Instituto de Estudios Inmunológicos y Fisiopatológicos (IIFP)

Evaluación de la suplementación temprana con Kluyveromyces marxianus CIDCA 9121 en la producción de pollos parrilleros

Thu, 01 Jan 2026 00:00:00 GMT

Objeto de conferencia XIX Congreso Argentino de Ciencia y Tecnología de Alimentos (Buenos Aires, 12 al 14 de noviembre de 2025); Libro de trabajos completos XIX CYTAL 2025 Las crecientes restricciones en el uso de antimicrobianos en producción animal han estimulado la búsqueda de aditivos alimentarios que permitan mejorar los resultados productivos sin seleccionar microorganismos resistentes. Este estudio plantea evaluar el efecto de la suplementación temprana con la levadura probiótica Kluyveromyces marxianus CIDCA 9121 sobre el rendimiento productivo y la salud intestinal de pollos parrilleros buscando minimizar el uso de antibióticos como promotores de crecimiento. El ensayo se realizó en los boxes de bioseguridad del CICVyA – INTA (Hurlingham, Buenos Aires). 180 pollitos Cobb 500 de 1 día de vida se distribuyeron al azar en 9 corrales, los cuales fueron asignados a 3 tratamientos (n=60): Control, Bacitracina (50g/Tn de alimento) y Km 9121. Todos los grupos recibieron alimento y agua ad libitum. El grupo Km 9121 fue suplementado con 109 UFC/día/pollo empleando levadura fresca por instilación en buche desde el día 1 al 7 de vida de los animales. Durante todo el ensayo se monitoreo el estado de los animales y se registró el peso vivo y consumo de alimento desde el nacimiento hasta el día 42 de vida. A los 21 y 35 días, se sacrificaron 5 aves por grupo y se cuantificaron ácidos grasos de cadena corta (AGCC) por HPLC en contenido intestinal de íleon y ciego. Se evaluó la presencia de alteraciones histopatológicas en hígado e intestino mediante observación al microscopio óptico con tinción hematoxilina y eosina. Además, se determinó actividad transaminasa glutámico-oxalacética sérica, niveles de triglicéridos y colesteloremia en un autoanalizador Abbot Laboratories Aeroset. La suplementación dietaria temprana con K. marxianus CIDCA 9121 en pollos de engorde demostró ser inocua, sin causar efectos negativos en el desarrollo de los animales. Aunque no se observaron diferencias en el peso final entre los grupos, la conversión alimenticia fue menor en el grupo levadura en comparación con el grupo control y el grupo bacitracina con valores de 1,87; 1,97 y 2,16 a los 42 días, respectivamente. La concentración total de AGCC en el contenido intestinal de ciego fue mayor que en íleon para todos los animales. Para todos los grupos, a los 21 y 35 días de vida se detectaron en ciego ácido acético y fórmico, mientras que acido isovalérico solo en los pollos del grupo control y bacitracina. Los análisis histopatológicos al día 35 revelaron aumento significativo en la longitud de las vellosidades y profundidad de criptas para pollos que recibieron levadura (p<0.05). Por otro lado, se detectaron vacuolizaciones o aumento de gotas lipídicas en hepatocitos del grupo control, alteraciones que no estuvieron presentes en hígados del grupo levadura. Además, a los 21 días de vida disminuyeron los niveles de transaminasas, triglicéridos y colesterol plasmáticos para el grupo levadura, diferencia que se atenuó a los 35 días. En conjunto, estos resultados sugieren que K. marxianus CIDCA 9121 podría utilizarse como aditivo alimentario en pollos parrilleros con el fin de mejorar los parámetros productivos y la salud de los animales.

Cocoon Syndrome as a Cause of Intestinal Failure and Indication for Combined Liver–Intestine–Kidney Transplantation

Wed, 01 Jan 2025 00:00:00 GMT

Articulo Intestinal Failure; vol. 6 Background: Cocoon Syndrome, or sclerosing encapsulating peritonitis, is a rare but severe complication of long-term peritoneal dialysis. It can lead to progressive fibrotic encapsulation of the intestines, causing intestinal obstruction due to a fibrous membrane encasing the abdominal organs. Case report: We present a complex case of a 29-year-old male with history of nephronophthisis and four years on peritoneal dialysis who developed Cocoon Syndrome, leading to intestinal perforation and total enterectomy, resulting in irreversible intestinal failure. After prolonged dependence on parenteral nutrition, he developed intestinal failure-associated liver disease, ultimately becoming a candidate for combined liver–intestine and kidney transplantation. Conclusion: This case illustrates how advanced Cocoon Syndrome can lead to severe and progressive complications that may ultimately require combined liver–intestine and kidney transplantation. It underscores the importance of early recognition and appropriate multidisciplinary management to improve patient outcomes.

