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Analysis of bovine tuberculosis transmission in Jalisco, Mexico through whole-genome sequencing

Abstract

Introduction

Bovine tuberculosis, caused by M. bovis, is endemic in Mexico and has had a big impact on public health. Jalisco is considered to be an important dairy region in the country, accounting for approximately 19% of the total milk production. Within Jalisco, the region of Altos Sur holds the largest proportion of the cattle inventory of the state.

Material and Methods

To determine the frequency of bovine tuberculosis in Altos Sur, Jalisco, as well as M. bovis genetic diversity, sampling of tissue (lymph nodes, lungs, and liver) from Holstein cattle was performed in four abattoirs belonging to three municipalities of this region (Tepatitlán de Morelos, San Miguel el Alto, and Arandas). Spoligotyping and whole-genome sequencing were carried out to assess the genetic relationships of M. bovis strains circulating in this area, as well as a comparison to isolates from other places in Mexico.

Results

Prevalence was 15.06%, and distribution similar among the three municipalities. The most frequent spoligotypes were SB0673, SB121, and SB0145. Whole-genome sequencing revealed three main clades (I, II, III), but isolates did not show clustering by region.

Conclusion

Phylogenetic analysis suggested ongoing transmission between herds of the different regions, and no unique source of infection was determined. This hinders efforts under the national program for the control and eradication of the disease, so serious attention must be paid to rural regions such as Altos Sur in order to improve its success.

Open access
The kinetics of γ-H2AX during radiotherapy of head and neck cancer potentially allow for prediction of severe mucositis

Abstract

Background

The aim of the study was to evaluate the changes in γ-H2AX expression in peripheral blood lymphocytes (PBL) according to severity of radiation-induced mucositis.

Patients and method

Fifty patients with head and neck cancer treated with radiotherapy (RT) or chemoradiation were included in the study. Blood samples were collected before treatment to measure baseline γ-H2AX levels. Second sample was taken 45 minutes after the first RT fraction and then once a week, 45 min after irradiation. In patients treated with chemoradiation the blood sample was taken the day after chemotherapy. Mucositis was evaluated once a week and reported according to CTCAE v4 and RTOG/EORTC scales. PBL were analyzed with flow cytometry and level of H2AX phosphorylation at every time point was evaluated.

Results

In 35 patients mild to moderate (grade 1–2) mucositis was observed and 15 patients developed severe (grade 3) mucositis. No cases of grade 4 mucositis were observed. The difference in baseline levels of γ-H2AX between groups with mild and severe mucositis was statistically insignificant (p = 0.25). The statistically significant difference in γ-H2AX level was observed in week 7 of treatment (p = 0.01). No significant differences in γ-H2AX level were found neither between group treated with concomitant chemoradiation or RT alone neither between groups with and without common comorbidities. In the analysis of the kinetics of γ-H2AX during treatment, a statistically significant difference (p = 0.0088) between groups with mild and severe mucositis was observed. After fourth week of treatment levels of γ-H2AX decreased significantly in the group with severe mucositis and increased in patients with mild side effects. The observed difference was not caused by the decrease in peripheral lymphocyte count, which was similar in both groups.

Conclusions

Presented results indicate that severity of radiation-induced mucositis does not correlate directly with γ-H2AX levels measured in vivo in PBL. Prediction of mucositis grade based on γ-H2AX level is not yet possible, either before treatment or early during treatment, but preliminary results, indicating significant differences in γ-H2AX kinetics between groups, encourage further studies.

Open access
Molecular heterogeneity in breast carcinoma cells with increased invasive capacities

Abstract

Background

Metastatic progression of breast cancer is still a challenge in clinical oncology. Therefore, an elucidation how carcinoma cells belonging to different breast cancer subtypes realize their metastatic capacities is needed. The aim of this study was to elucidate a similarity of activated molecular pathways underlying an enhancement of invasiveness of carcinoma cells belonging to different breast carcinoma subtypes.

Materials and methods

In order to reach this aim, parental and invasive (INV) MDA-MB-231 (triple-negative), T47D (hormone receptor-positive), and Au565 (Her2-positive) breast carcinoma cells were used and their molecular phenotypes were compared using a proteomic approach.

Results

Independently from breast cancer subtypes, INV cells have demonstrated fibroblast-like morphology accompanied by enhancement of invasive and migratory capacities, increased expression of cancer stem cell markers, and delayed tumor growth in in vivo animal models. However, the global proteomic analysis has highlighted that INV cells were different in protein expressions from the parental cells, and Her2-positive Au565-INV cells showed the most pronounced molecular differences compared to the triple-negative MDA-MB-231-INV and hormone receptor-positive T47D-INV cells. Although Au565-INV breast carcinoma cells possessed the highest number of deregulated proteins, they had the lowest overlapping in proteins commonly expressed in MDA-MB-231-INV and T47D-INV cells.

