Search Results (498)

Background: For almost two decades now, chimeric antigen receptor-natural killer cells (CAR-NK) have been investigated in pre-clinical breast cancer models, yet clinical evidence on efficacy remains scarce. This meta-analysis provides pooled evidence of pre-clinical CAR-NK effectiveness and safety in breast cancer and an overview of current clinical trials to support clinical translation. Methods: Following PRISMA guidelines and a registered protocol (PROSPERO, CRD420251131530), PubMed, Web of Science, and Scopus were searched up to 30 June 2025 for pre-clinical CAR-NK studies in breast cancer. Clinical studies were retrieved from clinicaltrials.gov and the International Clinical Trial Registry Platform (ICTRP) up to 1 March 2026. Pre-clinical studies without in vivo data or non-human CAR-NK cells were excluded. Primary outcomes were tumor burden (ratio of means, ROMs) and survival (median survival ratio, MSR). Data were analyzed in JASP™ and risk of bias (RoB) was assessed using SYRCLE’s tool. Results: Fourteen pre-clinical studies (38 CAR-NK treatment groups targeting EGFR, HER2, tissue factor, CD70, mesothelin, or folate receptor, with peripheral blood as the primary NK source, and a 5–10 million cell dose) and 11 early-phase clinical studies (targeting HER2, TROP2, PD-L1, MUC1, or NKG2D ligands under ongoing investigation) were included. In pooled pre-clinical analysis, CAR-NK significantly reduced tumor burden against untreated and unmodified/mock controls (ROM 0.311 [0.22–0.44] and 0.42 [0.33–0.53], p < 0.001, respectively). Survival was also prolonged significantly (MSR 1.47 [1.15–1.87], p = 0.010 vs. untreated; 1.30 [1.09–1.60], p = 0.007 vs. unmodified/mock NK cells). Subgroup analyses indicated improved efficacy with peripheral blood source and 5–10 M dosing. No treatment-related toxicities were reported. CAR-NK persistence was generally higher than unmodified/mock NK cells. Discussion and Conclusions: Significant heterogeneity was observed in ROM analysis which the multi-level meta-analysis configured as intra-study interventional variability. There was moderate RoB in pre-clinical studies. Published results from clinical trials remain limited, highlighting early stages of investigation. Overall, CAR-NK therapy demonstrated consistent pre-clinical efficacy and safety, supporting further translational and clinical evaluation in breast cancer. Full article

Background/Objectives: Blood transfusion is a crucial component in the treatment of individuals with sickle cell disease [SCD]; nonetheless, multiple transfusions can lead to considerable complications, notably alloimmunization. However, the prevalence of alloimmunization and its predictors remain incompletely explained. This review aimed to determine its global prevalence and identify associated risk factors. Method: Our protocol was registered in PROSPERO [ID: CRD420251167042] in accordance with the PRISMA 2020 criteria. A thorough literature search was conducted across PubMed, Embase, Web of Science, Scopus, and the Cochrane Library to identify studies reporting the prevalence of alloimmunization in adults with confirmed sickle cell disease who have received blood transfusions. This search included all publications up to 16 April 2026. Two reviewers independently screened and extracted data, and the Newcastle–Ottawa Scale was used to evaluate the study’s quality. After the Freeman–Tukey transformation, a random-effects model was used to estimate the pooled prevalence. We examined disparities among groups and geographies, study designs, and matching procedures to determine their differences. We additionally employed meta-regression to identify potential predictors. Results: Nine studies [n = 1711; 1978–2026] met the inclusion criteria. The overall rate of alloimmunization was 28.9% [95% CI 22.4–35.4; I² = 88.5%]. The most prevalent antibodies were those of the Rh and Kell systems, with anti-E antibodies being the most frequent, followed by anti-C and anti-K antibodies. A higher number of transfusions and the HbSβ⁰ genotype were both persistent risk factors, while older age at first transfusion appeared protective. Extended antigen matching dramatically reduced prevalence, though approximately 9% of individuals remained affected. Conclusions: Alloimmunization continues to challenge transfusion management in adults with SCD. Broader implementation of extended antigen matching and genotype-informed transfusion strategies may help mitigate this risk. Full article