Montelukast activates β4-containing BK channels and emerges as a pharmacological tool to reduce hippocampal excitability

Thu, 01 Jan 2026 00:00:00 GMT

Articulo Pharmacological Research; vol. 229 Large-conductance Ca²⁺- and voltage-activated K⁺ (BK) channels are critical regulators of neuronal excitability and have been implicated in multiple epileptic syndromes. Their functional diversity arises from the co-assembly of pore-forming α-subunits with auxiliary β subunits, among which β4 is highly expressed in distinct regions of the central nervous system, including hippocampal Dentate Gyrus Granule Cells (DGGCs). Here, we identify montelukast (MTK), a clinically approved cysteinyl-leukotriene receptor antagonist, as a direct activator of BK channels, with markedly enhanced efficacy in the presence of the β1 and β4 subunits. MTK acts at submicromolar concentrations and facilitates channel opening by altering the energetics of the pore domain, independent of voltage-sensor activation or Ca²⁺ binding to the cytosolic gating ring. In mouse hippocampal slices, MTK reduces intrinsic excitability of DGGCs by decreasing input resistance and enhancing the afterhyperpolarization, effects fully reversed by the BK channel blocker paxilline. Experiments using physiological DGGCs action potential voltage waveforms confirm that MTK enhances subthreshold and evoked BK currents in α/β4 channels as selective targets. Altogether, our findings suggest that MTK, beyond its known anti-inflammatory properties, may modulate neural excitability through direct BK channel activation, offering a novel therapeutic strategy for seizure suppression.

Harnessing outer membrane vesicles derived from Bordetella pertussis to overcome key limitations of acellular pertussis vaccines

Tue, 02 Sep 2025 00:00:00 GMT

Articulo Frontiers in Immunology; vol. 16 Acellular pertussis (aP) vaccines have markedly reduced the global burden of severe pertussis. However, their limited ability to elicit mucosal and durable immunity has been linked to waning protection and sustained Bordetella pertussis circulation. Selective pressure exerted by widespread aP vaccination has contributed to the emergence and regional dissemination of pertactin-deficient (PRN^-) strains, raising additional concerns regarding vaccine effectiveness. In this context, we investigated whether incorporating outer membrane vesicles (OMVs) derived from B. pertussis into the aP vaccine could enhance its immunological profile, specifically by promoting Th1/Th17 polarization, inducing tissue-resident memory (TRM) T cells, and broadening protective coverage to include PRN^- isolates, while maintaining aP-induced immunity against lower respiratory tract colonization. Using a murine intranasal challenge model with a two-dose vaccination schedule, we assessed the safety, immunogenicity, and protective capacity of the OMV+aP vaccine prototype (combined) versus aP vaccine. The combined formulation was well tolerated and induced robust systemic and mucosal responses, characterized by higher IgG2a/IgG1 ratios, increased Th1/Th17 cytokine production (IFN-γ, IL - 17, and IL - 22), and elevated anti-B. pertussis IgA titers. Flow cytometric analyses revealed lung- and nasal-resident CD4⁺ TRM cells in the combined immunized mice, which were absent in those receiving aP alone. Functionally, OMV+aP formulation conferred superior protection in pulmonary and nasal compartments, significantly reducing lung bacterial loads (including against PRN^- strains) and uniquely diminishing nasal colonization even under high-dose challenge conditions. Passive transfer experiments confirmed the role of cellular and humoral immunity in bacterial clearance. These results demonstrate that OMVs synergize with aP to enhance immune response magnitude and quality, addressing key gaps in current aP vaccines and offering a next-generation strategy to prevent both disease and transmission.