Conclusions

We can conclude that hormone receptor-positive cells with increased invasiveness acquire the molecular characteristics of triple-negative breast cancer cells, whereas Her2-positive INV cells specifically changed their own molecular phenotype with very limited partaking in the involved pathways found in the MDA-MB-231-INV and T47D-INV cells. Since hormone receptor-positive invasive cells share their molecular properties with triple-negative breast cancer cells, we assume that these types of metastatic disease can be treated rather equally with an option to add anti-hormonal agents. In contrast, Her2-positive metastasis should be carefully evaluated for more effective therapeutic approaches which are distinct from the triple-negative and hormone-positive metastatic breast cancers.

Open access
Percutaneous mechanical thrombectomy in patients with high-risk pulmonary embolism and contraindications for thrombolytic therapy

Abstract

Background

High-risk pulmonary embolism is associated with a high early mortality rate. We report our experience with percutaneous mechanical thrombectomy in patients with high-risk pulmonary embolism and contraindications for thrombolytic therapy.

Patients and methods

This was a retrospective analysis of consecutive patients with high-risk pulmonary embolism and contraindications to thrombolytic therapy. They were treated with percutaneous mechanical thrombectomy which included thrombectomy and additional thrombus aspiration when needed. Clinical parameters and survival to discharge were measured.

Results

From November 2005 to September 2015 we treated 25 patients with a mean age of 62.6 ± 12.7 years, 64% were men. Mean simplified Pulmonary Embolism Severity Index was 2.9. Mean maximum lactate levels were 7.8 ± 6.6 mmol/L, vasopressors were used in 77%, and 59% needed mechanical ventilation. Mechanical treatment included thrombus fragmentation complemented with aspiration (56%) and aspiration using Aspirex®S catheter (44%). Local (5 patients; 20%) and systemic (3 patients; 12%) thrombolytics were used as a salvage therapy. We observed nonsignificant improvements in systemic blood pressure (100 ± 41 mm Hg vs 119 ± 34; p = 0.100) and heart frequency (99 ± 35 min-1 vs 87 ± 31 min-1; p = 0.326) before and after treatment, respectively. Peak systolic tricuspid pressure gradient was significantly lower after treatment (57 ± 14 mm Hg vs 31 ± 3 mm Hg; p = 0.018). Overall the procedure was technically successful in 20 patients (80%) and 17 patients (68%) survived to hospital discharge.

Conclusions

In patients with high-risk pulmonary embolism who cannot receive thrombolytic therapy, percutaneous mechanical thrombectomy is a promising alternative to reduce pulmonary artery pressure.

Open access
Seroprevalence, clinical, and pathological characteristics of canine leishmaniasis in a central region of Colombia

Abstract

Introduction

Leishmaniasis is a zoonotic disease which is caused by protozoan parasites of the genus Leishmania. Canids are the most important reservoir of the parasites; however, limited data are available on the species of Leishmania prevalent in these animals and their impact on human health. The objective of this study was to estimate the seroprevalence of leishmaniasis in dogs from an inter-Andean region of Colombia during July 2016–July 2017, and to describe the clinical and histopathological features of the disease.

Material and Methods

A total of 155 dogs were subjected to clinical examination and a serological test for detection of antibodies against Leishmania. Necropsy was carried out on positive animals and tissue samples were processed by routine histopathology.

Results

Altogether 19 dogs were positive in the serological test, establishing a 12% seroprevalence of Leishmania. Clinical examination and necropsy revealed exfoliative and ulcerative dermatitis with haemorrhagic borders on the ears, head, nose, and legs. Histopathology revealed severe multifocal dermatitis with abundant Leishmania amastigotes within the cytoplasm of phagocytic cells, depletion of lymphocytes in lymphoid tissues, interstitial pneumonia, and interstitial nephritis. Tissue samples were positive for Leishmania by PCR.

Conclusions

The macro- and microscopic changes correlated with the presence of Leishmania as established by serological test and PCR.

Open access
Short-term heart-rate variability in healthy small and medium-sized dogs over a five-minute measuring period

Abstract

Introduction

Five-minute heart-rate variability (HRV) measurement is a useful tool for assessing the autonomic nervous system (ANS) balance in humans, but there are no studies on healthy dogs. The aim of the study was, therefore, to provide the reference ranges in small and medium-sized breeds for short-term HRV time and frequency domain (TFD) analyses.

Material and Methods

A total of 79 healthy dogs were included in the study between 2015 and 2019. Grouping by age with the breakpoint at six years and subgrouping by reproductive status and sex was imposed. All the dogs were included after physical and cardiological examinations and blood analyses. The TFD of HRV were analysed from a five-minute-long digital ECG recording after removal of non-sinus complexes.

Results

There were no statistically significant differences in any TFD parameters between age, reproductive status or sex groups. A mild increase in all time domain parameters and the high-frequency (HF) band was observed in older dogs, and the low frequency (LF):HF ratio decreased in these dogs. In males, the time domain parameters and HF band increased slightly.

Conclusions

The normal ranges for HRV derived from short-term ECG recording in the usual clinical environment now have proposed reference ranges. Our findings suggest that accommodation time, age, sex, or reproductive status do not influence the results derived from these recordings, indicating that this method is reliable for assessing the ANS function in small and medium-sized dog breeds.

