Pannexin 3 deletion reduces fat accumulation and inflammation in a sex-specific manner

Background: Pannexin 3 (PANX3) is a channel-forming glycoprotein that enables nutrient-induced inflammation in vitro, and genetic linkage data suggest that it regulates body mass index. Here, we characterized inflammatory and metabolic parameters in global Panx3 knockout (KO) mice in the context of forced treadmill running (FEX) and high-fat diet (HFD). Methods: C57BL/6N (WT) and KO mice were randomized to either a FEX running protocol or no running (SED) from 24 until 30 weeks of age. Body weight was measured biweekly, and body composition was measured at 24 and 30 weeks of age. Male WT and KO mice were fed a HFD from 12 to 28 weeks of age. Metabolic organs were analyzed for a panel of inflammatory markers and PANX3 expression. Results: In females there were no significant differences in body composition between genotypes, which could be due to the lack of PANX3 expression in female white adipose tissue, while male KOs fed a chow diet had lower body weight and lower fat mass at 24 and 30 weeks of age, which was reduced to the same extent as 6 weeks of FEX in WT mice. In addition, male KO mice exhibited significantly lower expression of multiple pro-inflammatory genes in white adipose tissue compared to WT mice. While on a HFD body weight differences were insignificant, multiple inflammatory genes were significantly different in quadriceps muscle and white adipose tissue resulting in a more anti-inflammatory phenotype in KO mice compared to WT. The lower fat mass in male KO mice may be due to significantly fewer adipocytes in their subcutaneous fat compared to WT mice. Mechanistically, adipose stromal cells (ASCs) cultured from KO mice grow significantly slower than WT ASCs. Conclusion: PANX3 is expressed in male adult mouse adipose tissue and may regulate adipocyte numbers, influencing fat accumulation and inflammation

Reading tea leaves worldwide: decoupled drivers of initial litter decomposition mass‐loss rate and stabilization

The breakdown of plant material fuels soil functioning and biodiversity. Currently, process understanding of global decomposition patterns and the drivers of such patterns are hampered by the lack of coherent large‐scale datasets. We buried 36,000 individual litterbags (tea bags) worldwide and found an overall negative correlation between initial mass‐loss rates and stabilization factors of plant‐derived carbon, using the Tea Bag Index (TBI). The stabilization factor quantifies the degree to which easy‐to‐degrade components accumulate during early‐stage decomposition (e.g. by environmental limitations). However, agriculture and an interaction between moisture and temperature led to a decoupling between initial mass‐loss rates and stabilization, notably in colder locations. Using TBI improved mass‐loss estimates of natural litter compared to models that ignored stabilization. Ignoring the transformation of dead plant material to more recalcitrant substances during early‐stage decomposition, and the environmental control of this transformation, could overestimate carbon losses during early decomposition in carbon cycle models

Synthesis and in vitro leishmanicidal activity of novel [1,2,3]triazolo[1,5-a] pyridine salts

Leishmaniasis remains a significant worldwide problem; it is of great interest to develop new drugs to fight this disease. Recently we described some [1,2,3] triazolo[1,5-a] pyridine compounds with significant leishmanicidal activity. The importance of water solubility in drug action made us realise that we could transform non charged triazolopyridines into charged analogues that could increase the degree of water solubility. With this objective we report here the synthesis of novel [1,2,3] triazolo[1,5-a] pyridinium salts 2-7 from triazolopyridines 1, and the study of their in vitro leishmanicidal activity. The activity was tested on Leishmania infantum, Leishmania braziliensis and Leishmania donovani parasites, using promastigote and intracellular amastigote forms. The cytotoxicity of the tested compounds on J774.2 macrophage cells was also measured. Five of the tested compounds (2b, 4a, 4c, 6, 7d) showed selectivity indexes higher than those of the reference drug Glucantime for the three Leishmania species. Moreover, the data on infection rate and on amastigotes showed that these compounds are the most active against the three Leishmania species. The changes in the excretion product profiles of parasites treated with the compounds were also consistent with substantial cytoplasmic alterations. On the other hand, the most active compounds were potent inhibitors of Fe-SOD in the three parasite species considered whereas their impact on human CuZn-SOD was low

Differential blockade, comparative study of different ropivacaine concentrations (0.75%; 0.2%; 0.12%) for ultrasound guided sciatic and femoral nerve blocks in calves: Prospective cross-over study