Revisión del uso de membrana amniótica para el tratamiento de queratoconjuntivitis seca canina

Wed, 01 Jan 2025 00:00:00 GMT

Objeto de conferencia IV Jornadas de Investigación UNAJ (Florencio Varela, 31 de octubre 2024); https://rid.unaj.edu.ar/handle/123456789/3375.2; 4tas. Jornadas de Investigación / UNAJ : investigaciones interdisciplinarias en el territorio en el actual contexto Desde hace más de siglo son conocidos los efectos antiinflamatorios, inmunomoduladores y regeneradores de heridas de la membrana amniótica (AM). El uso de membrana amniótica en humanos (hAM) y en canes (cAM) es de gran utilidad en oftalmología. La utilización clínica de AM está mucho más difundida en humanos, asimismo existen miles de publicaciones científicas en humanos y solo una decena en caninos. Hasta hace pocos años el uso de AM era exclusivamente quirúrgico principalmente en úlceras corneales persistentes y neurotróficas. Desarrollos de innovación en la unión europea, y en nuestro país, en el banco de tejidos AmniosBMA, han permitido avanzar en el uso no quirúrgico de la AM, ampliando las patologías de la superficie ocular a tratar, entre ellas, ojo seco severo, conocido como queratoconjuntivitis sicca (QCS), defectos epiteliales de córnea, úlceras corneales neurotróficas, úlceras persistentes, enfermedades mucosinequiantes, etc. Estas patologías, de no ser diagnosticadas y tratadas correctamente, llevan a la ceguera. La analogía clínica en humanos del ojo seco severo canino es el Síndrome de Sjögren (SS) que se caracteriza por infiltrado linfoplasmocitario de las glándulas exocrinas, que son las que producen lágrimas y saliva, lo que lleva a la destrucción de las estas glándulas y sequedad de las mucosas. Las consecuencias de la QCS son una disminución de lágrimas que lleva a una inflamación crónica de la conjuntiva y de la córnea, dolor ocular, queratitis, úlcera y pérdida de la visión. El diagnóstico se realiza por examen con lámpara de hendidura, test de Schirmer, Rosa de Bengala o Lisamina verde. La osmolaridad de la lágrima y la citología de impresión pueden ser útiles para detectar la enfermedad de QCS en etapas tempranas. El tratamiento convencional de la QCS en caninos y humanos suele ser diferente; en canes incluye lágrimas artificiales, ciclosporina, corticoides y antibióticos y en humanos se utiliza ciclosporina, insulina tópica y suero autólogo. El presente trabajo tuvo como objetivo resumir el estado del arte del diagnóstico de ojo seco canino, su tratamiento y se describieron algunos trabajos que utilizan AM para el tratamiento de patologías oculares caninas. Se llevó a cabo la revisión de trabajos publicados. No se encontraron artículos específicos sobre el tratamiento del ojo seco canino con cAM, por lo cual se concluye que es un campo interesante de explorar y con gran potencial de aplicación.

Hereditary angioedema diagnosis evaluation score (HADES): A new clinical scoring system for predicting hereditary angioedema with C1 inhibitor deficiency

Thu, 01 May 2025 00:00:00 GMT

Articulo Journal of Allergy and Clinical Immunology: Global; vol. 4, no. 2 Background: Diagnosis of hereditary angioedema (HAE) poses challenges because of its rarity and its overlapping symptoms with allergic and gastrointestinal conditions, resulting in misdiagnosis. Objective: We developed a predictive score using clinical variables for suspected HAE patients with C1 inhibitor deficiency (HAE-C1INH) to increase suspicion of HAE and thus improve diagnosis. Methods: The HADES (HAE diagnosis evaluation score) study used a nationwide retrospective cohort of individuals with suspected HAE-C1INH in Argentina. A questionnaire was designed to collect relevant clinical information on possible predictors for HAE. Blood samples were analyzed for C1-INH/ C1q levels and C1-INH function. A predictive score was developed from the odds ratios derived from multivariate logistic regression analysis. Results: The study included 2423 individuals (1642 suspected index cases and 781 family cases). Only patients with confirmed HAE types I or II (n 5 499) were included in the final analysis; acquired angioedema/F12 gene variants were excluded. Eight clinical variables were identified as independent predictors of HAE: age at onset <20 years, recurrent limb edema, abdominal pain, vomiting, trauma as a trigger, absence of wheals, family history of angioedema, and recurrent edema lasting >24 hours. The predictive score demonstrated favorable performance in identifying HAE cases within the index population (range, 0- 18.5), with low scores (1.5-6.5) associated with high sensitivity (100%) and negative predictive value (100%), and high scores (>15) associated with high specificity (99.4%) and positive predictive value (75.0%). Conclusions: The predictive HADES offers a simple and efficient method for improving testing for suspicion of HAE by using clinical parameters. Further validation studies are required to confirm its reliability and accuracy. (J Allergy Clin Immunol Global 2025;4:100414.)

