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.

Nuclear pore density controls heterochromatin reorganization during senescence.

During oncogene-induced senescence (OIS), heterochromatin is lost from the nuclear periphery and forms internal senescence-associated heterochromatin foci (SAHFs). We show that an increased nuclear pore density during OIS is responsible for SAHF formation. In particular, the nucleoporin TPR is necessary for both formation and maintenance of SAHFs. Loss of SAHFs does not affect cell cycle arrest but abrogates the senescence-associated secretory phenotype-a program of inflammatory cytokine gene activation. Our results uncover a previously unknown role of nuclear pores in heterochromatin reorganization in mammalian nuclei and demonstrate the importance of heterochromatin organization for a specific gene activation program.

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.

Condensin II mutation causes T-cell lymphoma through tissue-specific genome instability.

Chromosomal instability is a hallmark of cancer, but mitotic regulators are rarely mutated in tumors. Mutations in the condensin complexes, which restructure chromosomes to facilitate segregation during mitosis, are significantly enriched in cancer genomes, but experimental evidence implicating condensin dysfunction in tumorigenesis is lacking. We report that mice inheriting missense mutations in a condensin II subunit (Caph2nes) develop T-cell lymphoma. Before tumors develop, we found that the same Caph2 mutation impairs ploidy maintenance to a different extent in different hematopoietic cell types, with ploidy most severely perturbed at the CD4+CD8+ T-cell stage from which tumors initiate. Premalignant CD4+CD8+ T cells show persistent catenations during chromosome segregation, triggering DNA damage in diploid daughter cells and elevated ploidy. Genome sequencing revealed that Caph2 single-mutant tumors are near diploid but carry deletions spanning tumor suppressor genes, whereas P53 inactivation allowed Caph2 mutant cells with whole-chromosome gains and structural rearrangements to form highly aggressive disease. Together, our data challenge the view that mitotic chromosome formation is an invariant process during development and provide evidence that defective mitotic chromosome structure can promote tumorigenesis.

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.

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.

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.

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.

Gut microbial ecology of lizards: insights into diversity in the wild, effects of captivity, variation across gut regions and transmission.

Animals maintain complex associations with a diverse microbiota living in their guts. Our understanding of the ecology of these associations is extremely limited in reptiles. Here, we report an in-depth study into the microbial ecology of gut communities in three syntopic and viviparous lizard species (two omnivores: Liolaemus parvus and Liolaemus ruibali and an herbivore: Phymaturus williamsi). Using 16S rRNA gene sequencing to inventory various bacterial communities, we elucidate four major findings: (i) closely related lizard species harbour distinct gut bacterial microbiota that remain distinguishable in captivity; a considerable portion of gut bacterial diversity (39.1%) in nature overlap with that found on plant material, (ii) captivity changes bacterial community composition, although host-specific communities are retained, (iii) faecal samples are largely representative of the hindgut bacterial community and thus represent acceptable sources for nondestructive sampling, and (iv) lizards born in captivity and separated from their mothers within 24 h shared 34.3% of their gut bacterial diversity with their mothers, suggestive of maternal or environmental transmission. Each of these findings represents the first time such a topic has been investigated in lizard hosts. Taken together, our findings provide a foundation for comparative analyses of the faecal and gastrointestinal microbiota of reptile hosts.

Thubunaea eleodori sp. nov. (Nematoda: Physalopteridae) from Liolaemus eleodori (Sauria: Liolaemidae) from Argentina.

Thubunaea eleodori sp. nov. is described from the stomach of Liolaemus eleodori (Sauria: Liolaemidae) from San Guillermo National Park, Province of San Juan, Argentina. T. eleodori is most similar to those species lacking spicules, T. cnemidophorus, T. fitsimonsi, T. parkeri, T. schukurovi, and T. smogorzhewskii. T. eleodori is separated from these species based on the papilla pattern. T. eleodori has 12 pedunculate papillae and 14 sessile papillae, T. smogorzhewskii lacks pedunculate papillae, T. fitsimonsi and T. parkeri lack sessile papillae, and T. cnemidophorus has14-16 pedunculate papillae and 12 sessile papillae. T. eleodori represents the first member of the genus to be reported from Argentina.

Thermal biology in two syntopic lizards, Phymaturus extrilidus and Liolaemus parvus, in the Puna region of Argentina.

