Cobalamin cbiP mutant shows decreased tolerance to low temperature and copper stress in Listeria monocytogenes.

BACKGROUND: Listeria monocytogenes is a foodborne pathogen that causes listeriosis in humans. This pathogen activates multiple regulatory mechanisms in response to stress, and cobalamin biosynthesis might have a potential role in bacterial protection. Low temperature is a strategy used in the food industry to control bacteria proliferation; however, L. monocytogenes can grow in cold temperatures and overcome different stress conditions. In this study we selected L. monocytogenes List2-2, a strain with high tolerance to the combination of low temperature + copper, to understand whether the cobalamin biosynthesis pathway is part of the tolerance mechanism to this stress condition. For this, we characterized the transcription level of three cobalamin biosynthesis-related genes (cbiP, cbiB, and cysG) and the eutV gene, a transcriptional regulator encoding gene involved in ethanolamine metabolism, in L. monocytogenes strain List2-2 growing simultaneously under two environmental stressors: low temperature (8 °C) + copper (0.5 mM of CuSO4 × 5H2O). In addition, the gene cbiP, which encodes an essential cobyric acid synthase required in the cobalamin pathway, was deleted by homologous recombination to evaluate the impact of this gene in L. monocytogenes tolerance to a low temperature (8 °C) + different copper concentrations. RESULTS: By analyzing the KEGG pathway database, twenty-two genes were involved in the cobalamin biosynthesis pathway in L. monocytogenes List2-2. The expression of genes cbiP, cbiB, and cysG, and eutV increased 6 h after the exposure to low temperature + copper. The cobalamin cbiP mutant strain List2-2ΔcbiP showed less tolerance to low temperature + copper (3 mM) than the wild-type L. monocytogenes List2-2. The addition of cyanocobalamin (5 nM) to the medium reverted the phenotype observed in List2-2ΔcbiP. CONCLUSION: These results indicate that cobalamin biosynthesis is necessary for L. monocytogenes growth under stress and that the cbiP gene may play a role in the survival and growth of L. monocytogenes List2-2 at low temperature + copper.

Installing oncofertility programs for common cancers in limited resource settings (Repro-Can-OPEN Study): An extrapolation during the global crisis of Coronavirus (COVID-19) pandemic.

PURPOSE: The state of limited resource settings that Coronavirus (COVID-19) pandemic has created globally should be taken seriously into account especially in healthcare sector. In oncofertility, patients should receive their fertility preservation treatments urgently even in limited resource settings before initiation of anticancer therapy. Therefore, it is very crucial to learn more about oncofertility practice in limited resource settings such as in developing countries that suffer often from shortage of healthcare services provided to young patients with cancer. METHODS: As an extrapolation during the global crisis of COVID-19 pandemic, we surveyed oncofertility centers from 14 developing countries (Egypt, Tunisia, Brazil, Peru, Panama, Mexico, Colombia, Guatemala, Argentina, Chile, Nigeria, South Africa, Saudi Arabia, and India). Survey questionnaire included questions on the availability and degree of utilization of fertility preservation options in case of childhood cancer, breast cancer, and blood cancer. RESULTS: All surveyed centers responded to all questions. Responses and their calculated oncofertility scores showed different domestic standards for oncofertility practice in case of childhood cancer, breast cancer, and blood cancer in the developing countries under limited resource settings. CONCLUSIONS: Medical practice in limited resource settings has become a critical topic especially after the global crisis of COVID-19 pandemic. Understanding the resources necessary to provide oncofertility treatments is important until the current COVID-19 pandemic resolves. Lessons learned will be valuable to future potential worldwide disruptions due to infectious diseases or other global crises.

Is there a link between influenza and type I diabetes? Increased incidence of TID during the pandemic H1N1 influenza of 2009 in Chile.

