Global availability of parenteral nutrition: Pre- and post-COVID-19 pandemic surveys.

OBJECTIVE: Parenteral nutrition represents a therapeutic option for patients with type 3 intestinal failure. If used exclusively, parenteral nutrition has to be complete to provide all essential nutrients. The aim was to assess the availability of parenteral nutrition in all parts of the world, to better comprehend the global situation, and to prepare an action plan to increase access to parenteral nutrition. METHODS: An international survey using an electronic questionnaire was conducted in August 2019 and repeated in May 2022. An electronic questionnaire was sent to 52 members or affiliates of the International Clinical Nutrition Section of the American Society for Parenteral and Enteral Nutrition. Questions addressed the availability of parenteral nutrition admixtures and their components, reimbursement, and prescribing pre- and post-COVID-19 pandemic. All participating countries were categorized by their economic status. RESULTS: Thirty-six country representatives responded, answering all questions. Parenteral nutrition was available in all countries (100%), but in four countries (11.1%) three-chamber bags were the only option, and in six countries a multibottle system was still used. Liver-sparing amino acids were available in 18 (50%), kidney-sparing in eight (22.2%), and electrolyte-free in 11 (30.5%) countries (30.5%). In most countries (n = 28; 79.4%), fat-soluble and water-soluble vitamins were available. Trace elements solutions were unavailable in four (11.1%) countries. Parenteral nutrition was reimbursed in most countries (n = 33; 91.6%). No significant problems due to the coronavirus pandemic were reported. CONCLUSIONS: Despite the apparent high availability of parenteral nutrition worldwide, there are some factors that may have a substantial effect on the quality of parenteral nutrition admixtures. These shortages create an environment of inequality.

Thermal niches and activity periods in syntopic Phymaturus and Liolaemus lizards from the Andes, Argentina.

Explanations for differences in thermal biology within and between species of lizards employ concepts of phylogenetic inertia and plasticity. We compared the thermal biology of three liolaemid species in the Andean highlands in Argentina: two allopatric congeners (Phymaturus williamsi and P. aguanegra) each in syntopy with Liolaemus parvus. We predicted intra and inter-generic differences in ecophysiological traits and periods of activity at both sites, ecotypic differences between the (labile) Liolaemus populations, but predicted no interspecific differences between the (putatively conservative) Phymaturus. We determined the operative temperatures (T e), field body temperatures (T b), preferred temperatures (T pref), effectiveness of thermoregulation (E), and activity periods. As expected, P. williamsi differed from L. parvus in T b, T pref, and activity periods, likely as result of niche segregation. Contrary to predictions, the Phymaturus populations exhibited differentiation in T b and T pref, while L. parvus populations differed in T pref and E. Accordingly, Phymaturus species tend to be effective thermoregulators whereas L. parvus populations behave as good thermoregulators or thermoconformers depending on thermal conditions in fluctuating habitats. Phymaturus may be less evolutionarily conservative than previously suggested. The suite of co-evolving traits affecting thermal ecology may not be collectively conservative nor labile but rather a continuum between both evolutionary paths.

Effects of habitat perturbation on lizard assemblages in the center-west of the Arid Chaco region, Argentina.

Several human disturbances contribute to the decrease of vertebrate species' richness and abundance, altering the processes of an ecosystem. We evaluate richness, diversity and relative abundance of species for lizard assemblages at sites with different degrees of perturbation in the center-west of the Arid Chaco region in Argentina. Between 2015 and 2018, six lizard assemblages were sampled monthly -using pitfall traps- in three areas of the Chaco, with a perturbed and an unperturbed (control) replica at each of the areas: (1) Chaco Mountain plain, (2) Chaco Mountain slope, and (3) Chaco Plains, and habitat characteristics of each study site were recorded. We captured 1446 lizards, belonging to 12 species. The perturbed area at the Chaco Mountain plain showed the greatest richness, diversity and abundance of species. In the perturbed Chaco Plains, species abundance decreased by about 50% with respect to the control site. Liolaemus chacoensis was the dominant species at all sites. Some species could be negatively affected by a total loss of arboreal strata, tree trunks and fallen leaves. Structural parameters of lizard assemblages were related to the habitat characteristics; therefore, these results provide information for the conservation and management of lands and lizard assemblages in the Arid Chaco.

