Immunohistochemistry and real-time Polymerase Chain Reaction: importance in the diagnosis of intestinal tuberculosis in a Peruvian population.

INTRODUCTION: The diagnosis of intestinal tuberculosis is challenging even nowadays. This study aims to report the positivity rates of new diagnostic methods such as immunohistochemistry and Real-Time Polymerase Chain Reaction in patients with intestinal tuberculosis, as well as describe the pathological and endoscopic features of intestinal tuberculosis in our population. METHODS: This was a retrospective observational study conducted in patients diagnosed with intestinal tuberculosis, between 2010 to 2023 from the Hospital Nacional Daniel Alcides Carrion and a Private Pathology Center, both located in Peru. Clinical data was obtained, histologic features were independently re-evaluated by three pathologists; and immunohistochemistry and real-time Polymerase Chain Reaction evaluation were performed. The 33 patients with intestinal tuberculosis who fulfilled the inclusion criteria were recruited. RESULTS: Immunohistochemistry was positive in 90.9% of cases, while real-time Polymerase Chain Reaction was positive in 38.7%. The ileocecal region was the most affected area (33.3%), and the most frequent endoscopic appearance was an ulcer (63.6%). Most of the granulomas were composed solely of epithelioid histiocytes (75.8%). Crypt architectural disarray was the second most frequent histologic finding (78.8%) after granulomas, but most of them were mild. CONCLUSION: Since immunohistochemistry does not require an intact cell wall, it demonstrates higher sensitivity compared to Ziehl-Neelsen staining. Therefore, it could be helpful for the diagnosis of paucibacillary tuberculosis.

Discovery of Potent, Reversible, and Competitive Cruzain Inhibitors with Trypanocidal Activity: A Structure-Based Drug Design Approach.

A virtual screening conducted with nearly 4 000 000 compounds from lead-like and fragment-like subsets enabled the identification of a small-molecule inhibitor (1) of the Trypanosoma cruzi cruzain enzyme, a validated drug target for Chagas disease. Subsequent comprehensive structure-based drug design and structure-activity relationship studies led to the discovery of carbamoyl imidazoles as potent, reversible, and competitive cruzain inhibitors. The most potent carbamoyl imidazole inhibitor (45) exhibited high affinity with a Ki value of 20 nM, presenting both in vitro and in vivo activity against T. cruzi. Furthermore, the most promising compounds reduced parasite burden in vivo and showed no toxicity at a dose of 100 mg/kg. These carbamoyl imidazoles are structurally attractive, nonpeptidic, and easy to prepare and synthetically modify. Finally, these results further advance our understanding of the noncovalent mode of inhibition of this pharmaceutically relevant enzyme, building strong foundations for drug discovery efforts.

Autoantibodies in breast cancer sera are not epiphenomena and may participate in carcinogenesis.

BACKGROUND: The objective of this work was to demonstrate that autoantibodies in breast cancer sera are not epiphenomena, and exhibit unique immunologic features resembling the rheumatic autoimmune diseases. METHODS: We performed a comprehensive study of autoantibodies on a collection of sera from women with breast cancer or benign breast disease, undergoing annual screening mammography. All women in this study had suspicious mammography assessment and underwent a breast biopsy. We used indirect immunofluorescence, the crithidia assay for anti-dsDNA antibodies, and multiple ELISAs for extractable nuclear antigens. RESULTS: Autoantibodies were detected in virtually all patients with breast cancer, predominantly of the IgG1 and IgG3 isotypes. The profile detected in breast cancer sera showed distinctive features, such as antibodies targeting mitochondria, centrosomes, centromeres, nucleoli, cytoskeleton, and multiple nuclear dots. The majority of sera showing anti-mitochondrial antibodies did not react with the M2 component of pyruvate dehydrogenase, characteristic of primary biliary cirrhosis. Anti-centromere antibodies were mainly anti-CENP-B. ELISAs for extractable nuclear antigens and the assays for dsDNA were negative. CONCLUSIONS: The distinctive autoantibody profile detected in BC sera is the expression of tumor immunogenicity. Although some of these features resemble those in the rheumatic autoimmune diseases and primary biliary cirrhosis, the data suggest the involvement of an entirely different set of epithelial antigens in breast cancer. High titer autoantibodies targeting centrosomes, centromeres, and mitochondria were detected in a small group of healthy women with suspicious mammography assessment and no cancer by biopsy; this suggests that the process triggering autoantibody formation starts in the pre-malignant phase and that future studies using validated autoantibody panels may allow detection of breast cancer risk in asymptomatic women. Autoantibodies developing in breast cancer are not epiphenomena, but likely reflect an antigen-driven autoimmune response triggered by epitopes developing in the mammary gland during breast carcinogenesis. Our results support the validity of the multiple studies reporting association of autoantibodies with breast cancer. Results further suggest significant promise for the development of panels of breast cancer-specific, premalignant-phase autoantibodies, as well as studies on the autoantibody response to tumor associated antigens in the pathogenesis of cancer.