Background: Gallbladder carcinoma (GBC) represents one of the most aggressive malignancies of the hepatobiliary system, evolving along a continuum from chronic inflammation to preneoplastic lesions and invasive cancer. This progression is frequently associated with gallstones and chronic cholecystitis and shares common pathogenic mechanisms with systemic inflammatory and metabolic disorders. Despite its relatively low incidence, GBC is characterized by poor prognosis, largely due to late-stage diagnosis and limited understanding of its molecular underpinnings. Methods: We conducted an observational study including 60 adult patients with radiologically suspected gallbladder cancer (GBC). Patients with disseminated disease, ongoing oncologic treatment, or synchronous malignancies were excluded. Fasting venous blood samples were collected to evaluate tumor markers and biochemical parameters, including carcinoembryonic antigen (CEA) and carbohydrate antigen CA 19-9. Surgical specimens were analyzed histopathologically and staged according to the European Society for Medical Oncology TNM classification system. Statistical analysis was performed using SPSS software (version 26.0), with appropriate parametric or non-parametric tests applied based on data distribution, and a p-value < 0.05 considered statistically significant. Results: Based on histological findings, patients were stratified into benign gallbladder disease (GBD) and GBC groups. CA 19-9 demonstrated higher mean serum levels with lower variability compared to CEA, suggesting superior sensitivity and diagnostic stability for gallbladder adenocarcinoma. In contrast, CEA levels exhibited greater fluctuation, limiting its reliability as a standalone biomarker. Importantly, the combined use of CA 19-9 and CEA improved diagnostic accuracy, supporting a multimarker approach for better clinical stratification. Our findings highlight the diagnostic value of CA 19-9 as a robust biomarker in GBC and support the integration of combined biomarker panels. Beyond tumor markers, the study identified a strong interplay between systemic inflammation and metabolic comorbidities, with obesity and hypertension significantly associated with chronic gallbladder pathology, and diabetes mellitus contributing to increased risk of acute inflammatory episodes. Elevated inflammatory markers, leukocytosis, and cholestatic enzyme alterations further supported the presence of a systemic inflammatory milieu. Multivariate analysis revealed that C-reactive protein (CRP), as a marker of systemic inflammation, was significantly influenced by a combination of clinical and biochemical variables, including age, hemoglobin, hypertension, amylase, CA 19-9, and CEA, explaining over 50% of its variability and up to 85% in advanced fibrotic changes. Additionally, platelet counts were significantly reduced in adenocarcinoma and correlated specifically with CA 19-9 levels, suggesting a potential link between tumor burden, inflammation, and platelet dynamics. Conclusions: Therefore, the observed associations between chronic inflammation, metabolic dysregulation, and tumor marker expression suggest a potential link between gallbladder carcinogenesis and systemic cardiometabolic pathways, opening new perspectives for early detection and targeted therapeutic strategies. Full article

Norovirus is the leading cause of acute infectious gastroenteritis in the world and accounts for a significant proportion of outbreaks at the food-borne and person-to-person levels. Due to their low infectious dose, persistence in the environment, and broad genetic diversity, they can quickly spread and reappear in even the most diverse populations. This review integrates current knowledge on the epidemiology of noroviruses, genomic organization, structural biology, virus–host interactions, and replication mechanisms, with a focus on factors that determine virus evolution and strain dominance. Literature has been systematically searched in the PubMed and Scopus databases to incorporate recent experimental and epidemiological findings. Analysis of global surveillance data indicates ongoing genetic diversification of circulating strains, with periodic replacement of major variants, particularly the GII.4 lineage. Variability of the capsid and recognition of histo-blood Group Antigens strongly affects the host’s susceptibility, viral attachment and immune escape. The capsid consists of most of the viral protein complexes. The structural proteins VP1 and VP2 are responsible for determining the contours of the capsid and antigenic specificity. Non-structural proteins are responsible for coordinating the genome replication and the modification of host cell pathways to favor the production of the virus. Eliminating these gaps by means of integrated genomic surveillance and functional studies will provide insight into the evolution of norovirus and help to develop broadly effective vaccines and antiviral strategies. Full article

