Regional moderate hyperthermia for mild-to-moderate COVID-19 (TherMoCoV study): a randomized controlled trial.

Background: To prevent COVID-19 progression, low-cost alternatives that are available to all patients are needed. Diverse forms of thermotherapy have been proposed to prevent progression to severe/critical COVID-19. Objective: The aim of this study is to evaluate the efficacy and safety of local thermotherapy to prevent disease progression in hospitalized adult patients with mild-to-moderate COVID-19. Methods: A multicenter, open-label, parallel-group, randomized, adaptive trial is used to evaluate the efficacy and safety of local thermotherapy to prevent disease progression in hospitalized adult patients with mild-to-moderate COVID-19. Eligible hospitalized adult patients with symptoms of COVID-19 with ≤5 days from symptom onset, meeting criteria for mild or moderate COVID-19, were randomly assigned to the intervention consisting of local thermotherapy via an electric heat pad in the thorax (target temperature range 39.5­42°C) continuously for 90 min, twice daily, for 5 days, or standard care. The main outcome was the proportion of patients who progressed to severe-to-critical COVID-19 or death. Patients were randomized in a 1:1 ratio through a centralized computer-generated sequence of minimization with a random component of 20%. Participants and medical staff were not blinded to the intervention. Results: One-hundred and five participants (thermotherapy n = 54, control n = 51) with a median age of 53 (IQR: 41­64) years were included for analysis after the early cessation of recruitment due to the closure of all temporal COVID-19 units (target sample size = 274). The primary outcome of disease progression occurred in 31.4% (16/51) of patients in the control group vs. 25.9% (14/54) of those receiving thermotherapy (risk difference = 5.5%; 95%CI: −11.8­22.7, p = 0.54). Thermotherapy was well tolerated with a median total duration of thermotherapy of 900 (IQR: 877.5­900) min. Seven (13.7%) patients in the control group and seven (12.9%) in the thermotherapy group had at least one AE (p = 0.9), none of which were causally attributed to the intervention. No statistically significant differences in serum cytokines (IL-1ß, IL-6, IL-8, IL-10, IL-17, and IFN-γ) were observed between day 5 and baseline among groups. Conclusion: Local thermotherapy was safe and well-tolerated. A non-statistically significant lower proportion of patients who experienced disease progression was found in the thermotherapy group compared to standard care. Local thermotherapy could be further studied as a strategy to prevent disease progression in ambulatory settings.Clinical Trial registration: www.clinicaltrials.gov, identifier: NCT04363541.

Bases para un tratamiento con hipertermia moderada en leishmaniosis cutánea / Bases for a treatment of cutaneous leishmaniasis with moderate hyperthermia

Resumen Introducción: La hipertermia regional entre 38 y 39.5 °C ha sido empleada para tratar procesos inflamatorios y, ocasionalmente, infecciones cutáneas. En zonas endémicas de leishmaniosis se aplican compresas calientes como tratamiento antiparasitario. Objetivo: Conocer las bases del tratamiento térmico de la leishmaniosis para regularlo adecuadamente. Métodos: Parásitos Leishmania mexicana cultivados in vitro fueron incubados por periodos variables de 37 °C y después a 25 °C.. Los parásitos se tiñeron con anexina V-FITC y yoduro de propidio para detectar inducción de apoptosis y su viabilidad. Se realizaron curvas de crecimiento postratamiento e identificación del ciclo celular con anticuerpos anticiclinas. Resultados: Después de 30 minutos de exposición a una temperatura de 37 °C, un porcentaje variable de parásitos perdieron su característica forma ovalada y se tornaron esféricos, sin refringencia y con núcleos condensados, cambios que sugirieron apoptosis, la cual fue confirmada mediante tinción con anexina V-FITC. La cantidad de parásitos en proceso de apoptosis fue proporcional al tiempo de exposición. Los parásitos en los que se observó apoptosis se tiñeron con anticuerpos anticiclinas. Conclusiones: La elevación constante, regulada y fisiológica de la temperatura por más de 30 minutos induce apoptosis de parásitos Leishmania mexicana cultivados in vitro, cuando se encuentran en fase activa en el ciclo celular.

