Amyloid ß-Peptide Causes the Permanent Activation of CaMKIIα through Its Oxidation.

Alzheimer's disease (AD) is characterised by the presence of extracellular amyloid plaques in the brain. They are composed of aggregated amyloid beta-peptide (Aß) misfolded into beta-sheets which are the cause of the AD memory impairment and dementia. Memory depends on the hippocampal formation and maintenance of synapses by long-term potentiation (LTP), whose main steps are the activation of NMDA receptors, the phosphorylation of CaMKIIα and the nuclear translocation of the transcription factor CREB. It is known that Aß oligomers (oAß) induce synaptic loss and impair the formation of new synapses. Here, we have studied the effects of oAß on CaMKIIα. We found that oAß produce reactive oxygen species (ROS), that induce CaMKIIα oxidation in human neuroblastoma cells as we assayed by western blot and immunofluorescence. Moreover, this oxidized isoform is significantly present in brain samples from AD patients. We found that the oxidized CaMKIIα is active independently of the binding to calcium/calmodulin, and that CaMKIIα phosphorylation is mutually exclusive with CaMKIIα oxidation as revealed by immunoprecipitation and western blot. An in silico modelling of the enzyme was also performed to demonstrate that oxidation induces an activated state of CaMKIIα. In brains from AD transgenic models of mice and in primary cultures of murine hippocampal neurons, we demonstrated that the oxidation of CaMKIIα induces the phosphorylation of CREB and its translocation to the nucleus to promote the transcription of ARC and BDNF. Our data suggests that CaMKIIα oxidation would be a pro-survival mechanism that is triggered when a noxious stimulus challenges neurons as do oAß.

Evaluation of the Antiviral Activity against Infectious Pancreatic Necrosis Virus (IPNV) of a Copper (I) Homoleptic Complex with a Coumarin as Ligand.

The aquatic infectious pancreatic necrosis virus (IPNV) causes a severe disease in farmed salmonid fish that generates great economic losses in the aquaculture industry. In the search for new tools to control the disease, in this paper we show the results obtained from the evaluation of the antiviral effect of [Cu(NN1)2](ClO4) Cu(I) complex, synthesized in our laboratory, where the NN1 ligand is a synthetic derivate of the natural compound coumarin. This complex demonstrated antiviral activity against IPNV at 5.0 and 15.0 µg/mL causing a decrease viral load 99.0% and 99.5%, respectively. The Molecular Docking studies carried out showed that the copper complex would interact with the VP2 protein, specifically in the S domain, altering the process of entry of the virus into the host cell.

Mechanosensory organ regeneration in zebrafish depends on a population of multipotent progenitor cells kept latent by Schwann cells.

BACKGROUND: Regenerating damaged tissue is a complex process, requiring progenitor cells that must be stimulated to undergo proliferation, differentiation and, often, migratory behaviors and morphological changes. Multiple cell types, both resident within the damaged tissue and recruited to the lesion site, have been shown to participate. However, the cellular and molecular mechanisms involved in the activation of progenitor cell proliferation and differentiation after injury, and their regulation by different cells types, are not fully understood. The zebrafish lateral line is a suitable system to study regeneration because most of its components are fully restored after damage. The posterior lateral line (PLL) is a mechanosensory system that develops embryonically and is initially composed of seven to eight neuromasts distributed along the trunk and tail, connected by a continuous stripe of interneuromastic cells (INCs). The INCs remain in a quiescent state owing to the presence of underlying Schwann cells. They become activated during development to form intercalary neuromasts. However, no studies have described if INCs can participate in a regenerative event, for example, after the total loss of a neuromast. RESULTS: We used electroablation in transgenic larvae expressing fluorescent proteins in PLL components to completely ablate single neuromasts in larvae and adult fish. This injury results in discontinuity of the INCs, Schwann cells, and the PLL nerve. In vivo imaging showed that the INCs fill the gap left after the injury and can regenerate a new neuromast in the injury zone. Further, a single INC is able to divide and form all cell types in a regenerated neuromast and, during this process, it transiently expresses the sox2 gene, a neural progenitor cell marker. We demonstrate a critical role for Schwann cells as negative regulators of INC proliferation and neuromast regeneration, and that this inhibitory property is completely dependent on active ErbB signaling. CONCLUSIONS: The potential to regenerate a neuromast after damage requires that progenitor cells (INCs) be temporarily released from an inhibitory signal produced by nearby Schwann cells. This simple yet highly effective two-component niche offers the animal robust mechanisms for organ growth and regeneration, which can be sustained throughout life.