Discovery of senolytics using machine learning.

Cellular senescence is a stress response involved in ageing and diverse disease processes including cancer, type-2 diabetes, osteoarthritis and viral infection. Despite growing interest in targeted elimination of senescent cells, only few senolytics are known due to the lack of well-characterised molecular targets. Here, we report the discovery of three senolytics using cost-effective machine learning algorithms trained solely on published data. We computationally screened various chemical libraries and validated the senolytic action of ginkgetin, periplocin and oleandrin in human cell lines under various modalities of senescence. The compounds have potency comparable to known senolytics, and we show that oleandrin has improved potency over its target as compared to best-in-class alternatives. Our approach led to several hundred-fold reduction in drug screening costs and demonstrates that artificial intelligence can take maximum advantage of small and heterogeneous drug screening data, paving the way for new open science approaches to early-stage drug discovery.

Senolytic treatment preserves biliary regenerative capacity lost through cellular senescence during cold storage.

Liver transplantation is the only curative option for patients with end-stage liver disease. Despite improvements in surgical techniques, nonanastomotic strictures (characterized by the progressive loss of biliary tract architecture) continue to occur after liver transplantation, negatively affecting liver function and frequently leading to graft loss and retransplantation. To study the biological effects of organ preservation before liver transplantation, we generated murine models that recapitulate liver procurement and static cold storage. In these models, we explored the response of cholangiocytes and hepatocytes to cold storage, focusing on responses that affect liver regeneration, including DNA damage, apoptosis, and cellular senescence. We show that biliary senescence was induced during organ retrieval and exacerbated during static cold storage, resulting in impaired biliary regeneration. We identified decoy receptor 2 (DCR2)-dependent responses in cholangiocytes and hepatocytes, which differentially affected the outcome of those populations during cold storage. Moreover, CRISPR-mediated DCR2 knockdown in vitro increased cholangiocyte proliferation and decreased cellular senescence but had the opposite effect in hepatocytes. Using the p21KO model to inhibit senescence onset, we showed that biliary tract architecture was better preserved during cold storage. Similar results were achieved by administering senolytic ABT737 to mice before procurement. Last, we perfused senolytics into discarded human donor livers and showed that biliary architecture and regenerative capacities were better preserved. Our results indicate that cholangiocytes are susceptible to senescence and identify the use of senolytics and the combination of senotherapies and machine-perfusion preservation to prevent this phenotype and reduce the incidence of biliary injury after transplantation.

Toll-like receptor 2 orchestrates a tumor suppressor response in non-small cell lung cancer.

Targeting early-stage lung cancer is vital to improve survival. However, the mechanisms and components of the early tumor suppressor response in lung cancer are not well understood. In this report, we study the role of Toll-like receptor 2 (TLR2), a regulator of oncogene-induced senescence, which is a key tumor suppressor response in premalignancy. Using human lung cancer samples and genetically engineered mouse models, we show that TLR2 is active early in lung tumorigenesis, where it correlates with improved survival and clinical regression. Mechanistically, TLR2 impairs early lung cancer progression via activation of cell intrinsic cell cycle arrest pathways and the proinflammatory senescence-associated secretory phenotype (SASP). The SASP regulates non-cell autonomous anti-tumor responses, such as immune surveillance of premalignant cells, and we observe impaired myeloid cell recruitment to lung tumors after Tlr2 loss. Last, we show that administration of a TLR2 agonist reduces lung tumor growth, highlighting TLR2 as a possible therapeutic target.

Reversal of mitochondrial malate dehydrogenase 2 enables anaplerosis via redox rescue in respiration-deficient cells.

Inhibition of the electron transport chain (ETC) prevents the regeneration of mitochondrial NAD+, resulting in cessation of the oxidative tricarboxylic acid (TCA) cycle and a consequent dependence upon reductive carboxylation for aspartate synthesis. NAD+ regeneration alone in the cytosol can rescue the viability of ETC-deficient cells. Yet, how this occurs and whether transfer of oxidative equivalents to the mitochondrion is required remain unknown. Here, we show that inhibition of the ETC drives reversal of the mitochondrial aspartate transaminase (GOT2) as well as malate and succinate dehydrogenases (MDH2 and SDH) to transfer oxidative NAD+ equivalents into the mitochondrion. This supports the NAD+-dependent activity of the mitochondrial glutamate dehydrogenase (GDH) and thereby enables anaplerosis-the entry of glutamine-derived carbon into the TCA cycle and connected biosynthetic pathways. Thus, under impaired ETC function, the cytosolic redox state is communicated into the mitochondrion and acts as a rheostat to support GDH activity and cell viability.