Background/Objectives: The GMZ2.6c malaria vaccine candidate is a multi-stage P. falciparum chimeric protein that contains a fragment of the sexual stage Pfs48/45-6c protein genetically fused to GMZ2, which is an asexual stage vaccine construct consisting of conserved domains of Glutamate-Rich Protein (GLURP) and Merozoite Surface Protein-3 (MSP-3). Previous studies showed that GMZ2.6c is widely recognized by antibodies from individuals living in endemic areas of Brazil and that levels of anti-GMZ2.6c increase with malaria exposure and may contribute to immunity against the parasite. As cell-mediated responses are crucial for parasite control and protection, identifying antigens that elicit antigen-specific T cell recall in naturally exposed populations is the key to vaccine development. This study aimed to evaluate the cellular immune response against GMZ2.6c and its components (MSP-3, GLURP, and Pfs48/45) and to identify promiscuous T cell epitopes in individuals exposed to malaria in the Brazilian Amazon, considering the impact of active P. falciparum infection on antigen-specific T cell recall. Methods: This study was carried out using peripheral blood mononuclear cells (PBMCs) from individuals with active P. falciparum infection (PFI) and non-infected individuals exposed to malaria (NI) from Cruzeiro do Sul and Mâncio Lima, Acre State, and Guajará, Amazonas State. The PBMCs were stimulated with GMZ2.6c and its components, and cellular activation, CD4⁺ and CD8⁺ memory T cell subsets, and cytokine production were evaluated by flow cytometry. IFN-γ-secreting T cells were quantified by ELISpot using predicted T cell epitopes. Results: The individuals infected by P. falciparum displayed more CD8⁺ T cell activation in response to MSP-3 and Pfs48/45 and an increase in CD4⁺ T_CM cells and a reduction in CD4⁺ T_EM cells following stimulation with Pfs48/45 and GMZ2.6c. The PBMCs from both groups showed elevated production of IL-6 and TNF after stimulation with GMZ2.6c, MSP-3, and Pfs48/45, but only the non-infected individuals had high levels of IL-10. T cell epitope prediction identified sequences within MSP-3, GLURP, and Pfs48/45 that elicited IFN-γ responses in both the non-infected and P. falciparum-infected individuals. Conclusions: Individuals exhibit cellular immune responses to MSP-3 and Pfs48/45 that are recalled following GMZ2.6c stimulation. P. falciparum infection may modulate immune response, inducing a prominent pro-inflammatory response. Conversely, in the absence of the parasite, the individuals displayed balanced Th1/Th2 cytokine production. Several promiscuous T cell epitopes were able to recall IFN-γ responses. Further studies are needed to fully ascertain the potential of GMZ2.6c as a protective candidate vaccine against malaria. Full article

Background: Prostate-specific antigen (PSA) remains the primary biomarker for prostate cancer detection; however, its limited specificity leads to unnecessary biopsies. Systemic inflammatory indices derived from routine blood tests have been proposed as potential adjunctive diagnostic tools. This study aimed to evaluate the diagnostic performance of these indices in patients undergoing prostate biopsy for suspected prostate cancer. Methods: A retrospective observational study was conducted, including 307 patients who underwent transrectal ultrasound-guided prostate biopsy for elevated PSA between January 2021 and January 2023. Patients were classified as benign prostatic hyperplasia (BPH) or prostate cancer (PCa) based on histopathological findings. Inflammatory indices, including neutrophil-to-lymphocyte ratio (NLR), systemic immune–inflammation index (SII), Systemic Inflammation Response Index (SIRI), and aggregate index of systemic inflammation (AISI), were calculated. Logistic regression and receiver operating characteristic (ROC) analyses were performed to assess diagnostic performance. Results: PCa was diagnosed in 65.1% of patients. Several inflammatory indices were significantly higher in the PCa group. Among them, AISI showed diagnostic performance comparable to PSA. The integration of inflammatory indices into multivariable models improved predictive accuracy, with the combined model demonstrating the highest discriminative ability. In contrast, these markers showed limited capacity in distinguishing tumour aggressiveness. Conclusions: Systemic inflammatory indices are associated with prostate cancer presence and may enhance diagnostic performance when used alongside PSA. However, their role remains complementary, as they provide limited value in risk stratification and should not be considered standalone diagnostic tools. Full article

