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The present study was conducted to investigate the internal and external biosecurity levels and their association with Mycoplasma gallisepticum infection in broiler farms in Khartoum State, Sudan. Sixty-six broiler farms were selected. A questionnaire of 77 questions on internal and external biosecurity measures was administered during each farm visit. A total of 360 blood samples were collected and used for serological examination to detect antibodies against M. gallisepticum. The overall score of biosecurity was 0.57, and the average score of internal biosecurity (0.75) was higher (p<0.01) than the average score of external biosecurity (0.51). Poor internal biosecurity practices were observed in 23.4% of broiler farms, whereas most farms (93.6%) recorded poor external biosecurity. A significant (p<0.01) positive correlation was encountered between the external biosecurity and the number of houses (0.62), as well as the number of workers (rs= 0.54). Both internal and external biosecurity displayed significant positive correlations with the number of birds (rs= 0.64, 0.35; p< 0.05). Fifty percent of the farms were positive for M. gallisepticum. The disease incidence correlated positively with the spread of manure on neighboring farms (rs= 0.44, p<0.05). A negative correlation was observed between the presence of Mycoplasma and the distance between farms (rs= -0.50, p<0.01) as well as farm enclosure (rs= -0.44, p<0.05). It is concluded that broiler farms in Khartoum State exhibited low biosecurity measures and a high seroprevalence of M. gallisepticum infection. Further efforts are necessary to improve external biosecurity practices. [ABSTRACT FROM AUTHOR]

Background: Toxoplasmosis is a common disease caused by an apicomplexian protozoa, Toxoplasma gondii; one of the potential sources of human zoonotic infection is meat handling and consumption. Objectives: This study aims to determine the prevalence of T. gondii in the blood of butchers and slaughtered cattle by using the B1 gene and demonstrate the role of slaughtered cattle in transmitting toxoplasmosis to humans. Methods: The study involved 100 blood samples collected randomly from butchers (male) and 100 from cattle before slaughtering from September 2022 to September 2023 and examined by real-time PCR technique. Results: The results revealed total infection rates of 28% and 12% in butchers and cattle, respectively, with the highest rate recorded in butchers aged 18-30 years old and the lowest at age >40 years. While in cattle, the highest rate is seen at age (2-4 years) and the lowest at age (1-2 years) without significant differences. Calves reported a higher infection rate (14.28%) compared with cows (8.10%), with a significant difference (P>0.05). Conclusion: Cattle are an important source of zoonotic transmission of T. gondii infection to humans by handling meat and consuming raw or uncooked meat. We recommended a control program by awareness of butchers during meat handling, avoiding consuming raw or uncooked meat, and appropriate heat treatments for beef. [ABSTRACT FROM AUTHOR]

Introduction: Night shift work disrupts circadian rhythms and has been associated with immune system alterations and various health conditions. However, there is limited data regarding its impact on psoriasis. The aim of our study was to compare psoriasis severity and the hormonal and immunological profile in patients with a night shift work to those with a daytime occupation. Methods: In this case-control study, we enrolled psoriatic patients aged >18 years engaged in night shift work and a control group of psoriatic patients with a daytime occupation. A further categorization was performed by the duration of night shift work: < or ≥7 days a month and < or ≥8 years. Disease severity was evaluated by PASI, BSA, and DLQI, and blood samples were taken to measure various hormonal and immunological markers. Univariable and multivariable analysis were performed to assess differences between the two groups. Results: A total of 40 night shift workers were included, along with 36 patients in the control group. Patients who worked night shifts at least 7 days a month had significantly higher PASI scores (11.2 ± 6.6 vs. 8.5 ± 6.6; p = 0.04) and higher IL-8 serum (115.33 ± 463.65 pg/mL vs. 19.98 ± 29.78 pg/mL; p = 0.006) compared to patients who did not. Night shifts workers for at least 8 years had higher BMI (28.65 ± 4.56 vs. 25.32 ± 5.50, p = 0.010), and females had higher testosterone levels (0.46 ± 0.53 ng/mL vs. 0.23 ± 0.13 ng/mL; p = 0.055). Conclusion: Night shift might increase psoriasis severity and have an impact on chronic inflammation, obesity, and hormonal imbalances. [ABSTRACT FROM AUTHOR]

BNP is a sensitive and widely used biomarker for an early diagnosis of heart failure. Currently, most commercial BNP detection products use EDTA plasma samples. The aim of this study was to evaluate the clinical performance of the BNP test by using whole blood samples compared to plasma samples, and to evaluate the effect of the anticoagulant type on the BNP test result. Methods: In total, 106 patients with different BNP levels from the Dahua Hospital volunteered for this study. Clinically homogenous samples, including EDTA anticoagulant plasma, EDTA whole blood, and heparin anticoagulant plasma, were collected and analyzed by using i-Reader S automatic immuno-analyzer and its supporting reagent kits. Pearson’s correlation and weighted least squares linear regression analysis, Bland-Altman plotting, and Kappa test were used for statistical analysis. Results: Correlation analysis showed that BNP concentrations, measured from EDTA anticoagulated plasma samples, had a good linear regression relationship with BNP from whole blood samples, with a slope of 0.9477, r = 0.9978, p < 0.05. A similar correlation was observed between EDTA anticoagulated plasma samples and heparin anticoagulant plasma, with a slope of 0.8413, r = 0.9793, p < 0.05. The BNP concentration measured from the heparin plasma samples were lower than of the EDTA plasma samples. Bland-Altman analysis for assessing BNP concentration agreement showed there was no outlier ratio between EDTA whole blood and EDTA plasma within the range of the detection system, as well as no outlier between EDTA anticoagulated and heparin anticoagulant plasma. Kappa coefficient of BNP concentration between homologous EDTA anticoagulated and heparin anticoagulant plasma was 0.8553 (p < 0.001), and for EDTA anticoagulated plasma and homologous whole blood it was 0.8941 (p < 0.001). Conclusions: The diagnostic performance of EDTA anticoagulated whole blood samples did not differ significantly from EDTA anticoagulated plasma samples for the BNP test. This study showed no big significant difference between EDTA anticoagulated and heparin anticoagulated plasma measurements within 2 hours. The type of anticoagulant should be carefully chosen when performing the BNP test if BNP samples were in vitro for a long time. [ABSTRACT FROM AUTHOR]

Heavy metals have been observed as competitor for receptor sites in hormones thereby resulting in infertility problems. Herbal cosmetics have been observed to contain heavy metals which interfere with the receptor sites of hormones. Three herbal cosmetics sold in Port-Harcourt were evaluated for heavy metals and rabbits exposed to them for 30 -90days. The aim is to determine the effect of heavy metals on the sex hormones of the rabbits. Three herbal oil namely All things natural (Emi herbal oil), Kakiva (Kakiva herbal oil) and Amal (botanical herbal oil) as well as forty -eight rabbits were purchase from a market in and animal house in Port Harcourt respectively. The rabbits were exposed to the oil for 30, 60 and 90 days and scarified for blood samples. The sex hormones of the rabbits evaluated using standard methods. Data were analyzed using statistical package for social sciences (SPSS) to compare means. There were no significant differences in the sex hormones in 30days and 60days. However, there was a significantly lowered progesterone and estrogen after exposure 90days. It is therefore pertinent to conclude that herbal cosmetics contain heavy metals which may alter the sex hormones pattern which leads to luteal phase dysfunction. [ABSTRACT FROM AUTHOR]

Background: The clinically significant Factor V Leiden (FVL) point mutation (1691 G/A) causes replacement of Arg with Gln (glutamine), preventing activated protein C from inactivating Factor V leading to a lengthened clotting process. Individuals with the Factor V Leiden mutations have an increased risk for venous thrombosis. The aim of this study is to compare an unlabeled probe high-resolution melting analysis (HRMA) assay for Factor V Leiden mutation to a TaqMan hydrolysis assay (fluorogenic 5' nuclease PCR hydrolysis assay). HRMA is a post-PCR, homogenous, closed-tube system for the detection of sequence variants. Post-PCR, the amplicons are heated gradually until the melting temperature is reached and the fluorescent dye unbinds from the amplicon and exhibits low fluorescence. A melt-curve analysis is generated that is characteristic of a particular sequence variant. Therefore, HRMA allows for comparison of one base changes in genetic sequences based on their differences in melting rate. Methods: Blood samples were collected in EDTA tubes and DNA extracted using the Roche MagNaPure. Reactions of both HRMA and TaqMan were carried out on 3 controls (1691 G/G, 1691 G/A, and 1691 G/G and G/A) and 20 samples. Results: The genotypes for 3 reference controls purchased from Coriell (F5 1691 G/G, FVL 1691 G/A, and Heterozygote 1691 G/G and G/A) were confirmed by both the HRMA and TaqMan FVL assays. All 20 samples were confirmed to be F5 1691 G/G by both HRMA and TaqMan assays. Conclusions: Comparing the results of the unlabeled probe HRMA FVL assay with a real-time TaqMan probe end point genotyping assay resulted in 100% sensitivity and 100% specificity for both assays. [ABSTRACT FROM AUTHOR]