MYC sensitises cells to apoptosis by driving energetic demand.

The MYC oncogene is a potent driver of growth and proliferation but also sensitises cells to apoptosis, which limits its oncogenic potential. MYC induces several biosynthetic programmes and primary cells overexpressing MYC are highly sensitive to glutamine withdrawal suggesting that MYC-induced sensitisation to apoptosis may be due to imbalance of metabolic/energetic supply and demand. Here we show that MYC elevates global transcription and translation, even in the absence of glutamine, revealing metabolic demand without corresponding supply. Glutamine withdrawal from MRC-5 fibroblasts depletes key tricarboxylic acid (TCA) cycle metabolites and, in combination with MYC activation, leads to AMP accumulation and nucleotide catabolism indicative of energetic stress. Further analyses reveal that glutamine supports viability through TCA cycle energetics rather than asparagine biosynthesis and that TCA cycle inhibition confers tumour suppression on MYC-driven lymphoma in vivo. In summary, glutamine supports the viability of MYC-overexpressing cells through an energetic rather than a biosynthetic mechanism.

Ozonetherapy In The Treatment Of Nocturnal Tinnitus.

This systematic review was carried out in order to highlight the implementation of ozone therapy as a treatment for nocturnal tinnitus. It is important to note that ozone treatment has resolved nocturnal tinnitus with 100% efficacy in previous studies, producing in turn a symptomatic improvement in the patient. From these results, a high degree of efficacy can be inferred in overcoming nocturnal tinnitus treated with ozone therapy in patients with a severe index of perception of tinnitus.

Season-sex interaction induces changes in the ecophysiological traits of a lizard in a high altitude cold desert, Puna region.

Functional traits are those characteristics of organisms that influence the ability of a species to develop in a habitat and persist in the face of environmental changes. The traits are often affected by a multiplicity of species-dependent and external factors. Our objective was to investigate thermal biology of Liolaemus ruibali in a high altitude cold desert at the arid Puna region, Argentina. We address the following question: do sex and seasonal variations in environmental temperature induce changes in the ecophysiological traits? We measured and compared the operative temperatures between fall and spring; and between sexes and seasons, we compared the ecophysiological traits of lizards, microenvironmental temperatures and thermoregulatory behavior. Air and operative temperatures were different between seasons. We found an effect of season-sex interaction on field body temperatures, preferred temperatures, panting threshold and thermal quality. The voluntary and critical temperatures presented seasonal variation in relation to changes in environmental temperatures, suggesting thermal acclimatization. We note behavioral changes between seasons, with the substrate being the main resource for gaining heat in spring. We conclude that Liolaemus ruibali is an efficient thermoregulator; it is a eurythermic lizard and presents phenotypic plasticity in different ecophysiological and behavioral traits induced by sex and seasonality. In addition, we predict that this population could buffer the effects of projected global warming scenarios.

In Vivo Modeling of Patient Genetic Heterogeneity Identifies New Ways to Target Cholangiocarcinoma.

Intrahepatic cholangiocarcinoma (ICC) is an aggressive malignancy of the bile ducts within the liver characterized by high levels of genetic heterogeneity. In the context of such genetic variability, determining which oncogenic mutations drive ICC growth has been difficult, and developing modes of patient stratification and targeted therapies remains challenging. Here we model the interactions between rare mutations with more common driver genes and combine in silico analysis of patient data with highly multiplexed in vivo CRISPR-spCas9 screens to perform a functional in vivo study into the role genetic heterogeneity plays in driving ICC. Novel tumor suppressors were uncovered, which, when lost, cooperate with the RAS oncoprotein to drive ICC growth. Focusing on a set of driver mutations that interact with KRAS to initiate aggressive, sarcomatoid-type ICC revealed that tumor growth relies on Wnt and PI3K signaling. Pharmacologic coinhibition of Wnt and PI3K in vivo impeded ICC growth regardless of mutational profile. Therefore, Wnt and PI3K activity should be considered as a signature by which patients can be stratified for treatment independent of tumor genotype, and inhibitors of these pathways should be levied to treat ICC. SIGNIFICANCE: This work shows that, despite significant genetic heterogeneity, intrahepatic cholangiocarcinoma relies on a limited number of signaling pathways to grow, suggesting common therapeutic vulnerabilities across patients.