Background/objectives: Current treatments for advanced hepatocellular carcinoma (HCC) are tyrosine kinase inhibitors (TKI) and immune checkpoint inhibitors (ICI). When HCC is resistant to TKI or ICI, subsequent available treatments are limited. Dendritic cells (DC) can activate antigen-specific cytotoxic T-cells and may be employed as a subsequent treatment when HCC is resistant to TKI or ICI treatments. Methods: Fifty advanced HCC patients with resistance to ICI or TKI treatment were invited to have autologous DC therapy. DCs were propagated from peripheral blood monocytes and pulsed with tumor lysate. All the patients received ≥3 courses of DC intravenously. For the outcome analysis, 17 patients who were resistant to TKI or other traditional treatments were grouped into A, and 33 patients who were resistant to ICI treatment were grouped into B. Results: For group A patients, the median (interquartile) progression-free and overall survivals were 6 (3–16.3) and 19 (8–24) months, respectively. The 1-, 2- and 3-year overall survivals after DC therapy were 58.2%, 21.8%, and 14.6%, respectively. For group B patients, the median (interquartile) progression-free and overall survivals were 5.0 (4–8) and 9 (5–14.5) months, respectively. The 1-, 2- and 3-year overall survivals after DC therapy were 27.6%, 8.6% and 4.3%, respectively. Taking together 50 patients, the objective response rate was 6.0% and disease control rate was 72.0%. Only three patients (6.0%) had grade I-II hepatitis. Conclusions: DC therapy is a safe treatment for advanced HCC patients. DC can serve as a subsequent therapy to extend the patients’ lives when TKI or ICI treatments are ineffective as advanced HCC treatments. Full article

Background and Objectives: Healthcare workers are at heightened risk of SARS-CoV-2 infection. Understanding the duration and protective value of vaccine-induced immunity is critical to inform booster strategies. This study investigates longitudinal dynamics of anti-SARS-CoV-2 receptor-binding domain IgG (anti-RBD IgG) antibodies and their association with infection risk among vaccinated healthcare workers. Materials and Methods: A prospective cohort study was conducted at Vilnius University Hospital Santaros Klinikos, Lithuania. A total of 1778 healthcare workers who completed a primary COVID-19 vaccination series were followed. Blood samples were collected every three months to measure anti-RBD IgG levels. Participants also received up to three booster doses. COVID-19 was identified by PCR, antigen tests, or positive anti-nucleocapsid IgG. For serologically detected cases, infection timing was assigned to the interval between study visits. Antibody dynamics were analyzed across vaccination stages, time, age groups, and circulating SARS-CoV-2 variants. Results: Anti-RBD IgG titers peaked in the first quarter after primary vaccination (mean 7904 AU/mL), declined sharply by quarters 2–3, and rose substantially after booster doses. Following the first booster, titers increased to ~12,598 AU/mL in quarter 1 and continued rising through quarter 3. The highest levels were observed after the second booster (24,456 AU/mL in Q1), followed by gradual decline. A high-titer plateau persisted from quarters 6 to 9 (~21,000 AU/mL), followed by decline in quarters 10–11 and partial rebound in Q12. Approximately 49.6% of participants experienced COVID-19 during follow-up. Antibody response patterns were similar across age groups, with only minor transient differences. Conclusions: COVID-19 booster doses significantly enhance and prolong humoral immunity in healthcare workers compared with the primary vaccination series. However, antibody waning over time emphasizes the need for timely boosters, particularly during periods of variant circulation. These findings support continued booster vaccination and monitoring of long-term immune protection, although anti-RBD IgG should be interpreted as a surrogate marker of humoral rather than overall immunity. Full article