TleA, a Tsh-like autotransporter identified in a human enterotoxigenic Escherichia coli strain.

Enterotoxigenic Escherichia coli (ETEC), a leading cause of acute diarrhea, colonizes the intestine by means of adhesins. However, 15 to 50% of clinical isolates are negative for known adhesins, making it difficult to identify antigens for broad-coverage vaccines. The ETEC strain 1766a, obtained from a child with watery diarrhea in Chile, harbors the colonization factor CS23 but is negative for other known adhesins. One clone, derived from an ETEC 1766a genomic library (clone G10), did not produce CS23 yet was capable of adhering to Caco-2 cells. The goal of this study was to identify the gene responsible for this capacity. Random transposon-based mutagenesis allowed the identification of a 4,110-bp gene that codes for a homologue of the temperature-sensitive hemagglutinin (Tsh) autotransporter described in avian E. coli strains (97% identity, 90% coverage) and that is called TleA (Tsh-like ETEC autotransporter) herein. An isogenic ETEC 1766a strain with a tleA mutation showed an adhesion level similar to that of the wild-type strain, suggesting that the gene does not direct attachment to Caco-2 cells. However, expression of tleA conferred the capacity for adherence to nonadherent E. coli HB101. This effect coincided with the detection of TleA on the surface of nonpermeabilized bacteria, while, conversely, ETEC 1766a seems to secrete most of the produced autotransporter to the medium. On the other hand, TleA was capable of degrading bovine submaxillary mucin and leukocyte surface glycoproteins CD45 and P-selectin glycoprotein ligand 1 (PSGL-1). These results suggest that TleA promotes colonization of the intestinal epithelium and that it may modulate the host immune response.

Immunoproteomic analysis to identify Shiga toxin-producing Escherichia coli outer membrane proteins expressed during human infection.

Shiga-toxin producing Escherichia coli (STEC) is the etiologic agent of acute diarrhea, dysentery, and hemolytic-uremic syndrome (HUS). There is no approved vaccine for STEC infection in humans, and antibiotic use is contraindicated, as it promotes Shiga toxin production. In order to identify STEC-associated antigens and immunogenic proteins, outer membrane proteins (OMPs) were extracted from STEC O26:H11, O103, O113:H21, and O157:H7 strains, and commensal E. coli strain HS was used as a control. SDS-PAGE, two-dimensional-PAGE analysis, Western blot assays using sera from pediatric HUS patients and controls, and matrix-assisted laser desorption ionization-tandem time of flight analyses were used to identify 12 immunogenic OMPs, some of which were not reactive with control sera. Importantly, seven of these proteins have not been previously reported to be immunogenic in STEC strains. Among these seven proteins, OmpT and Cah displayed IgG and IgA reactivity with sera from HUS patients. Genes encoding these two proteins were present in a majority of STEC strains. Knowledge of the antigens produced during infection of the host and the immune response to those antigens will be important for future vaccine development.

Prophylactic treatment with the c-Abl inhibitor, neurotinib, diminishes neuronal damage and the convulsive state in pilocarpine-induced mice.

The molecular mechanisms underlying seizure generation remain elusive, yet they are crucial for developing effective treatments for epilepsy. The current study shows that inhibiting c-Abl tyrosine kinase prevents apoptosis, reduces dendritic spine loss, and maintains N-methyl-d-aspartate (NMDA) receptor subunit 2B (NR2B) phosphorylated in in vitro models of excitotoxicity. Pilocarpine-induced status epilepticus (SE) in mice promotes c-Abl phosphorylation, and disrupting c-Abl activity leads to fewer seizures, increases latency toward SE, and improved animal survival. Currently, clinically used c-Abl inhibitors are non-selective and have poor brain penetration. The allosteric c-Abl inhibitor, neurotinib, used here has favorable potency, selectivity, pharmacokinetics, and vastly improved brain penetration. Neurotinib-administered mice have fewer seizures and improved survival following pilocarpine-SE induction. Our findings reveal c-Abl kinase activation as a key factor in ictogenesis and highlight the impact of its inhibition in preventing the insurgence of epileptic-like seizures in rodents and humans.