Cytoplasmic innate immune sensing by the caspase-4 non-canonical inflammasome promotes cellular senescence.

Cytoplasmic recognition of microbial lipopolysaccharides (LPS) in human cells is elicited by the caspase-4 and caspase-5 noncanonical inflammasomes, which induce a form of inflammatory cell death termed pyroptosis. Here we show that LPS-mediated activation of caspase-4 also induces a stress response promoting cellular senescence, which is dependent on the caspase-4 substrate gasdermin-D and the tumor suppressor p53. Furthermore, we found that the caspase-4 noncanonical inflammasome is induced and assembled in response to oncogenic RAS signaling during oncogene-induced senescence (OIS). Moreover, targeting caspase-4 expression in OIS showed its critical role in the senescence-associated secretory phenotype and the cell cycle arrest induced in cellular senescence. Finally, we observed that caspase-4 induction occurs in vivo in mouse models of tumor suppression and ageing. Altogether, we are showing that cellular senescence is induced by cytoplasmic LPS recognition by the noncanonical inflammasome and that this pathway is conserved in the cellular response to oncogenic stress.

Comparative thermal ecophysiology in Pristidactylus scapulatus populations from the Puna region of Argentina.

Reptiles are important models for understanding fundamental aspects of physiological ecology and for assessing how environmental change can impact biodiversity. Abiotic factors (micro-environmental temperatures, operative temperatures, thermal quality) may vary geographically along an altitudinal and latitudinal gradient, and therefore the different thermal resources available for thermoregulation also vary. Comparative analyses among populations provide an opportunity to understand how variation in abiotic factors can affect different ecophysiological traits of a species at different geographical points. Our objective was to carry out a comparative study between two populations of Pristidactylus scapulatus in the Puna region of Argentina, providing the first data available on thermal ecophysiology, thermoregulatory efficiency and locomotor performance of the species. We determined field body temperature, micro-environmental temperatures and operative temperatures. In the laboratory, we measured preferred temperatures and calculated the index of thermoregulatory efficiency. In addition, we recorded critical temperature (minimum and maximum) and we estimated the thermal sensitivity of locomotion by measuring sprint speed at different body temperatures; based on these data we calculated the optimal temperature for performance, the optimal performance breadth and thermal safety margin. Air temperatures and operative temperatures were different between sites. However, we only found differences between populations in the minimum critical temperatures, with these being lower at higher latitude. We note that P. scapulatus populations adjust optimal temperatures of performance to field body temperatures and preferred temperatures, which could reduce the costs of thermoregulation in lizards with limited daily activity and who inhabit sites with variable and unpredictable environment temperatures. We conclude that Pristidactylus scapulatus has thermal sensitivity in locomotor performance, is a moderate thermoregulator with respect to the environment and is a eurithermic lizard, which has thermal flexibility in the cold.

A sensitive and affordable multiplex RT-qPCR assay for SARS-CoV-2 detection.

With the ongoing COVID-19 (Coronavirus Disease 2019) pandemic, caused by the novel coronavirus SARS-CoV-2 (Severe Acute Respiratory Syndrome Coronavirus 2), there is a need for sensitive, specific, and affordable diagnostic tests to identify infected individuals, not all of whom are symptomatic. The most sensitive test involves the detection of viral RNA using RT-qPCR (quantitative reverse transcription PCR), with many commercial kits now available for this purpose. However, these are expensive, and supply of such kits in sufficient numbers cannot always be guaranteed. We therefore developed a multiplex assay using well-established SARS-CoV-2 targets alongside a human cellular control (RPP30) and a viral spike-in control (Phocine Herpes Virus 1 [PhHV-1]), which monitor sample quality and nucleic acid extraction efficiency, respectively. Here, we establish that this test performs as well as widely used commercial assays, but at substantially reduced cost. Furthermore, we demonstrate >1,000-fold variability in material routinely collected by combined nose and throat swabbing and establish a statistically significant correlation between the detected level of human and SARS-CoV-2 nucleic acids. The inclusion of the human control probe in our assay therefore provides a quantitative measure of sample quality that could help reduce false-negative rates. We demonstrate the feasibility of establishing a robust RT-qPCR assay at approximately 10% of the cost of equivalent commercial assays, which could benefit low-resource environments and make high-volume testing affordable.

