Search Results (351)

Cardiorenal Syndrome (CRS) types depend on the primary impaired organ, Heart (Types I/II) or Kidney (Types III/IV), and are chronic (Types II/IV) or acute (Types I/III). Type V associates to a systemic disease. Diagnosis of CRS type via biochemical indices (e.g., B-type-natriuretic-peptide (BNP), neutrophil-gelatinase-associated-lipocalin (NGAL)) has not been documented absolutely, until now. Here we used advanced imaging facilities, atomic force microscope (AFM) and scanning electron microscope (SEM) for the biopsy of peripheral blood smears coming from CRS patients, aiming to investigate the possible existence of cellular indices associated with CRS pathology. Standard biochemical and hematological indices were comparatively recorded. Cylindrical micro/nano-metric Vesicles (mnVs) were observed in all CRS patients. In CRS patients with Types III/IV, the AFM/SEM data revealed an increased mnVs population and activated platelets that may represent their potential parent cells. In these patients, increased basic uraemic indices and BNP and NGAL levels were also observed. On the contrary, in patients with CRS Types I/II/V, the AFM/SEM data showed a comparatively smaller mnVs population, accompanied by lower levels of BNP and NGAL. According to our results, a higher mnVs population was observed in CRS Types III/IV, possibly released from platelets. The mnVs population may be associated with basic uraemic indices and increased BNP and NGAL levels. Full article

Canine monocytic ehrlichiosis, caused by Ehrlichia canis, is an important tick-borne disease of dogs in tropical and subtropical regions. This cross-sectional study investigated the occurrence, associated risk factors, and molecular confirmation of E. canis infection among clinically suspected dogs in Andhra Pradesh, India, and evaluated the diagnostic performance of blood smear microscopy relative to PCR. Peripheral blood samples were collected from 442 clinically suspected dogs presented to 90 veterinary dispensaries across the state. Dogs were selected based on clinical features suggestive of ehrlichiosis, including inappetence, pale mucous membranes, tick infestation, fever, lymphadenopathy, weakness, and hemorrhagic signs. Samples were examined by Giemsa-stained blood smear microscopy and subsequently tested by PCR. Representative virB9 gene amplicons were sequenced and compared with reference sequences available in GenBank. Microscopy detected E. canis in 37/442 dogs (8.37%), whereas PCR detected infection in 51/442 dogs (11.54%). Logistic regression analysis indicated that female dogs, mongrels, kennel dogs, and dogs with tick infestation had significantly higher odds of infection, while dogs with a history of tick-control measures had lower odds. Sequence comparison and phylogenetic placement based on the partial virB9 fragment supported molecular confirmation of the detected E. canis isolates and showed similarity with previously reported Indian and international isolates. These findings provide regional evidence on the occurrence and epidemiology of canine ehrlichiosis in clinically suspected dogs and support PCR as a more reliable confirmatory method than microscopy. Full article

Background: Accurate segmentation of white blood cells (WBCs) in peripheral blood smear images is a fundamental step in computational hematology, enabling downstream tasks such as classification, morphological assessment, and quantitative analysis. However, reliable segmentation remains challenging due to staining variability, complex cellular morphology, overlapping structures, and limited availability of high-quality annotations. Aim and Methods: The aim of this study is to develop a robust and fully automated segmentation framework for white blood cells (WBCs) in microscopic blood smear images, providing a reliable foundation for subsequent computational analysis. We propose TransNet–SAM2, a hybrid deep learning architecture that integrates hierarchical transformer-based feature extraction with a foundation-model-based decoder for prompt-free segmentation. Specifically, a Swin Transformer backbone is employed to capture multi-scale contextual representations, which are subsequently aligned and fused through a feature adaptation module. The fused features are directly injected into the SAM2 mask decoder, replacing conventional prompt-based conditioning and enabling fully automatic segmentation. In addition, a weakly supervised self-training strategy is incorporated to utilize partially annotated data, improving model generalization while reducing annotation requirements. The proposed framework is evaluated using a clinically curated dataset from Dicle University, the publicly available Raabin-WBC dataset, and an additional external leukemic blast validation dataset (ALL-IDB) to assess robustness under both routine and atypical hematological conditions. Results: TransNet-SAM2 achieved a Dice coefficient of 0.95 ± 0.01 and IoU of 0.90 on internal testing, significantly outperforming U-Net (0.89), Mask R-CNN (0.90), and SAM2 (0.92) (p < 0.05). In cross-dataset evaluation (Dicle training, Raabin-WBC testing), the framework maintained strong performance (Dice: 0.91, IoU: 0.84), demonstrating robustness to domain shifts. Ablation studies confirmed each component’s contribution, with the full model improving Dice by 6% over a CNN baseline. Qualitative analysis showed accurate boundary delineation even with cell overlap and background clutter. Conclusions: These findings indicate that the proposed framework provides a promising and scalable framework for WBC segmentation. While the current study focuses on segmentation, future work will investigate integration with classification and radiomics pipelines, as well as validation on more diverse clinical datasets, including bone marrow and leukemia samples. Full article