This research paper demonstrates a metamaterial (MTM) based sensing technique to detect various blood samples by analyzing their dielectric properties. The performance of this MTM-based sensor is evaluated with the help of mimicked human blood samples that closely resemble the dielectric properties of actual human blood samples. Moreover, the ISM band frequency of 2.4 GHz is chosen as one of the reference resonance frequencies due to its various industrial and medical applications. The resonating patch is developed on the FR-4 substrate with a dimension of 10 × 20 mm2 that provides sharp reference resonances of 2.4 and 4.72 GHz for the spectra of the transmission coefficient with a good quality factor (Q-factor). The MTM sensor can detect the mimicked blood samples with a maximum frequency deviation of up to 650 MHz at 2.4 GHz and up to 850 MHz at 4.72 GHz, with maximum sensitivity of 0.917 and 0.707, respectively. The measured results using the prototype of the sensor support the simulation result with good agreement, indicating high sensing capability. Due to its high sensitivity, figure of merit (FoM), and frequency shifting with dielectric property changes in blood samples, the developed MTM-based sensor can be implemented effectively for quick sensing of infected blood samples and biomedical applications. [ABSTRACT FROM AUTHOR]

Blood doping remains a significant problem that threatens the integrity of sport. The current indirect method for detecting blood doping involves the longitudinal measurement of an athlete's haematological variables and identification of suspicious results that are indicative of doping (i.e., the athlete biological passport). While this has played a significant role in the protection of clean sport, improvements are needed. The development of a transcriptomic test, that can be used to detect the use of blood doping has been discussed and researched for decades and yet, an anti-doping test that can be adopted by the World Anti-Doping Agency (WADA) is yet to be established. With recent advancements in our understanding, as well as in methods of sample collection, the possibility of a transcriptomic test that can be used by WADA, is imminent. There are, however, several practical considerations that must first be made, that will be highlighted in this perspective article. [ABSTRACT FROM AUTHOR]

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Yield stress, τy, is a key rheological property of complex materials such as gels, dense suspensions, and dense emulsions. While there is a range of established techniques to measure τy in the order of tens to thousands of pascals, the measurement of low τy, specifically below 1 Pa, remains underexplored. In this article, we present the measurement of low apparent τy using a Hele-Shaw microfluidic extensional flow device (MEFD). Using the MEFD, we observe a gradient in shear stress, τ, such that τ is lower near the center or stagnation point, and higher away from the stagnation point. For a yield stress fluid, we observe that, below a certain flow rate, τ exceeds τy only in the outer region, leading to stagnation or unyielding of the fluid in the inner region. We use scaling analysis based on a Hele-Shaw linear extensional flow to deduce τy by measuring the size of the unyielded region, S. We validate this scaling relationship using Carbopol solutions with concentrations ranging between 0.015 to 0.3%, measuring τy as low as ∼10 mPa to ∼1 Pa, and comparing it with τy measured using a standard rheometer. While the experimental lower limit of our technique is 5 mPa, modifying the geometry or improving the image analysis can reduce this limit to the order of 10-4 Pa. The MEFD facilitates rapid measurement of τy, allowing for its real-time assessment. We further report τy of human blood samples between 30 to 80 mPa with their hematocrit ranging between 14 to 63%. Additionally, we determine τy for a mucus simulant (∼0.7 Pa), and lactic drink (∼7 mPa) to demonstrate the versatility of the MEFD technique. [ABSTRACT FROM AUTHOR]

Background: Effective testing for malaria, including the detection of infections at very low densities, is vital for the successful elimination of the disease. Unfortunately, existing methods are either inexpensive but poorly sensitive or sensitive but costly. Recent studies have shown that mid-infrared spectroscopy coupled with machine learning (MIRs-ML) has potential for rapidly detecting malaria infections but requires further evaluation on diverse samples representative of natural infections in endemic areas. The aim of this study was, therefore, to demonstrate a simple AI-powered, reagent-free, and user-friendly approach that uses mid-infrared spectra from dried blood spots to accurately detect malaria infections across varying parasite densities and anaemic conditions. Methods: Plasmodium falciparum strains NF54 and FCR3 were cultured and mixed with blood from 70 malaria-free individuals to create various malaria parasitaemia and anaemic conditions. Blood dilutions produced three haematocrit ratios (50%, 25%, 12.5%) and five parasitaemia levels (6%, 0.1%, 0.002%, 0.00003%, 0%). Dried blood spots were prepared on Whatman™ filter papers and scanned using attenuated total reflection-Fourier Transform Infrared (ATR-FTIR) for machine-learning analysis. Three classifiers were trained on an 80%/20% split of 4655 spectra: (I) high contrast (6% parasitaemia vs. negative), (II) low contrast (0.00003% vs. negative) and (III) all concentrations (all positive levels vs. negative). The classifiers were validated with unseen datasets to detect malaria at various parasitaemia levels and anaemic conditions. Additionally, these classifiers were tested on samples from a population survey in malaria-endemic villages of southeastern Tanzania. Results: The AI classifiers attained over 90% accuracy in detecting malaria infections as low as one parasite per microlitre of blood, a sensitivity unattainable by conventional RDTs and microscopy. These laboratory-developed classifiers seamlessly transitioned to field applicability, achieving over 80% accuracy in predicting natural P. falciparum infections in blood samples collected during the field survey. Crucially, the performance remained unaffected by various levels of anaemia, a common complication in malaria patients. Conclusion: These findings suggest that the AI-driven mid-infrared spectroscopy approach holds promise as a simplified, sensitive and cost-effective method for malaria screening, consistently performing well despite variations in parasite densities and anaemic conditions. The technique simply involves scanning dried blood spots with a desktop mid-infrared scanner and analysing the spectra using pre-trained AI classifiers, making it readily adaptable to field conditions in low-resource settings. In this study, the approach was successfully adapted to field use, effectively predicting natural malaria infections in blood samples from a population-level survey in Tanzania. With additional field trials and validation, this technique could significantly enhance malaria surveillance and contribute to accelerating malaria elimination efforts. [ABSTRACT FROM AUTHOR]