Termorregulación de la lagartija Liolaemus chacoensis (Squamata: Liolaemidae) durante su ciclo reproductivo, en el Chaco occidental, Argentina

La regulación de la temperatura corporal en las lagartijas juega un rol clave para la supervivencia de las especies. Su condición fisiológica, en referencia a su dependencia de la temperatura ambiente, los convierte en un excelente modelo de estudio. Asimismo, los cambios en el ciclo reproductivo provocan modificaciones en los parámetros termofisiológicos. Este estudio es el primero en abordar la relación de la termorregulación con la actividad reproductiva y el sexo, para una especie de Liolaemus de la ecorregión del Chaco. El objetivo de este trabajo fue determinar cómo influye el período reproductivo, de reclutamiento y el pos-reproductivo, así como también el efecto del sexo y la longitud-hocico-cloaca sobre la termorregulación de Liolaemus chacoensis. Los individuos fueron capturados a mano en el sector Sur de Valle Fértil, provincia de San Juan, Argentina. Se capturaron un total de 44 individuos durante noviembre 2014, febrero 2015 y marzo 2015. Se registró la temperatura corporal (Tc), la temperatura del sustrato (Ts) y del aire (Ta). También, se utilizaron modelos biofísicos para registrar la temperatura operativa. Posteriormente, en el laboratorio se evaluó la temperatura seleccionada (Tsel). Luego se calculó el índice de eficiencia termorregulatoria. Los resultados indicaron que la Tc de L. chacoensis se relacionó positivamente con la Ta, presentando un comportamiento heliotérmico que se mantuvo durante los períodos y el sexo. Asimismo, la Tc fue mayor para el período reproductivo y de reclutamiento con respecto al pos-reproductivo. En particular, solo el sexo tuvo efecto sobre la Tsel, sin embargo para el rango intercuartil de Tsel tuvo efecto tanto el período del ciclo reproductivo como el sexo. El índice de eficiencia termorregulatoria indicó que L. chacoensis es un termoconforme absoluto. Sin embargo, en el período reproductivo tanto machos como hembras cambian hacia la termorregulación activa. Estos resultados muestran que los parámetros termofisiológicos, varían en los diferentes períodos del ciclo reproductivo y la selección activa de micrositios sería un mecanismo para el mantenimiento de la temperatura corporal de la especie.

Thermoregulation of Liolaemus chacoensis lizard (Squamata: Liolaemidae) during its reproductive cycle, in the Western Chaco, Argentina. Regulation of body temperature in lizards plays a key role in the survival of species. Their physiological condition, in reference to their dependence on the ambient temperature makes them an excellent study model. Also, changes in the reproductive cycle cause changes in thermophysiological parameters. This study is the first to address the relationship of thermoregulation with reproductive activity and sex, for a species of Liolaemus from the Chaco ecoregion. The aim of this work was to determine how the reproductive, recruitment and post-reproductive periods influences, as well as the effect of sex and snout-vent length on the thermoregulation of Liolaemus chacoensis. Individuals were captured by hand in the Southern sector of Valle Fertil, San Juan province, Argentina. A total of 44 individuals were captured during the months of November 2014, February 2015 and March 2015. Body (Tb), substrate (Ts) and air (Ta) temperatures were recorded. Also, biophysical models were used to record operative temperatures. Later selected temperatures (Tsel) were evaluated at the laboratory. Finally, thermoregulatory efficiency index was calculated. Results indicated that L. chacoensis Tb was positively related to Ta, presenting a heliothermic behavior that was maintained during periods and sex. Also, Tb was higher in reproductive and recruitment periods than in post-reproductive period. In particular, only sex had an effect on Tsel, however for the interquartile range of Tsel both the period of reproductive cycle and sex had an effect. The thermoregulatory efficiency index indicated that L. chacoensis is an absolute thermoconformist. However, in the reproductive period both males and females change towards active thermoregulation. These results show that thermophysiological parameters vary in different periods of reproductive cycle and the active microsite selection would be a mechanism in maintenance of body temperature of this species.