Projected impact of climate change on human health in low- and middle-income countries: a systematic review.

Low- and middle-income countries (LMICs) contribute relatively little to global carbon emissions but are recognised to be among the most vulnerable parts of the world to health-related consequences of climate change. To help inform resilient health systems and health policy strategies, we sought to systematically analyse published projections of the impact of rising global temperatures and other weather-related events on human health in LMICs. A systematic search involving multiple databases was conducted in accordance with Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines to identify studies with modelled projections of the future impact of climate change on human health. Qualitative studies, reviews and meta-analyses were excluded. The search yielded more than 2500 articles, of which 70 studies involving 37 countries met criteria for inclusion. China, Brazil and India were the most studied countries while the sub-Saharan African region was represented in only 9% of studies. Forty specific health outcomes were grouped into eight categories. Non-disease-specific temperature-related mortality was the most studied health outcome, followed by neglected tropical infections (predominantly dengue), malaria and cardiovascular diseases. Nearly all health outcomes studied were projected to increase in burden and/or experience a geographic shift in prevalence over the next century due to climate change. Progressively severe climate change scenarios were associated with worse health outcomes. Knowledge gaps identified in this analysis included insufficient studies of various high burden diseases, asymmetric distribution of studies across LMICs and limited use of some climate parameters as independent variables. Findings from this review could be the basis for future research to help inform climate mitigation and adaptation programmes aimed at safeguarding population health in LMICs.

Structure-Metabolism Relationships of Benzimidazole Derivatives with anti-Trypanosoma cruzi Activity for Chagas Disease.

This study introduces further insights from the hit-to-lead optimization process involving a series of benzimidazole derivatives acting as inhibitors of the cruzain enzyme, which targets Trypanosoma cruzi, the causative parasite of Chagas disease. Here, we present the design, synthesis and biological evaluation of 30 new compounds as a third generation of benzimidazole analogues with trypanocidal activity, aiming to enhance our understanding of their pharmacokinetic profiles and establish a structure-metabolism relationships within the series. The design of these new analogues was guided by the analysis of previous pharmacokinetic results, considering identified metabolic sites and biotransformation studies. This optimization resulted in the discovery of two compounds (42 e and 49 b) exhibiting enhanced metabolic stability, anti-Trypanosoma cruzi activity compared to benznidazole (the reference drug for Chagas disease), as well as being non-cruzain inhibitors, and demonstrating a satisfactory in vitro pharmacokinetic profile. These findings unveil a new subclass of aminobenzimidazole and rigid compounds, which offer potential for further exploration in the quest for discovering novel classes of antichagasic compounds.

Structure-activity relationships of novel N-imidazoylpiperazines with potent anti-Trypanosoma cruzi activity.

Background: Chagas disease is caused by the parasite Trypanosoma cruzi, and the lack of effective and safe treatments makes identifying new classes of compounds with anti-T. cruzi activity of paramount importance. Methods: Hit-to-lead exploration of a metabolically stable N-imidazoylpiperazine was performed. Results: Compound 2, a piperazine derivative active against T. cruzi, was selected to perform the hit-to-lead exploration, which involved the design, synthesis and biological evaluation of 39 new derivatives. Conclusion: Compounds 6e and 10a were identified as optimized compounds with low micromolar in vitro activity, low cytotoxicity and suitable preliminary absorption, distribution, metabolism and excretion and physicochemical properties. Both compounds reduced parasitemia in mouse models of Chagas disease, providing a promising opportunity for further exploration of new antichagasic compounds.

Targeted Protein Degradation for Infectious Diseases: from Basic Biology to Drug Discovery.

Targeted protein degradation (TPD) is emerging as one of the most innovative strategies to tackle infectious diseases. Particularly, proteolysis-targeting chimera (PROTAC)-mediated protein degradation may offer several benefits over classical anti-infective small-molecule drugs. Because of their peculiar and catalytic mechanism of action, anti-infective PROTACs might be advantageous in terms of efficacy, toxicity, and selectivity. Importantly, PROTACs may also overcome the emergence of antimicrobial resistance. Furthermore, anti-infective PROTACs might have the potential to (i) modulate "undruggable" targets, (ii) "recycle" inhibitors from classical drug discovery approaches, and (iii) open new scenarios for combination therapies. Here, we try to address these points by discussing selected case studies of antiviral PROTACs and the first-in-class antibacterial PROTACs. Finally, we discuss how the field of PROTAC-mediated TPD might be exploited in parasitic diseases. Since no antiparasitic PROTAC has been reported yet, we also describe the parasite proteasome system. While in its infancy and with many challenges ahead, we hope that PROTAC-mediated protein degradation for infectious diseases may lead to the development of next-generation anti-infective drugs.