UNLABELLED: Pandemic H1N1 2009 had the highest incidence in the middle-high income area of Santiago and affected mostly school age patients. Influenza A virus (IAVs) causes systemic and most commonly non-systemic infection. Interestingly, it is able to replicate only in the presence of trypsin-like enzymes, as lung and pancreas. HYPOTHESIS: IAVs infection may trigger beta cell destruction and increase the incidence of T1DM. METHODS: A retrospective observational study of new T1DM pediatric patients from database of Clinica Las Condes between 1995 and 2012. RESULTS: From 58 patients, 44.7% were diagnosed between 2009 and 2010, coincident with the H1N1 virus outbreak. There were no differences in clinical neither metabolic parameters between those patients from the 2009-2010 period and the rest. From those patients with available antibody panel, it was negative in 30% of the 2009-2010 group vs. 12.5% of the rest of the cohort (p < 0.05). Only one 5.8 year old boy had history of H1N1 virus infection three months prior to the DM1 onset with negative antibodies. CONCLUSIONS: The temporal coincidence suggests a possible link between T1DM and H1N1 virus, might be thought to be through direct cytopathic damage. Unfortunately we could only confirm H1N1 previous infection in only one case. Prospective studies in new T1DM cases are necessary to test this hypothesis.

Effect of modified waste introduction methods over short-term and long-term use of onsite sanitation systems.

Insufficiently treated wastes contained within onsite sanitation systems (OSS) commonly used by disadvantaged and developing communities contribute to public and environmental health concerns, calling for practical alternative solutions. At the basic level, an improved understanding of the evolution of chemical and physical constituents under different waste introduction methods and for short-term and long-term operation is needed. While receiving non-dilute waste under mixed, unmixed, toilet paper exclusion, and urine diversion (UD) regimes, self-flushing OSS simulated using anaerobic digesters (ADs) were compared during three operational stages: (1) 0-1 month service for unsheltered encampments; (2) 1-3 month disaster relief scenario; and (3) ≥ 3 months representing refugee camps and long-term household use. Although stratification was found to promote suitable conditions for short-term use of self-flushing toilets, mixing increased beneficial biodegradation of organic constituents. Urine-containing ADs demonstrated a shift from sulfide to ammonia odor accompanied by high pH (> 8) after ~ 240 d. E. coli reduction following elevated nitrogen and dissolved solids levels pointed to decreased pathogen survival in ADs with urine. The benefits of bacterial disinfection, reduction of sulfurous odors, and heightened organics degradation in mixed, urine-containing ADs suggest this format as more desirable for prolonged use of self-flushing OSS over unmixed or urine-diverting formats.

Compressive myelopathy associated with ectasia of the vertebral and spinal arteries in a dog.

A 4-year-old dog was presented for acute, progressive tetraparesis and cervical hyperesthesia. Symmetrical tubular structures coursing along the lateroventral aspects of the spinal cord at the fourth and fifth cervical vertebrae were identified in magnetic resonance images. At necropsy, vertebral arteries and their spinal branches were severely ectatic bilaterally, and the cervical spinal cord was compressed. Histologically, the ectatic branches of the vertebral and ventral spinal arteries were surrounded by fibrosis with scant mononuclear cell infiltrates and hemorrhage. Spinal branches of the vertebral arteries had focally severe reduction in the tunica media. A thrombus was in an arterial branch. Smaller vessels in adjacent tissue had fibrinoid degeneration. Axonal degeneration was detected in the affected spinal cord and nerve roots. The segmental degenerative radiculomyelopathy in this dog was attributed to anomalous ectasia of the vertebral and ventral spinal arteries.

Phenotypic characterization of PIGN-associated paroxysmal dyskinesia in Soft-coated wheaten terriers and preliminary response to acetazolamide therapy.

A hereditary movement disorder in Soft coated wheaten terriers (SCWT) has been associated with a mutation in PIGN which encodes an enzyme involved in synthesis of glycosylphosphatidylinositol (GPI). The objective of this study was to describe and classify the clinical phenotype and assess therapeutic response. Twenty-five SCWT and related dogs homozygous for PIGN:c.398C>T with paroxysmal dyskinesia were available for inclusion. Medical records and video recordings of 17 dogs were evaluated in a retrospective case series. Affected dogs had episodes of involuntary, hyperkinetic movements and dystonia. Median age of onset was 2.5 years. A typical episode consisted of rapid, irregular hyperflexion and extension of the pelvic limbs with some degree of truncal dystonia. A mild episode consisted of spontaneous flexion of one pelvic limb while walking which could resemble a lameness. Episodes lasted several minutes to several hours and occurred up to 10 times/day or more. They were not associated with exercise or fasting but were sometimes triggered by excitement or stress. Acetazolamide therapy improved nine of 11 dogs, in seven cases abolishing episodes. Five of 17 dogs treated with other agents had mild improvement with clonazepam (n = 2), levetiracetam (n = 1), or phenobarbital (n = 2). Paroxysmal dyskinesias must be differentiated from seizure disorders since they often respond to different therapies. The SCWT phenotype consisted predominantly of hyperkinesia, and can respond dramatically to acetazolamide. GPI anchors proteins to the cell surface including carbonic anhydrase IV which modulates synaptic pH in the brain. Altered activity of this enzyme may be the target of acetazolamide therapy.