Simple Summary: The metabolic profile implies simultaneous determination of carbohydrate, fat, protein, and mineral metabolism parameters as well as endocrinological parameters in the blood of cows. Blood is exposed to a variety of preanalytical factors during sampling, transport to the laboratory, and laboratory preparation for analysis, which may cause hemolysis of the sample. As hemolysis affects the values of the metabolic profile, the analyzed blood parameters may falsely increase or decrease, and the metabolic status of the cows may be misinterpreted. Preventing hemolysis is important because severe hemolysis requires discarding the sample and resampling, which is very resource-intensive. In this paper, three levels of hemolysis were determined for each blood parameter tested: (a) a hemolysis level that does not affect the values of the parameters and allows the results to be issued without restriction; (b) a hemolysis level that affects the values of the parameters but remains within the acceptable biological variability and permits the results to be issued along with a note in the form of a correction formula; and (c) a hemolysis level at which the obtained values of the parameters or the entire sample must be discarded. The results are presented graphically using interferograms, which can be easily implemented in every laboratory after validation. Hemolysis is a common cause of errors in laboratory tests as it affects blood parameters and leads to a positive or negative bias. This study aims to examine the relationship between the level of hemolysis (expressed as cell-free hemoglobin concentration, g/L) and the variability of metabolic and endocrine parameters and to determine the threshold level of hemolysis that causes an analytically and clinically significant bias for the twenty most frequently examined blood parameters in cows. Paired blood samples of 10 mL each were obtained from 30 cows. One was subjected to mechanical trauma and plasma was extracted directly from the other. Hemolyzed and non-hemolyzed samples from the same animal were mixed to obtain final samples with cell-free hemoglobin concentrations of 0, 1, 2, 4, 6, 8, and 10 g/L. Metabolic and endocrine parameters were measured in the samples and their deviation and the linear equation between the level of hemolysis and the deviation were determined. The following threshold values of hemolysis were determined, which correspond to the acceptable analytical (lower value) and clinical (upper value) levels of parameter variability: BHB 0.96 and 4.81; NEFA 0.39 and 3.31; GLU 0.38 and 3.90; ALB 1.12 and 6.11; TPROT 1.40 and 6.80; UREA 6.62 and 20.1; TBIL 0.75 and 5.65; AST 0.11 and 2.18; GGT 1.71 and 8.90, LDH 0.01 and 0.11, ALP 0.97 and 2.95; TGC 1.56 and 15.5; CHOL 1.29 and 8.56; Ca 5.68 and 25.7; P 0.57 and 8.43; Mg 1.10 and 8.47; INS 1.15 and 3.89; T3 8.19 and 15.6; T4 8.97 and 18.5; and CORT 2.78 and 11.22 g/L cell-free hemoglobin. Three decision levels are available for each metabolic and endocrine parameter: if hemolysis is below the lower (analytical) threshold value, results can be reported without restriction; if hemolysis is between the lower and upper thresholds, the results can be issued with guidance in the form of corrective linear equations; and if hemolysis is above the upper (clinical) threshold, the results and sample must be discarded. This method contributes to an optimal approach to hemolysis interference with metabolic profile parameters in blood samples from cows. [ABSTRACT FROM AUTHOR]

A pre-transfusion crossmatch test is crucial for ensuring safe blood transfusions by identifying the compatibility between donor and recipient blood samples. Conventional tube methods for crossmatching have limitations, including subjectivity in result interpretation and the potential for human error. In this study, we evaluated the diagnostic performance of a new crossmatch test using Microscanner C3, which can overcome these shortcomings. The crossmatch test results using the method were obtained in 323 clinical samples. The sensitivity, specificity, positive predictive value, negative predictive value, and concordance rate of the crossmatch test using Microscanner C3 were 98.20%, 100.00%, 100.00%, 98.11%, and 99.07%, respectively. The diagnostic performance of the new system offers a promising alternative to conventional tube methods for pre-transfusion crossmatch testing. Microscanner C3 could also increase the automation, standardization, and accuracy of crossmatch tests. The crossmatch test using Microscanner C3 is thought to increase the efficiency and reliability in identifying blood samples suitable for transfusion, thereby improving patient safety and optimizing the use of blood products in clinical settings. [ABSTRACT FROM AUTHOR]

Background. Bovine trypanosomosis produces significant economic losses due to anemia, loss of body condition, and emaciation. The disease is transmitted biologically by tsetse flies and mechanically by biting flies such as Tabanus and Stomoxys. Therefore, this research is aimed at estimating the prevalence of bovine trypanosomosis and the apparent density of its vectors in the Etang Special District. Methods. A cross‐sectional study was employed from November 2016 to April 2017 for the prevalence and entomological survey. Trypanosoma species were identified using buffy coat and Giemsa staining techniques. Besides, the entomological surveys were conducted using NGU, pyramidal, biconical, and monoconical traps. The vectors were identified to their genus level based on their morphological features like size, color, wing venation, and proboscis. Result. A total of 457 bovine blood samples were collected and tested, of these 16 (3.50%) animals were positive for trypanosomosis. Similarly, 13 T. vivax (81.25%) and 3 T. congolense (18.75%) were the trypanosome species detected. The prevalence in the young (2.56%) and adult (3.99%) age groups was not statistically significant (P > 0.05). However, there were significant (P < 0.05) differences in prevalence between body condition scores, where poor is 6.31%, medium is 1.30%, and good is 0%. Moreover, the difference in mean PCV values between the parasitemic (20.97) and aparasitemic (28.58) groups was statistically significant (P < 0.05). Glossina flies were not found, although 1756 Tabanus and 52 Stomoxys biting flies were gathered. Accordingly, the overall apparent density of Glossina flies was zero (0), with biting flies (20.54) recorded per trap per day. Moreover, the apparent density of Tabanus and 52 Stomoxys was 39.01 and 1.18, respectively. Conclusion. This study confirmed that trypanosomosis and abundant mechanical vectors continue to be problems in the study area, resulting in cattle productivity losses. As a result, strategic management and prevention methods for trypanosomosis and associated vectors should be prioritized. Further investigation of vector needs to be conducted to clear out tsetse presence. [ABSTRACT FROM AUTHOR]

Hybridisation between Australian dingoes and domestic dogs is a controversial area of interest and research. An ongoing canine sterilisation programme in rural and remote Western Australia provided an opportunity to assess the dingo ancestry of camp dogs and opportunities for hybridisation. Blood samples were collected from 345 individual community dogs at 21 locations. Dogs were screened using 23 microsatellite loci and ancestry percentage assigned using an iterative Bayesian assignment algorithm. A single individual was a dingo, 96% were domestic dogs and 3.5% were hybrids. Camp dog and dingo hybridisation in these areas is of little concern in terms of conserving dingo purity. Hybridisation of dingoes occurs as domestic dogs and dingoes are the same species, Canis familiaris. An assessment of camp dogs or dogs that reside in rural and remote communities found very little dingo ancestry. Based on these samples camp dogs are not of high concern when it comes to conserving dingo purity in Western Australia. Image by Dr Nicole Laing. [ABSTRACT FROM AUTHOR]

In the current study, we examined the association between eveningness and testosterone levels in men. Specifically, we differentiated between free and total testosterone fractions, with free testosterone being recognized as the most bioavailable form of this hormone. We collected blood samples from 298 men aged 18–44 to assess total and free testosterone. Additionally, we measured sleep timing variables using the Munich Chronotype Questionnaire. The main result of the current study indicated that evening chronotype was associated with higher levels of free testosterone, but was unrelated to total testosterone. Sleep loss was unrelated to the both testosterone fractions. We expanded prior findings by utilizing a more comprehensive testosterone assay what indicated that evening chronotype is primarily associated with the most bioavailable form of testosterone (i.e. free testosterone) in adult men. [ABSTRACT FROM AUTHOR]

Introduction: The aim of this study is to assess the outcomes of parathyroid gland reimplantation with PR-FaST technique in patients undergoing thyroid surgery, focusing on graft functionality over a 5-year follow-up period. Materials and Methods: We analyzed data from 131 patients who underwent parathyroid reimplantation using the PR-FaST technique during thyroid surgery due to inadvertent parathyroid removal or evident vascular damage. Postoperative evaluations included serum calcium (Ca), magnesium (Mg), and phosphorus (P) analyses on the 1st and 2nd postoperative days, at 10 days, and at 1, 3, 6 months, 1 year, and 5 years of follow-up. Additionally, the mean values of serum intact parathyroid hormone (iPTH) concentration were measured from blood samples collected from both the reimplanted arm (iPTH RA) and non-reimplanted arm (iPTH NRA) within the same period. Results: Among 131 patients, at 10 days post-surgery, only 46 patients (35.1%) out of 131 exhibited graft viability (iPTH ratio >1.5). This percentage increased to 72.8% (94 patients) after 1 month and further to 87.8% (108 patients) after 3 months post-surgery. At 1 year, 84.7% of patients showed good graft functionality. After 5 years, the percentage remained stable, with graft viability observed in 81.3% of patients. Only 91 of the initial 131 patients completed follow-up up to 5 years, with a dropout rate of 30.5 %. Conclusions: Parathyroid reimplantation using the PR-FaST technique is a viable option for patients undergoing thyroidectomy and has been shown to be a reproducible and effective technique in most patients, with sustained graft functionality and parathyroid hormone production over a 5-year follow-up period. [ABSTRACT FROM AUTHOR]

