Sodium and lithium exert differential effects on the central carbon metabolism of Debaryomyces hansenii through the glyoxylate shunt regulation.

Debaryomyces hansenii is a halotolerant/halophilic yeast usually found in salty environments. The yeast accumulated sodium at high concentrations, which improved growth in salty media. In contrast, lithium was toxic even at low concentrations and its presence prevented cell proliferation. To analyse the responses to both cations, metabolite levels, enzymatic activities and gene expression were determined, showing that NaCl and LiCl trigger different cellular responses. At high concentrations of NaCl (0.5 or 1.5 M) cells accumulated higher amounts of the intermediate metabolites glyoxylate and malate and, at the same time, the levels of intracellular oxoglutarate decreased. Additionally, 0.5 M NaCl increased the activity of the enzymes isocitrate lyase and malate synthase involved in the synthesis of glyoxylate and malate respectively and decreased the activity of isocitrate dehydrogenase. Moreover, transcription of the genes coding for isocitrate lyase and malate synthase was activated by NaCl. Also, cells accumulated phosphate upon NaCl exposure. None of these effects was provoked when LiCl (0.1 or 0.3 M) was used instead of NaCl. Lithium induced accumulation of higher amounts of oxoglutarate and decreased the concentrations of glyoxylate and malate to non-detectable levels. Cells incubated with lithium also showed higher activity of the isocitrate dehydrogenase and neither increased isocitrate lyase and malate synthase activities nor the transcription of the corresponding genes. In summary, we show that sodium, but not lithium, up regulates the shunt of the glyoxylic acid in D. hansenii and we propose that this is an important metabolic adaptation to thrive in salty environments.

Heterologous expression reveals unique properties of Trk K+ importers from nonconventional biotechnologically relevant yeast species together with their potential to support Saccharomyces cerevisiae growth.

In the model yeast Saccharomyces cerevisiae, Trk1 is the main K+ importer. It is involved in many important physiological processes, such as the maintenance of ion homeostasis, cell volume, intracellular pH, and plasma-membrane potential. The ScTrk1 protein can be of great interest to industry, as it was shown that changes in its activity influence ethanol production and tolerance in S. cerevisiae and also cell performance in the presence of organic acids or high ammonium under low K+ conditions. Nonconventional yeast species are attracting attention due to their unique properties and as a potential source of genes that encode proteins with unusual characteristics. In this work, we aimed to study and compare Trk proteins from Debaryomyces hansenii, Hortaea werneckii, Kluyveromyces marxianus, and Yarrowia lipolytica, four biotechnologically relevant yeasts that tolerate various extreme environments. Heterologous expression in S. cerevisiae cells lacking the endogenous Trk importers revealed differences in the studied Trk proteins' abilities to support the growth of cells under various cultivation conditions such as low K+ or the presence of toxic cations, to reduce plasma-membrane potential or to take up Rb+ . Examination of the potential of Trks to support the stress resistance of S. cerevisiae wild-type strains showed that Y. lipolytica Trk1 is a promising tool for improving cell tolerance to both low K+ and high salt and that the overproduction of S. cerevisiae's own Trk1 was the most efficient at improving the growth of cells in the presence of highly toxic Li+ ions.

Exploring Biocontrol of Unwanted Fungi by Autochthonous Debaryomyces hansenii Strains Isolated from Dry Meat Products.