Abstract Introduction: Regional hyperthermia at between 38 and 39.5 °C has been used to treat inflammatory processes and, occasionally, skin infections. In areas where leishmaniasis is endemic, hot compresses are applied as anti-parasitic treatment. Objective: To identify the bases of leishmaniasis thermal treatment in order to properly regulate it. Methods: In vitro-cultured Leishmania mexicana parasites were incubated for variable periods at 37 and then, to 25 °C. The parasites were then stained with Annexin V-FITC to detect apoptosis induction and with propidium iodide for viability. Post-treatment growth curves and cell cycle identification with anti-cyclin antibodies were performed. Results: After 30 minutes of exposure to a temperature of 37 °C, a variable proportion of parasites lost their characteristic oval shape and became spherical, without refringence and with condensed nuclei, with these changes suggesting apoptosis, which was confirmed by Annexin V-FITC staining. The number of parasites that underwent apoptosis was proportional to exposure time. Parasites in which apoptosis was observed were stained with anti-cyclin antibodies. Conclusions: Constant, regulated and physiological elevation of temperature for more than 30 minutes induces apoptosis of in vitro-cultured L. mexicana parasites when they are in an active phase of the cell cycle.

Lactancia materna y COVID-19 / Breastfeeding and COVID-19

Resumen La pandemia de enfermedad por coronavirus 2019 (COVID-19) ha afectado a todas las dimensiones de la atención en salud, entre ellas el aseguramiento de la lactancia materna exclusiva y su promoción. El riesgo de contagio y las consecuencias de la pandemia han provocado preocupación entre las futuras madres o las que se ya encuentran lactando debido al riesgo de una posible transmisión del virus a través de la leche materna. Aunque aún no se ha detectado el coronavirus 2 del síndrome respiratorio agudo grave (SARS-CoV-2) activo en la leche materna. El miedo al contagio ha favorecido las políticas de aislamiento madre-hijo. Hasta el momento no existe evidencia de transmisión vertical y el riesgo de transmisión horizontal en el lactante es similar al de la población general. En lactantes con COVID-19 la lactancia materna incluso puede cambiar favorablemente el curso clínico de la enfermedad.

Abstract The COVID-19 pandemic has affected the health attention in all dimensions, one of them, the exclusive breastfeeding assurance and her promotion. The high risk of contagion and the pandemic consequences have raised a number of concerns in future mothers or those who are breastfeeding because of the risk of a possible transmission of the virus through breast milk. Although SARS-CoV2 has no evidence of being active on breast milk, the fear of contagion has favored mother-child isolation policies. At this point, there are no evidence of vertical transmission and the risk of horizontal transmission in the infant is similar to the general population. Breastfeeding in newborn with COVID-19, can even favorably change the clinical course of the disease.

Exploring the rationale for thermotherapy in COVID-19.

Increased transmissibility of the pandemic severe acute respiratory coronavirus 2 (SARS-CoV-2) has been noted to occur at lower ambient temperatures. This is seemingly related to a better replication of most respiratory viruses, including SARS-CoV-2, at lower-than-core body temperatures (i.e., 33 °C vs 37 °C). Also, intrinsic characteristics of SARS-CoV-2 make it a heat-susceptible pathogen. Thermotherapy has successfully been used to combat viral infections in plants which could otherwise result in great economic losses; 90% of viruses causing infections in plants are positive-sense single-stranded ribonucleic acid (+ssRNA) viruses, a characteristic shared by SARS-CoV-2. Thus, it is possible to envision the use of heat-based interventions (thermotherapy or mild-temperature hyperthermia) in patients with COVID-19 for which moderate cycles (every 8-12 h) of mild-temperature hyperthermia (1-2 h) have been proposed. However, there are potential safety and mechanistic concerns which could limit the use of thermotherapy only to patients with mild-to-moderate COVID-19 to prevent disease progression rather than to treat patients who have already progressed to severe-to-critical COVID-19. Here, we review the characteristics of SARS-CoV-2 which make it a heat-susceptible virus, potential host mechanisms which could be enhanced at higher temperatures to aid viral clearance, and how thermotherapy could be investigated as a modality of treatment in patients with COVID-19 while taking into consideration potential risks.

Inflammatory Response in Preterm and Very Preterm Newborns with Sepsis.