Tacrolimus is immunosuppressive drugs that have a narrow therapeutic range, and a large inter-patient variability in their pharmacokinetics. Serial monitoring of tacrolimus after transplantation is important to ensure the concentration in biological fluids is maintained at the therapeutic range. Blood sampling for therapeutic drug monitoring of tacrolimus is mostly by venipuncture, making it inconvenient and invasive due to repeated sampling. Thus, microsampling has been adopted as an alternative for sample collection as it is less invasive and more convenient for patients. This article aims to review validated bioanalysis methods for tacrolimus to compare the methods using venous sample and microsampling methods. Related studies about tacrolimus analysis method in blood samples were screened from several databases and summarized with PRISMA flow diagram. A total of 12 studies were considered eligible and reviewed in this article. This systematic review provides a narrative of sample quantification of tacrolimus that collected by venipuncture and microsampling method, its bioanalytical method employed to perform the analysis, and clinical validation or application to patients with organ transplants. Studies with microsampling methods have been validated and applied to the patients. Some studies that compared both methods concluded there is no significant difference between tacrolimus concentrations by venipuncture or microsampling methods. Therefore, microsampling can be considered an option for future development of sampling method in tacrolimus routine monitoring. [ABSTRACT FROM AUTHOR]

The study presents the importance of forensic bacteriology, its use in forensic cases, the methods for bacteriological sampling from corpses, the types and species of bacteria isolated from human and pig corpses, which are most commonly used in forensic biology. The microbial changes that occur after death remain unclear. Postmortem microbiology is a relatively new field of research. After death, the failure of the immune system and other physical barriers leads to the proliferation and spread of microbes. In order for bacteriological information to be accepted within the scope of forensic bacteriology, the court must find suspicion to be present about whether the bacteria seen on the body will contribute to solving the case. Experts must be appointed to examine the issue in line with this suspicion, and these experts must prepare and submit their reports to the court at the requested time. When considering the literature studies, forensic bacteriology has been suggested to be a scientific discipline in the developmental stage and to only be able to provide circumstantial evidence in forensic cases as opposed to primary evidence. According to the literature review, most bacterial studies isolated from corpses were conducted in Romania. Although bacterial samples were isolated from various parts of the corpses, bacteria were mostly isolated from their blood samples. According to literature searches from various scientific journal databases, no study has occurred with a list of the bacteria isolated from corpses. This study is thought to be able to fill this important gap. [ABSTRACT FROM AUTHOR]

An unidentified compound in putrefied postmortem blood samples showed identical accurate mass and chromatographic behavior as 3,4-methylenedioxyamphetamine (MDA) and led to false-positive preliminary screening results. The aim of the study was to identify this unknown interference. Postmortem blood samples were analyzed after protein precipitation on a QExactive Focus high-resolution mass spectrometer (Thermo Fisher, Germany) coupled to a RP C18 column (Macherey-Nagel, Germany). Based on the analysis of mass spectrometry (MS) adducts and isotope ratios using fullscan (m/z 134–330) information, the empiric formula of the protonated molecule [M + H]+ of the unknown compound was found to be C10H14O2N (+ 0.6 ppm). Product ion spectra recorded using normalized collision energy 22% showed a base peak of C8H9O1 (+ 1.5 ppm) and a low-abundant water loss to C7H9 (+ 1.9 ppm), neutral losses of C2H2O and NH3 were found. Based on fullscan and MS-MS information and under consideration of the observed order of neutral losses, the compound was presumptively identified as N -acetyltyramine. This assumption was supported by SIRIUS software showing a SIRIUS score of 99.43% for N -acetyltyramine. Finally, the putative structure annotation was confirmed by a reference compound. The described false-positive MDA findings could be attributed to the presence of N -acetyltyramine in putrefied blood samples. Being an isomer of MDA, N -acetyltyramine could not be distinguished by high-resolution data of the protonated molecules. The presented results once again highlight that false-positive findings may occur even in hyphenated high-resolution mass spectrometry (HRMS) when using full-scan information only. [ABSTRACT FROM AUTHOR]

Simple Summary: Early pregnancy diagnosis is of major importance to the economics of dairy herds. The detection of placental proteins, termed pregnancy-associated glycoproteins (PAGs), represents a diagnostic tool to identify both pregnant and non-pregnant cows. However, as milk production correlates negatively with plasma levels of PAGs during early pregnancy, detection of PAGs may be hindered by high milk production. This study evaluates a newly developed commercial PAGs-based pregnancy test using whole blood or plasma samples during early pregnancy (28–55 days of gestation) in high-producing dairy cows. This study evaluated a commercial pregnancy-associated glycoproteins (PAGs)-based pregnancy test using whole blood or plasma samples during early pregnancy (28–55 days of gestation) in high-producing dairy cows. Transrectal ultrasonography was used as the gold standard method. The study population constituted of 284 cows. False positive diagnoses were recorded from Day 60 to 89 and from Day 60 to 99 postpartum in blood and plasma samples, respectively. In early pregnancy screening, correct positive diagnoses were recorded in 75% and 100% of blood and plasma samples, respectively. High milk production was associated with negative results in blood samples and with the lowest test line intensity in plasma samples. False positive or negative diagnoses were recorded in 0% of both types of samples in cows previously diagnosed as pregnant and showing signs of estrus. In conclusion, the use of plasma was more effective than the use of blood in early pregnancy diagnosis. In cows previously diagnosed as pregnant and showing signs of estrus, both types of samples showed the same results. Because of large individual variations, normal single pregnancies could not be differentiated from twin pregnancies, from pregnancies with a recently dead conceptus, or from pregnancies that experienced subsequent pregnancy loss. [ABSTRACT FROM AUTHOR]

Currently, tandem mass spectrometry-based newborn screening (NBS), which examines targeted biomarkers, is the first approach used for the early detection of maple syrup urine disease (MSUD) in newborns, followed by confirmatory genetic mutation tests. However, these diagnostic approaches have limitations, demanding the development of additional tools for the diagnosis/screening of MUSD. Recently, untargeted metabolomics has been used to explore metabolic profiling and discover the potential biomarkers/pathways of inherited metabolic diseases. Thus, we aimed to discover a distinctive metabolic profile and biomarkers/pathways for MSUD newborns using untargeted metabolomics. Herein, untargeted metabolomics was used to analyze dried blood spot (DBS) samples from 22 MSUD and 22 healthy control newborns. Our data identified 210 altered endogenous metabolites in MSUD newborns and new potential MSUD biomarkers, particularly L-alloisoleucine, methionine, and lysoPI. In addition, the most impacted pathways in MSUD newborns were the ascorbate and aldarate pathways and pentose and glucuronate interconversions, suggesting that oxidative and detoxification events may occur in early life. Our approach leads to the identification of new potential biomarkers/pathways that could be used for the early diagnosis/screening of MSUD newborns but require further validation studies. Our untargeted metabolomics findings have undoubtedly added new insights to our understanding of the pathogenicity of MSUD, which helps us select the appropriate early treatments for better health outcomes. [ABSTRACT FROM AUTHOR]

We investigated the pharmacokinetic pathway of berberine and its metabolites in vitro, in Caco-2 cells, and in human participants following the administration of dihydroberberine (DHB) and micellar berberine (LipoMicel®, LMB) formulations. A pilot trial involving nine healthy volunteers was conducted over a 24 h period; blood samples were collected and subjected to Ultra High-Performance Liquid Chromatography–High Resolution Mass Spectrometry (UHPLC-HRMS) analyses to quantify the concentrations of berberine and its metabolites. Pharmacokinetic correlations indicated that berberrubine and thalifendine follow distinct metabolic pathways. Additionally, jatrorrhizine sulfate appeared to undergo metabolism differently compared to the other sulfated metabolites. Moreover, berberrubine glucuronide likely has a unique metabolic pathway distinct from other glucuronides. The human trial revealed significantly higher blood concentrations of berberine metabolites in participants of the DHB treatment group compared to the LMB treatment group—except for berberrubine glucuronide, which was only detected in the LMB treatment group. Similarly, results from in vitro investigations showed significant differences in berberine metabolite profiles between DHB and LMB. Dihydroberberine, dihydroxy-berberrubine/thalifendine and jatrorrhizine sulfate were detected in LMB-treated cells, but not in DHB-treated cells; thalifendine and jatrorrhizine-glucuronide were detected in DHB-treated cells only. While DHB treatment provided higher blood concentrations of berberine and most berberine metabolites, both in vitro (Caco-2 cells) and in vivo human studies showed that treatment with LMB resulted in a higher proportion of unmetabolized berberine compared to DHB. These findings suggest potential clinical implications that merit further investigation in future large-scale trials. [ABSTRACT FROM AUTHOR]