Open access
Utility of urinary markers in the assessment of renal dysfunction in familial glomerulonephritis in Dobermann dogs

Abstract

Introduction

Dobermann dogs are reportedly predisposed to familial glomerulonephropathy. Proteinuria is a hallmark of canine familial glomerular diseases. The identification of glomerular abnormalities in breeds so predisposed is of great importance in improving breeding policy. Therefore, markers that allow the detection and localisation of renal damage are needed. The purpose of this study was to investigate the urinary concentrations of immunoglobulin G (uIgG), retinol-binding protein (uRBP), and Tamm–Horsfall protein (uTHP) in a family of Dobermanns with proteinuria and compare these concentrations with the corresponding values in healthy controls.

Material and Methods

Ten dogs of the Dobermann breed with proteinuria (five with a urine protein-to-creatinine ratio (UPC) of 0.5–1 and five with a UPC >1) and twelve healthy dogs were enrolled. An ELISA was performed to measure uIgG, uRBP, and uTHP, and these proteins were quantified in relation to urinary creatinine (uCrea).

Results

uIgG/uCr and uRBP/uCr were significantly higher in the family of Dobermanns than in the healthy dogs. A significant difference in the uTHP/uCr value was found only in dogs with a UPC of >1.

Conclusions

IgG seems to facilitate the diagnosis of primary hereditary glomerulopathy in Dobermanns. Moreover, in affected dogs, proteinuria characterisation seems to be a promising alternative option for the detection and localisation of renal lesions.

Open access
Safety margin assessment after microwave ablation of liver tumors: inter- and intrareader variability

Abstract

Background

The aim of the study was to evaluate the inter- and intrareader variability of the safety margin assessment after microwave ablation of liver tumors using post-procedure computed tomography (CT) images as well as to determine the sensitivity and specificity of identification remnant tumor tissue.

Patients and methods

A retrospective analysis of 58 patients who underwent microwave ablation (MWA) of primary or secondary liver malignancies (46 hepatocellular carcinoma, 9 metastases of a colorectal cancer and 3 metastases of pancreatic cancer) between September 2017 and June 2019 was conducted. Three readers estimated the minimal safety margin in millimeters using side-by-side comparison of the 1-day pre-ablation CT and 1-day post-ablation CT and judged whether ablation was complete or incomplete. One reader estimated the safety margin again after 6 weeks. Magnetic resonance imaging (MRI) after 6 weeks was the gold standard.

Results

The intraclass correlation coefficient (ICC) for estimation of the minimal safety margin of all three readers was 0.357 (95%-confidence interval 0.194–0.522). The ICC for repeated assessment (reader 1) was 0.774 (95%-confidence interval 0.645–0.860). Sensitivity and specificity of the detection of complete tumor ablation, defined as no remnant tumor tissue in 6 weeks follow-up MRI, were 93%/82%/82% and 33%/17%/83%, respectively.

Conclusions

In clinical practice, the safety margin after liver tumor ablation is often assessed using side-by-side comparison of CT images. In the study, we were able to show, that this technique has a poor reliability (ICC 0.357). From our point of view, this proves the necessity of new technical procedures for the assessment of the safety distance.

Open access
Laboratory Tests in Diagnosis of Mastocytosis: Literature Review and Case Report

Abstract

Mastocytosis is a heterogeneous group of disorders characterized by abnormal growth and accumulation of mast cells (MCs) in the skin and/or other organ systems. Mastocytosis is a rare disease. The annual incidence is 5–10 cases per 1 million people. However, the majority of cases stay undiagnosed due to the lack of specific tests and a wide variety of clinical features of the disease. In mastocytosis, somatic mutations of KIT gene lead to autocrine dysregulation and constitutive c-KIT activation in the absence of its ligand SCF. Clinical symptoms of the disease are determined by MC mediator release and/or infiltration of tissues by MCs. According to the World Health Organisation classification updated in 2016 mastocytosis is divided to cutaneous mastocytosis (CM), indolent systemic mastocytosis (ISM), smoldering systemic mastocytosis (SSM), SM with an associated hematologic (non-MC-lineage) neoplasm (SMAHN), aggressive SM (ASM), MC leukemia (MCL) and MC sarcoma (MCS). The CM and ISM prognosis is excellent with (almost) normal life expectancy, unlike aggressive forms (ASM and MCL) with poor prognosis. In this paper the key aspects of clinical features and diagnostic criteria of mastocytosis are discussed. We present a case report of a patient with mastocytosis in the skin following psoralen plus ultraviolet A (PUVA) therapy with good response.

Open access
Diffuse Cutaneous Mastocytosis (Pigmented Maculopapular Cutaneous Mastocytosis) in a Cat

Abstract

Diffuse cutaneous mastocytosis was diagnosed in a 6-year-old, indoor, neutered female domestic European shorthair cat. Marked pruritus located mainly on the head and neck was noticed in the cat and in this area the animal had developed alopecia, crusts, and plaques. Histologically, monomorphic mast cells were found in the superficial dermis and around the hair follicles. Mast cells were well differentiated, with central nuclei and granular cytoplasm, with metachromatic granules which stained positively with Toluidine blue stain. The animal was successfully treated with oclacitinib at a dose of 1 mg/kg, twice a day per os.

Open access