The effect of age and sex on glycated hemoglobin in dogs

We investigated the effect of age and sex on canine glycated hemoglobin (HbA1c) using a validated capillary electrophoresis assay. Aliquots of EDTA blood samples collected for routine health checks were used. HbA1c was measured using the Capillarys 2 flex-piercing system (Sebia). We included 58 clinically and hematologically healthy, normoglycemic dogs (29 males, 29 females), allocated to 3 age groups: young (14 dogs <1-y-old), adult (31 dogs 1–7.9-y-old), and senior (13 dogs ≥8-y-old). The mean (± SD) HbA1c was not significantly different ( p = 0.428) between the age groups (young: 1.68 ± 0.54%; adult: 1.59 ± 0.41%; senior: 1.80 ± 0.57%). The HbA1c was not significantly correlated with age (rho = 0.144, p = 0.280). The median (range) HbA1c was not significantly different ( p = 0.391) between male [1.7% (0.5–2.5%)] and female [1.5% (1.0–2.7%)] dogs. Age and sex do not appear to affect canine HbA1c; however, a study of geriatric dogs would be needed to fully exclude an effect of age on HbA1c.

Significance Aberrant Of CD133 Expression and Their Clinical Impact in Acute Lymphoblastic Leukemia among Sudanese Patients in Khartoum State

Background: Acute lymphoblastic leukemia (ALL) is a malignant disease that arises from several genetic mutations in a single B- or T-lymphoid precursor, resulting in altered blast cell survival, proliferation, and maturation. Objectives: This current study was aimed to determine the significance of aberrant CD133 and 45 expressions in Sudanese Patients with Acute lymphoblastic Leukemia, and their clinical significance in Khartoum state. Materials and Methods: One hundred Sudanese participants were enrolled in this study as follows: 88 Patients diagnosed with ALL as a case and 12 healthy controls donors were randomly selected. 2.5-5 mL of EDTA blood/bone marrow for flowcytometry from each patient and controls subject. Results: Eighty-eight newly diagnosed Sudanese patients with acute lymphocytic leukemia (ALL) were involved in this study, their age average is (15.7) and their stander deviation (SD) is 17.4. The frequency of aberrant markers concerning control groups was significantly associated with all patients in CD 45 with a P value (0.0001), while there was no difference between cases and control in the frequency of CD 133 antigen, there was no significant difference between age groups in the expression of different aberrant antigens. The study was significantly more expressed CD133 in males than females. Also no significant difference in hematological parameters between patients with or without expression of Aberrant CD 45 and 133 markers, while we found significantly high TWBCs in T. cell leukemia. Conclusion: Aberrant myeloid antigens CD45 was significantly associated with childhood and adult ALL and may be considered as important prognostic factors, while CD 133 were not associated with ALL.

Comparison of the composition of lymphocyte subpopulations in non-relapse and relapse patients with squamous cell carcinoma of the head and neck before, during radiochemotherapy and in the follow-up period: a multicenter prospective study of the German Cancer Consortium Radiation Oncology Group (DKTK-ROG)