Typing of HLA susceptibility alleles as complementary tool in diagnosis of controversial cases of pediatric celiac disease

Sat, 01 Feb 2025 00:00:00 GMT

Articulo Frontiers in Nutrition; vol. 12 Objectives: Diagnosis of celiac disease (CeD), an immune-mediated disorder, is based on clinical presentation, a panel of serological markers, and the histopathological findings in duodenal biopsies. Commonly, pediatric CeD patients fulfill these criteria for diagnosis. However, lack of correlation between serology tests and histology, or no accessible biopsies because of clinical conditions or during the COVID pandemic, are conditions that led to inconclusive diagnoses. Since the majority of CeD patients carry HLA-DQ2 and/ or DQ8 alleles, HLA testing is used as a complementary tool in diagnosis though is costly and not broadly available for gastroenterology centers. Methods: We performed a retrospective study to assess the performance of HLA testing when applied to selected groups of patients who could not be definitely diagnosed following the common algorithm. Eighty patients underwent testing for CeD-related HLA-DQ2 and DQ8 alleles. Results: HLA typing contributed to diagnosis in 34 patients with positive serology but normal mucosa or those who presented negative serology or slightly positive serology (less than 3 times ULN) and duodenal histopathological changes. In patients with normal histology and negative or slightly positive serology, or those who did not undergo intestinal biopsy (39 in total), HLA typing contributed to CeD diagnosis in 23 cases, only 16 patients were admitted for a clinical follow-up program. Conclusion: HLA-DQ typing supported the diagnosis in 57 of 80 children (71.2%) with previously inconclusive results, providing a beneficial approach for diagnosing celiac disease (CeD) in selected cases.

Role of germinal center and CD39ʰⁱᵍʰCD73⁺ B cells in the age-related tonsillar involution

Mon, 01 Jan 2024 00:00:00 GMT

Articulo Immunity & Ageing; vol. 21 Background The tonsils operate as a protection ring of mucosa at the gates of the upper aero-digestive tract. They show similarities with lymph nodes and participate as inductive organs of systemic and mucosal immunity. Based on the reduction of their size since puberty, they are thought to experience involution in adulthood. In this context, we have used tonsillar mononuclear cells (TMC) isolated from patients at different stages of life, to study the effect of ageing and the concomitant persistent inflammation on these immune cells. Results We found an age-dependent reduction in the proportion of germinal center B cell population (BGC) and its T cell counterpart (T follicular helper germinal center cells, TfhGC). Also, we demonstrated an increment in the percentage of local memory B cells and mantle zone T follicular helper cells (mTfh). Furthermore, younger tonsils rendered higher proportion of proliferative immune cells within the freshly isolated TMC fraction than those from older ones. We demonstrated the accumulation of a B cell subset (CD20⁺CD39ʰⁱᵍʰCD73⁺ cells) metabolically adapted to catabolize adenosine triphosphate (ATP) as patients get older. To finish, tonsillar B cells from patients at different ages did not show differences in their proliferative response to stimulation ex vivo, in bulk TMC cultures. Conclusions This paper sheds light on the changing aspects of the immune cellular landscape, over the course of time and constant exposure, at the entrance of the respiratory and digestive systems. Our findings support the notion that there is a re-modelling of the immune functionality of the excised tonsils over time. They are indicative of a transition from an effector type of immune response, typically oriented to reduce pathogen burden early in life, to the development of an immunosuppressive microenvironment at later stages, when tissue damage control gets critical provided the time passed under immune attack. Noteworthy, when isolated from such histologic microenvironment, older tonsillar B cells seem to level their proliferation capacity with the younger ones. Understanding these features will not only contribute to comprehend the differences in susceptibility to pathogens among children and adults but would also impact on vaccine developments intended to target these relevant mucosal sites.