Anti-inflammatory mechanisms and pharmacological actions of phycocyanobilin in a mouse model of experimental autoimmune encephalomyelitis: A therapeutic promise for multiple sclerosis.

Cytokines, demyelination and neuroaxonal degeneration in the central nervous system are pivotal elements implicated in the pathogenesis of multiple sclerosis (MS) and its nonclinical model of experimental autoimmune encephalomyelitis (EAE). Phycocyanobilin (PCB), a chromophore of the biliprotein C-Phycocyanin (C-PC) from Spirulina platensis, has antioxidant, immunoregulatory and anti-inflammatory effects in this disease, and it could complement the effect of other Disease Modifying Treatments (DMT), such as Interferon-ß (IFN-ß). Here, our main goal was to evaluate the potential PCB benefits and its mechanisms of action to counteract the chronic EAE in mice. MOG35-55-induced EAE was implemented in C57BL/6 female mice. Clinical signs, pro-inflammatory cytokines levels by ELISA, qPCR in the brain and immunohistochemistry using precursor/mature oligodendrocytes cells antibodies in the spinal cord, were assessed. PCB enhanced the neurological condition, and waned the brain concentrations of IL-17A and IL-6, pro-inflammatory cytokines, in a dose-dependent manner. A down- or up-regulating activity of PCB at 1 mg/kg was identified in the brain on three (LINGO1, NOTCH1, and TNF-α), and five genes (MAL, CXCL12, MOG, OLIG1, and NKX2-2), respectively. Interestingly, a reduction of demyelination, active microglia/macrophages density, and axonal damage was detected along with an increase in oligodendrocyte precursor cells and mature oligodendrocytes, when assessed the spinal cords of EAE mice that took up PCB. The studies in vitro in rodent encephalitogenic T cells and in vivo in the EAE mouse model with the PCB/IFN-ß combination, showed an enhanced positive effect of this combined therapy. Overall, these results demonstrate the anti-inflammatory activity and the protective properties of PCB on the myelin and support its use with IFN-ß as an improved DMT combination for MS.

Chemoinformatics Studies on a Series of Imidazoles as Cruzain Inhibitors.

Natural products based on imidazole scaffolds have inspired the discovery of a wide variety of bioactive compounds. Herein, a series of imidazoles that act as competitive and potent cruzain inhibitors was investigated using a combination of ligand- and structure-based drug design strategies. Quantitative structure-activity relationships (QSARs) were generated along with the investigation of enzyme-inhibitor molecular interactions. Predictive hologram QSAR (HQSAR, r2pred = 0.80) and AutoQSAR (q2 = 0.90) models were built, and key structural properties that underpin cruzain inhibition were identified. Moreover, comparative molecular field analysis (CoMFA, r2pred = 0.81) and comparative molecular similarity indices analysis (CoMSIA, r2pred = 0.73) revealed 3D molecular features that strongly affect the activity of the inhibitors. These findings were examined along with molecular docking studies and were highly compatible with the intermolecular contacts that take place between cruzain and the inhibitors. The results gathered herein revealed the main factors that determine the activity of the imidazoles studied and provide novel knowledge for the design of improved cruzain inhibitors.

Aspectos epidemiológicos del síndrome de inmunosupresión, inflamación y catabolismo persistente en pacientes crónicos críticamente enfermos / Epidemiological aspects of the syndrome of immunosuppression, inflammation and persistent catabolism in critical chronic ill patients / Aspectos epidemiológicos da síndrome de imunossupressão, inflamação e catabolismo persistente em pacientes crônico em estado crítico