[Paracoccidioidomicosis and multidrug-resistant tuberculosis (TBC-MDR) in patient coinfected with HIV and hepatitis C]. / Paracoccidioidomicosis y TBC-MR en portador de VIH/VHC.

A case of an adult male patient diagnosed with HIV and Hepatitis C co infection is presented. He had granu-lomatuos hepatitis and blood smear positive to Paracoccidioides brasiliensis concomitant to the detection of MDR Mycobacterium tuberculosis in sputum further complicated with reactivation of cytomegalovirus (possible pancreatitis and retinitis). Difficulties in diagnostic and therapeutic approach in a patient with multiple infections are reviewed.

Effect of semen dilution to low-sperm number per dose on motility and functionality of cryopreserved bovine spermatozoa using low-density lipoproteins (LDL) extender: comparison to Triladyl and Bioxcell.

Artificial insemination with doses containing low-sperm numbers has been utilized to optimize the use of elite bulls. Hen egg yolk is widely used as a cryoprotective agent in semen freezing extender protecting the spermatozoa. Its action is due to the presence of low-density lipoproteins (LDL) in the hen egg yolk. The objectives of the present study were to evaluate the effects of the semen dilution to low-sperm number/dose on sperm motility and integrity of sperm plasma membrane in the cryopreservation process, using two commercial extenders (Triladyl, Bioxcell and LDL extender prepared in our laboratory, 97% purity. Fifteen ejaculates were collected from five fertile crossbred bulls (Bos taurusxBos indicus). After collection, sperm motility was examined by Computer-Assisted Semen Analysis (Hamilton Thorne), morphological sperm characteristics were evaluated by differential interference microscopy and the integrity of plasma membranes was determined using the hypo-osmotic swelling test. The semen was subsequently divided into three aliquots and diluted with the three extenders into 120 x 10(6), 60 x 10(6) and 20 x 10(6)sperm/mL, corresponding to 30 x 10(6), 15 x 10(6) and 5 x10(6) sperm/dose, respectively. This study revealed that LDL extender was more effective in preservation of motility and integrity of the plasma membrane of spermatozoa than Bioxcell and Triladyl (p<0.05), but no significant difference was observed between Triladyland Bioxcell. Therefore we can conclude that LDL extender could be used instead of Triladyl or Bioxcellat low semen concentration per dose for elite bulls, it also could be envisaged for the industry of sex-stored semen.

Personality traits and temporomandibular disorders in a group of children with bruxing behaviour.

The aim of the present study was to compare the personality pattern, the anxiety level and the temporomandibular disorders (TMD) between bruxist and non-bruxist children with mixed dentition. Fifty-two subjects, with a mean age of 9.45 years (range 8-11) were evaluated and classified as bruxist (n = 26), according to the American Academy of Sleep Medicine (AASM) and the presence of dental wear clinically visible, under the same conditions of artificial light and position. The control children (n = 26) did not present dental wear and did not accomplished all the AASM criteria. The personality pattern and the anxiety of the bruxist children were studied by means of the Children's Personality Questionnaire (CPQ) and the Conners' Parents Rating Scales (CPRS), respectively, and compared with the personality traits and the anxiety level of a non-bruxist population. The TMD were also evaluated using the Research Diagnostic Criteria for temporomandibular disorders (RDC/TMD) criteria. The data were analysed with the student's t-test, Fisher's exact test and chi-squared tests. A multivariated analysis was performed using a logistic regression with the stepwise likelihood ratio method. Compared with the controls, the bruxist children had significantly higher tension personality and were more anxiety prone. The bruxist children presented more TMD-related signs and symptoms than children in the control group. A strong correlation was found among bruxism, TMD, the high anxiety level and the high tension personality trait.