Introduction: this study was designed to measure ethyl alcohol concentrations in blood samples of different matrices (Li-He plasma, K2-EDTA plasma and serum) with the DRI® method on the ILab Taurus analyzer. The aim of the protocol was to compare the values between the matrices K2-EDTA and serum with Li-He plasma. Furthermore, 18 real samples from the emergency room were used to compare the results obtained with the enzymatic method on both Li-He and K2-EDTA plasma in comparison with the GC/HS reference method. Methods: serum, Li-He and K2-EDTA samples were obtained from left-over routine samples to obtain different pools (serum, Li-He plasma and K2-EDTA plasma pools). Samples with different expected concentrations of ethanol (range 0.00-2.22 g/L) were prepared in situ (60 samples for each matrix) by precise additions of a 10% ethanol solution. All these samples were stored at -20°C until use. Results: in the expected concentration range between 0.42 and 0.85 g/L and around 0.8 and 1.5 g/L [which incorporates most of the legally permitted levels of BAC (Blood Alcohol Concentration) in Europe including Italy] the CV% obtained are all <5.0%. The GC/HS correlation with ILab Taurus for both plasma matrices, LI-He and K2-EDTA evaluated in 18 genuine samples was excellent. Discussion: all the obtained results show that for all the matrices evaluated in the study, the analytical performances are highly satisfactory. [ABSTRACT FROM AUTHOR]

Introduction: Early diagnosis of neonatal sepsis is very essential part of newborn care to prevent mortality and decrease morbidity in newborns. Aim: The aim of this study is to correlate an increase in C‑reactive protein (CRP) titre and a decrease in platelet count with an increased incidence of neonatal septicemia, for an effective subtle approach in neonatal septicemia. Materials and Methods: A retrospective study was conducted in the neonatal intensive care unit (NICU) of a tertiary care hospital from Jan 2022 to July 2023. Neonates admitted to the NICU with suspected sepsis were screened for sepsis. Screening was done by taking blood culture samples before administration of antibiotics, serum samples for CRP and blood samples for platelet count. Result: A total of 270 newborns with suspected sepsis were included in the study. Blood culture positivity was seen in 27.7 (27/75) cases. About 32.9% (89/270) of the neonates with suspected sepsis and 61.3% (46/75) neonates with confirmed sepsis had raised CRP; 32.2% (87/270) neonates with suspected sepsis and 64% (48/75) with confirmed sepsis had decreased platelet count. Both an increase in CRP and a decrease in platelets were seen in 61.3% (46/75) of confirmed cases. Conclusion: In our study, both raised CRP and decreased platelet count were seen in around 60% of confirmed cases of sepsis. So, CRP titre and platelet count can be used as early, rapid diagnostic markers for confirmed sepsis. [ABSTRACT FROM AUTHOR]

The current study examined the levels of antioxidants superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px), glutathione (GSH), and Malondialdehyde (MDA) and reproductive hormones (progesterone, estrogen, and cortisol) in pregnant local breed ewes and after lambing in the Basrah governorate. Twenty ewes were used in this study from several fields in the Basrah governorate. Blood sample collection was done in three different periods, pre-lambing, immediately after lambing, and two weeks after lambing. The result showed a significant decrease (p<0.05) in SOD, CAT, and GSH-px activity before and during lambing compared to after lambing. The results also confirmed a significant increase (p<0.05) in the level of GSH pre and during lambing compared to after lambing. While evaluating, the level of MDA revealed a significant increase (p<0.05) in its level during lambing compared to two weeks after lambing. Hormonal analysis showed a significant increase (p<0.05) in the levels of cortisol and estradiol pre and during lambing compared to after lambing, with a significant decrease (p<0.05) in the levels of progesterone after lambing compared to before lambing. This study concluded that during lambing, ewes were under stress factors, and an internal mechanism represented by enzymatic antioxidants like SOD, CAT, and GSH-Px acted actively during this period to neutralize the side effects of stress. [ABSTRACT FROM AUTHOR]

Hereditary transthyretin cardiac amyloidosis (ATTRv-CA) has a long latency phase before clinical onset, creating a need to identify subclinical disease. We hypothesized circulating transthyretin (TTR) and retinol binding protein 4 (RBP4) levels would be associated with TTR carrier status and correlated with possible evidence of subclinical ATTRv-CA. TTR and RBP4 were measured in blood samples from V122I TTR carriers and age-, sex- and race-matched non-carrier controls (1:2 matching) among Dallas Heart Study participants (phases 1 (DHS-1) and 2 (DHS-2)). Multivariable linear regression models determined factors associated with TTR and RBP4. There were 40 V122I TTR carriers in DHS-1 and 54 V122I TTR carriers in DHS-2. In DHS-1 and DHS-2, TTR was lower in V122I TTR carriers (p <.001 for both), and RBP4 in DHS-2 was lower in V122I TTR carriers than non-carriers (p =.002). Among V122I TTR carriers, TTR was negatively correlated with markers of kidney function, and limb lead voltage (p <.05 for both) and TTR and RBP4 were correlated with atrial volume in DHS-2 (p <.05). V122I TTR carrier status is independently associated with lower TTR and RBP4 in comparison with non-carriers. These findings support the hypothesis that TTR and RBP4 may correlate with evidence of subclinical ATTRv-CA. [ABSTRACT FROM AUTHOR]

1. A reduction in crude protein (CP) in feed for broiler chickens necessitates elevated free amino acid (AA) levels to meet the requirement of each AA. This study investigated adaptations following a change to diets with increasing free AA concentrations and possible reasons for the limitation caused by the inclusion of more free AA. 2. Male Ross 308 broiler hatchlings received a starter diet (164 g CP/kg containing 80 g/kg soy protein isolate (SPI)) until d 7. From d 7–22, birds received a diet almost identical to the starter diet or two other diets, where 50% or 100% of digestible AA in SPI were substituted with a free AA mixture. Birds were allocated to metabolism units located in the same barn to determine performance (n = 7 units) and blood traits (n = 14 birds). Total excreta collection was performed on d 7–8, 8–9, 9–10, 11–12, 14–15 and 21–22. Blood samples were collected on d 7, 8, 9, 11, 14 and 21. 3. Average daily weight gain (ADG) and average daily feed intake (ADFI) was unaffected at 50% AA substitution but decreased at 100% AA substitution on d 7–22 (p ≤ 0.001). The 100% substitution led to a decline in ADG and ADFI consistently on all days (p ≤ 0.037) except on d 11–12. A 50% AA substitution resulted in lower ADFI on d 7–8 and 14–15 (p ≤ 0.032). Nitrogen utilisation efficiency (NUE) was on a level of ~ 0.74 and was only affected by treatment up to d 11–12 (p ≤ 0.008). Concentrations of 10, 9, 8, 10 and 4 plasma free AA were affected on d 8, 9, 11, 14 and 21, respectively (p ≤ 0.037). 4. Following a change to diets containing high levels of free AA, NUE and free AA concentrations in the circulation became more balanced within 3 to 7 d. The results suggested that peptide-bound and free AA did not cause different NUE, particularly 3 and 7 d after the diet change. [ABSTRACT FROM AUTHOR]