Advances and challenges in the diagnosis and management of left ventricular noncompaction in adults: A literature review.

In the realm of cardiovascular health, isolated left ventricular noncompaction (LVNC) stands out for its distinct morphological features and the clinical challenges it presents, particularly in adults. This literature review explores the intricacies of LVNC, aiming to unravel its epidemiological spread, diagnostic hurdles, and therapeutic strategies. Despite technological advancements in cardiac imaging that have improved the recognition of LVNC, a significant gap persists alongside a fragmented understanding of its pathogenesis. The studies scrutinized reveal a broad spectrum of prevalence rates influenced by diverse diagnostic tools and demographic variables. This variation underscores the complexity of accurately identifying LVNC and the resultant implications for clinical management. The review succinctly addresses the need for precise guidelines to navigate the diagnosis of LVNC and outlines the imperative for tailored clinical management approaches that cater to the wide array of patient presentations, from asymptomatic cases to those with severe cardiac dysfunction. By highlighting the critical gaps in current literature-namely the absence of standardized diagnostic criteria and a comprehensive pathogenic model-the review sets the stage for future research directions. These endeavors are essential for enhancing diagnostic accuracy, refining management protocols, and ultimately improving patient outcomes in this complex subset of cardiomyopathy, thus contributing significantly to the advancement of cardiovascular medicine.

Factors that influence a career choice in primary care among medical students from high-, middle-, and low-income countries: a systematic review.

OBJECTIVE: To determine which factors influence a medical student's decision to choose a career in primary care; and to establish if these factors are similar or different among students in high-, middle- and low-income countries. METHODS: An extensive search was done of PubMed, Google Scholar, and Virtual Library of Health for articles on primary care careers published in 2003-2013 in English, Spanish, and/or Portuguese. Initially, 600 records were identified; 74 full-text articles were assessed for eligibility and 55 were selected (42 from high-income countries; 13 from middle- and low-income). These were assessed to identify intrinsic and extrinsic factors that influence career choice among medical students from high-, middle-, and low-income countries. RESULTS: A comparison framework with common and specific factors that influence career choice in primary care among medical students from high-, middle- and low-income was developed. Factors were classified as extrinsic or intrinsic, and as facilitators or barriers. Several factors common to all countries were identified: facilitators were exposure to rural location, role models, working conditions; barriers were low income, prestige, and medical school environment. Some factors specific to middle- and low-income countries were: understanding of rural needs and intellectual challenge. Other factors specific to high-income countries were: attitude towards social problems, voluntary work, influence of family, and length of residency. CONCLUSIONS: Further studies on the subject are needed, especially in low- and middle-income countries. Identifying factors as barriers or facilitators for career choice will promote a better understanding of the reasons behind the shortage of primary care professionals and will contribute to policy building, improved training, and recruitment and retention of these professionals.

[Single-photon emission computed tomography for the assessment of ventricular perfusion and function]. / Perfusión y función ventricular en SPECT cardiaco y coronariografía del mismo día según localización en enfermedad de un vaso.

BACKGROUND: Single-photon emission computed tomography (SPECT) can be used as a non-invasive tool for the assessment of coronary perfusion. AIM: To assess ventricular perfusion and function by SPECT in patients with single vessel coronary artery disease. MATERIAL AND METHODS: Among patients with indications for a coronary artery angiography, those with significant lesions in one vessel, were selected for the study. Within 24 hours, cardiac SPECT examinations on basal conditions and after high doses of dipyridamole, were performed. SPECT data from 38 patients with a low probability of coronary artery disease was used for comparisons. RESULTS: Ten patients aged 61 ± 8 years (seven men) were studied. Visual analysis of SPECT revealed signs suggestive of ischemia in eight patients. The remaining two patients did not have perfusion disturbances. SPECT detected eight of ten abnormal vessels reported in the coronary artery angiography. There were two false negative results Summed stress, summed rest and summed difference scores were 9.78 ± 6.51, 3.22 ± 5.07 and 6.33 ± 4.97, respectively. The ejection fractions under stress and at rest were 53 ± 11.7% and 61 ± 15.7% respectively (p < 0.01). The figures for the control group were 69.1 ± 13.5% and 75.2 ± 12.04% respectively (significantly different from patients). Two patients had a summed motion score above 14.9. Likewise, two patients had a summed thickening score above 10.9. CONCLUSIONS: SPECT detected 80% of coronary lesions found during coronary artery angiography. Visual analysis of perfusion is highly reliable for diagnosis. Quantitative parameters must be considered only as reference parameters.

c-Abl kinase at the crossroads of healthy synaptic remodeling and synaptic dysfunction in neurodegenerative diseases.