The exploration of alternatives to the use of chemical preservatives in food is a topic that has attracted great attention. The implementation of regulations associated with the reduction of these elements directly affects the production of cured meat products, with the premise of looking for more "natural" alternatives. From a previously identified collection of 24 strains of Debaryomyces hansenii, isolated from dry meat products of the "Valle de los Pedroches" (Córdoba), a screening was carried out to determine which strains had inhibitory potential against a battery of fungi belonging to the genera Aspergillus, Penicillium, and Candida. After a series of general trials, four strains showing the greatest potential were selected by a streak inhibition assay performed at several concentrations of NaCl. The inhibitory activity of the selected D. hansenii strains was later evaluated by measuring their fungal antagonistic diffusible and volatile compound production following radial inhibition and mouth-to-mouth approaches, respectively. Growth aspects, sporulation, and morphology changes were also considered during these assays. The results support ideas already raised in previous studies, such as the presence of D. hanseniii could imply a reduction of pathogenic fungi in food. Autochthonous yeast strains inhibited not only the mycelial growth, but also sporulation, which strengthens the biocontrol activity of this yeast. Our results show that, under certain conditions, all tested D. hansenii strains were able to alter the growth/development of fungi, being especially evident in the cases of Penicillium expansum and Aspergillus niger. Finally, our research can facilitate the future comparison of results in this area, since we contributed to standardize the methodology described to date, we quantified the number of yeast cells and spores used during the experiments, we homogenized growth conditions for both, yeasts, and molds, and applied an image analyzer software to quantify the growth of the studied microorganisms in solid media.

Debaryomyces hansenii Is a Real Tool to Improve a Diversity of Characteristics in Sausages and Dry-Meat Products.

Debaryomyces hansenii yeast represents a promising target for basic and applied biotechnological research It is known that D. hansenii is abundant in sausages and dry-meat products, but information regarding its contribution to their characteristics is blurry and contradictory. The main goal in this review was to define the biological contribution of D. hansenii to the final features of these products. Depending on multiple factors, D. hansenii may affect diverse physicochemical characteristics of meat products. However, there is general agreement about the significant generation of volatile and aromatic compounds caused by the metabolic activities of this yeast, which consequently provide a tendency for improved consumer acceptance. We also summarize current evidence highlighting that it is not possible to predict what the results would be after the inoculation of a meat product with a selected D. hansenii strain without a pivotal previous study. The use of D. hansenii as a biocontrol agent and to manufacture new meat products by decreasing preservatives are examples of exploring research lines that will complement current knowledge and contribute to prepare new and more ecological products.

Overlapping responses between salt and oxidative stress in Debaryomyces hansenii.

Debaryomyces hansenii is a halotolerant yeast of importance in basic and applied research. Previous reports hinted about possible links between saline and oxidative stress responses in this yeast. The aim of this work was to study that hypothesis at different molecular levels, investigating after oxidative and saline stress: (i) transcription of seven genes related to oxidative and/or saline responses, (ii) activity of two main anti-oxidative enzymes, (iii) existence of common metabolic intermediates, and (iv) generation of damages to biomolecules as lipids and proteins. Our results showed how expression of genes related to oxidative stress was induced by exposure to NaCl and KCl, and, vice versa, transcription of some genes related to osmotic/salt stress responses was regulated by H2O2. Moreover, and contrary to S. cerevisiae, in D. hansenii HOG1 and MSN2 genes were modulated by stress at their transcriptional level. At the enzymatic level, saline stress also induced antioxidative enzymatic defenses as catalase and glutathione reductase. Furthermore, we demonstrated that both stresses are connected by the generation of intracellular ROS, and that hydrogen peroxide can affect the accumulation of in-cell sodium. On the other hand, no significant alterations in lipid oxidation or total glutathione content were observed upon exposure to both stresses tested. The results described in this work could help to understand the responses to both stressors, and to improve the biotechnological potential of D. hansenni.

Inoculation with a terroir selected Debaryomyces hansenii strain changes physico-chemical characteristics of Iberian cured pork loin.

Debaryomyces hansenii Lr1, previously isolated from naturally fermented cured pork loin ("lomo embuchado"), was used to inoculate Iberian pork loin under four different conditions. In all cases, specifically inoculated D. hansenii yeasts grew on the surface of the product and affected its physico-chemical and sensory characteristics. Inoculated pork loin kept higher water activity and pH and possessed lower sodium content than control samples at the end of the 90 days ripening period. Moreover, inoculation with Lr1 yeast did not change the general profile of fatty acids but modified the levels of volatile and aroma compounds by decreasing aldehydes and increasing esters and alcohol compounds. Principal Component Analysis (PCA) of the different physico-chemical parameters that were determined showed a clear separation between the samples indicating that they were different. A tendency in consumers acceptance to prefer cured pork loin specifically inoculated with the higher amounts of yeasts used in this study was observed.