Building a network for multicenter, prospective research of central nervous system infections in South America: Process and lessons learned.

Multicenter collaborative networks are essential for advancing research and improving clinical care for a variety of conditions. Research networks are particularly important for central nervous system infections, which remain difficult to study due to their sporadic occurrence and requirement for collection and testing of cerebrospinal fluid. Establishment of long-term research networks in resource-limited areas also facilitates diagnostic capacity building, surveillance for emerging pathogens, and provision of appropriate treatment where needed. We review our experience developing a research network for encephalitis among twelve hospitals in five Peruvian cities since 2009. We provide practical suggestions to aid other groups interested in advancing research on central nervous system infections in resource-limited areas.

Factores extrínsecos implicados en la pigmentación de las resinas compuestas dentales / Extrinsic factors involved in the pigmentation of dental composite resins

RESUMEN Las resinas compuestas dentales fueron introducidas en la primera mitad del siglo XX como una avance mecánico- estético en el campo de la odontología restauradora, en respuesta a los silicatos, que hasta aquel entonces era el material restaurador de primera elección, dentro de su composición, posee una matriz orgánica formada por Bis- GMA y otros monómeros de dimetacrilato (TEGMA, UDMA, siloranos), que son susceptibles a la pigmentación de sustancias extrínsecas, en diferente grado , las bebidas más comunes como vino tinto, té, café y bebidas azucaradas Objetivo: Revisar y analizar la literatura disponible que responda a la pregunta ¿Cuáles son los factores extrínsecos más comunes que causan pigmentación de las resinas compuestas?

ABSTRACT Dental composite resins were introduced in the first half of the 20th century as a mechanical-aesthetic advance in the field of restorative dentistry, as a response to silicates, which at that time was the first choice restorative material, composite resin contains a matrix of organic made up of Bis-GMA and other dimethacrylate monomers (TEGMA, UDMA, silorans) doing them susceptible to pigmentation with extrinsic substances, in this case drinks, such as red wine, tea and coffee are the main cause of the pigmentation of resins. Objective: To Review and analyze the available literature that answers the question: what are the most common extrinsic factors that cause pigmentation of composite resins?

Cobalamin cbiP mutant shows decreased tolerance to low temperature and copper stress in Listeria monocytogenes

BACKGROUND: Listeria monocytogenes is a foodborne pathogen that causes listeriosis in humans. This pathogen activates multiple regulatory mechanisms in response to stress, and cobalamin biosynthesis might have a potential role in bacterial protection. Low temperature is a strategy used in the food industry to control bacteria proliferation; however, L. monocytogenes can grow in cold temperatures and overcome different stress conditions. In this study we selected L. monocytogenes List2-2, a strain with high tolerance to the combination of low temperature +copper, to understand whether the cobalamin biosynthesis pathway is part of the tolerance mechanism to this stress condition. For this, we characterized the transcription level of three cobalamin biosynthesis related genes ( cbiP , cbiB, and cysG ) and the eutV gene, a transcriptional regulator encoding gene involved in ethanolamine metabolism, in L. monocytogenes strain List2-2 growing simultaneously under two environmental stressors: low temperature (8 °C) +copper (0.5 mM of CuSO4 ×5H2O). In addition, the gene cbiP , which encodes an essential cobyric acid synthase required in the cobalamin pathway, was deleted by homologous recombination to evaluate the impact of this gene in L. monocytogenes tolerance to a low temperature (8 °C) +different copper concentrations. RESULTS: By analyzing the KEGG pathway database, twenty-two genes were involved in the cobalamin biosynthesis pathway in L. monocytogenes List2-2. The expression of genes cbiP , cbiB, and cysG, and eutV increased 6 h after the exposure to low temperature +copper. The cobalamin cbiP mutant strain List2-2Δ cbiP showed less tolerance to low temperature +copper (3 mM) than the wild type L. monocytogenes List2-2. The addition of cyanocobalamin (5 nM) to the medium reverted the phenotype observed in List2-2Δ cbiP . CONCLUSION: These results indicate that cobalamin biosynthesis is necessary for L. monocytogenes growth under stress and that the cbiP gene may play a role in the survival and growth of L. monocytogenes List2-2 at low temperature +copper.