The hairy-legged vampire bat, Diphylla ecaudata, is a relatively rare, poorly known, and specialized species that feeds primarily on the blood of birds. We studied a large colony of D. ecaudata rosting with the common hematophagous bats Desmodus rotundus, in a brick house in São Paulo, a large metropolis in southeastern Brazil. The colony size varied from 68 individuals in July 2011 to 21 in February 2014, and females were slightly larger than males. This is the first record of a large group of D. ecaudata sheltering in a man-made structure in Brazil and the first largest group of the species recorded in the country. As a rabies control measure, some animals were subjected to fluorescent antibody and mouse inoculation tests. Blood samples were collected from 24 animals, and the sera were tested for the presence of rabies antibodies using a simplified fluorescent inhibition microtest. No bat tested positive for rabies. Using 0.5 IU/ml as the cut-off, the prevalence of rabies antibodies was 58.8% in D. ecaudata and 83.0% in D. rotundus. The expressive levels of rabies antibodies indicate that the rabies virus actively circulates between the two species. El murciélago vampiro patas peludas, Diphylla ecaudata, es una especie especializada, relativamente rara y poco conocida, que se alimenta principalmente de sangre de aves. Estudiamos una gran colonia de D. ecaudata cohabitando con el murciélago vampiro común Desmodus rotundus en una casa de ladrillos en la ciudad de São Paulo, una gran metrópolis en el sureste de Brasil. El tamaño de la colonia varió de 68 individuos en julio de 2011 a 21 en febrero de 2014, y las hembras fueron ligeramente más numerosas que los machos. Este es el primer registro de un grupo grande de D. ecaudata ocupando una estructura artificial en Brasil y el primer registro de un grupo tan grande de la especie en el país. Como medida de control de la rabia, algunos animales fueron sometidos a pruebas de anticuerpos fluorescentes y de inoculación utilizadas en ratones. Se recolectaron muestras de sangre de 24 animales y los sueros se analizaron para detectar la presencia de anticuerpos contra la rabia mediante una microprueba de inhibición fluorescente simplificada. Ningún murciélago dio positivo por rabia. Utilizando 0,5 UI/ml como punto de corte, la prevalencia de anticuerpos contra la rabia fue del 58,8% en D. ecaudata y del 83,0% en D. rotundus. Los niveles de expresión de anticuerpos contra la rabia indican que el virus de la rabia circula activamente entre las dos especies. [ABSTRACT FROM AUTHOR]

Sexual abuse and rape are among the crimes that are increasing day by day in our country. According to 2019 TUIK data, 13.1% of children brought to security units are victimized by sexual crimes. According to data published by the World Population Review website in 2020, 1071 people were raped in Türkiye that year. The provability of crimes is a very important factor for the perpetrator to be punished. The field of forensic sciences tries to serve justice by examining the evidence found at the crime scene. In cases where the possible body fluids of the perpetrator and the victim are mixed, it is very difficult to make an identification because there is no method for the separation of these mixed samples. In this study, we aimed to differentiate mixed body fluids (blood and saliva) by microRNA method. The samples collected from volunteers for this study were prepared as two-donor mixtures in four different groups in five different dilutions. A total of 36 mixtures were prepared from fluids collected from 4 female and 4 male individuals. The miR-451A biomarker was used to indicate the presence of blood and the miR-203A-3p marker was used to indicate the presence of saliva in the mixture samples. The samples were first centrifuged and then RNA isolation was performed using miRNeasy Mini Kit. The concentration and purity of the obtained RNAs were determined by spectrophotometer. Then cDNA (complementary DNA) was synthesized from RNAs with Taqman microRNA Reverse Transcription Kit.U6 gene region was preferred as the reference gene and analyzes were performed by RT-PCR. The presence of blood samples in the mixture was shown in all dilutions, the best level was seen in 1x1 ratio. When all groups were analyzed, it was seen that the mixtures with male donors were expressed at a higher rate than the other mixtures. [ABSTRACT FROM AUTHOR]

As the beef industry moves towards efficient animal production to improve sustainability in agriculture, new production and management approaches are emerging. Among the many facets of the beef industry, cow–calf operations have the most opportunity for efficiency improvement, including improvements in fertility. This project accounts for measures and methods of (1) twinning reproductive technologies and (2) twin calf perinatal care and pre-weaning rearing. The overall objective was to produce twin calves using two reproductive technologies—embryo transfer and artificial insemination. The subobjectives were to determine accuracy of twin pregnancies embryo/fetal losses using ultrasonography, evaluate parturition and dystocia, and determine the effects of different twin-raising methods on neonatal behavior and growth. A fixed-time artificial insemination (FTAI) protocol was applied to 77 multiparous Angus-cross cows from a commercial beef herd in northcentral South Dakota during the summer of 2019. Cows were assigned to two different treatments groups: only artificially inseminated (AI) or received an embryo transfer following artificial insemination (ET + AI). They were estrous-synchronized, artificially inseminated (AI) with black Angus semen at day 0, and received and embryo transfer (ET) at day 7. Ultrasound examination detected 56% pregnancy risk for both groups, with sensitivity, specificity, and accuracy of 75%, 100%, and 90.5%, respectively, for bilateral twin detection. Calves were born during spring 2020. Twin calves (n = 34) and singleton calves (n = 11) were assigned to one of three raising methods: (1) twin born and twin raised (TT; n = 16), (2) twin born and single raised (TS; n = 18), and (3) single born and single raised (S; n = 11). Neonatal nursing behavior and birth weights were recorded, and adjusted day 200 and day 280 were calculated measures of vitality and growth. Blood samples were collected at age 24 h for colostrum intake measures (total serum protein, IgG1, and IgM). Twin calves were born 20% (p < 0.05) lighter in body weight than singletons; however, weights did not differ at day 280 between TT and S calves. TS calves had the shortest average latency to stand, but immunoglobulin concentrations did not differ among treatments. At weaning, cows that had birthed and raised twins produced more kilograms of live weight per pregnancy than cows birthing and raising singletons. Using ET + AI proved to increase twinning rate, and growth was maintained when raising both twins with their dam. [ABSTRACT FROM AUTHOR]

This study was designed to determine the prevalence of autoimmune hypothyroidism (Hashimoto's) and its impact on men and women in Thi-Qar Governorat. The current study included (100) people who were in two groups, with (50) people in each group. The first group includes patients with hypothyroidism (Hashimoto's), and the second group includes (Healthy) people who do not suffer from any thyroid disease. Blood samples were collected for the period from (January 1 to March 30/ 2023), for use in measuring the level of thyroid hormones and measuring the level of autoantibodies. The results of the current study showed that female patients had the highest incidence of autoimmune hypothyroidism (Hashimoto’s), at a rate of (68%), while for males, the incidence rate was (32%). As for the age group, the highest incidence rate was for ages over 40 years, and in both cases of the disease The results also showed a relationship between (Sex, Family History, Smoking) and the risk of developing autoimmune hypothyroidism (Hashimoto’s). The results of the current study also showed an increase in the average concentration of thyroid-stimulating hormone (TSH) in patients with hypothyroidism (Hashimoto’s), corresponding to a decrease in the average concentration of each of the hormones (T3, T4, FT4) compared to the healthy group. An increase in the level of autoantibodies (AntiTPOAb, Anti-TgAb) was also observed in patients compared to healthy people. [ABSTRACT FROM AUTHOR]

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Aims: The aim of this study was to identify septicemia in intensive care unit (ICU) patients, determine the antibiotic resistance profiles and minimum inhibitory concentrations (MICs) of all isolates by using the Vitek-2 compact system and study some immunological parameters (IgM, IgG and hs-CRP) in ICU patients, with and without septicemia. Methodology and results: Ninety blood samples from both sexes and different age groups (ranging from 21-90 years) were collected from different hospitals in Mosul city. Twenty-four blood samples were collected from healthy people and used as a control group for the period between July and December of 2022. Samples were subjected to bacterial diagnosis, antibiotic sensitivity test using Vitek2 and measurement of some immunological parameters. The results showed that 24/90 (26.67%) of the blood samples exhibited positive cultures, with Gram-positive bacteria predominating, 14/24 (58.34%), while Gram-negative bacteria represented 10/24 (41.66%) of the positive samples. Antimicrobial susceptibility testing showed that vancomycin, linezolid, imipenem, and rifampicin were mostly effective against most Gram-positive septicemia isolates, whereas meropenem, imipenem and colistin were mostly effective against most Gram-negative bacterial isolates. The ELISA results revealed elevated levels of IgM, IgG and hs-CRP in the serum of septicemia patients compared to the healthy control group, with a significant difference in P-value (0.001). In addition, the mean levels of IgM, IgG and hs-CRP in the serum of non-septicemia patients compared to the healthy control group indicated a highly significant difference in P-value (0.001). Conclusion, significance and impact of study: IgM, IgG and hs-CRP levels were significantly different in sepsis patients infected with Gram-positive and Gram-negative bacteria compared to the healthy control group. [ABSTRACT FROM AUTHOR]