Background and Objectives: The incidence of colorectal cancer (CRC) in developed Western countries is constantly growing. CRC represents the third most common cancer and the second leading cancer-related cause of death worldwide. Innate and adaptive immunity play a pivotal role in the tumor response, but many of these interactions are still not well understood. Granulysin (GNLY) is an effector, cytolytic molecule, present in human cytotoxic granules of different lymphocyte subpopulations, mainly in cytotoxic T cells and NK cells. Pore-forming proteins GNLY, perforin and granzymes play a key role in cell-mediated immune responses against tumors and infections. Materials and Methods: We aimed to analyze perforin and GNLY-mediated cytotoxicity in the peripheral blood of patients with CRC by flow cytometry. Simultaneously, the cells were labeled with monoclonal antibodies against perforin, GNLY and different surface antigens (CD3, CD4, CD8 and CD56). Phenotypes of lymphocyte subpopulation and expression of perforin and GNLY were analyzed using intracellular and surface immunofluorescence. Results: Total perforin and GNLY expressions in peripheral blood mononuclear cells (PBMC) were significantly lower than in the control group. Statistically significant differences were observed in the distribution of perforin and GNLY expression in different stages of tumors classified according to Dukes’, indicating that the percentage of total perforin and GNLY was significantly diminished in accordance with tumor progression. Perforin and GNLY expression were significantly reduced in NK and NKT cells, accompanied by reduced cytolytic potential in patients with CRC and a consequent reduction in their ability to eliminate tumors and infected cells. Conclusions: The determination of cytotoxic potential may provide a valuable assessment of a patient’s immune status and represent a novel therapeutic target. Patients with CRC exhibit markedly impaired perforin- and GNLY-mediated cytotoxicity that correlates with disease progression. Assessment and restoration of cytolytic potential may therefore serve as indicators of immune competence and promising therapeutic strategies to improve perioperative and oncologic outcomes. Full article

Background/Objectives: The main goal of this study is to identify predictors associated with Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) seropositivity in children, including demographics, history of coronavirus disease 2019 (COVID-19) infection of the child and the household members, prevention practices, and maternal vaccination. Methods: This retrospective cross-sectional study within the Caribbean Consortium for Research in Environmental and Occupational Health (CCREOH)/MeKiTamara cohort included 300 mother-child dyads recruited in Paramaribo and Nickerie, Suriname (February–April 2023). The total immunoglobulin G (IgG) anti-spike domain 1 (S1) and anti-nucleoprotein (NP) were quantified in dried blood spot (DBS) eluates from children using indirect enzyme-linked immunosorbent assays (ELISAs). Demographic information, COVID-19 prevention measures, history of viral infection of the child and the household members, and COVID-19 vaccination questionnaire data were recorded. Predictors of SARS-CoV-2 seroprevalence were determined using binary logistic regression. Results: Among 278 seropositive children in 2023, 73.4% were in the 5–6-year-old age group, 54.7% were female, 36.3% were of Asian descent, and 69.8% were recruited in Paramaribo. Seroprevalence increased from 33.8% in 2021–2022 to 93.3% in 2023, with a mean follow-up of 21.5 months. Of the 100 children previously tested by Polymerase Chain Reaction (PCR) or antigen test, 25 had confirmed COVID-19, as reported by mothers. Children from unvaccinated mothers were 6.11 times more likely to be seropositive (p = 0.022). Conclusions: This study shows a significant increase in SARS-CoV-2 seropositivity in Surinamese children aged 3–6 years between collection periods, indicating multiple exposures. Future public health interventions and policies should account for maternal vaccination status to reduce children’s exposure to COVID-19 during future outbreaks. Full article