Resumen: Los pacientes crónicos críticamente enfermos mantienen un ambiente inflamatorio persistente, inmunidad reducida y consumo progresivo de reservas fisiológicas. Se caracterizan por ingresos hospitalarios con estadías más prolongadas, mayor mortalidad y costos. Objetivo general: Describir las características del síndrome de inflamación, inmunosupresión y catabolismo persistente en pacientes críticos crónicamente enfermos hospitalizados en la Unidad de Cuidados Intensivos del Hospital General La Villa. Con el fin de conocer los aspectos epidemiológicos. Material y métodos: Estudio clínico prospectivo, descriptivo y observacional, analítico. Se ingresaron 25 pacientes con criterios de PICS acorde a la escala NUTRIC y cuadro clínico de enero-abril de 2018. Se realizaron pruebas de estadística descriptiva para variables cualitativas y para cuantitativas se utilizaron medidas de tendencia central. Pruebas inferenciales para contrastar variables cualitativas y para cuantitativas tablas de 2 x 2 para el cálculo de la odds ratio. Resultados: 25 pacientes cumplieron criterios de PICS, incidencia de 37.9%, la distribución por sexos fue 60% hombres y 40% mujeres. Edad promedio fue 48.8. Los diagnósticos más comunes fueron choque hipovolémico (28%), choque séptico (16%), 68% de los pacientes tenían un riesgo nutricional bajo y 32% riesgo alto, 84% de los pacientes requirieron algún tipo de soporte, el más común fue ventilación mecánica, Las complicaciones más comunes fueron las infecciosas (80%) principalmente NAVM. La mortalidad fue de 20%. Conclusiones: Los resultados en este estudio no difieren de los reportados en la bibliografía a nivel internacional, el síndrome de PICS se presenta en pacientes con mayor edad, mayor estancia hospitalaria y carencias nutricionales superiores en quienes las reservas biológicas no son suficientes para evitar la inmunosupresión que conlleva y genera el círculo vicioso que finalmente conduce a la muerte no sólo dentro de la UCI.

Abstract: Critically ill chronic patients maintain a persistent inflammatory environment, reduced immunity and progressive consumption of physiological reserves. They are characterized by hospital admissions with longer stays, higher mortality and costs. General objective: To describe the characteristics of the syndrome of inflammation, immunosuppression and persistent catabolism in critically ill critically ill patients hospitalized in the Intensive Care Unit of the La Villa General Hospital. In order to know the epidemiological aspects. Material and methods: Prospective, descriptive and observational clinical, analytical study. Patients were admitted with PICS criteria according to the NUTRIC score and clinical features from January-April 2018. Descriptive statistics tests were performed for qualitative variables, quantitative measures were used for quantitative central. Inferential tests to compare qualitative variables and for quantitative 2 x 2 tables for calculating the odds ratio. Results: 25 patients met criteria of PICS, incidence of 37.9%, the distribution by sex was 60% men and 40% women. Average age was 48.8. The most common diagnoses hypovolemic shock (28%), septic shock (16%), 68% of patients had a low nutritional risk and 32% high risk, 84% of patients required some kind of support the most common was ventilation mechanical, the most common complications were infectious (80%) mainly VAP. The mortality was 20%. Conclusions: The results in this study do not differ from those reported in the literature at the international level, the syndrome of PICS is presented in patients with older age, longer hospital stay and higher nutritional deficiencies in which biological reserves are not sufficient to avoid immunosuppression that leads to and generates the vicious circle that ultimately leads to death not only within the ICU.

Resumo: Os pacientes crônicos em estado crítico mantêm um ambiente inflamatório persistente, imunidade reduzida e consumo progressivo de reservas fisiológicas. Eles são caracterizados por internações hospitalares com estadias prolongadas, maior mortalidade e custos. Objetivo geral: Descrever as características da síndrome de inflamação, imunossupressão e catabolismo persistente em pacientes críticos cônicos internados na Unidade de Terapia Intensiva do Hospital Geral La Villa, com a finalidade de conhecer os aspectos epidemiológicos. Métodos: Estudo clínico prospectivo, analítico, descritivo e observacional. Foram admitidos 25 pacientes com critérios da PICS segundo a escala NUTRIC e quadro clínico de janeiro a abril de 2018. Realizou-se testes estatísticos descritivos para variáveis qualitativas e medidas de tendência central foram utilizadas para variáveis quantitativas. Testes inferenciais para comparar variáveis qualitativas e para quantitativas tabelas 2 x 2 para calcular o Odds Ratio. Resultados: 25 pacientes preencheram os critérios da PICS, incidência de 37.9%. A distribuição por sexo foi de 60% homens e 40% mulheres sendo que a idade média foi de 48.8 anos. Os diagnósticos mais comuns foram: choque hipovolêmico (28%) e choque séptico (16%) sendo que 68% dos pacientes apresentaram baixo risco nutricional e 32% risco elevado. 84% dos pacientes necessitaram de algum tipo de suporte sendo o mais comum a ventilação mecânica. As complicações mais comuns foram infecciosas (80%) principalmente a PAVM. A mortalidade foi de 20%. Conclusões: Os resultados deste estudo não diferem dos relatados na literatura a nível internacional. A síndrome PICS ocorre em pacientes com idade avançada, maior tempo de internação e deficiências nutricionais graves em que as reservas biológicas não são suficientes para evitar imunossupressão. Isso implica e gera o círculo vicioso que finalmente leva à morte não apenas dentro da UTI.