The role of tubular cells in the pathogenesis of Fabry nephropathy

Mon, 01 Jan 2024 00:00:00 GMT

Articulo Frontiers in Cardiovascular Medicine; vol. 11 The pathophysiology of Fabry nephropathy (FN) is induced by galactosidase A deficiency with a chronic exposure of glycolipids to every lineage of renal cells. Tissue damage is attributed to the activation of molecular pathways, resulting in tissue fibrosis and chronic kidney disease. Podocytes have been the primary focus in clinical pathophysiological research because of the striking accumulation of large glycolipid deposits observable in histology. Yet, the tubular interstitium makes up a large portion of the whole organ, and therefore, its role must be further considered in pathogenic processes. In this review, we would like to propose Fabry tubulopathy and its ensuing functional effects as the first pathological signs and contributing factors to the development of FN. We will summarize and discuss the current literature regarding the role of tubular cells in Fabry kidney pathophysiology. Starting from clinical and histological evidence, we will highlight the data from animal models and cell cultures outlining the pathophysiological pathways associated with tubular interstitial injury causing renal fibrosis in Fabry nephropathy.

Activation of smooth muscle BK channels by hydrochlorothiazide requires cell integrity and the presence of BK β1-subunits

Thu, 30 Nov 2017 00:00:00 GMT

Articulo Acta Pharmacologica Sinica; vol. 39 Thiazide-like diuretics are one of the most commonly used drugs to treat arterial hypertension, with their efficacy being linked to their chronic vasodilatory effects. Previous studies have suggested that activation of the large conductance voltage- and Ca2+-dependent K+ (BK) channel (Slo 1, MaxiK channel) is responsible for the thiazide-induced vasodilatory effect. However, direct electrophysiological evidence supporting this claim is lacking. BK channels can be associated with small accessory β-subunits (β1-β4) that confer specific biophysical and pharmacological characteristics to the current phenotype. The β1-subunit is primarily expressed in smooth muscle cells (SMCs). The effect of hydrochlorothiazide (HCTZ) on BK channel activity was measured using patch-clamp electrophysiology on native SMCs from human umbilical artery (HUASMCs) and HEK293T cells expressing the BK channel (with and without the β1-subunit). HCTZ significantly activated the BK current when evaluated using the whole-cell and cell-attached configurations. However, HCTZ did not affect the unitary conductance and open probability of the BK channel in the inside-out configuration, suggesting an indirect mechanism requiring cell integrity. The increase in BK channel activity due to HCTZ was concentration dependent, with an EC50 of 28 μmol/L, and membrane potential did not influence the concentration relationship. Moreover, our data c learly demonstrated that the HCTZ-induced activation of BK channels required the presence of β1-subunits. A β1-subunit-dependent mechanism that requires SMC integrity leads to HCTZ-induced BK channel activation.

Direct evidence for local IgE production in the human colonic mucosa

Sun, 02 May 2021 00:00:00 GMT

Articulo Allergy; vol. 76, no. 5 Understanding the biology of IgE in humans has become a matter of interest that remains incompletely understood due to the rarity of peripheral IgE+ cells. The increased incidence of allergic diseases and food-induced anaphylaxis overtime demands an urgent development of disease-modifying therapies that reverse the synthesis of IgE and the induction of IgE-mediated allergic reactions. Although some reports showed specific IgE in stools and IgE+ cells in the gastrointestinal tract of allergic patients, the microanatomical location of the class-switch recombination (CSR) to ε chain is largely unknown. This isotype is produced through CSR mechanism by activated IgM-producing B cells (direct switch) or following IgG+ B memory cell-switch to ε chain (sequential switch) in a Th2 milieu with the induction of the cytidine deaminase (AID). It has been demonstrated that this mechanism occurs prior to germinal center (GC) formation in secondary lymphoid tissues such as lymph nodes and tonsils, but it is controversial whether the IgE isotype switching can occur in the human intestinal mucosa and which niches could be involved. Our study aimed to investigate the local IgE synthesis in the stroma of juvenile colonic polyps (JP) from patients with rectal bleeding and the relationship between IgE production and food sensitization, a risk factor for food allergy.