Background Radiochemotherapy (RCT) has been shown to induce changes in immune cell homeostasis which might affect antitumor immune responses. In the present study, we aimed to compare the composition and kinetics of major lymphocyte subsets in the periphery of patients with non-locoregional recurrent (n = 23) and locoregional recurrent (n = 9) squamous cell carcinoma of the head and neck (SCCHN) upon primary RCT. Methods EDTA-blood of non-locoregional recurrent SCCHN patients was collected before (t0), after application of 20–30 Gy (t1), in the follow-up period 3 (t2) and 6 months (t3) after RCT. In patients with locoregional recurrence blood samples were taken at t0, t1, t2 and at the time of recurrence (t5). EDTA-blood of age-related, healthy volunteers (n = 22) served as a control (Ctrl). Major lymphocyte subpopulations were phenotyped by multiparameter flow cytometry. Results Patients with non-recurrent SCCHN had significantly lower proportions of CD19+ B cells compared to healthy individuals before start of any therapy (t0) that dropped further until 3 months after RCT (t2), but reached initial levels 6 months after RCT (t3). The proportion of CD3+ T and CD3+/CD4+ T helper cells continuously decreased between t0 and t3, whereas that of CD8+ cytotoxic T cells and CD3+/CD56+ NK-like T cells (NKT) gradually increased in the same period of time in non-recurrent patients. The percentage of CD4+/CD25+/FoxP3+ regulatory T cells (Tregs) decreased directly after RCT, but increased above initial levels in the follow-up period 3 (t2) and 6 (t3) months after RCT. Patients with locoregional recurrence showed similar trends with respect to B, T cells and Tregs between t0 and t5. CD4+ T helper cells remained stably low between t0 and t5 in patients with locoregional recurrence compared to Ctrl. NKT/NK cell subsets (CD56+/CD69+, CD3−/CD56+, CD3−/CD94+, CD3−/NKG2D+, CD3−/NKp30+, CD3−/NKp46+) increased continuously up to 6 months after RCT (t0-t3) in patients without locoregional recurrence, whereas in patients with locoregional recurrence, these subsets remained stably low until time of recurrence (t5). Conclusion Monitoring the kinetics of lymphocyte subpopulations especially activatory NK cells before and after RCT might provide a clue with respect to the development of an early locoregional recurrence in patients with SCCHN. However, studies with larger patient cohorts are needed. Trial registration Observational Study on Biomarkers in Head and Neck Cancer (HNprädBio), NCT02059668. Registered on 11 February 2014, https://clinicaltrials.gov/ct2/show/NCT02059668.

Colorectal Cancer Study of Austria (CORSA): A Population-Based Multicenter Study

The COloRectal cancer Study of Austria (CORSA) is comprised more than 13,500 newly diagnosed colorectal cancer (CRC) patients, patients with high- and low-risk adenomas as well as population-based controls. The recruitment for the CORSA biobank is performed in close cooperation with the invited two-stage CRC screening project “Burgenland PREvention trial of colorectal Disease with ImmunologiCal Testing” (B-PREDICT). Annually, more than 150,000 inhabitants of the Austrian federal state Burgenland aged between 40 and 80 are invited to participate using FIT-tests as an initial screening. FIT-positive tested participants are offered a diagnostic colonoscopy and are asked to take part in CORSA, sign a written informed consent, complete questionnaires concerning dietary and lifestyle habits and provide an ethylenediaminetetraacetic acid (EDTA) blood sample as well as a stool sample. Additional CRC cases have been recruited at four hospitals in Vienna and a hospital in lower Austria. A major strength of CORSA is the population-based controls who are FIT-positive and colonoscopy-confirmed to be free of polyps and/or CRC.

A Low Aldosterone/Renin Ratio and High Soluble ACE2 Associate with COVID-19 Severity

Background The severity of COVID-19 after SARS-CoV-2 infection is unpredictable. Angiotensin-converting enzyme-2 (ACE2) is the receptor responsible for coronavirus binding, while subsequent cell entry relies on priming by the serine protease TMPRSS2 (transmembrane protease, serine 2). Although renin-angiotensin-aldosterone-system (RAAS) blockers have been suggested to upregulate ACE2, their use in COVID-19 patients is now considered safe. The aim of our study was to investigate parameters that determine COVID-19 severity, focusing on RAAS-components and variation in the genes encoding for ACE2 and TMPRSS2.MethodsAdult patients hospitalized due to SARS-CoV-2 infection between May 2020 and October 2020 in the Haga Teaching Hospital were included, and soluble ACE2 (sACE2), renin, aldosterone (in heparin plasma), and polymorphisms in the ACE2 and TMPRSS2 genes (in DNA obtained from EDTA blood) were determined. Measurements and main resultsOf the 188 patients that were included, 60 were defined as severe COVID-19 (ICU and/or death). These patients more often used antidiabetic drugs, were older, had higher renin and sACE2 levels, lower aldosterone levels, and a lower aldosterone/renin ratio. In addition, they displayed the TMPRSS2-rs2070788 AA genotype less frequently. No ACE2 polymorphism-related differences were observed. Multivariate regression analysis revealed independent significance for age, sACE2, the aldosterone/renin ratio, and the TMPRSS2 rs2070788 non-AA genotype as predictors of COVID-19 severity, together yielding a C-index of 0.79. Findings were independent of the use of RAAS blockers. Conclusion High sACE2, a low aldosterone/renin ration, and having the TMPRSS2 rs2070788 non-AA genotype are novel independent determinants that may help to predict COVID-19 disease severity. Trial registration: retrospectively registered