Lepra en mujeres: estigma y dolor / No disponible

La lepra o enfermedad de Hansen es una enfermedad infecciosa crónica incapacitante. Esta enfermedad permanece en el olvido de la ciencia y de la investigación en el campo de la salud; las mujeres que la padecen sufren doblemente la exclusión, por su condición de enfermas y de mujeres. Ser mujer hará que tengan menos probabilidades que los hombres de ser diagnosticadas a tiempo, aumentando el riesgo de desarrollar una discapacidad de por vida. Lo que hace que se observe, dentro de la propia exclusión de la enfermedad, una triple discriminación en las mujeres y niñas enfermas de lepra, por su género, por las discapacidades que resultan de la enfermedad y por sufrir el impacto del estigma, lo que acentúa aún más su pobreza

Leprosy or Hansen's disease is a chronic disabling infectious disease. This disease remains in the oblivion of science and research in the field of health; women who suffer from it suffer double exclusion, because they are sick and women. Being a woman will make them less likely than men to be diagnosed early, increasing the risk of developing a disability for life. What makes observed, within the exclusion of the disease itself, a triple discrimination in women and girls suffering from leprosy, by their gender, by the disabilities that result from the disease and by suffering the impact of stigma, which it further accentuates their poverty

Epidemiological and Clinical Characteristics of Children and Adolescents with Leprosy Admitted Over 16 Years at a Rural Hospital in Ethiopia: A Retrospective Analysis.

AIM: To analyse differences in children and adolescents aged ≤18 years admitted to the leprosy ward in a rural Ethiopian hospital >16 years. METHODS: We retrospectively collected data from leprosy admission registry books on patients with leprosy who were admitted to a referral hospital from September 2000 to September 2016. RESULTS: There were 2129 admissions for leprosy during the study period: 180 (8.4%) patients were s ≤ 18 years old. Of these, 98 (54.4%) were male and 82 (45.6%) were female. The proportion of new diagnoses in children and adolescents was 31.7%, significantly higher than in adults (11.7%; p < 0.001). There were also significant differences in the prevalence of lepromatous ulcers (46.9 vs. 61.7%), leprosy reaction (29.4 vs. 13.0%) and neuritis (16.9 vs.5.3%) between these age groups. CONCLUSIONS: There were more new diagnoses, leprosy reactions and neuritis, and fewer lepromatous ulcers, in children and adolescents compared with adults, with younger patients being referred more frequently to reference centres.

Diffuse Multibacillary Leprosy of Lucio and Latapí with Lucio's Phenomenon, Peru.

Diffuse multibacillary leprosy of Lucio and Latapí is mainly reported in Mexico and Central America. We report a case in a 65-year-old man in Peru. He also had Lucio's phenomenon, characterized by vascular thrombosis and invasion of blood vessel walls by leprosy bacilli, causing extensive skin ulcers.

Debaryomyces hansenii Strains from Valle De Los Pedroches Iberian Dry Meat Products: Isolation, Identification, Characterization, and Selection for Starter Cultures.

Yeasts, filamentous fungi, and bacteria colonize the surface of fermented sausages during the ripening process. The source of this microbiota is their surrounding environment, and is influenced by the maturing conditions and starter cultures. Debaryomyces hansenii was previously isolated from several dry-cured meat products and associated with the lipolytic and proteolytic changes that occur in these products, influencing their taste and flavor. Therefore, this study isolated the yeast microbiota present in the casing from different meat products ("lomo," "chorizo," and "salchichón") from the Valle de los Pedroches region in southern Spain. D. hansenii was by far the most abundant species in each product, as all 22 selected isolates were identified as D. hansenii by biochemical and/or molecular methods. In contrast, no yeasts were found in the meat batter. These data constitute the first study of the yeasts present in "lomo" sausages and particularly the highly appreciated Valle de los Pedroches "lomo" sausages. Furthermore, the resistance of these isolates to different pHs, temperatures, and saline stress was studied, together with their catabolic characteristics. Based on the results, certain isolates are proposed as valuable candidate starter cultures that could improve both the manufacture and the flavor of such dry-cured meat products, and provide an understanding of new mechanisms involved in stress tolerance. Applied mediumscale industrial tests are currently in progress.