Neutrophils are central executors of innate immunity. Yet murine models are inherently limited by low baseline neutrophil counts. NSG mice are among the most widely used models for xenotransplantation and studies on the humanized immune system. Although G-CSF is known to stimulate granulopoiesis, the dose- and schedule-dependent effects of intraperitoneal G-CSF administration have not been systematically characterized in this immunodeficient background. Male NSG mice received intraperitoneal G-CSF according to one of five regimens (n = 6 per group): group 0 served as the saline control, group 1 received a single dose of 250 µg/kg G-CSF administered at 48 h; group 2 received a single dose of 250 µg/kg G-CSF administered at 24 h; group 3 received three doses of 250 µg/kg administered G-CSF at 0 h, 24 h, and 48 h and group 4 received a single dose of 500 µg/kg G-CSF administered at 48 h. All animals were sacrificed at 72 h. Circulating neutrophils were then quantified by flow cytometry, bone marrow neutrophil proportions by panoptic smear analysis, and splenic neutrophil abundance by Ly6G immunofluorescence. Systemic neutrophil activation was assessed via plasma neutrophil elastase (NE) activity and cell-free DNA (cfDNA) levels. Repeated G-CSF administration (Group 3) induced an approximately 13-fold expansion of circulating neutrophils, approaching the human physiological range, with significant increases also observed in bone marrow and a trend towards increased neutrophil abundance in the spleen. A single dose of 250 µg/kg administered at 24 h (group 2), produced significant neutrophil expansion in peripheral blood and bone marrow but not in the spleen, while all other single-dose regimens failed to induce significant expansion in any compartment. NE activity and cfDNA concentrations and a selected cytokine panel remained unaltered across all groups. This systematic comparison establishes repeated intraperitoneal G-CSF administration as a reproducible strategy to achieve human-like neutrophil levels in NSG mice without inducing systemic inflammation. This provides a validated protocol with direct utility in translational models of neutrophil-dependent diseases. Full article

Abortion in cattle entails substantial economic loss, and rapid identification of abortigenic pathogens is critical for timely on-farm response and reduction in human exposure risk. In 2024, two Holstein cows from a small farm in Inner Mongolia aborted in close succession without an obvious cause. Vulvar swabs from both cows, one afterbirth sample, and whole blood from one aborted fetus were collected. Shotgun metagenomic sequencing was performed, followed by host-read removal, taxonomic profiling with Kraken2, de novo assembly of Brucella-aligned reads, and whole-genome comparison. Serological tests, Gram-stained smears, and Brucella genus- and species-specific qPCR assays were used as orthogonal verification. Putative resistance and virulence determinants were screened against CARD and VFDB. Brucella reads were detected in all samples, with the highest relative abundance in the 138-afterbirth (96%). qPCR assays detected Brucella DNA and B. abortus-specific signals in all four samples. A draft Brucella genome was assembled from the 138-afterbirth sample and was phylogenetically placed within B. abortus, showing relatedness to previously circulating Chinese lineages. Cows 138 and 198 were RBT-positive with SAT titres of 1:100 (++). No acquired Brucella resistance genes were identified in CARD. Within 72 h of sample receipt, B. abortus was reported to the farm and local authorities and emergency biosecurity measures were implemented. This field investigation shows that metagenomic sequencing, when combined with conventional serology, microscopy, and targeted qPCR, can support rapid etiological investigation when culture is delayed, hazardous, or biosafety level 3 facilities are unavailable. Full article

To elucidate hematopoietic ontogeny in Monopterus albus, systematic cytological analysis was performed using Wright–Giemsa-stained peripheral blood smears and hematopoietic organ imprints. Peripheral blood smears revealed five major cell types: erythrocytes (60.05 ± 8.08%), lymphocytes (13.50 ± 4.27%), thrombocytes (8.95 ± 3.47%), neutrophils (6.40 ± 2.17%), and monocytes (5.32 ± 1.63%), with occasional basophils and no eosinophils. Microscopic analysis revealed that M. albus hematopoiesis progresses through three distinct developmental phases—primitive, juvenile (with granulocytic series exhibiting promyelocyte/myelocyte/metamyelocyte substages), and mature stages—across erythroid, monocytic, lymphoid, and granulocytic lineages. The head kidney served as the primary hematopoietic organ, containing pan-stage cells and supporting all lineages, whereas the spleen lacked myeloblasts but participated in non-granulocytic hematopoiesis, and the liver was restricted to juvenile-stage erythrocyte, lymphocyte, and neutrophil development. Notably, erythrocytes in the peripheral blood with elongated nucleus were observed in the peripheral blood. Full article