Our ability to learn and remember depends on the active formation, remodeling, and elimination of synapses. Thus, the development and growth of synapses as well as their weakening and elimination are essential for neuronal rewiring. The structural reorganization of synaptic complexes, changes in actin cytoskeleton and organelle dynamics, as well as modulation of gene expression, determine synaptic plasticity. It has been proposed that dysregulation of these key synaptic homeostatic processes underlies the synaptic dysfunction observed in many neurodegenerative diseases. Much is known about downstream signaling of activated N-methyl-D-aspartate and α-amino-3-hydroxy-5-methyl-4-isoazolepropionate receptors; however, other signaling pathways can also contribute to synaptic plasticity and long-lasting changes in learning and memory. The non-receptor tyrosine kinase c-Abl (ABL1) is a key signal transducer of intra and extracellular signals, and it shuttles between the cytoplasm and the nucleus. This review focuses on c-Abl and its synaptic and neuronal functions. Here, we discuss the evidence showing that the activation of c-Abl can be detrimental to neurons, promoting the development of neurodegenerative diseases. Nevertheless, c-Abl activity seems to be in a pivotal balance between healthy synaptic plasticity, regulating dendritic spines remodeling and gene expression after cognitive training, and synaptic dysfunction and loss in neurodegenerative diseases. Thus, c-Abl genetic ablation not only improves learning and memory and modulates the brain genetic program of trained mice, but its absence provides dendritic spines resiliency against damage. Therefore, the present review has been designed to elucidate the common links between c-Abl regulation of structural changes that involve the actin cytoskeleton and organelles dynamics, and the transcriptional program activated during synaptic plasticity. By summarizing the recent discoveries on c-Abl functions, we aim to provide an overview of how its inhibition could be a potentially fruitful treatment to improve degenerative outcomes and delay memory loss.

Comparison of Two Tuberculosis Infection Tests in a South American Tertiary Hospital: STANDARD F TB-Feron FIA vs. QIAreachTM QuantiFERON-TB.

INTRODUCTION: Tuberculosis (TB) is one of the most prevalent respiratory diseases in the world. In 2020 there were at least 9.9 million new infections, with 1.5 million deaths. Approximately 10% of people infected with Mycobacterium tuberculosis develop the disease during the first 2 to 5 years after infection. In South America, the diagnosis of Latent Tuberculosis Infections (LTBI) continues to be performed through the Mantoux tuberculin skin test (TST). OBJECTIVE: The objective of our study was to compare the sensitivity of a new immunofluorescence IGRA test against a widely available IGRA kit on the market. MATERIAL AND METHOD: Close contact with infectious TB patients, HIV patients, or immunocompromised for another cause were recruited. Two interferon-gamma release assay (IGRA) diagnostic kits were used and compared with TST. RESULTS: 76 patients were recruited, 93.42% were Chilean nationality, and 98.68% of the patients did not have immunosuppression. The sensitivity of the new technique was 88.89%, and the specificity was 92.50% in the study population compared to the IGRA previously used. In the subgroup older than 36 years, the sensitivity was 95.65%, and the specificity was 89.47%. CONCLUSION: IGRA techniques are a new resource in clinical laboratories to make an accurate diagnosis of LTBI in the region of the Americas. In our population, the greatest benefit of this new IGRA would be observed in people over 36 years of age, where the sensitivity of the technique was like that of the currently available test.

Education's impact on primary care dentists' confidence in treating patients with special health care needs.