The halotolerant Debaryomyces hansenii, the Cinderella of non-conventional yeasts.

Debaryomyces hansenii is a halotolerant yeast with a high biotechnological potential, particularly in the food industry. However, research in this yeast is limited by its molecular peculiarities. In this review we summarize the state of the art of research in this microorganisms, describing both pros and cons. We discuss (i) its halotolerance, (ii) the molecular factors involved in saline and osmotic stress, (iii) its high gene density and ambiguous CUG decoding, and (iv) its biotechnological and medical interests. We trust that all the bottlenecks in its study will soon be overcome and D. hansenii will become a fundamental organism for food biotechnological processes. Copyright © 2016 John Wiley & Sons, Ltd.

Epidemiology of Leprosy in Spain: The Role of the International Migration.

BACKGROUND: Although incidence of leprosy in Spain has declined steadily over the years, the fivefold increase in immigration since the turn of the century--much of it from countries where leprosy is still prevalent--has been linked to an uptick in registered cases. OBJECTIVE: To describe the epidemiologic trends of incident leprosy cases detected in Spain among Spanish- and foreign-born population groups. METHODS: Observational, retrospective study of suspected leprosy cases in Spain, as reported through the System of Compulsory Notification of Diseases from 2003 to 2013, with results disaggregated by country of birth. We collected statistical data on leprosy burden for other countries from WHO to estimate the expected number of imported cases. RESULTS: Of the 168 leprosy cases registered during the study period, 40 (24.6%) were in Spanish patients, while 128 (76.2%) were detected in legally resident immigrants. We identified a significantly higher number of imported leprosy cases during the 2008-2010 and 2011-2013 trienniums compared to the reference biennium 2003-2004 (OR 5.38, 95% CI 1.83-14.88 and OR 4.80, 95% CI 1.41-16.33, respectively). Most imported cases were diagnosed in Latin American immigrants (71.9%), especially Brazilians, but also Paraguayans, Bolivians and other nationalities from South and Central America. However, registered incidence was lower than expected for each year. For example, in 2003, the expected new cases in immigrants was 47.12, compared to only four cases that were actually detected (a 91% difference). Likewise, we expected to find 49.6 incident cases among immigrants in 2009, but only 15 new cases were reported (60% fewer than expected). CONCLUSION: Imported cases of leprosy are responsible for most leprosy incidence in Spain, and we cannot rule out some under-diagnosis. Clinicians should be made more aware of the potential for leprosy incidence among patients from countries where the disease is endemic.

Dolor neuropático en pacientes afectados por la lepra / No disponible

Algunos pacientes con lepra pueden sufrir dolor crónico o dolor neuropático, tras el éxito del tratamiento. La prevalencia oscila entre un 17% y un 70% de los antiguos pacientes leprosos. El dolor neuropático se asocia con un deterioro de la calidad de vida de los pacientes y conlleva un elevado agravamiento de la morbilidad psicológica propia de los pacientes con lepra. El manejo de los pacientes con dolor neuropático puede requerir los servicios de salud durante muchos años y un elevado consumo de analgésicos

Some leprosy patients may suffer chronic pain or neuropathic pain after successful treatment. The prevalence ranges between 17% and 70 % of former leprosy patients. Neuropathic pain is associated with deterioration in the quality of life of patients and involves high worsening own psychological morbidity of patients with leprosy. The management of patients with neuropathic pain may require health services for many years and a high consumption of analgesic drugs

Leprosy ulcers in a rural hospital of Ethiopia: pattern of aerobic bacterial isolates and drug sensitivities.