The response of the adaptive immune system is usually less intense in premature neonates than term neonates. The primary objective of this study was to determine whether immunological parameters vary between preterm (PT) neonates (≥32 weeks of gestational age) and very preterm (VPT) neonates (<32 weeks of gestational age). A cross-sectional study was designed to prospectively follow PT and VPT neonates at risk of developing sepsis. Plasma concentrations of IFN-γ, TNF-α, IL-6, IL-4, and IL-10 were detected using flow cytometry. C-reactive protein (C-RP) and the complex SC5b-9 were detected in the plasma using commercial kits. A total of 83 patients were included. The laboratory results and clinical histories showed that 26 patients had sepsis; 14 were VPT, and 12 were PT. The levels of C-RP, SC5b-9 (innate immune response mediators), and IL-10 or IL-4 (anti-inflammatory cytokines) were elevated during sepsis in both groups. IFN-γ, TNF-α, and IL-6 (proinflammatory cytokines) were differentially elevated only in PT neonates. The VPT neonates with sepsis presented increases in C-RP, SC5b-9, and anti-inflammatory cytokines but not in proinflammatory cytokines, whereas PT neonates showed increases in all studied mediators of inflammation.

Polymorphic variation of hypoxia inducible factor-1 A (HIF1A) gene might contribute to the development of knee osteoarthritis: a pilot study.

BACKGROUND: Osteoarthritis (OA) is a multifactorial degenerative condition of the whole joint with a complex pathogenesis whose development and progression is significantly mediated by interactions between the joint cartilage and articular tissues, particularly, proinflammatory mediators and oxidative stress, which results in cartilage deterioration and subchondral bone destruction. HIF-1 alpha regulates oxygen homeostasis in hypoxic tissues such as joint cartilage; efficiency of transcriptional activity of the HIF1A gene is strongly influenced by the presence of polymorphic variants. Given the loss of articular cartilage and with intention to restore damaged tissue, WISP-1 participates in the development of subchondral bone; further, its expression is highly increased in chondrocytes of OA patients. The aim of this study was to evaluate gene frequencies of HIF1A and WISP1 polymorphisms in Mexican patients suffering from knee OA. METHODS: We determined HIF1A rs11549465 (P582S), rs11549467 (A588T), and rs2057482 (C191T), and WISP1 rs2929970 (A2364G) polymorphisms in 70 Mexican patients with knee OA and compare them to those present in 66 ethnically matched healthy controls. Genotyping for these polymorphisms was performed by Real-Time PCR using TaqMan probes. RESULTS: Gene frequencies exhibited a significant increase of the CC genotype of rs11549465 polymorphism in knee OA patients as compared with those present in controls (P = 0.003 OR = 5.7, 95% CI = 1.7-21.6); CT genotype and T allele showed decreased frequency in the knee OA group vs. the controls (P = 0.003 OR = 0.2, CI = 0.05-0.6; and P = 0.004 OR = 0.2, CI = 0.05-0.65, respectively). Allele frequencies of the other polymorphic variants were similar in both patients and controls. CONCLUSIONS: These results suggest that the presence of the rs11549465 SNP (HIF1A) plays a role protective in the loss of articular cartilage in our population, and offers the possibility to further study the molecular mechanisms within cartilage and subchondral bone.

Antileishmanial activity of quinazoline derivatives: synthesis, docking screens, molecular dynamic simulations and electrochemical studies.

A series of quinazoline-2,4,6-triamine were synthesized and evaluated in vitro against Leishmania mexicana. Among them, N(6)-(ferrocenmethyl)quinazolin-2,4,6-triamine (H2) showed activity on promastigotes and intracellular amastigotes, as well as low cytotoxicity in mammalian cells. Docking and electrochemical studies showed the importance of both the ferrocene and the heterocyclic nucleus to the observed activity. H2 is readily oxidized electrochemically, indicating that the mechanism of action probably involves redox reactions.

Activity of hydroxyurea against Leishmania mexicana.