Body temperature is the most important ecophysiological variable affecting reptiles' life history. Moreover, thermoregulation in ectotherms implies a struggle to reach preferred temperatures in natural conditions due to the influence of biotic and abiotic factors. Our objective was to evaluate and compare the thermal biology of two syntopic species, Phymaturus extrilidus and Liolaemus parvus, in the Puna region of San Juan, Argentina. We determined body temperature (Tb), micro-environmental temperatures (Ta and Ts) and operative temperatures (Te) in the field. In the laboratory, we measured preferred temperatures (Tpref) and calculated the index of thermoregulatory efficiency (E). Neither body temperatures in the field nor preferred temperatures varied between seasons and sexes. Body temperatures were lower than preferred temperatures for both species. Nevertheless, regardless of the low thermal offer available in habitat, both species did achieve body temperatures higher than operative temperatures during activity. Thermoregulatory effectiveness was moderate in P. extrilidus (E=0.65), while L. parvus presented greater thermoregulatory efficiency (E=0.78). We conclude that under the rigorous climate conditions of the Puna, Phymaturus extrilidus and Liolaemus parvus are able to actively and efficiently thermoregulate, maintaining body temperatures close to the preferred and higher than those of its habitat. Differences in thermal characteristics between Phymaturus extrilidus and Liolaemus parvus are a consequence of differential limitations imposed on each species by the environment and of forces inherent to their life histories.

Senescence impairs successful reprogramming to pluripotent stem cells.

Somatic cells can be reprogrammed into induced pluripotent stem (iPS) cells by overexpressing combinations of factors such as Oct4, Sox2, Klf4, and c-Myc. Reprogramming is slow and stochastic, suggesting the existence of barriers limiting its efficiency. Here we identify senescence as one such barrier. Expression of the four reprogramming factors triggers senescence by up-regulating p53, p16(INK4a), and p21(CIP1). Induction of DNA damage response and chromatin remodeling of the INK4a/ARF locus are two of the mechanisms behind senescence induction. Crucially, ablation of different senescence effectors improves the efficiency of reprogramming, suggesting novel strategies for maximizing the generation of iPS cells.

Histone demethylase JMJD3 contributes to epigenetic control of INK4a/ARF by oncogenic RAS.

The INK4a/ARF tumor suppressor locus, a key executor of cellular senescence, is regulated by members of the Polycomb group (PcG) of transcriptional repressors. Here we show that signaling from oncogenic RAS overrides PcG-mediated repression of INK4a by activating the H3K27 demethylase JMJD3 and down-regulating the methyltransferase EZH2. In human fibroblasts, JMJD3 activates INK4a, but not ARF, and causes p16(INK4a)-dependent arrest. In mouse embryo fibroblasts, Jmjd3 activates both Ink4a and Arf and elicits a p53-dependent arrest, echoing the effects of RAS in this system. Our findings directly implicate JMJD3 in the regulation of INK4a/ARF during oncogene-induced senescence and suggest that JMJD3 has the capacity to act as a tumor suppressor.

Physiological and microbial adjustments to diet quality permit facultative herbivory in an omnivorous lizard.

While herbivory is a common feeding strategy in a number of vertebrate classes, less than 4% of squamate reptiles feed primarily on plant material. It has been hypothesized that physiological or microbial limitations may constrain the evolution of herbivory in lizards. Herbivorous lizards exhibit adaptations in digestive morphology and function that allow them to better assimilate plant material. However, it is unknown whether these traits are fixed or perhaps phenotypically flexible as a result of diet. Here, we maintained a naturally omnivorous lizard, Liolaemus ruibali, on a mixed diet of 50% insects and 50% plant material, or a plant-rich diet of 90% plant material. We compared parameters of digestive performance, gut morphology and function, and gut microbial community structure between the two groups. We found that lizards fed the plant-rich diet maintained nitrogen balance and exhibited low minimum nitrogen requirements. Additionally, lizards fed the plant-rich diet exhibited significantly longer small intestines and larger hindguts, demonstrating that gut morphology is phenotypically flexible. Lizards fed the plant-rich diet harbored small intestinal communities that were more diverse and enriched in Melainabacteria and Oscillospira compared with mixed diet-fed lizards. Additionally, the relative abundance of sulfate-reducing bacteria in the small intestine significantly correlated with whole-animal fiber digestibility. Thus, we suggest that physiological and microbial limitations do not sensu stricto constrain the evolution of herbivory in lizards. Rather, ecological context and fitness consequences may be more important in driving the evolution of this feeding strategy.

Gestational diabetes induces alterations in the function of neonatal endothelial colony-forming cells.

BACKGROUND: Children born to mothers with gestational diabetes mellitus (GDM) experience increased risk of developing hypertension, type 2 diabetes mellitus, and obesity. Disrupted function of endothelial colony-forming cells (ECFCs) may contribute to this enhanced risk. The goal of this study was to determine whether cord blood ECFCs from GDM pregnancies exhibit altered functionality. METHODS: ECFCs isolated from the cord blood of control and GDM pregnancies were assessed for proliferation, senescence, and Matrigel network formation. The requirement for p38MAPK in hyperglycemia-induced senescence was determined using inhibition and overexpression studies. RESULTS: GDM-exposed ECFCs were more proliferative than control ECFCs. However, GDM-exposed ECFCs exhibited decreased network-forming ability in Matrigel. Aging of ECFCs by serial passaging led to increased senescence and reduced proliferation of GDM-exposed ECFCs. ECFCs from GDM pregnancies were resistant to hyperglycemia-induced senescence compared with those from controls. In response to hyperglycemia, control ECFCs activated p38MAPK, which was required for hyperglycemia-induced senescence. In contrast, GDM-exposed ECFCs showed no change in p38MAPK activation under equivalent conditions. CONCLUSION: Intrauterine exposure of ECFCs to GDM induces unique phenotypic alterations. The resistance of GDM-exposed ECFCs to hyperglycemia-induced senescence and decreased p38MAPK activation suggest that these progenitor cells have undergone changes that induce tolerance to a hyperglycemic environment.