Energy balance and physical activity in obese children attending day-care centres.

OBJECTIVE: To evaluate energy balance and physical activity in obese preschool children attending day-care centres. DESIGN: Cross-sectional study. SUBJECTS: Twenty-four 3-5 years old obese children selected at random from four different day-care centres in Santiago City, Chile. Total energy expenditure (TEE) was measured by doubly labelled water and physical activity as recorded by TRITRAC R3D motion sensor. Energy intake was assessed by measuring food intake while at the centre, along with the recording of additional food intake in the home. RESULTS: Obese children had a 5.4% higher weighed energy intake than their energy requirements 2001. Energy intake during the week was 7716+/-1092 kJ/day and 7401+/-1023 kJ/day in boys and girls, respectively. Minimal activity was higher in the day-care centres (62%) compared to 52% during the weekend at home. Light activity was higher during the weekend at home than during the week (25 versus 20%), but moderate-intense activity was similar (3-5%). CONCLUSIONS: Reduction in dietary energy provided at the day-care centres helps to balance energy requirements during the week. Obese children had marked sedentary characteristics with regards to physical activity, although children are more active at home in weekends. This situation suggests that educators and parents need to improve children's physical activity levels and nutrition habits.

From GWAS to function: lessons from blood cells.

Haematopoiesis, or the process of formation of mature blood cells from committed progenitors, represents an accessible and well-studied paradigm of cell differentiation and lineage specification. Genetic association studies provide a powerful approach to discover new genes, biological pathways and mechanisms underlying haematopoietic development. Here, we highlight recent findings of genomewide association studies (GWAS) linking 145 genomic loci to traits affecting the formation of red and white cells and platelets in European and other ancestries. We present strategies to address the main challenges in GWAS discoveries, particularly to find functional and regulatory effects of genetic variants, and to identify genes through which these genetic variants affect haematological phenotypes. We argue that studies of haematological trait variation provide an ideal paradigm for understanding the function of GWAS-associated variants owing to the accessible nature of cells, simple cellular phenotype and focused efforts to characterize the genetic and epigenetic factors influencing the regulatory landscape in highly pure mature cell populations.

Decreased morbidity and mortality of autologous hematopoietic transplants for children with malignant central nervous system tumors: the 'Head Start' trials, 1991-2009.

Since 1991, three sequential prospective clinical trials have been conducted by the 'Head Start' (HS) Consortium in which young children with newly-diagnosed malignant central nervous system (CNS) tumors were treated with induction chemotherapy followed by single-cycle marrow-ablative chemotherapy and autologous hematopoietic rescue as a means of improving disease cure rate and quality of survival through avoidance (<6 years old at diagnosis) or reduction (6-10 years old) of brain irradiation. Bone Marrow (HS I) or filgrastim-mobilized peripheral hematopoietic cells (HS II and III) were obtained following recovery from the first and/or second induction cycles. Radiotherapy was administered following all chemotherapy only for patients with residual tumor following completion of induction or with age greater than 6 years at diagnosis. Two hundred and twenty-six children were enrolled on three consecutive HS trials with primary malignant CNS tumors and underwent marrow-ablative chemotherapy. The 100-day treatment-related mortality (TRM) steadily declined as did grade IV transplant-related oropharyngeal mucositis. Factors most likely associated with the decrease in TRM and morbidity are increasing experience with the marrow-ablative chemotherapy regimen combined with improved leukapheresis and post-reinfusion supportive care techniques, contributing toward improved overall survival.

Perspectives on the fellowship training in cardiac, thoracic, and vascular anesthesia and critical care in Europe from program directors and educational leads around europe

ABSTRACT:This article reviews fellowship training in adult cardiac, thoracic, and vascular anesthesia and critical care from the perspective of European program initiators and educational leaders in these subspecialties together with current training fellows. Currently, the European Association of Cardiothoracic Anaesthesiology (EACTA) network has 20 certified fellowship positions each year in 10 hosting centers within 7 European countries, with 2 positions outside Europe (São Paulo, Brazil). Since 2009, 42 fellows have completed the fellowship training. The aim of this article is to provide an overview of the rationale, requirements, and contributions of the fellows, in the context of the developmental progression of the EACTA fellowship in adult cardiac, thoracic, and vascular anesthesia and critical care from inception to present. A summary of the program structure, accreditation of host centers, requirements to join the program, teaching and assessment tools, certification, and training requirements in transesophageal electrocardiography is outlined. In addition, a description of the current state of EACTA fellowships across Europe, and a perspective for future steps and challenges to the educational program, is provided. (AU)

Analysis of a novel functional polymorphism within the promoter region of the serotonin transporter gene (5-HTT) in Brazilian patients affected by bipolar disorder and schizophrenia.