BACKGROUND: Plantar ulcers, which commonly occur in leprosy patients, tend to recur increasing physical disability. The aim of this study is to identify both the bacteriological profile of these ulcers and the antibiotic susceptibility of the isolated bacteria. MATERIALS AND METHODS: 68 leprosy patients with chronic ulcers attending the in-patient department of Gambo General Hospital, West Arsi, were included in this study. Proper sample collection, inoculation on culture media, and final identification using biochemical methods were undertaken. RESULTS: 66 patients (97.1%) had a positive culture. A total of 81 microorganisms were isolated. Multiple organisms (two or more) were isolated in 15 (22.7% out of positive culture) patients. The main isolation was Proteus spp (30.9%), followed by Escherichia coli (21.0%), Staphylococcus aureus (18.5%) and Pseudomonas aeruginosa (9.9%). In the total number of the isolated bacteria, the antibiotics with less resistance were gentamicin (18.5%), fosfomycin (22.2%) cefoxitin (24.7%), ceftriaxone (25.9%) ciprofloxacin (25.9%), and amoxicillin-clavulanic acid (28.49%). CONCLUSION: The bacteriological study of plantar ulcers of leprosy patients revealed Enterobacteriaceae and S. aureus as the main pathogens involved in such infections. The results of this study may guide empirical therapy in a rural area hospital where culture and susceptibility testing facilities are scarce.

Prevalence and characteristics of neuropathic pain in leprosy patients treated years ago.

The aim of this study was to determine the prevalence of neuropathic pain, now recognized as another late complication of leprosy, and its characteristics among leprosy patients. A cross-sectional study was carried out of people treated for leprosy up to at least 5 years ago in a rural area of Ethiopia. Seventy-four patients were interviewed using the Neuropathic Pain Symptom Inventory (NPSI) questionnaire. In total, 78.9% of the patients were female with a mean age of 42.9. The mean time from initial diagnosis to the time of the study was 28.0 years, and 73.0% of patients were diagnosed over 20 years ago. Fifty-two (70.3%) reported having symptoms suggestive of neuropathic pain and the majority described the pain as burning (88.5%), electric (80.8%), stabbing (76.9%), cutting (76.9%), tingling (65.4%), squeezing (57.7%), and/or pressure (53.8%). The pain caused a severe or moderate impact on daily life in 75% and 57.7% of cases, respectively, and 92.3% suffered from disrupted sleep. Eighty percent of patients with pain (42/52) took some medication for pain relief. Neuropathic pain is common in patients treated for leprosy and in more than half of them, it causes disruption in their daily life and sleep, limiting their quality of life even more.

The burden of leprosy in children and adolescents in rural southern Ethiopia.

BACKGROUND: Childhood leprosy has an important bearing on the epidemiology of disease and reflects the level of control in a community. There is limited information about this disease in rural Ethiopia. SUBJECTS AND METHODS: Data were collected retrospectively from leprosy patient records and treatment cards in a rural mission hospital in southern Ethiopia from 1999 to 2011. RESULTS: Over the 13-year period, 298 patients with leprosy were registered for treatment. Of these, 22 (7.4%) were children (under 15 years) and 66 (21.1%) were adolescents (from 15 to 18 years). The male:female ratio was 2.6:1 in children and 1.7:1 in adolescents. Slit skin examination was positive in eight of 15 (36.4%) child patients and in 26 of 53 (41.3%) adolescents. Multibacillary leprosy was the most common type in both age groups, encountered in 95.5% of children and in 84.1% of adolescents. Six (27.3%) children and 18 (28.6%) adolescents had deformities of the hand, feet or eyes (WHO grade II), detected either at the time of diagnosis or during follow-up. Sixty-one per cent of children were transferred to their health institutions for treatment and follow-up; seven children (38.9%) completed the treatment and one (5.9%) defaulted while on therapy. Of 63 adolescent patients, 31 (49.2%) completed the recommended therapy, 28 (44.4%) were transferred out, and four (6.3%) defaulted on therapy. CONCLUSION: Childhood leprosy continues to be a common problem in rural southern Ethiopia. Multibacillary disease and disabilities remain common in children. Early detection and treatment of cases including the study of contacts should reduce the burden of leprosy in the community.