Plant diversity in live fences and pastures, two examples from the Mexican humid tropics.

This study analyzes the potential uses of live fences and pastures as reservoirs of plant diversity for two regions with different management histories, Los Tuxtlas (LT) and Uxpanapa (UX), Veracruz, México. We studied two habitats, live fences and pastures, analyzed their species richness, diversity, structure and plant composition and classified species according to plant regeneration modes (light-demanding and shade tolerant), seed dispersal syndrome and their local uses. We recorded 62 species of trees at LT and 48 at UX. Live fences were more diverse than pastures in both regions. The LT site showed to analyze the relationship a higher diversity of plants in regeneration stages than the one at UX. However, UX had higher diversity of adult plants in the pastures than LT. Composition and structure of live fences were different between regions, as well as within live fences and pastures, 53 % of species were light-demanding and 40 % were shade tolerant; 70 % of the species were dispersed by birds. Differences between sites are associated with the modifications in live fences structure, which changed according to managerial practices and the use of local species; this may influence plant regeneration modes as well as the visits of avian dispersal agents. In LT, all species found in live fences were useful to humans, whereas in UX, less than half were used by the local population. Our results underline the importance of live fences and isolated trees in pasture habitats as potential sites to host native and useful species from tropical rain forests in livestock landscapes.

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.

Description of new andean species of the genus Phymaturus (Iguania: Liolaemidae) from Northwestern Argentina.

As a result of several field trips and studies of collections of Phymaturus samples from Andean areas of central western Argentina (San Juan province), we discovered two populations that exhibit a particular character combination not seen in other species formally recognized in the literature. Based on a detailed analysis of an extended list of morphological characters (93), including scalation, color pattern, gular and nuchal folds, precloacal pores, and morphometric data, we conclude that these populations represent independent lineages that deserve to be considered as new species. According to the most recent revision of the genus and considering the descriptions made in another recent contribution, the taxonomic composition of the genus was increased to 38 species. In this study we provide the formal description of two additional new taxa, including their diagnosis and detailed comparisons with other members of their species group. The two new species belong to the palluma group, and can be assigned to the Puna subclade because they present the typical dorsal "spray" pattern. Other characters described in this study suggest their close phylogenetic relationship with other species of this subclade inhabiting the southern Puna region of Argentina, such as Phymaturus punae. Within the Puna subclade, Phymaturus aguanegra sp. nov. differs from all other members (P. antofagastensis, P. denotatus, P. laurenti, P punae, P extrilidus, P mallimaccii and P paihuanense) exhibiting the following combination of diagnostic characters: a complete melanism over the dorsum of neck, the presence of enlarged scales at the base of tail in males, having strongly keeled tarsal scales, lacking enlarged scales on the anterior margin of the antehumeral fold and centre of chest, females without flank coloration, a vertebral dark gray stripe usually present on the dorsum, females exhibiting a tricolor dorsal pattern, with two types of brown and scattered ferriferous oxide spots, and the absence of a scapular spot. Phymaturus williamsi sp. nov. differs from all other members of the Puna clade because: exhibits an "aggregate" dorsal pattern, unlike the homogeneous spray of most Puna species, lacks enlarged scales on the anterior margin of the antehumeral fold and in the centre of chest, flank coloration in females is absent, females of Phymaturus williamsi sp. nov. lack white transversal stripes on the dorsal pattern, preocular scale in contact with canthal scale in Phymaturus williamsi sp. nov., rostral scale can be divided in Phymaturus williamsi sp. nov. and shows the largest number of scales counted around midbody within the Puna subclade (x= 213.4; 186-235).

The rediscovery of Oplurus bibronii Guichenot, 1848 a valid species of the liolaemid genus Phymaturus (Iguania: Liolaemidae).

Oplurus bibronii was a species described more than 160 years ago from the highlands of Ovalle, Chile. The species was earlier synonymized with Phymaturus palluma and since then its taxonomic status has not been re-examined. In February of 2011, we were successful in capturing specimens of Phymaturus in the highlands of Ovalle and additionally we examined high quality digital pictures of the type series of O. bibronii. Our analysis shows that the type series is composed of two different species, one of which overlaps their diagnostic characters with the characters of the specimens collected in the highlands of Ovalle, and therefore they correspond to Phymaturus bibronii (new combination). In addition, our review of the holotype, and some paratypes and topotypes of the recently described P. paihuanense shows that this is a junior synonym of P. bibronii.

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.