totaling US$1119.74/procedure. CONCLUSION: The economic results obtained may support hospital managers in the decision-making process regarding the adoption of the High Intensity Focused Ultrasound for the treatment of prostate cancer.

Interaction between starch and dietary compounds: New findings and perspectives to produce functional foods.

Due to the increased prevalence of overweight, obesity, diabetes, colon cancer, cardiovascular diseases, and metabolic syndrome, dietary approaches to reduce starch digestion and regulate glucose homeostasis have gained attention. Starch is a polysaccharide in most daily food consumed as bakery products, snacks, breakfast cereals, and pasta, which are often vilified. However, it is also present in beans, lentils, and oatmeal, which are considered healthy food products. The difference relays on the food matrix and the thermal process that can produce interactions between starch and dietary compounds (protein, lipid, non-starch polysaccharide, and bioactive compounds) or among starch chains (retrogradation). Such interactions produce structural changes so the digestive enzymes cannot hydrolyze them; additionally, the physical barrier of some macromolecules (proteins, hydrocolloids) restricts starch gelatinization and accessibility of the digestive enzymes to hydrolyze the starch. The interactions mentioned above and the use of some macromolecules as physical barriers could be explored as a pathway to develop functional foods. This review analyzes the interactions between starch and dietary compounds influenced by the processing of some food matrices to better understand their potential for developing functional foods.

Impact of plasma-activated water on the supramolecular structure and functionality of small and large starch granules.

Hydrothermal (HMT) and water agitation (WA) treatments using plasma-activated water (PAW) were employed as sustainable methods to modify the molecular and functional performance of small (rice) and large (potato) starch granules. HMT-PAW and WA-PAW treatments resulted in etched and damaged granular surfaces that rearranged the long and short-range crystallinity of the modified starches. Both treatments seemed to predominantly occur in the amorphous region of the rice starch and the crystalline regions of the potato starch, changing the crystallinity values from 22.9 and 14.8 % to 31.8 and 10.4 %, respectively. Thus, the level of the arrangement of chains reached after PAW treatment decreased the ability of rice starch granules to swell (16 to 9 %) and leach out starch molecules from the granules (4.5 to 1.3 %), decreasing the viscosity and pasting profiles as indicated by n and k values. Opposite behavior was observed in the modified potato starches since starch components leached out to a higher extent (1.7 to 5.4 %). The results showed that HMT and WA treatments using PAW are feasible eco-friendly methods for modifying starch granules without chemical reagents. These modified starches could be suitable as functional ingredients or biopolymeric matrices for the food and packaging industry.

A bvrR/bvrS Non-Polar Brucella abortus Mutant Confirms the Role of the Two-Component System BvrR/BvrS in Virulence and Membrane Integrity.

Brucella abortus is a bacterial pathogen causing bovine brucellosis worldwide. This facultative extracellular-intracellular pathogen can be transmitted to humans, leading to a zoonotic disease. The disease remains a public health concern, particularly in regions where livestock farming is present. The two-component regulatory system BvrR/BvrS was described by isolating the attenuated transposition mutants bvrR::Tn5 and bvrS::Tn5, whose characterization led to the understanding of the role of the system in bacterial survival. However, a phenotypic comparison with deletion mutants has not been performed because their construction has been unsuccessful in brucellae and difficult in phylogenetically related Rhizobiales with BvrR/BvrS orthologs. Here, we used an unmarked gene excision strategy to generate a B. abortus mutant strain lacking both genes, called B. abortus ∆bvrRS. The deletion was verified through PCR, Southern blot, Western blot, Sanger sequencing, and whole-genome sequencing, confirming a clean mutation without further alterations at the genome level. B. abortus ∆bvrRS shared attenuated phenotypic traits with both transposition mutants, confirming the role of BvrR/BvrS in pathogenesis and membrane integrity. This B. abortus ∆bvrRS with a non-antimicrobial marker is an excellent tool for continuing studies on the role of BvrR/BvrS in the B. abortus lifestyle.

If You're Not Confused, You're Not Paying Attention: Ochrobactrum Is Not Brucella.