Experimental Assessment of Intestinal Damage in Controlled Donation After Circulatory Death for Visceral Transplantation

Thu, 12 Jan 2023 00:00:00 GMT

Articulo Transplant International; vol. 36, art. 10803 There is an urgent need to address the shortage of potential multivisceral grafts in order to reduce the average time in waiting list. Since donation after circulatory death (DCD) has been successfully employed for other solid organs, a thorough evaluation of the use of intestinal grafts from DCD is warranted. Here, we have generated a model of Maastricht III DCD in rodents, focusing on the viability of intestinal and multivisceral grafts at five (DCD5) and twenty (DCD20) minutes of cardiac arrest compared to living and brain death donors. DCD groups exhibited time-dependent damage. DCD20 generated substantial intestinal mucosal injury and decreased number of Goblet cells whereas grafts from DCD5 closely resemble those of brain death and living donors groups in terms intestinal morphology, expression of tight junction proteins and number of Paneth and Globet cells. Upon transplantation, intestines from DCD5 showed increased ischemia/reperfusion damage compared to living donor grafts, however mucosal integrity was recovered 48 h after transplantation. No differences in terms of graft rejection, gene expression and absorptive function between DCD5 and living donor were observed at 7 post-transplant days. Collectively, our results highlight DCD as a possible strategy to increase multivisceral donation and transplantation procedures.

Acute cellular rejection in small-bowel transplantation impairs NCR+ innate lymphoid cell subpopulation 3/interleukin 22 axis

Mon, 01 Jun 2020 00:00:00 GMT

Articulo Transplant Immunology; vol. 60 Acute cellular rejection (ACR) remains as one of the main causes of graft loss and death in intestinal transplant (ITx) patients. ACR promotes intestinal injury, disruption of the mucosal barrier, bacterial translocation, and organ dysfunction. As epithelial regeneration is critical in reversing these consequences, the functional axis between the innate lymphoid cell subpopulation 3 (ILC3) and interleukin 22 plays an essential role in that process. Natural-cytotoxic-receptor–positive (NCR+) ILC3 cells have been demonstrated to induce intestinal-stem-cell proliferation along with an IL-22–dependent expansion of that population in several intestinal pathologies, though thus far not after ITx. Therefore, we intended to determine the impact of chronic immunosuppression and ACR on ILC3 cells and interleukin-22 (IL-22) production in the lamina propria after that intervention. Materials and methods We compared biopsies from healthy volunteers with biopsies from ITx recipients without or with mild-to-moderate ACR, using flow cytometry and the quantitative-PCR. Results NCR+ ILC3 cells were found to be unaffected by immunosuppression at different time points posttransplant when patients did not experience ACR, but were diminished upon the occurrence of ACR independently of the post-ITx time. Moreover, IL-22–expression levels were notably reduced in ACR. Conclusion The NCR+-ILC3/IL-22 axis is impaired during ACR contributing to a delay in or lack of a complete and efficient epithelial regeneration. Thus, our findings reveal that IL-22 analogues could potentially be used as a new complementary therapeutic approach, in conjunction with immunosuppressant drugs, in order to promote mucosal regeneration upon ACR.

Exploring the pathophysiologic cascade leading to osteoclastogenic activation in gaucher disease monocytes generated via CRISPR/Cas9 technology

Sun, 01 Jan 2023 00:00:00 GMT

Articulo International Journal of Molecular Sciences; vol. 24, no. 13 Gaucher disease (GD) is caused by biallelic pathogenic variants in the acid β-glucosidase gene (GBA1), leading to a deficiency in the β-glucocerebrosidase (GCase) enzyme activity resulting in the intracellular accumulation of sphingolipids. Skeletal alterations are one of the most disabling features in GD patients. Although both defective bone formation and increased bone resorption due to osteoblast and osteoclast dysfunction contribute to GD bone pathology, the molecular bases are not fully understood, and bone disease is not completely resolved with currently available specific therapies. For this reason, using editing technology, our group has developed a reliable, isogenic, and easy-to-handle cellular model of GD monocytes (GBAKO-THP1) to facilitate GD pathophysiology studies and high-throughput drug screenings. In this work, we further characterized the model showing an increase in proinflammatory cytokines (Interleukin-1β and Tumor Necrosis Factor-α) release and activation of osteoclastogenesis. Furthermore, our data suggest that GD monocytes would display an increased osteoclastogenic potential, independent of their interaction with the GD microenvironment or other GD cells. Both proinflammatory cytokine production and osteoclastogenesis were restored at least, in part, by treating cells with the recombinant human GCase, a substrate synthase inhibitor, a pharmacological chaperone, and an anti-inflammatory compound. Besides confirming that this model would be suitable to perform high-throughput screening of therapeutic molecules that act via different mechanisms and on different phenotypic features, our data provided insights into the pathogenic cascade, leading to osteoclastogenesis exacerbation and its contribution to bone pathology in GD.