INTRODUCTION: Special care dentistry (SCD) is still developing in XX. This study aimed to clarify whether primary care dentists are treating patients with special health care needs (SHCN), to know if they have had previous education on SCD (on an undergraduate or postgraduate level), whether their training level impacts their confidence when treating patients with SHCN, and to assess their opinion on SCD as a relevant topic in undergraduate education. METHODS: A survey was answered by 149 primary care dentists working for the National Health Service of the XX region in XX, including information on their daily clinical practice, undergraduate, and postgraduate training in SCD, and their opinions on them. RESULTS: Most interviewees would like to complement their training and believed that SCD should be formally incorporated into undergraduate programs. There was a significant association between confidence in treating patients with SHCN and the rating of their undergraduate training, and between confidence and the number of hours of continuous development courses. CONCLUSION: Most primary care dentists treat patients with SHCN regularly. Therefore, including training in the undergraduate curriculum and afterward becomes necessary to increase their confidence when facing this challenging group of patients.

Microplastics in water, feed and tissues of European seabass reared in a recirculation aquaculture system (RAS).

Plastic particles (PLs) are ubiquitous in aquatic ecosystems, and aquaculture production is susceptible to contamination from external or endogenous sources. This study investigated PL presence in water, fish feed and body sites of 55 European seabass produced in a recirculating aquaculture system (RAS). Fish morphometric parameters and health status biomarkers were determined. A total of 372 PLs were recovered from water (37.2 PL/L), 118 PLs from feed (3.9 PL/g), and 422 from seabass (0.7 PL/g fish; all body sites analysed). All 55 specimens had PLs in at least two of the four body sites analysed. Concentrations were higher in the gastrointestinal tract (GIT; 1.0 PL/g) and gills (0.8 PL/g) than in the liver (0.8 PL/g) and muscle (0.4 PL/g). PL concentration in GIT was significantly higher than in muscle. Black, blue, and transparent fibres made of man-made cellulose/rayon and polyethylene terephthalate were the most common PLs in water and seabass, while black fragments of phenoxy resin were the most common in feed. The levels of polymers linked to RAS components (polyethylene, polypropylene, and polyvinyl chloride) were low, suggesting a limited contribution to the overall PL levels found in water and/or fish. The mean PL size recovered from GIT (930 µm) and gills (1047 µm) was significantly larger than those found in the liver (647 µm) and dorsal muscle (425 µm). Considering all body sites, PLs bioconcentrated in seabass (BCFFish >1), but their bioaccumulation did not occur (BAFFish <1). No significant differences were observed in oxidative stress biomarkers between fish with low (<7) and high (≥7) PL numbers. These findings suggest that fish produced in RAS are mainly exposed to MPs through water and feed. Further monitoring under commercial conditions and risk assessment are warranted to identify potential threats to fish and human health and define mitigating measures.

c-Abl tyrosine kinase down-regulation as target for memory improvement in Alzheimer's disease.

Background: Growing evidence suggests that the non-receptor tyrosine kinase, c-Abl, plays a significant role in the pathogenesis of Alzheimer's disease (AD). Here, we analyzed the effect of c-Abl on the cognitive performance decline of APPSwe/PSEN1ΔE9 (APP/PS1) mouse model for AD. Methods: We used the conditional genetic ablation of c-Abl in the brain (c-Abl-KO) and pharmacological treatment with neurotinib, a novel allosteric c-Abl inhibitor with high brain penetrance, imbued in rodent's chow. Results: We found that APP/PS1/c-Abl-KO mice and APP/PS1 neurotinib-fed mice had improved performance in hippocampus-dependent tasks. In the object location and Barnes-maze tests, they recognized the displaced object and learned the location of the escape hole faster than APP/PS1 mice. Also, APP/PS1 neurotinib-fed mice required fewer trials to reach the learning criterion in the memory flexibility test. Accordingly, c-Abl absence and inhibition caused fewer amyloid plaques, reduced astrogliosis, and preserved neurons in the hippocampus. Discussion: Our results further validate c-Abl as a target for AD, and the neurotinib, a novel c-Abl inhibitor, as a suitable preclinical candidate for AD therapies.

Evaluation of metabolic syndrome after hematopoietic stem cell transplantation in children and adolescents.