Bacteria of the genus Brucella are facultative intracellular parasites that cause brucellosis, a severe animal and human disease. Recently, a group of taxonomists merged the brucellae with the primarily free-living, phylogenetically related Ochrobactrum spp. in the genus Brucella. This change, founded only on global genomic analysis and the fortuitous isolation of some opportunistic Ochrobactrum spp. from medically compromised patients, has been automatically included in culture collections and databases. We argue that clinical and environmental microbiologists should not accept this nomenclature, and we advise against its use because (i) it was presented without in-depth phylogenetic analyses and did not consider alternative taxonomic solutions; (ii) it was launched without the input of experts in brucellosis or Ochrobactrum; (iii) it applies a non-consensus genus concept that disregards taxonomically relevant differences in structure, physiology, population structure, core-pangenome assemblies, genome structure, genomic traits, clinical features, treatment, prevention, diagnosis, genus description rules, and, above all, pathogenicity; and (iv) placing these two bacterial groups in the same genus creates risks for veterinarians, medical doctors, clinical laboratories, health authorities, and legislators who deal with brucellosis, a disease that is particularly relevant in low- and middle-income countries. Based on all this information, we urge microbiologists, bacterial collections, genomic databases, journals, and public health boards to keep the Brucella and Ochrobactrum genera separate to avoid further bewilderment and harm.

Experimental model of nephropathy associated with diabetes mellitus in mice.

PURPOSE: Nontransmissible chronic diseases, such as diabetes mellitus (DM) and nephropathy, affect a significant portion of the population, often treated due to injuries that require healing and regeneration. To create an experimental model of associated comorbidities, for healing and regeneration studies, protocols for induction of nephropathy by ischemia and reperfusion (I/R) and induction of DM by injection of streptozotocin (STZ) were associated. METHODS: Sixty-four mice (Mus musculus), female, adult, Swiss strain, weighing approximately 20 g, were divided into four groups: G1: control (n = 24), G2: nephropathy group (N) (n = 7), G3, DM (n = 9), and G4: N+DM (n = 24). Arteriovenous stenosis (I/R) of the left kidney was performed as the first protocol. The animals received a hyperlipidemic diet for 7 days after the injection of STZ (150 mg/kg, via i.p.) and an aqueous glucose solution (10%) for 24 h. The animals in the G3 and G4 groups were observed for 14 days before receiving the diet and STZ. The evolution of nephropathy was observed using a urine test strip and the DM, through the analysis of blood glucose with a reagent strip on a digital monitor. RESULTS: The ischemic induction protocols of nephropathy and DM with STZ, associated, were sustainable, low-cost, and without deaths. There were alterations compatible with initial renal alterations, in the first 14 days, such as increased urinary density, pH alteration, presence of glucose, proteins and leukocytes, when compared to the control group. DM was confirmed by the presence of hyperglycemia 7 days after induction and its evolution after 14 days. The animals in the G4 group showed constant weight loss when compared to the other groups. It was possible to observe morphological alterations in the kidneys submitted to I/R, regarding coloration, during surgery and after the end of the observation period, in the volume and size of the left kidney, when compared to the contralateral kidney. CONCLUSIONS: It was possible to induce nephropathy and DM associated in the same animal, in a simple way, confirmed with rapid tests, without losses, providing a basis for future studies.

Phenotypes controlled by the Brucella abortus two component system BvrR/BvrS are differentially impacted by BvrR phosphorylation.

Brucella abortus is a zoonotic pathogen whose virulence depends on its ability to survive intracellularly at the endoplasmic reticulum derived compartment. The two-component system BvrR/BvrS (BvrRS) is essential for intracellular survival due to the transcriptional control of the type IV secretion system VirB and its transcriptional regulator VjbR. It is a master regulator of several traits including membrane homeostasis by controlling gene expression of membrane components, such as Omp25. BvrR phosphorylation is related to DNA binding at target regions, thereby repressing or activating gene transcription. To understand the role of BvrR phosphorylation we generated dominant positive and negative versions of this response regulator, mimicking phosphorylated and non-phosphorylated BvrR states and, in addition to the wild-type version, these variants were introduced in a BvrR negative background. We then characterized BvrRS-controlled phenotypes and assessed the expression of proteins regulated by the system. We found two regulatory patterns exerted by BvrR. The first pattern was represented by resistance to polymyxin and expression of Omp25 (membrane conformation) which were restored to normal levels by the dominant positive and the wild-type version, but not the dominant negative BvrR. The second pattern was represented by intracellular survival and expression of VjbR and VirB (virulence) which were, again, complemented by the wild-type and the dominant positive variants of BvrR but were also significantly restored by complementation with the dominant negative BvrR. These results indicate a differential transcriptional response of the genes controlled to the phosphorylation status of BvrR and suggest that unphosphorylated BvrR binds and impacts the expression of a subset of genes. We confirmed this hypothesis by showing that the dominant negative BvrR did not interact with the omp25 promoter whereas it could interact with vjbR promoter. Furthermore, a global transcriptional analysis revealed that a subset of genes responds to the presence of the dominant negative BvrR. Thus, BvrR possesses diverse strategies to exert transcriptional control on the genes it regulates and, consequently, impacting on the phenotypes controlled by this response regulator.