Background/Objectives: The association between ABO/Rh blood groups and knee osteoarthritis (OA) remains controversial, with inconsistent findings reported across different populations. This study aimed to evaluate the distribution of ABO and Rh(D) blood groups in patients undergoing primary total knee arthroplasty (TKA) for primary knee OA and to compare these distributions with a regional external reference population. Methods: This retrospective, single-center, observational study reviewed hospital records of patients who underwent primary TKA between January 2011 and October 2024. After applying predefined exclusion criteria (different ethnic background, age < 50 years, secondary knee OA, and missing blood group data), 4969 patients with primary knee OA were included. ABO/Rh(D) data were obtained from the institutional electronic hospital information system and transfusion/laboratory records. The external reference population consisted of a previously published dataset of regional blood donors (10,867 unique donors). Observed blood group frequencies in the study cohort were compared with expected frequencies derived from the reference distribution using chi-square goodness-of-fit tests for (1) 8-category ABO/Rh(D) distribution, (2) ABO-only distribution, and (3) Rh(D)-only distribution. Results: Among 4969 patients, 4096 (82.4%) were female and 873 (17.6%) were male. Mean age was 66.8 ± 7.0 years (range, 50–94) in females and 68.8 ± 7.3 years (range, 50–88) in males. The most frequent blood groups were A Rh (+) (39.3%), O Rh (+) (30.0%), and B Rh (+) (14.5%). The sex-specific ABO/Rh distribution did not differ significantly (p = 0.052). Compared with the regional reference distribution, the overall 8-category ABO/Rh(D) distribution showed a borderline difference (χ² (7) = 14.04, p = 0.050; Cramér’s V = 0.020). However, neither the ABO-only distribution (χ² (3) = 5.26, p = 0.153; Cramér’s V = 0.019) nor the Rh(D)-only distribution (χ² (1) = 0.11, p = 0.737; Cramér’s V = 0.005) differed significantly from the regional reference. The observed deviations were numerically small and not suggestive of a clinically meaningful difference. Conclusions: In this large single-center cohort of patients undergoing primary TKA for primary knee OA, the ABO and Rh(D) blood group distributions were largely comparable to those of the regional population. Although the overall 8-category ABO/Rh(D) comparison showed a borderline difference, separate ABO-only and Rh(D)-only analyses were not significant. These findings do not support a strong association between blood group status and surgically treated primary knee OA in this population. Full article

Rhipicephalus microplus infestations are a major concern for cattle production in neotropical regions. Anti-tick vaccines represent a sustainable alternative to chemical acaricides; however, improving vaccine efficacy remains challenging. This study evaluated the protective efficacy of polypeptide-based formulations of pBm86 and pSubolesin administered as co-immunization or as a cocktail in naturally infested cattle. Twelve calves were assigned to three groups: G1: 100 μg pBm86 + 50 μg pSubolesin co-immunized; G2: same formulation in cocktail; and G3: control. The cocktail significantly reduced adult female tick numbers, oviposition, and larvae hatching (p < 0.05), achieving a vaccine efficacy of 75%. In contrast, co-immunization achieved 50% efficacy. Vaccinated cattle developed significantly increased antigen-specific IgG levels against Subolesin than Bm86. IgG antibodies recognized native tick proteins by Western blot, and their efficacy was confirmed in vitro using blood-feeding assays. These results indicate that the cocktail formulation showed improved efficacy compared with co-immunization in this study, possibly by affecting complementary biological processes in ticks. Finally, this study supports the development of multi-antigen, polypeptide-based vaccines as a promising and sustainable approach for the control of R. microplus under natural field conditions. Full article