Leishmania mexicana is a protozoan parasite that causes a disease in humans with frequent relapses after treatment. It is also highly resistant to the currently available drugs. For this reason, there is an urgent need for more effective antileishmanial drugs. Hydroxyurea, an anticancer drug, is toxic to replicating eukaryotic cells and has been proven to be effective in arresting the Leishmania major cell cycle. In this study, hydroxyurea was tested in an in vitro model of intracellular Leishmania infection in macrophages. The parasite density in infected macrophages was measured by microscopy after incubation for various times and treatment with hydroxyurea at different concentrations. Viable parasites that could be transformed into promastigotes by shifting the temperature to 26 degrees C were counted every other day after the replacement of hydroxyurea with fresh medium. Meglumine antimoniate, the standard drug treatment for Leishmania mexicana, was used as a reference drug under the same experimental conditions. Hydroxyurea completely eliminated Leishmania parasites when it was used at a dosage of 10 or 100 microg/ml. Differences in the length of treatment needed to achieve elimination were as follows: the 10-microg/ml doses required 9 days, while 3 days was sufficient when 100 microg/ml was used. Hydroxyurea had a 50% effective dose of 0.015 microg/ml in vitro, which was observed on day 6 after exposure. Hydroxyurea is highly effective in killing intracellular amastigotes in vitro.

Low serum levels of dehydroepiandrosterone and cortisol in human diffuse cutaneous leishmaniasis by Leishmania mexicana.

Low levels of dehydroepiandrosterone (DHEA) and cortisol hormones produced by the suprarenal cortex have been associated with diseases involving chronic inflammation, low interferon (IFN)-gamma, and high interleukin (IL)-6. Diffuse cutaneous leishmaniasis (DL), a long-lasting intracellular parasitic infectious disease, can spread unknown levels of DHEA and cortisol. Serum concentrations of both were measured in 5 patients with DL, in 15 patients with localized lesions produced by Leishmania (LL), and in 20 healthy volunteers. Leishmania mexicana mexicana was identified as the causal agent in patients with DL and LL. Hormone levels were lower in DL compared with controls and LL. Furthermore, we detected a lower percentage of IFN-gamma-positive cells with higher levels of IL-6 and higher titers of anti-Leishmania antibodies in patients with DL, whereas patients with LL were similar to controls. These data suggest that patients with DL may be good candidates for DHEA and cortisol supplementation.

Antígenos de Giardia lamblia que son reconocidos por anticuerpos de lecha materna / Antigens of Giarda lamblia that are recognized by antibodies from human milk

Existen evidencias de que anticuerpos específicos participan en la eliminación del parásito Giardia lamblia, como lo indican los mayores índices de giardiosis en pacientes con deficiencias de IgA, y la protección contra esta parasitosis que la leche de animales inmunizados confiere a las crías. Con el fin de identificar los antígenos de este protozoario que son reconocidos por los anticuerpos presentes en las secreciones, se cuantificaron por ELISA las IgG e IgA anti-giardia en 104 leches maternas colectadas en el Departamento de Nutrición del Hospital del Niño, de Villahermosa, Tabasco; se determinó el sitio de reconocimiento en el trofozoíto por inmunofluorescencia indirecta (IFI) y se identificó el peso molecular relativo (PMr) de las proteínas de G. lamblia reconocidas por inmuno-electrotransferencia (Western-blot). En la prueba de ELISA, 89 por ciento de las muestras tuvo anticuerpos de clase IgA por arriba del punto de corte y hubo una estrecha correlación entre los niveles de IgA e IgG en la leche. En la IFI se observó que la membrana del trofozoíto reaccionó con intensidad a los anticuerpos anti-giardia de las muestras de leche estudiadas, algunas de las cuales tiñeron intensamente al disco suctor. Los antígenos identificados por Western-blot fueron 27 bandas con PMr de 26 a 185 kDa. Las seis bandas reconocidas con mayor frecuencia fueron las de 135, 127, 123, 112, 84 y 75 kDa. Otras con menor intensidad y frecuencia correspondieron a 185, 45, 42, 36 y 26 kDa. Se observaron dos patrones de bandas: múltiple, únicas (84 o_ 75 kDa). Este estudio identificó el peso molecular y la localización de las proteínas de G. lamblia hacia las que están dirigidos los anticuerpos secretorios específicos de la leche humana y que pudieran ser de importancia en la prevención y eliminación de la giardiosis