Background: Dirofilaria spp. is an etiological agent of dirofilariasis, a mosquito-borne parasitic disease of increasing zoonotic concern in Europe. The aim of this study was to assess the occurrence of Dirofilaria spp. in dogs from Southern Poland using retrospective data from a commercial veterinary diagnostic laboratory (Vetlab, Katowice, Poland). Methods: Blood tests from 2060 dogs were analyzed between 1 August 2018 and 31 December 2022. All samples were collected by the clinicians during routine veterinary activity and examined by a specific test—microscopic (blood smear/blood smear and Knott’s test), molecular or both—from the Vetlab laboratory offer (test selected by clinician). Results: Out of all examined dogs, 19 (0.92%) tested positive for Dirofilaria. Positive samples originated from the Śląskie (n = 13), Opolskie (n = 3), and Małopolskie (n = 3) voivodeships. Co-infections with Babesia spp. and Anaplasma spp. were identified in two blood samples. Conclusions: This study demonstrates the presence of Dirofilaria spp. in dogs from Southern Poland, a region where data about dirofilariasis cases remain limited. Its overall occurrence was low in comparison to endemic areas in Central Poland. However, the presence of confirmed cases highlights the need for increased veterinary awareness, implementation of preventive measures, and further molecular epidemiological studies to better evaluate the risk of exposure to Dirofilaria in this region. Full article

Vector-borne protozoal infections—including babesiosis, theileriosis, hepatozoonosis, trypanosomosis, and leishmaniosis—impose a substantial burden on livestock and companion animal health worldwide and carry important zoonotic and public health implications. Accurate diagnosis is essential yet challenging, given the diversity of parasite genera, their markedly different tissue tropisms, and the uneven distribution of diagnostic resources across veterinary settings. This review provides an integrated overview of the principal diagnostic approaches available, structured around the biological logic that guides test selection in practice. Microscopic examination remains the first-line method; its strengths and limitations are discussed for intraerythrocytic parasites (Plasmodium spp., Babesia spp., Theileria spp., Cytauxzoon spp.—the latter two with additional extra-erythrocytic schizont stages in leukocytes and tissue macrophages, respectively), leukocyte-associated forms (Hepatozoon spp.), extracellular trypanosomes, and tissue-stage parasites, including emerging applications of artificial intelligence. Serological methods—enzyme-linked immunosorbent assay (ELISA), indirect fluorescence antibody test (IFAT), and point-of-care lateral flow assays—are evaluated for their role in exposure detection, population screening, and international trade certification, with attention to cross-reactivity and the active-versus-past-infection distinction. Molecular diagnostics, encompassing conventional PCR, qPCR, droplet digital PCR, isothermal amplification, and next-generation sequencing, are reviewed with respect to target selection, sensitivity, and point-of-care applicability. Finally, diagnostic challenges are contextualised within a One Health framework, highlighting the fragmentation of veterinary surveillance and the need for integrated, cross-sector approaches to detect emerging threats. Full article

Climate change significantly affects human physiology and contributes to increased morbidity and mortality, with heat stress representing one of the most severe consequences of thermal imbalance. The aim of this study was to analyze morphological changes to leukocytes on the peripheral blood smears of Wistar rats exposed to hyperthermia using the geometric morphometrics method. A total of forty Wistar albino rats were divided into three experimental groups according to water temperature exposure (37 °C, 41 °C, and 44 °C). Peripheral blood smears were prepared, stained, and digitally recorded using Motic Images Plus 2.0 software, after which selected images were analyzed using geometric morphometric programs (tpsDig, tpsUtil, and MorphoJ) to evaluate leukocyte shape variations. Comparative analysis demonstrated statistically significant morphological changes in polymorphonuclear cell shapes between the control group (37 °C) and rats exposed to 41 °C (p = 0.009). Significant differences were also identified in mononuclear cell morphology between the antemortem and postmortem groups (p = 0.00307). The findings indicate that exposure to elevated temperatures induces measurable alterations in white blood cell morphology, confirming that hyperthermia produces significant structural changes in polymorphonuclear cells and mononuclear cells detectable through geometric morphometric analysis. Full article