Background/Objectives: Thalassemia and sickle cell anemia (SCA) patients require regular blood transfusions, a necessity that increases the risk of alloimmunization and complicates subsequent transfusion management. Methods: This retrospective cohort study, conducted at King Saud Medical City (KSMC) and King Fahad Medical City (KFMC) between 2018 and 2022, evaluated the frequency and risk factors of alloimmunization among 144 transfusion-dependent patients in Riyadh, Saudi Arabia. Results: By reviewing clinical and transfusion records alongside antibody screening results, the study found an overall alloimmunization prevalence of 20.1%. Notably, females exhibited a significantly higher rate (13.2%) compared to males (6.8%; p = 0.003), and younger patients (<20 years) showed a higher prevalence than older cohorts (p = 0.004). Analysis of ABO blood groups revealed that group A patients had a significantly lower alloimmunization rate (7.5%) compared to non-A patients (23.1%; p = 0.018), a finding that raises hypotheses about differential immune responsiveness but requires confirmation in larger studies. Group B showed the highest rate (35.3%), though this did not reach statistical significance after correction for multiple comparisons. RhD status was not significantly associated with alloimmunization. The most frequent alloantibodies identified were anti-E (31.3%), anti-K (12.5%), anti-D (10.4%), and anti-C (10.4%). Logistic regression further identified gender as a significant predictor (OR = 0.270; 95% CI: 0.113–0.646). Conclusions: Given that alloimmunization rates in Riyadh are moderately high—particularly among females and specific blood groups—and that the antibody profile (anti-E, anti-K, anti-C, anti-D) mirrors patterns seen in populations with recipient–donor ethnic mismatches, implementing extended blood group phenotyping for at least Rh (C, c, E, e) and Kell antigens prior to the first transfusion, and incorporating these findings into donor selection protocols, is critical to mitigating these risks. Full article

Spotted fever group (SFG) rickettsioses are tick-borne infectious diseases caused by more than 30 Rickettsia species. As ticks may harbor and transmit multiple pathogens during a single blood meal, sensitive and specific molecular detection methods are essential for early diagnosis. Conventional nested PCR is commonly used but is time-consuming and prone to cross-contamination due to multiple amplification steps. This study evaluated a dual-target one-step nested PCR assay developed as a rapid alternative to conventional nested PCR for SFG Rickettsia detection. Gene-specific primers targeting the Rickettsia outer membrane protein A (ompA) gene and the 17 kDa antigen gene were designed, with a Plasmodium falciparum thrombospondin-related anonymous protein (TRAP) gene included as an internal amplification control. Primer specificity was verified in silico, and assay performance was assessed using synthetic DNA templates. The dual-target one-step nested PCR achieved detection limits of 10 gene copies for the 17 kDa gene and 1000 gene copies for ompA, compared with 10 and 100,000 gene copies, respectively, using conventional nested PCR. Screening of 184 tick specimens identified one positive sample (0.54%) for the Rickettsia 17 kDa gene. Overall, the dual-target one-step nested PCR demonstrated comparable sensitivity to conventional nested PCR while reducing assay time and contamination risk, indicating its potential as a reliable tool for SFG Rickettsia detection. Full article

Infection with SARS-CoV-2, the virus responsible for COVID-19, is associated with cytokine storm, an excessive immune response. Interleukin-6, a multifunctional cytokine, is involved in the COVID-19 immune response. Functional polymorphisms in the interleukin-6 gene promoter, namely rs1800796, rs1800797, and rs2069845, may contribute to individual susceptibility to or severity of COVID-19. In this study, 106 healthy SARS-CoV-2-negative individuals (controls) and 106 patients with severe COVID-19 (COVID-19 group), confirmed by qPCR or rapid antigen tests, were genotyped using fluorescent probes for polymorphisms. All participants were from southern Brazil. Groups were matched for sex and body mass index, with a median age of 56–57 years. The COVID-19 group exhibited blood biomarker concentrations consistent with severe disease. No significant differences were detected in genotypic or allelic frequencies between groups, and all polymorphisms conformed to the Hardy–Weinberg equilibrium. The control group minor allelic frequencies for rs1800796 (allele C, 11.3%; 95% CI, 7–16), rs1800797 (allele A, 28.3%; 95% CI, 22–34), and rs2069845 (allele G, 36.8%; 95% CI, 30–50) were similar to those of African, American, and European populations. The polymorphisms investigated were not associated with severe COVID-19 in this cohort. Full article