Kisspeptin is a neuropeptide that governs the reproductive axis upstream to GnRH. Present study was undertaken to compare the effect of different doses of kisspeptin on release of luteinizing hormone (LH) in Murrah buffaloes during pre-pubertal period. Twenty four pre-pubertal buffalo heifers were selected and divided into 4 groups (Control, T1, T2 and T3), having 6 animals in each group. Kisspeptin-10 was administered at the dose of 2 µg/kg body weight, 1.5 µg/ kg body weight, 1µg/kg body weight respectively in T1, T2 and T3 group along with administration of equal volume of NSS in control group. Blood sampling was done at every 20 minutes interval beginning with zero minutes up to 2 hours and analyzed for LH concentration. Results indicated that LH concentration was significantly (p<0.05) higher in T1 group. The peak level of LH was recorded at 20 minutes post administration and falls to basal level at 60 minutes post administration. Although increased LH concentration was observed in other treatment groups but it was lower than T1 group. No LH peak or rise in LH level was observed in control group. The result of this study indicates that during the pre-pubertal period, administration of Kisspeptin causes dose dependent increase in LH concentration and peak level was seen at 2 µg/kg body weight of dose. [ABSTRACT FROM AUTHOR]

Forty-two Japanese Black cattle on two farms in Kagoshima prefecture, Japan were used in this study. The rearing style of farm A was in a dark barn with a large roof to block sunlight (n=21). The rearing style of farm B was grazing, and exposed to direct sunlight (n=21). Blood sampling was performed twice on the same cattle in August 2022 (summer) and February 2023 (winter). As the results, the serum 25(OH)D3 concentrations were significantly lower in cattle of farm A than in cattle of farm B (P<0.01), and were significantly lower in winter season than in summer season (P<0.01). These results showed that there were differences in blood 25(OH)D3 concentrations between the farms or seasons. [ABSTRACT FROM AUTHOR]

Background: Home blood self-collection devices can enable remote monitoring, but their implementation requires validation. Our objectives were to explore (i) the impact of sampling sites and topical analgesia on capillary blood volume and pain perception and (ii) the safety, acceptability, and failure of capillary self-collection among adults and children using the Tasso-SST device. Methods: We conducted a two-phase study. The investigational phase consisted of two on-site cross-sectional studies in healthy adult participants (≥ 12 years) and children (1–17 years) with their accompanying parent. Adults received 4 capillary samplings, where puncture sites and topical analgesia were randomized in a factorial design, and a venipuncture; children (and one parent) had one capillary sampling. The two co-primary outcomes were blood volume and pain. The implementation phase was conducted in two multicentre trials in participants choosing remote visits; blood volume, collection failure, adverse events, and satisfaction were documented. Results: In the investigational phase, 90 participants and 9 children with 7 parents were enrolled; 15 adults and 2 preschoolers participated in the implementation phase. In the adult investigational study, the device collected a median (25%, 75%) of 450 (250, 550) μl of blood with no significant difference between the puncture site, topical analgesia, and its interaction. Using topical analgesia reduced pain perception by 0.61 (95% CI: 0.97, 0.24; P <0.01) points on the 11-point scale; the pain reduction varied by puncture site, with the lower back showing the most significant decrease. Overall, combining all studies and phases, the median volume collected was 425 (250, 500) μl, and the device failure rate was 4.4%; minor adverse effects were reported in 8.9% of the participants, all were willing to use the device again. Conclusion: Capillary blood self-collection, yielding slightly less than 500 μl, proves to be a safe and relatively painless method for adults and children, with high satisfaction and low failure rates. The puncture site and topical analgesia do not affect blood volume, but topical analgesia on the lower back could reduce pain. [ABSTRACT FROM AUTHOR]

Cell-free DNA (cfDNA), a burgeoning class of molecular biomarkers, has been extensively studied across a variety of biomedical fields. As a key component of liquid biopsy, cfDNA testing is gaining prominence in disease detection and management due to the convenience of sample collection and the abundant wealth of genetic information it provides. However, the broader clinical application of cfDNA is currently impeded by a lack of standardization in the preanalytical procedures for cfDNA analysis. A number of fundamental challenges, including the selection of appropriate preanalytical procedures, prevention of short cfDNA fragment loss, and the validation of various cfDNA measurement methods, remain unaddressed. These existing hurdles lead to difficulties in comparing results and ensuring repeatability, thereby undermining the reliability of cfDNA analysis in clinical settings. This review discusses the crucial preanalytical factors that influence cfDNA analysis outcomes, including sample collection, transportation, temporary storage, processing, extraction, quality control, and long-term storage. The review provides clarification on achievable consensus and offers an analysis of the current issues with the goal of standardizing preanalytical procedures for cfDNA analysis. [ABSTRACT FROM AUTHOR]

This study investigates how missing data samples in continuous blood glucose data affect the prediction of postprandial hypoglycemia, which is crucial for diabetes management. We analyzed the impact of missing samples at different times before meals using two datasets: virtual patient data and real patient data. The study uses six commonly used machine learning models under varying conditions of missing samples, including custom and random patterns reflective of device failures and arbitrary data loss, with different levels of data removal before mealtimes. Additionally, the study explored different interpolation techniques to counter the effects of missing data samples. The research shows that missing samples generally reduce the model performance, but random forest is more robust to missing samples. The study concludes that the adverse effects of missing samples can be mitigated by leveraging complementary and informative non-point features. Consequently, our research highlights the importance of strategically handling missing data, selecting appropriate machine learning models, and considering feature types to enhance the performance of postprandial hypoglycemia predictions, thereby improving diabetes management. [ABSTRACT FROM AUTHOR]

Conventional venipuncture is invasive and challenging in low and middle‐income countries. Conversely, point‐of‐care devices paired with fingersticks, although less invasive, suffer from high variability and low blood volume collection. Recently approved microsampling devices address some of these issues but remain cost‐prohibitive for resource‐limited settings. In this work, a cost‐effective microsampling device is described for the collection of liquid blood with minimal invasiveness and sufficient volume retrieval for laboratory analyses or immediate point‐of‐care testing. Inspired by the anatomy of sanguivorous leeches, the single‐use device features a storage compartment for blood collection and a microneedle patch hidden within a suction cup. Finite Element Method simulations, corroborated by mechanical analyses, guide the material selection for device fabrication and design optimization. In piglets, the device successfully collects ≈195 µL of blood with minimal invasiveness. Additionally, a tailor‐made lid and adapter enable safe fluid transportation and integration with commercially available point‐of‐care systems for on‐site analyses, respectively. Taken together, the proposed platform holds significant promise for enhancing healthcare in the pediatric population by improving patient compliance and reducing the risk of needlestick injuries through concealed microneedles. Most importantly, given its cost‐effective fabrication, the open‐source microsampling device may have a meaningful impact in resource‐limited healthcare settings. [ABSTRACT FROM AUTHOR]

Background: Circulating tumor cells (CTCs) are considered as a useful biomarker for early cancer diagnosis, which play a crucial role in metastatic process. Unfortunately, the tumor heterogeneity and extremely rare occurrence rate of CTCs among billions of interfering leukocytes seriously hamper the sensitivity and purity of CTCs isolation. Methods: To address these, we firstly used microfluidic chips to detect the broad-spectrum of triple target combination biomarkers in CTCs of 10 types of cancer patients, including EpCAM, EGFR and Her2. Then, we constructed hybrid engineered cell membrane-camouflaged magnetic nanoparticles (HE-CM-MNs) for efficient capture of heterogeneous CTCs with high-purity, which was enabled by inheriting the recognition ability of HE-CM for various CTCs and reducing homologous cell interaction with leukocytes. Compared with single E-CM-MNs, HE-CM-MNs showed a significant improvement in the capture efficiency for a cell mixture, with an efficiency of 90%. And the capture efficiency of HE-CM-MNs toward 12 subpopulations of tumor cells was ranged from 70 to 85%. Furthermore, by using HE-CM-MNs, we successfully isolated heterogeneous CTCs with high purity from clinical blood samples. Finally, the captured CTCs by HE-CM-MNs could be used for gene mutation analysis. Conclusions: This study demonstrated the promising potential of HE-CM-MNs for heterogeneous CTCs detection and downstream analysis. [ABSTRACT FROM AUTHOR]