Background. Acquired platelet dysfunction with eosinophilia (APDE) is a transient bleeding disorder initially thought to occur exclusively in Southeast Asia. There are no uniformly agreed diagnostic criteria, and its full clinical features have not been defined. Methods. A literature search was conducted through MEDLINE, EMBASE, and Google Scholar for publications on APDE in order to explore patient demography, epidemiology, diagnostic criteria, and laboratory findings of the disease. Results. Ten retrospective, observational studies, five case series, and 21 case reports were identified with a total of 431 patients. Diagnostic criteria varied, with a two-tier approach for the diagnosis of impaired platelet function. In recent years, cases of APDE have been reported extensively outside of the Malay peninsula. Male patients (243/390, 62.3%) predominated. Their ages ranged from 11 months to 30 years, with only 40 (9.3%) subjects aged 18 years or older. Eosinophilia was absent in 10 to 33% of subjects in a few observational studies. Thrombocytopenia was present in 42 (9.7%) subjects. Parasitic infestation was less common in the new millennium. Spontaneous recovery within six months was the trend, and serious complications were extremely rare. Conclusions. APDE is no longer restricted to Southeast Asia. A uniform set of diagnostic criteria is needed. Clinician awareness followed by reliable but easily available laboratory tests is essential to confirm diagnosis. It is proposed that rapid diagnosis can be accomplished by screening the blood smear for grey platelets by a trained examiner followed by confirmatory platelet aggregometry. National or international collaboration and prospective studies are required to delineate the core and variable clinical and laboratory features of APDE. Full article

Accurate and efficient classification of hematological malignancies from peripheral blood smear (PBS) images remains challenging due to the scarcity of annotated datasets, staining variability, and subtle morphological differences among blood cancer subtypes. To address these limitations, this study proposes an Advanced Lightweight Deep Learning (ALDL) framework for the multi-class classification of Acute Lymphoblastic Leukemia (ALL) across four clinically significant stages: Benign, Pro-B, Pre-B, and Early Pre-B. The framework integrates EfficientNetV2-S with Convolutional Block Attention Modules (CBAM) to enhance spatial and channel-wise feature refinement. At the same time, Focal Loss is employed to mitigate class imbalance by prioritizing hard-to-classify samples. A robust preprocessing pipeline, including CLAHE contrast enhancement, Reinhard stain normalization, and data augmentation, improves feature visibility and dataset generalization. Lesion segmentation is performed using RGB-based thresholding and watershed overlay, followed by lesion-level cropping to ensure consistency across inputs. Experimental evaluations on the ALL-DB dataset demonstrate the superior performance of the proposed method, achieving an average accuracy of 96.11%, an F1-score of 95.99%, and an AUC of 0.9875. Comparative analyses against MobileNetV3, ResNet50, DenseNet121, VGG16, and InceptionV3 confirm that the proposed segmentation-guided EfficientNetV2-S + CBAM + Focal Loss framework consistently outperforms conventional CNN architectures across both 70:30 and 60:40 train–test splits. Furthermore, a detailed investigation of color spaces (RGB, HSV, LAB, and HED) indicates that RGB yields the most reliable segmentation and classification results. At the same time, HED enhances lesion visualization at the expense of higher computational cost. The proposed ALDL framework demonstrates strong potential for real-world application as a computer-aided diagnostic (CAD) system for early leukemia detection, offering improved diagnostic reliability, reduced error rates, and practical scalability for clinical environments. Full article

Background: Accurate and early detection of hematological disorders from microscopic peripheral blood smear images remains a technically challenging task due to inherent imaging limitations, including noise contamination, low contrast, staining variability, and significant cellular overlap. Conventional deep learning-based object detection frameworks often exhibit limited robustness under such conditions and demonstrate reduced sensitivity to small-scale morphological structures, particularly platelets and abnormal cell variants. Methods: To address these challenges, this study proposes a hybrid detection framework that integrates a fuzzy logic-driven image preprocessing module with the YOLOv11 object detection architecture. The proposed preprocessing pipeline employs adaptive fuzzy membership functions to normalize pixel intensity distributions, suppress high-frequency noise, and enhance edge-defined cellular boundaries. This transformation produces a structurally optimized feature representation, improving downstream feature extraction and localization performance. The proposed framework was evaluated on a curated dataset of 3000 annotated microscopic blood smear images spanning five hematological classes. Results: Experimental results show that the fuzzy logic module improves mAP@0.5 by +3.4% and mAP@0.5:0.95 by +3.6%, confirming its effectiveness in enhancing both classification and localization accuracy. Conclusions: These findings demonstrate the robustness and practical applicability of the proposed hybrid approach under challenging imaging conditions. Full article