A Comparative Study of Erythrocyte Sedimentation Rate using Saline Diluted and Undiluted EDTA with TSC

Erythrocyte sedimentation rate (ESR) is the rate at which RBC sediment in a period of 1 hr. It is a common Haematology test that is a non-specific measure of inflammation and it became a common screening test worldwide for acute phase proteins and chronic diseases. The International Council for Standardization in Haematology (ICSH) recommended the Westergren method as the method of choice for ESR determination. TSC is almost exclusively used as the diluent of choice for setting up ESR, but some contemporary laboratories have resolved to use Normal saline (NS) as the diluent of choice while other set ESR using EDTA anticoagulated Blood (BLD) without any diluent. The objective of this study is to assess the comparison between saline diluted and undiluted EDTA with TSC as an anticoagulant in ESR detection and to find out any gender wise variations by using these anticoagulants. A total of 50 students were participated in this study. From each of the participants 5 ml of BLD was collected and it is then divided into 3 parts. 1.6 ml BLD to 0.4ml 3.8% TSC tube, 1.6 ml EDTA blood to 0.4 ml NS tube. 3rd tube with 2 ml EDTA BLD and set for ESR and obtained result within 1 hour. The Result is the mean +SD value of ESR were 19.48+5.7 mm/hr. in undiluted EDTA, 15.22+4.6 mm/hr. in saline diluted EDTA & 15.36+4.5 mm/hr. in TSC. The mean difference of ESR value between saline diluted EDTA with TSC BLD was 0 and it with undiluted EDTA was 4 mm /hr. The study indicates that there was a significant difference between ESR value with undiluted EDTA and TSC while diluted EDTA and TSC were there is no significant difference. The mean +SD of ESR value using undiluted, diluted EDTA and TSC in males were 16.20+3.3, 11.05+2.8, 11.29+2.6 and while it for females were 21.69+5.2, 17.36+3.8, 17.45+3.8 respectively. In conclusion, TSC is the best diluent to be used in contemporary lab to set ESR as compared to EDTA BLD. But we can use saline diluted EDTA as an alternative to citrate diluted BLD to set ESR and it also showed there is a gender wise variation in ESR using these anticoagulants.

Is Encephalitozoon cuniculi of Significance in Young Dogs With Neurological Signs?

Encephalitozoon cuniculi is a microsporidium belonging to the phylum Microspora. A few reports have described the clinical significance of E. cuniculi infection in young dogs. In American and Japanese household dog populations, the seroprevalence was found to be 21%, indicating its wide-spread existence. To evaluate the clinical significance of E. cuniculi in a cohort of young dogs with neurological signs, the presence of the organism and possible response to treatment were studied. Over a 1-year period, all young dogs (&lt;3 years old) that were referred with neurological signs were examined for the presence of E. cuniculi. Dogs were selected if serology revealed a clearly elevated IgM titer (&gt;100) and/or if an EDTA-blood sample and/or urine sample tested positive by polymerase chain reaction (PCR). Sixteen dogs with various neurological signs were included in this study. Additional work-up included magnetic resonance imaging and cerebrospinal fluid analysis, but these revealed no abnormalities or indication of infection. All dogs were treated with fenbendazole for 10–30 days. Neurological signs disappeared completely in five dogs, 11 dogs continued to show neurological signs, and five dogs deteriorated and were euthanized, after which necropsy was performed in three. At necropsy no evidence of an E. cuniculi infection was found. We concluded that, although IgM titers and PCR indicated an E. cuniculi infection, it is most likely of limited clinical significance in young dogs.