Pharmacodynamic understanding of the different local anesthetic concentrations allows adapting their use to diverse clinical/surgical procedures, such as intraoperative and/or postoperative analgesia. A crossover study was performed, where 6 calves (5 male and 1 female), weighing 120 ± 28 Kg, were subjected to combined sciatic and femoral nerve block using three ropivacaine concentrations. The treatments were: R0.75, using 0.75% ropivacaine; R0.2, 0.2% ropivacaine; and R0.12%, 0.12% ropivacaine. All treatments were performed with ultrasound and neurostimulation assistance, and a volume of 0.1 mL/kg of the respective local anesthetic solution was administered in each block point. The sites of mechanical nociceptive threshold (MNT) evaluation were based on the calf pelvic limb dermatomes. The proportion between desensitized areas, MNT elevation time and level of ataxia were registered. Elevation of MNT occurred in 100% of the tested areas in the R0.75 and R0.2 treatments, and in 82% of the R0.12 treatment. Mean MNT elevation times were 9.5 ± 0.7 h for R0.75, 6 ± 0.8 for R.02, and 2.4 ± 2.3 for R0.12, differing significantly between all treatments. No difference was observed between MNT elevation time and ataxia duration time, in each treatment. It is concluded that the duration of sensory-motor effects is dose-dependent, but there was not possible to detect block selectivity as the concentrations was reduced. More desensitized areas and extension were obtained with the use of higher concentrations

Avaliações macroscópica e histológica do reparo da cartilagem articular equina tratada com microperfurações do osso subcondral associadas ou não à injeção intra-articular de cartogenina

Resumo O objetivo deste estudo foi avaliar o reparo da cartilagem hialina equina, por meio de análises macroscópica (através de videoartroscopia) e histológica (através de fragmentos de biopsia), em defeitos condrais induzidos na tróclea lateral do fêmur tratados pela técnica de microperfurações subcondral associada ou não com administração intra-articular de cartogenina. Foram utilizados seis equinos pesando em média (±DP) 342±1,58 kg, com a idade aproximada de 7,2±1,30 anos e escore corporal de 7,1±0,75, que foram submetidos a videoartroscopia para indução da lesão condral de 1 cm2 na tróclea lateral do fêmur e realização da técnica de microperfuração do osso subcondral de ambos os joelhos. Foram realizadas quatro aplicações semanais com 20 μM de cartogenina intra-articulares em um dos joelhos (grupo tratado) e solução de ringer com lactato na articulação contralateral (grupo controle). Após o período de 60 dias, foram feitas as avaliações macroscópicas, através de videoartroscopias, e histológicas, através de biopsia. Não foram observadas diferenças significativas nos escores macroscópicos e histológicos para reparação condral entre animais dos grupos tratados e não tratados (P>0,05). De modo geral, a porcentagem média de cartilagem hialina no tecido de reparo (17,5%) foi condizente com a literatura internacional usando outros tipos de perfuração condral. Entretanto, não se observaram diferenças estatísticas entre grupos (P>0,05). A terapia com cartogenina, segundo protocolo utilizado, não produziu melhora do processo cicatricial em lesões condrais induzidas e tratadas com microperfurações na tróclea lateral do fêmur em equinos

Administração seriada de salina hipertônica 7,5% na terapia para sepse grave decorrente da síndrome da diarreia hemorrágica aguda em cães

RESUMO: Na clínica de animais de companhia é frequente cães com síndrome da diarreia hemorrágica aguda associada a quadros de sepse, o que acarreta alta mortalidade. Nesse contexto, objetivou-se, em um ensaio clínico controlado aleatorizado de centro único, estudar os efeitos tardios da solução salina hipertônica a 7,5% em aplicações seriadas, sobre variáveis hemodinâmicas, clínicas e laboratoriais em cães com quadro de sepse grave decorrente desta síndrome. Para tal, 12 cães foram aleatoriamente distribuídos em dois grupos de igual número, sendo um controle (CON) e o outro, solução salina hipertônica 7,5% (SSH). Variáveis clínicas e laboratoriais foram avaliadas imediatamente após a admissão do paciente (T0), 24 (T24), 48 (T48) e 72 (T72) horas após a admissão. O grupo SSH recebeu Ringer com lactato, antibioticoterapia, analgésico e SSH 7,5% em bolus (5mL kg-1 em 4 minutos) no T24 e no T48. O grupo CON recebeu a mesma terapia acima, porém ao invés da utilização de SSH a 7,5%, administrou-se bolus de solução de Ringer lactato na mesma dose e tempos utilizado. As avaliações em cada tempo foram realizadas anteriormente à administração dos bolus, nos dois grupos. Avaliaram-se hemograma completo e as variáveis clínicas (escala AVDN, grau de desidratação, frequência respiratória e temperatura retal), frequência cardíaca (FC), pressão arterial sistólica (PAS). Os dados paramétricos foram avaliados pelos testes Student Newman Keuls e teste t de Student, e os não paramétricos pelo teste de Friedman e Mann Whitney, com nível de significância de P≥0,05. Nas variáveis clínicas estudadas não se observou diferença entre os grupos e entre os momentos avaliados. Observou-se diferença significativa no grupo SSH no T72, com elevação da PAS e redução da FC, fato não observado no grupo CON, onde esses parâmetros não se alteraram. O hematócrito e a concentração de hemoglobina diminuíram em ambos os grupos com o tempo. As contagens dos leucócitos totais e dos monócitos apresentaram uma elevação significativa no grupo SSH, estando os leucócitos dentro da faixa de normalidade no T72. Não houve diferenças significativas em relação aos neutrófilos segmentados, porém no grupo SSH verificou-se aumento de 9,5 vezes no T72 comparado com o T24 (P=0,09), enquanto que este aumento foi de apenas 2,5 vezes no grupo CON (P=0,30). Observou-se ainda redução nas contagens de plaquetas e na concentração de globulinas no grupo COM, enquanto essas variáveis se mantiveram estáveis no grupo SSH. Conclui-se que a administração seriada de SSH 7,5% se mostrou promissora no tratamento de cães com síndrome da diarreia hemorrágica aguda, pois auxilia na estabilização dos leucócitos, plaquetas e globulinas de cães com sepse grave decorrente da síndrome da diarreia hemorrágica aguda