Cytauxzoon felis is a tick-borne protozoan parasite of considerable importance to domestic cats in the United States. Despite high mortality among clinically presenting cats, some cats may either recover from infection, becoming permanent chronic carriers of C. felis, or may not develop disease following exposure. C. felis in healthy (asymptomatic) cats is currently detected by qPCR or through observation of piroplasms in blood smears. The present study aims to compare the prevalence of C. felis in healthy populations of pet and free-ranging cats by using two different diagnostic tests: an indirect ELISA and a probe-based qPCR. Samples, along with demographic information, were collected collaboratively from healthy cats in the states of Oklahoma, Arkansas, and Missouri (USA) in either pet or free-range populations. Significantly more cats were detected to be C. felis positive by the ELISA than qPCR. Of the total of cats tested by ELISA and qPCR, 248 (35.37%) tested ELISA positive but qPCR negative, indicating formation of specific C. felis antibodies in cats with undetectable circulating C. felis DNA. The results of this study provide insights into the occurrence of C. felis in healthy cats that are exposed in the south-central United States and present a useful diagnostic strategy for detecting C. felis. Full article

Background/Objectives: Despite the availability of the National Cancer Screening Program in Korea, participation rates remain suboptimal. The literature demonstrates that cancer screening participation extends beyond individual-level knowledge and attitudes and is largely associated with trust. This study examines the role of trust—across cancer screening tests, health care providers, and health care organizations—as a central determinant of cancer screening participation among Koreans. Methods: A cross-sectional study was conducted with 369 Korean adults aged 40 years and older, recruited through convenience sampling from community centers in a metropolitan city. Data were collected using structured, paper-based questionnaires assessing socio-demographic factors and multilevel trust across specific screening tests, doctors, and health care organizations. Following descriptive statistics, bivariate and multivariate logistic regression analyses were performed to identify significant predictors of cancer screening utilization. Results: Koreans had relatively high trust in doctors and cancer screening tests. On an 11-point numeric rating scale ranging from 0 (not at all) to 10 (completely), the mean scores were 7.47 for the trust in doctors; colonoscopy had the highest trust score (M = 7.71), whereas the fecal occult blood test had the lowest (M = 7.14). Multivariate logistic regression revealed that trust and distrust were associated with the utilization of Pap smear, gastroscopy, upper gastrointestinal series, and colonoscopy in complex and sometimes paradoxical ways, and having a usual source of care was a consistent facilitator of cancer screening. Conclusions: These findings suggest that future research is needed to examine the complex interplay among trust, access to health care, and national policy in shaping cancer screening utilization in the Republic of Korea. Full article

Background: Multimodal large language models (MLLMs) show promise in medical imaging, but their performance is highly dependent on prompt engineering. This study systematically evaluates how different prompting strategies affect diagnostic accuracy in clinical laboratory image interpretation. Methods: We evaluated five MLLMs (ChatGPT-4o, Gemini 2.0 Flash, Claude 3.5 Sonnet, Grok-2, and Perplexity Pro (Claude 3.5 Sonnet)) using 177 proficiency testing images across three domains: blood smears (n = 78), urinalysis (n = 50), and parasitology (n = 49). Three prompting approaches were compared: (1) complex multi-choice prompts with 20 diagnostic options, (2) zero-shot open-ended prompts, and (3) two-step descriptive-reasoning prompts. Images were sourced from the Taiwan Society of Laboratory Medicine external quality assurance archives with expert consensus diagnoses. Results: Zero-shot prompting significantly outperformed complex multi-choice prompts across all models and domains (p < 0.001). With zero-shot prompts, Gemini achieved 78.5% overall accuracy (urinalysis: 92.0%; parasitology: 75.5%; blood smears: 64.1%), representing a 17% improvement over complex prompts. Two-step descriptive-reasoning prompts further improved blood smear accuracy by 8–12% for top-performing models, but showed minimal benefit in urinalysis and parasitology. The re-query mechanism (“please reconsider”) improved urinalysis accuracy by 7.6% but had a negligible effect on blood smears and parasitology. Conclusions: Prompting strategy critically determines MLLM diagnostic performance. Zero-shot approaches with minimal constraints consistently outperform complex multi-choice formats. The remarkable performance of general-purpose models in structured domains like urinalysis (>90% accuracy) demonstrates the considerable progress of multimodal AI. However, complex morphological tasks like blood smear interpretation require either specialized prompting techniques or domain-specific fine-tuning. These findings provide evidence-based guidance for optimizing AI integration in clinical laboratories. Full article