BACKGROUND: To determine the prevalence, characteristics, and risk factors associated with metabolic syndrome (MS) in patients undergoing hematopoietic stem cell transplantation (HSCT) in the Chilean National Program. PROCEDURES: Descriptive and cross-sectional study including 69 patients was conducted. Body mass index, pubertal development, waist circumference, arterial pressure (AP), and triglycerides, HDL-cholesterol, and glucose levels were recorded at the time of study entry. The National Cholesterol Education Program (Adult Treatment Panel III, as modified by the American Heart Association) criteria are often used to diagnose MS in adults; however, for children and adolescents we followed criteria according to De Ferranti and American Diabetes Association. Statistical analyses were performed with a chi-square test or Fisher's exact test according to sample size. RESULTS: Sixty-nine patients were studied. The median age at the time of diagnosis was 12.9 years, and the median time of follow-up post-transplant was 4 years. Forty-three patients were males, 54 patients had malignant diseases, and 59 patients received allogeneic transplants. Of the 69 patients, 32% had MS; the most common MS features were abdominal obesity (73%), hypertriglyceridemia (91%), and a low HDL-cholesterol level (96%). The most significant risk factor for MS was corticosteroid therapy use pre- (P < 0.03) and post-HSCT (P < 0.03), obesity and overweight associated with MS (P < 0.001). No patient developed cardiovascular complications. CONCLUSIONS: The prevalence of MS was 32%, which was significantly higher than in a healthy pediatric population. We recommend prolonged follow-up for transplant recipients, coupled with enforcement of preventive measures, such as early diagnosis and encouragement of a healthy lifestyle.

[Evolution of scintigraphic renal lesions in children after an episode of acute pyelonephritis]. / Una lesión en el cintigrama renal DMSA 6 meses post fase aguda de una pielonefritis representa siempre una cicatriz: un debate abierto.

BACKGROUND: Abnormal Dimercaptosuccinic acid (DMSA) renal scintigraphy performed six months after an acute pyelonephritis (AP) is generally interpreted as scarring. AIM: To perform a follow up of childhood patients showing scintigraphic renal lesions during the acute phase of pyelonephritis (within 7 days from the beginning of fever). MATERIAL AND METHODS: A scintigraphic control was carried out at 5-7 months and, in case of persistent lesions, an additional late scintigraphy at 10-13 months. All patients were followed clinically for one year and those with a relapse of urinary tract infection were excluded from the study. RESULTS: Eighty five patients with a median age of 8 months were included. Among these, the first scintigraphic control was normal in 59 (69%) and abnormal in 26 patients (31%). In five of these 26 patients (5/26:19%-5/85: 6%), a considerable regression of the lesions was obvious on the early control, and normalized completely on the late control. When expressing the results in kidney units, 107 showed lesions during the acute phase of infection; 69% was normal at the early control. Thirty three showed lesions persisting at the early control (31%) and 7 out of these 33 (21%) became normal on the late control (7/107: 7%). In total, 25% of the children included in the study (24% of the kidney units) remained with renal sequelae one year after the initial episode of AP. CONCLUSIONS: The persistence of scintigraphic lesions six months after an episode of AP, does not necessarily correspond to permanent scars, since normalization can sometimes be observed on late controls.

Identification of Exoenzymes Secreted by Entomopathogenic Fungus Beauveria pseudobassiana RGM 2184 and Their Effect on the Degradation of Cocoons and Pupae of Quarantine Pest Lobesia botrana.

Beauveria pseudobassiana RGM 2184 has shown 80% maximum efficacy against the pest Lobesia botrana in the autumn and winter seasons. This suggests that the strain possesses an interesting battery of enzymes that are cold-adapted to penetrate the thick and hydrophobic cocoon of L. botrana. In this study, screening of the proteolytic, lipolytic, and chitinolytic activity of enzyme extracts secreted by the RGM 2184 strain was carried out in various culture media. The enzyme extracts with the highest activity were subjected to zymography and mass spectrometry. These analyses allowed the identification of two proteases, two lipases, and three chitinases. Comparative analysis indicated that the degree of similarity between these enzymes was substantially reduced when the highest degree of taxonomic relatedness between RGM 2184 and the entomopathogenic fungus strain was at the family level. These results suggest that there is a wide variety of exoenzymes in entomopathogenic fungi species belonging to the order Hypocreales. On the other hand, exoenzyme extract exposure of cocoons and pupae of L. botrana provoked damage at 10 °C. Additionally, an analysis of the amino acid composition of the RGM 2184 exoenzyme grouped them close to the cold-adapted protein cluster. These results support the use of this strain to control pests in autumn and winter. Additionally, these antecedents can form a scaffold for the future characterization of these exoenzymes along with the optimization of the strain's biocontrol ability by overexpressing them.