Lepra en población infantiles / No disponible

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Salt and oxidative stress tolerance in Debaryomyces hansenii and Debaryomyces fabryi.

We report the characterization of five strains belonging to the halotolerant highly related Debaryomyces hansenii/fabryi species. The analysis performed consisted in studying tolerance properties, membrane characteristics, and cation incell amounts. We have specifically investigated (1) tolerance to different chemicals, (2) tolerance to osmotic and salt stress, (3) tolerance and response to oxidative stress, (4) reactive oxygen species (ROS) content, (5) relative membrane potential, (6) cell volume, (7) K(+) and Na(+) ion content, and (8) membrane fluidity. Unexpectedly, no direct relationship was found between one particular strain, Na(+) content and its tolerance to NaCl or between its ROS content and its tolerance to H(2)O(2). Results show that, although in general, human origin D. fabryi strains were more resistant to oxidative stress and presented shorter doubling times and smaller cell volume than food isolated D. hansenii ones, strains belonging to the same species can be significantly different. Debaryomyces fabryi CBS1793 strain highlighted for its extremely tolerant behavior when exposed to the diverse stress factors studied.

Uña y Lepra / No disponible

En la lepra, las uñas pueden verse afectadas hasta en tres de cada cuatro pacientes que presentan la enfermedad. Los cambios ungueales no son específicos de la lepra, pueden ser muy variados y afectar al tamaño, el grosor, la superficie, la consistencia, el color de la relación placa-lecho y el tejido ungueal en general. Los cambios de las uñas en los pacientes con lepra son muy variados como estrías longitudinales, melanoniquia longitudinal, braquioniquia, uña en raqueta, anoniquia, onicauxis, onicogrifosis, hapaloniquia, onicorrexis, líneas de Beau, pterigium unguis dorsal, onicolisis, palidez de la uña y hematoma subungueal (AU)

In leprosy, the nails may be affected by up to three quarters of patients with the disease. Nail changes are not specific leprosy, can vary widely and affect the size, thickness, surface area, consistency, colour and relative bed plate-nail tissue in general. Nail changes in leprosy patients are varied as longitudinal striations, longitudinal melanonychia, brachionychia, nail racket, anonychia, onychauxix, onychogryphosis, hapalonychia onychorrhexis, Beau lines, dorsal pterygium unguis, onycholysis, pallor and subungual hematoma (AU)

Gender differential on characteristics and outcome of leprosy patients admitted to a long-term care rural hospital in South-Eastern Ethiopia.

INTRODUCTION: In previous studies, women are less aware of causation and symptoms of leprosy and have less access to health care coverage than men, thus contributing to their delay in seeking for treatment. We assess the gender differences in leprosy cases admitted to a rural referral hospital in Ethiopia for 7 and a half years. METHODS: Retrospective data of the leprosy patients admitted to referral hospital were collected using leprosy admission registry books from September 2002 to January 2010. Variables were entered in an Excel 97 database. RESULTS: During the period of study, 839 patients with leprosy were admitted; 541 (64.5%) were male, and 298 (35.6%) female. Fifteen per cent of female patients, and 7.3% of male patients were paucibacillary leprosy cases while 84.8% of female patients and 92.7% of males were multibacillary leprosy cases (p<0.001). Female leprosy patients were younger than male ones (median: 36 versus 44 years) (p<0.001). In the multivariate analysis, age (odds ratio [OR]: 0.97; 95% confidence interval [CI]: 0.96-0.98; p<0.001), admission for cardiovascular diseases (OR: 7.6, 95% CI: 1.9-29.3; p=0.004), admission for gastroenteritis (OR: 14.0; 95% CI: 1.7-117; p=0.02), admission from out patients clinic (OR: 2.04; 95% CI: 1.1-4.01; p=0.02), and mortality as final outcome (OR: 3.1, 95% CI: 1.2-8.0; p=0.02) were independently associated with female gender. CONCLUSIONS: Female patients with leprosy admitted to hospital were younger, had a different profile of admission and a higher mortality rate than male ones.