Inter-laboratory comparison of gene expression biodosimetry for protracted radiation exposures as part of the RENEB and EURADOS WG10 2019 exercise

Large-scale radiation emergency scenarios involving protracted low dose rate radiation exposure (e.g. a hidden radioactive source in a train) necessitate the development of high throughput methods for providing rapid individual dose estimates. During the RENEB (Running the European Network of Biodosimetry) 2019 exercise, four EDTA-blood samples were exposed to an Iridium-192 source (1.36 TBq, Tech-Ops 880 Sentinal) at varying distances and geometries. This resulted in protracted doses ranging between 0.2 and 2.4 Gy using dose rates of 1.5–40 mGy/min and exposure times of 1 or 2.5 h. Blood samples were exposed in thermo bottles that maintained temperatures between 39 and 27.7 °C. After exposure, EDTA-blood samples were transferred into PAXGene tubes to preserve RNA. RNA was isolated in one laboratory and aliquots of four blinded RNA were sent to another five teams for dose estimation based on gene expression changes. Using an X-ray machine, samples for two calibration curves (first: constant dose rate of 8.3 mGy/min and 0.5–8 h varying exposure times; second: varying dose rates of 0.5–8.3 mGy/min and 4 h exposure time) were generated for distribution. Assays were run in each laboratory according to locally established protocols using either a microarray platform (one team) or quantitative real-time PCR (qRT-PCR, five teams). The qRT-PCR measurements were highly reproducible with coefficient of variation below 15% in ≥ 75% of measurements resulting in reported dose estimates ranging between 0 and 0.5 Gy in all samples and in all laboratories. Up to twofold reductions in RNA copy numbers per degree Celsius relative to 37 °C were observed. However, when irradiating independent samples equivalent to the blinded samples but increasing the combined exposure and incubation time to 4 h at 37 °C, expected gene expression changes corresponding to the absorbed doses were observed. Clearly, time and an optimal temperature of 37 °C must be allowed for the biological response to manifest as gene expression changes prior to running the gene expression assay. In conclusion, dose reconstructions based on gene expression measurements are highly reproducible across different techniques, protocols and laboratories. Even a radiation dose of 0.25 Gy protracted over 4 h (1 mGy/min) can be identified. These results demonstrate the importance of the incubation conditions and time span between radiation exposure and measurements of gene expression changes when using this method in a field exercise or real emergency situation.

Putative Novel Atypical BTV Serotype ‘36’ Identified in Small Ruminants in Switzerland

We identified a putative novel atypical BTV serotype ‘36’ in Swiss goat flocks. In the initial flock clinical signs consisting of multifocal purulent dermatitis, facial oedema and fever were observed. Following BTV detection by RT-qPCR, serotyping identified BTV-25 and also a putative novel BTV serotype in several of the affected goats. We successfully propagated the so-called “BTV-36-CH2019” strain in cell culture, developed a specific RT-qPCR targeting Segment 2, and generated the full genome by high-throughput sequencing. Furthermore, we experimentally infected goats with BTV-36-CH2019. Regularly, EDTA blood, serum and diverse swab samples were collected. Throughout the experiment, neither fever nor clinical disease was observed in any of the inoculated goats. Four goats developed BTV viremia, whereas one inoculated goat and the two contact animals remained negative. No viral RNA was detected in the swab samples collected from nose, mouth, eye, and rectum, and thus the experimental infection of goats using this novel BTV serotype delivered no indications for any clinical symptoms or vector-free virus transmission pathways. The subclinical infection of the four goats is in accordance with the reports for other atypical BTVs. However, the clinical signs of the initial goat flock did most likely not result from infection with the novel BTV-36-CH0219.

Detection of circulating microfilariae in canine EDTA blood using lens-free technology: preliminary results

Dirofilaria immitis causes life-threatening heart disease in dogs, thus screening of dog populations is important. Lens-free technology (LFT) is a low-cost imaging technique based on light diffraction that allows computerized recognition of small objects in holographic images. We evaluated an algorithm capable of recognizing microfilariae in canine whole blood using the LFT. We examined 3 groups of 10 EDTA blood specimens, from dogs with microfilaremia (group A), healthy dogs (B), and dogs with hematologic modifications other than microfilaremia (C). The LFT analyzer photographed repeated series of 5 images of all samples. The algorithm declared a sample positive if a microfilaria was detected on ≥1, ≥2, or ≥3 of the 5 images of a series. Microfilariae were detected visually in the images in 9 of 10 cases in group A; no microfilariae were seen in the images from groups B and C. Of the 30 cases, there were 14, 4, and only 3 false-positives with the 1 of 5, 2 of 5, and 3 of 5 image cutoffs, respectively. There were no false-negatives, regardless of cutoff. LFT seems useful for detecting microfilaria and could have application in clinical pathology.