Alpinone: A positive regulator molecule of immune antiviral response in Atlantic salmon kidney cells.

Alpinone is a flavonoid obtained from the resinous exudate of Heliotropium huascoense. This flavonoid shows antiviral activity against the infectious salmon anemia virus (ISAV), which causes severe disease in farmed Atlantic salmon. Here, we aim to elucidate mechanisms underlying the antiviral effects of the flavonoid. In this regard, we evaluated whether Alpinone can act upregulating the pattern-recognition receptor genes, i.e., the RIG-I-like, TLR3, and TLR9 genes, and the genes of the downstream signaling pathways. Transcriptional expression of the genes was analyzed using real-time PCR after 8, 24, and 48 h treatment of salmon kidney adherent cells with 15 µg/mL of Alpinone. First, we showed that Alpinone induced IFNa expression in the kidney adherent cells, indicating that this type of salmon cells is in part responsible for the effects previously reported in vivo. Upregulation of the IFN-induced myxovirus resistance (Mx) gene was also observed in the head kidney cells in response to the treatment. Overexpression reached a maximum level at 24 h post-treatment. Interestingly, Alpinone also induced upregulation of the cytosolic receptors of ssRNA, named Retinoic acid-inducible gene I (RIG-I) and Melanoma Differentiation-Associated protein 5 (MDA5), but there were no effects on the transcriptional expression of the TLR3 and TLR9 endosomal receptors. In addition, Alpinone upregulated the expression of genes encoding the main components of the RIG-I/MDA5 signaling pathways, such as the mitochondrial antiviral-signaling protein (MAVS), TNF Receptor Associated Factor 3 (TRAF3), TANK-binding kinase 1 (TBK1), I-kappaB kinase ε (IKKε), the transcription factors IRF-3, and IRF7. The increased expression of all these genes is consistent with the upregulation of IFNa and Mx mRNAs. Because BX795 completely prevents Alpinone-dependent upregulation of IFNa and IRF3, the flavonoid targets seem to be upstream of the kinases TBK1 and IKKε. Altogether, this study contributes to elucidating the mechanisms involved in Alpinone antiviral activity in fish. Alpinone can be used to counteract virus mechanisms of evasion where the onset of interferon-mediated response is prevented or delayed.

Effects of testosterone therapy in adult males with hypogonadism and T2DM: A meta-analysis and systematic review.

BACKGROUND AND AIMS: Testosterone supplementation therapy (TST) is a longstanding treatment for hypogonadal men with type 2 diabetes mellitus (T2DM), even though the benefits of TST are variable among trials. This meta-analysis was done to determine the specific role of TST in hypogonadal men with T2DM. METHODS: PubMed, Embase, and Google Scholar were queried to discover eligible randomized controlled trials (RCTs) and observational studies. To quantify the specific effects of TST, we estimated pooled mean differences (MDs) and relative risks with 95% confidence intervals (CIs). RESULTS: Our meta-analysis included 1596 hypogonadal T2DM subjects from 12 randomized controlled trials and one observational study. TST can significantly enhance glycemic control compared to placebo by decreasing homeostatic model assessment of insulin resistance (WMD = -1.55 [-2.65, -0.45]; p = 0.26; I2 = 20.2%), fasting glucose (WMD = -0.35 [-0.79, 0.10]; p = 0.07; I2 = 69.7%), fasting insulin (WMD = -2.88 [-6.12, 0.36]; p = In addition, TST can decrease cholesterol (WMD = -0.28 [-0.47, -0.09] p = 0.0008; I2 = 91%) and triglyceride (WMD = -0.23 [-0.43, -0.03] p = 0.03; I2 = 79.2%). Furthermore, Testosterone therapy is related to a significant rise in total testosterone levels (WMD = 5.08 [2.90, 7.26] p = 0.0002; I2 = 92.9%). Pooling of free testosterone levels indicated a larger increase in the patients who got TST than placebo (WMD = 81.21 [23.87, 138.54] p = 0.07; I2 = 70%). CONCLUSION: Our findings suggested that TST can enhance glycemic control and hormone levels and reduce total cholesterol, triglyceride, LDL cholesterol whereas increase HDL cholesterol in hypogonadal T2DM patients. Therefore, in these patients, we propose TST alongside anti-diabetic treatment.