It has been suggested that the serotonin transporter (5-hydroxytryptamine-transporter or 5-HTT) may be involved in the pathogenesis of affective disorders. Recently, Collier et al. (1996) found that the frequency of the low-activity short variant (s) of the 5-HTT-linked polymorphic region (5-HTTLPR) was higher among patients with affective disorders than in normal controls. However, since the observed level of significance was not high, they suggest that these findings should be replicated in independent samples. We have analyzed 86 unrelated patients (47 with bipolar disorder and 39 with schizophrenia) and 98 normal controls from the Brazilian population for the 5-HTTLPR. Statistical analysis revealed that the genotypes (LL, Ls, ss) as well as the estimated allele frequencies (L,s) did not differ significantly among the three studied groups or between bipolar and normal controls. In addition, although not statistically significant, the genotype ss in our sample was less frequent among our bipolar patients than in our normal controls (12.8% versus 16.3%) which is the opposite of what was found by Collier et al. (24% versus 18%) in the European study. Although it will be important to extend the present analysis in a larger sample, our preliminary results suggest that the 5-HTTLPR does not seem to play a major role in the genetics of bipolar and schizophrenic disorders at least in this group of Brazilian psychiatric patients.

Clinical and programmatic considerations in the treatment of MDR-TB in children: a series of 16 patients from Lima, Peru.

SETTING: Since 2000, the directly observed treatment, short-course (DOTS) strategy has been expanded in several countries to include treatment of multidrug-resistant tuberculosis (MDR-TB). This strategy is known as DOTS-Plus. Tuberculosis is a common cause of morbidity and mortality for children throughout the developing world. Children may also be infected with MDR-TB, yet most developing countries do not specifically address pediatric MDR-TB. OBJECTIVE: To present the intermediate outcomes of the first 16 children enrolled in the Peruvian DOTS-Plus program and to demonstrate the tolerability of second-line anti-tuberculosis drugs. RESULTS: Three children completed therapy and are cured, one child had bacteriologic and clinical failure after 12 months of therapy and died of respiratory insufficiency, and 12 have intermediate outcomes demonstrating favorable clinical, bacteriologic, and radiographic evidence of improvement after 9-19 months of therapy. CONCLUSIONS: Of the 16 pediatric DOTS-Plus patients, 15 have tolerated therapy well and have had favorable clinical evolution. However, the diagnosis of pediatric MDR-TB is often extremely delayed due to reliance on the adult case definition and should be changed to prevent progressive, chronic illness in such children. Programmatic changes could facilitate earlier diagnosis and treatment of pediatric MDR-TB in Peru and in other DOTS-Plus programs.

Variation of the antimicrobial activity of Pseudognaphalium vira vira (Asteraceae): isolation and X-ray structure of ent-3beta-hydroxy-16-kauren-19-oic acid.

The antimicrobial properties of the resinous exudate from twigs and leaves of a population of Pseudognaphalium vira vira were re-examined against five Gram positive bacteria. The observed decrease in the antimicrobial activity might reflect a change in the diterpenoid composition of the exudate. The title compound 2 was isolated in substantial amount, together with ent-16-kauren-19-oic acid (1), and its structure determined by X-ray diffraction analysis.

Antimicrobial study of the resinous exudate and of diterpenoids isolated from Eupatorium salvia (Asteraceae).

The antimicrobial properties of the resinous exudate from twigs and leaves of Eupatorium salvia were tested against five gram-negative and five gram-positive bacteria. Comparison of the antimicrobial activities of 7-hydroxy-8(17)-labden-15-oic acid (salvic acid) and of its acetate, both compounds isolated from the plant, with that of the crude extract suggested that the latter ester derivative was the major active component in the exudate. These results validate the vernacular medicinal use of Eupatorium salvia as an antiseptic agent.