Bone marrow adipocytes alteration in an in vitro model of Gaucher Disease

Sun, 01 Jan 2023 00:00:00 GMT

Articulo Molecular Genetics and Metabolism Reports; vol. 36 Gaucher disease (GD) is caused by biallelic pathogenic variants in GBA1 gene that encodes the lysosomal enzyme glucocerebrosidase. Up to now, specific treatment for GD cannot completely reverse bone complications. Bone is composed of different cell types; including osteoblasts, osteocytes and osteoclasts. Osteoblasts are present on bone surfaces and are derived from local mesenchymal stem cells (MSCs). Depending on environment conditions, MSCs could differentiate into osteoblasts and adipocytes. Mature adipocytes-secreted adipokines and free fatty acids affect both osteoblasts and osteoclasts formation/activity and therefore mediate skeletal homeostasis. The aim of this study was to evaluate possible alterations in GD adipocyte (GD Ad) that could contribute to bone complications. MSCs were grown in adipogenic media in order to evaluate expression of differentiation markers as PPAR-γ. PPAR-γ was observed into the nucleus of GD Ad, indicating that these cells are properly stimulated. However, these cells accumulate lesser lipid droplets (LDs) than Control Ad. In order to study lipid droplet metabolism, we evaluated the lipolysis of these structures by the measurement of free glycerol in culture supernatant. Our results indicated that GD Ad had an alteration in this process, evidenced by an increase in glycerol release. We have also evaluated two enzymes involved in LDs synthesis: fatty acid synthase (FASN) and stearoylcoenzyme A desaturase 1 (SCD1). The transcription of these genes was decreased in GD Ad, suggesting a dysfunction in the synthesis of LDs. In conclusion, our results show an alteration in LDs metabolism of GD Ad, independent of adipocyte differentiation process. This alteration would be caused by an increase in lipolysis in early stages of differentiation and also by a reduction of lipid synthesis, which could contribute with the skeletal imbalance in GD.

Clostridioides difficile: characterization of the circulating toxinotypes in an Argentinean public hospital

Sun, 01 Jan 2023 00:00:00 GMT

Articulo Revista Argentina de Microbiología; vol. 55, no. 1 Clostridioides difficile is a spore-forming anaerobe microorganism associated to nosocomial diarrhea. Its virulence is mainly associated with TcdA and TcdB toxins, encoded by their respective tcdA and tcdB genes. These genes are part of the pathogenicity locus (PaLoc). Our aim was to characterize relevant C. difficile toxinotypes circulating in the hospital setting. The tcdA and tcdB genes were amplified and digested with different restriction enzymes: EcoRI for tcdA; HincII and AccI for tcdB. In addition, the presence of the cdtB (binary toxin) gene, TcdA and TcdB toxins by dot blot and the cytotoxic effect of culture supernatants on Vero cells, were evaluated. Altogether, these studies revealed three different circulating toxinotypes according to Rupnik’s classification: 0, I and VIII, being the latter the most prevalent one. Even though more studies are certainly necessary (e.g. sequencing analysis), it is worth noting that the occurrence of toxinotype I could be related to the introduction of bacteria from different geographical origins.

Opportunities and challenges for newborn screening and early diagnosis of rare diseases in Latin America

Sat, 01 Jan 2022 00:00:00 GMT

Articulo Frontiers in Genetics; vol. 13 Rare diseases (RDs) cause considerable death and disability in Latin America. Still, there is no consensus on their definition across the region. Patients with RDs face a diagnostic odyssey to find a correct diagnosis, which may last many years and creates a burden for caregivers, healthcare systems, and society. These diagnostic delays have repercussions on the health and economic burden created by RDs and continue to represent an unmet medical need. This review analyzes barriers to the widespread adoption of newborn screening (NBS) programs and early diagnostic methods for RDs in Latin America and provides recommendations to achieve this critical objective. Increasing the adoption of NBS programs and promoting early diagnosis of RDs are the first steps to improving health outcomes for patients living with RDs. A coordinated, multistakeholder effort from leaders of patient organizations, government, industry, medical societies, academia, and healthcare services is required to increase the adoption of NBS programs. Patients’ best interests should remain the guiding principle for decisions regarding NBS implementation and early diagnosis for RDs.