Opioid misuse in the United States contributes to >70% of annual overdose deaths. To develop additional therapeutics that may prevent opioid misuse, further studies on the neurobiological consequences of opioid exposure are needed. Here we sought to characterize molecular neuroadaptations involving microRNA (miRNA) pathways in the brain and blood of adult male rats that selfadministered the opioid heroin. miRNAs are -18-24 nucleotide RNAs that regulate protein expression by preventing mRNA translation into proteins. Manipulation of miRNAs and their downstream pathways can critically regulate drug seeking behavior. We performed small-RNA sequencing of miRNAs and proteomics profiling on tissue from the orbitofrontal cortex (OFC), a brain region associated with heroin seeking, following 2 days of forced abstinence from self-administration of 0.03mg/kg/infusion heroin or sucrose. Heroin self-administration resulted in a robust shift of the OFC miRNA profile, regulating 77 miRNAs, while sucrose self-administration only regulated 9 miRNAs that did not overlap with the heroin-induced profile. Conversely, proteomics revealed dual regulation of seven proteins by both heroin and sucrose in the OFC. Pathway analysis determined that heroinassociated miRNA pathways are predicted to target genes associated with the term "prion disease," a termthat was also enriched in the heroin-induced protein expression dataset. Lastly, we confirmed that a subset of heroin-induced miRNA expression changes in the OFC are regulated in peripheral serum and correlate with heroin infusions. These findings demonstrate that peripheral blood samples may have biomarker utility for assessment of drug-induced miRNA pathway alterations that occur in the brain following chronic drug exposure. [ABSTRACT FROM AUTHOR]

Background: Artemisinin resistance in Plasmodium falciparum threatens global malaria elimination efforts. To contain and then eliminate artemisinin resistance in Eastern Myanmar a network of community-based malaria posts was instituted and targeted mass drug administration (MDA) with dihydroartemisinin-piperaquine (three rounds at monthly intervals) was conducted. The prevalence of artemisinin resistance during the elimination campaign (2013–2019) was characterized. Methods: Throughout the six-year campaign Plasmodium falciparum positive blood samples from symptomatic patients and from cross-sectional surveys were genotyped for mutations in kelch-13—a molecular marker of artemisinin resistance. Result: The program resulted in near elimination of falciparum malaria. Of 5162 P. falciparum positive blood samples genotyped, 3281 (63.6%) had K13 mutations. The prevalence of K13 mutations was 73.9% in 2013 and 64.4% in 2019. Overall, there was a small but significant decline in the proportion of K13 mutants (p < 0.001). In the MDA villages there was no significant change in the K13 proportions before and after MDA. The distribution of different K13 mutations changed substantially; F446I and P441L mutations increased in both MDA and non-MDA villages, while most other K13 mutations decreased. The proportion of C580Y mutations fell from 9.2% (43/467) before MDA to 2.3% (19/813) after MDA (p < 0.001). Similar changes occurred in the 487 villages where MDA was not conducted. Conclusion: The malaria elimination program in Kayin state, eastern Myanmar, led to a substantial reduction in falciparum malaria. Despite the intense use of artemisinin-based combination therapies, both in treatment and MDA, this did not select for artemisinin resistance. [ABSTRACT FROM AUTHOR]

Avian haemosporidian parasites are useful model organisms to study the ecology and evolution of parasite-host interactions due to their global distribution and extensive biodiversity. Detection of these parasites has evolved from microscopic examination to PCR-based methods, with the mitochondrial cytochrome b gene serving as barcoding region. However, standard PCR protocols used for screening and identification purposes have limitations in detecting mixed infections and generating phylogenetically informative data due to short amplicon lengths. To address these issues, we developed a novel genusspecific nested PCR protocol targeting avian haemosporidian parasites. The protocol underwent rigorous testing utilizing a large dataset comprising blood samples from Malagasy birds of three distinct Passeriformes families. Furthermore, validation was done by examining smaller datasets in two other laboratories employing divergent master mixes and different bird species. Comparative analyses were conducted between the outcomes of the novel PCR protocol and those obtained through the widely used standard nested PCR method. The novel protocol enables specific identification of Plasmodium, Haemoproteus (Parahaemoproteus), and Leucocytozoon parasites. The analyses demonstrated comparable sensitivity to the standard nested PCR with notable improvements in detecting mixed infections. In addition, phylogenetic resolution is improved by amplification of longer fragments, leading to a better understanding of the haemosporidian biodiversity and evolution. Overall, the novel protocol represents a valuable addition to avian haemosporidian detection methodologies, facilitating comprehensive studies on parasite ecology, epidemiology, and evolution. [ABSTRACT FROM AUTHOR]

This study was designed to investigate associations between transfer of passive immunity, selected illnesses (diarrhoea, pneumonia, and fatigue anorexia syndrome-FAS) and growth performance [body weight (BW) and average daily gain (ADG)] in preweaned lambs. A total of 347 lambs were blood sampled at 24±1 h of age after birth and the serum IgG concentration for each lamb was measured after colostrum intake using a commercial ELISA kit. Lambs were weighed on a scale immediately after birth (before colostrum intake) and reweighed on day 28 (end of neonatal period) and day 84 (age of weaning). Lambs ADG was calculated from 0 to 28 d of life, from 29 to 84d of life, and from 0 to 84 d of life. Growth performance (BW and ADG) of lambs with diarrhoea and FAS in the neonatal period and with pneumonia in the postneonatal period was significantly lower than that of healthy lambs of the same period (P<0.05). BW of lambs with serum IgG concentration at 24th hour after the birth (SIgGC-24)<600 mg/dL and <1000 mg/ml was significantly lower on days 28 and 84 than that of lambs with SIgGC-24 >600 mg/dL and >1000 mg/ml (P<0.001). Similarly, the ADG of lambs with SIgGC-24 <600 mg/dL and <1000 mg/ml was significantly lower on days from 0-28, 29-84 and 0-84 of age than those of lambs with SIgGC-24 >600 mg/dL and >1000 mg/ml (P<0.001). In conclusion, our findings show that low serum IgG concentration in lambs, as well as the presence of pneumonia, diarrhea and FAS, reduces growth performance in pre-weaning lambs. [ABSTRACT FROM AUTHOR]

Hyperserotonemia is one of the most studied endophenotypes in autism spectrum disorder (ASD), but there are still no unequivocal results about its causes or biological and behavioral outcomes. This systematic review summarizes the studies investigating the relationship between blood serotonin (5-HT) levels and ASD, comparing diagnostic tools, analytical methods, and clinical outcomes. A literature search on peripheral 5-HT levels and ASD was conducted. In total, 1104 publications were screened, of which 113 entered the present systematic review. Of these, 59 articles reported hyperserotonemia in subjects with ASD, and 26 presented correlations between 5-HT levels and ASD-core clinical outcomes. The 5-HT levels are increased in about half, and correlations between hyperserotonemia and clinical outcomes are detected in a quarter of the studies. The present research highlights a large amount of heterogeneity in this field, ranging from the characterization of ASD and control groups to diagnostic and clinical assessments, from blood sampling procedures to analytical methods, allowing us to delineate critical topics for future studies. [ABSTRACT FROM AUTHOR]

This study aimed to estimate the estrous cycle length of the Algerian Arbia goat in Northern Algeria. For this, eighteen (18) Arbia goats, aged between 2 and 6 years, were used in our work that took place in the experimental farm of the Saad Dahlab University (Blida, Algeria). Blood samples were taken from each goat twice a week (at a 2 or 3-day interval) for 3 months. The serum progesterone concentration was determined by Radio-Immuno-Assay. Smears of the vaginal mucosa were taken at the same time as the blood samples. The predominance of superficial cells on the smear of the vaginal mucosa as well as a serum progesterone level less than 1 ng/mL expressed the return to estrous which was considered the beginning of a new cycle. A negative correlation was observed between the percentage of superficial cells (SC) and serum progesterone (P4) levels in all goats. Our results showed a significant difference (P<0.05) between the means of different cycle lengths obtained among the females. In addition, normal cycles had an average of 20.11±1.85 days (17-25 days) representing 59.6% of cycles. Besides, a large number of short cycles (<17 days) with an average of 14.41±1.51 days were found representing 25.5% of recorded cycles. The number of long cycles (>25 days; with an average of 32.14±5.58 days), represented 14.9% of recorded cycles. Following these results, it can be concluded that the local goat in Northern Algeria had different types of cycles (normal, short, and long) with a large percentage of normal cycles. [ABSTRACT FROM AUTHOR]