SARS-CoV-2 vaccination in 361 non-transplanted patients with aplastic anemia and/or paroxysmal nocturnal hemoglobinuria

We thank the support of the Severe Aplastic Anemia and Infectious Disease working parties of the EBMT and the patients for participation

Plasticity of patient-matched normal mammary epithelial cells is dependent on autologous adipose-derived stem cells

Due to the increasing clinical application of adipose-derived stem cells (ADSC), e.g. lipotransfer for breast reconstruction, this study aimed to gain novel insights regarding ADSC influence on breast tissue remodeling and determine patient-dependent factors affecting lipotransfer as well as begin to address its oncological risks. The ADSC secretome was analyzed from five normal breast reduction patients and contained elevated levels of growth factors, cytokines and proteins mediating invasion. ADSC/ADSC secretomes were tested for their influence on the function of primary mammary epithelial cells, and tumor epithelial cells using cell culture assays. ADSC/ADSC secretomes significantly stimulated proliferation, transmigration and 3D-invasion of primary normal and tumor epithelial cells. IL-6 significantly induced an EMT and invasion. The ADSC secretome significantly upregulated normal epithelial cell gene expression including MMPs and ECM receptors. Our study supports that ADSC and its secretome promote favorable conditions for normal breast tissue remodeling by changing the microenvironment. and may also be important regarding residual breast cancer cells following surgery

Myogenic differentiation of primary myoblasts and mesenchymal stromal cells under serum-free conditions on PCL-collagen I-nanoscaffolds

Abstract Background The creation of functional skeletal muscle via tissue engineering holds great promise without sacrificing healthy donor tissue. Different cell types have been investigated regarding their myogenic differentiation potential under the influence of various media supplemented with growth factors. Yet, most cell cultures include the use of animal sera, which raises safety concerns and might lead to variances in results. Electrospun nanoscaffolds represent suitable matrices for tissue engineering of skeletal muscle, combining both biocompatibility and stability. We therefore aimed to develop a serum-free myogenic differentiation medium for the co-culture of primary myoblasts (Mb) and mesenchymal stromal cells derived from the bone marrow (BMSC) and adipose tissue (ADSC) on electrospun poly-ε-caprolacton (PCL)-collagen I-nanofibers. Results Rat Mb were co-cultured with rat BMSC (BMSC/Mb) or ADSC (ADSC/Mb) two-dimensionally (2D) as monolayers or three-dimensionally (3D) on aligned PCL-collagen I-nanofibers. Differentiation media contained either AIM V, AIM V and Ultroser® G, DMEM/Ham’s F12 and Ultroser® G, or donor horse serum (DHS) as a conventional differentiation medium. In 2D co-culture groups, highest upregulation of myogenic markers could be induced by serum-free medium containing DMEM/Ham’s F12 and Ultroser® G (group 3) after 7 days. Alpha actinin skeletal muscle 2 (ACTN2) was upregulated 3.3-fold for ADSC/Mb and 1.7-fold for BMSC/Mb after myogenic induction by group 3 serum-free medium when compared to stimulation with DHS. Myogenin (MYOG) was upregulated 5.2-fold in ADSC/Mb and 2.1-fold in BMSC/Mb. On PCL-collagen I-nanoscaffolds, ADSC showed a higher cell viability compared to BMSC in co-culture with Mb. Myosin heavy chain 2, ACTN2, and MYOG as late myogenic markers, showed higher gene expression after long term stimulation with DHS compared to serum-free stimulation, especially in BMSC/Mb co-cultures. Immunocytochemical staining with myosin heavy chain verified the presence of a contractile apparatus under both serum free and standard differentiation conditions. Conclusions In this study, we were able to myogenically differentiate mesenchymal stromal cells with myoblasts on PCL-collagen I-nanoscaffolds in a serum-free medium. Our results show that this setting can be used for skeletal muscle tissue engineering, applicable to future clinical applications since no xenogenous substances were used