Inflammation Is Present, Persistent and More Sensitive to Proinflammatory Triggers in Celiac Disease Enterocytes

Tue, 01 Feb 2022 00:00:00 GMT

Articulo International Journal of Molecular Sciences; vol. 23, no. 4 Celiac disease (CD) is a chronic inflammatory disease caused by a genetic predisposition to an abnormal T cell-mediated immune response to the gluten in the diet. Different environmental proinflammatory factors can influence and amplify the T cell-mediated response to gluten. The aim of this manuscript was to study the role of enterocytes in CD intestinal inflammation and their response to different proinflammatory factors, such as gliadin and viruses. Intestinal biopsies from CD patients on a gluten-containing (GCD-CD) or a gluten-free diet (GFD-CD) as well as biopsies from potential CD patients (Pot-CD) before the onset of intestinal lesions and controls (CTR) were used to investigate IL-1β and IL-6 mRNA levels in situ. Organoids from CD patients were used to test the levels of NF-κB, ERK, IL-6, and IL-1β by Western blot (WB), ELISA, and quantitative PCR. The Toll-like receptor ligand loxoribine (Lox) and gliadin peptide P31-43 were used as proinflammatory stimuli. In CD biopsies inflammation markers IL-1β and IL-6 were increased in the enterocytes, and also in Pot-CD before the onset of the intestinal lesion and in GFD-CD. The inflammatory markers pNF-κB, pERK, IL-1β, and IL-6 were increased and persistent in CD organoids; these organoids were more sensitive to P31-43 and Lox stimuli compared with CTR organoids. Taken together, these observations point to constitutive inflammation in CD enterocytes, which are more sensitive to inflammatory stimuli such as food components and viruses.

Few amino acid mutations in H6 Influenza A virus from South American Lineage increase viral replication efficiency in poultry

Sat, 01 Jan 2022 00:00:00 GMT

Articulo Frontiers in Microbiology; vol. 13 In chickens, infections due to influenza A virus (IAV) can be mild to severe and lethal. The study of IAV infections in poultry has been mostly limited to strains from the North American and Eurasian lineages, whereas limited information exists on similar studies with strains from the South American lineage (SAm). To better evaluate the risk of introduction of a prototypical SAm IAV strain into poultry, chickens were infected with a wild-type SAm origin strain (WT557/H6N2). The resulting virus progeny was serially passaged in chickens 20 times, and the immunopathological effects of the last passage virus, 20Ch557/H6N2, in chickens were compared to those of the parental strain. A comparison of complete viral genome sequences indicated that the 20Ch557/H6N2 strain contained 13 amino acid differences compared to the wild-type strain. Five of these mutations are in functionally relevant regions of the viral surface glycoproteins hemagglutinin (HA) and neuraminidase (NA). However, despite higher and more prolonged virus shedding in chickens inoculated with the 20Ch557/H6N2 strain compared to those that received the WT557/H6N2 strain, transmission to naïve chickens was not observed for either group. Analyses by flow cytometry of mononuclear cells and lymphocyte subpopulations from the lamina propria and intraepithelial lymphocytic cells (IELs) from the ileum revealed a significant increase in the percentages of CD3CTCRgdC IELs in chickens inoculated with the 20Ch557/H6N2 strain compared to those inoculated with the WT557/H6N2 strain.

ARGENVAC será una herramienta para convivir con el virus de COVID-19

Sat, 01 Jan 2022 00:00:00 GMT

Comunicacion ARGENVAC: a tool to live with SARS-CoV-2 Revista de la Sociedad Odontológica de La Plata; año XXXII, no. 62 En este momento de la pandemia, mentalizarse que vamos a convivir con el virus es un aprendizaje al que debemos habituarnos luego de más de 2 años de la emergencia del SARS-CoV-2. El mundo ha cambiado en muchos aspectos y estamos frente a un enclave donde convivir con el virus será lo que nos permita retomar hábitos de vida más naturales, aunque seguramente diferentes a los pre-pandemia. La crisis sanitaria ocasionada por la COVID-19 nos ha enseñado que el ser humano es un ser social que necesita interacción entre pares para poder tener un estado saludable. Con un virus tan transmisible y peligroso, sobre todo para aquellos que son grupo de riesgo, la única herramienta que nos permite alcanzar y mantener este estado son las vacunas, y principalmente los planes de vacunación.