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The International Journal of... Nov 2023Tablets are the most widely available dosage form for the treatment of TB; however, adult tablets fail to meet the needs of young children who cannot swallow these...
Tablets are the most widely available dosage form for the treatment of TB; however, adult tablets fail to meet the needs of young children who cannot swallow these tablets or require dose titration. We tested a new, simple device (XTEMP-R) and the methodology for converting tablets of TB drugs into a homogeneous suspension for home use by children and caregivers. XTEMP-R is a new device used for converting tablets into liquid preparations. Four TB drugs - pretomanid, delamanid, clofazimine and bedaquiline - were dispersed in the device utilizing water and simple syrup. The reproducibility of accurately delivering aliquots from the suspension upon preparation and upon redispersion after storing for 2 days was studied. Suspensions of each of the drugs tested were easily prepared in about 10 min and were visually uniform in consistency. Dosages in 2 and 5 mL were assessed in suspension, and those in 5 mL tested upon redispersion after 2 days. The observed range for these dosages spanned from 94.6% to 101.1% of the theoretical concentration for the suspensions under examination. The cleaned device had no detectable residual drug. XTEMP-R can be used at home by caregivers to prepare doses of suspensions accurately for children and patients who cannot swallow tablets.
Topics: Child; Adult; Humans; Child, Preschool; Reproducibility of Results; Tuberculosis; Tablets; Suspensions; Drug Stability
PubMed: 37880886
DOI: 10.5588/ijtld.23.0165 -
Clinical Biochemistry Oct 2023The use of whole blood in rapid cytokine release assays (CRAs) is becoming an established technique for screening immune responses following natural infection or...
INTRODUCTION
The use of whole blood in rapid cytokine release assays (CRAs) is becoming an established technique for screening immune responses following natural infection or vaccination, especially in the context of the SARS-CoV-2 pandemic. Establishing an accurate capillary blood sampling method to replace the need for venipuncture could make CRAs more accessible. In this study, capillary blood was collected via two different methods alongside traditional venipuncture to investigate whether the method of blood draw affects cytokine quantification when performing CRAs.
METHODS
Adults previously vaccinated with SARS-CoV-2 vaccines donated three blood samples: one by venipuncture, one by finger prick, and one by a microneedle device. Whole blood was aliquoted and incubated overnight with SARS-CoV-2 peptides or left unstimulated. Cytokine release in plasma was measured by multiplex array.
RESULTS
In unstimulated samples, little to no cytokines were detected in blood collected via venipuncture or by microneedle devices. Conversely, capillary blood collected by finger prick showed detectable levels of all cytokines analysed, with significantly inflated levels of TNFα, IL-10 (p < 0.0001), IL-2, GM-CSF, and IL-13 (p < 0.01), and 53% of these samples were also positive for IFN-γ. Following peptide stimulation, 25% of samples collected via finger prick showed dysregulated production of IFN-γ, TNFα, IL-2, and IL-10, with lower cytokine production than unstimulated controls. Contrastingly, this was seen in just 4% of venous blood samples and in none of the microneedle samples.
CONCLUSIONS
Capillary blood draw via a microneedle device results in highly comparable immune responses to those seen via venipuncture at baseline and following peptide stimulation, suggesting this is a viable method for rapid whole blood CRAs. Conversely, differential cytokine production is observed following capillary blood draw via finger prick.
PubMed: 37742868
DOI: 10.1016/j.clinbiochem.2023.110648 -
Plant Disease Nov 2023Soybean ( L.) is produced in over 70,000 ha in the Altillanura Region, eastern Colombia (Agronet 2023). From 2018 to 2020, foliar symptoms like green stem and foliar...
Soybean ( L.) is produced in over 70,000 ha in the Altillanura Region, eastern Colombia (Agronet 2023). From 2018 to 2020, foliar symptoms like green stem and foliar retention of soybean, which in Brazil can cause up to 100% soybean yield losses (Meyer et al. 2017), were observed in soybean fields in Colombia. During 2020, samples from symptomatic plants in reproductive stages (R1-R8) were collected from different commercial soybean fields in the Altillanura Region. Over 200 samples were processed, using an incubation method described in Coyne et al. (2014). Nematodes were recovered from photosynthetic leaf tissues and enlarged nodes/buds with population densities ranging from 13 to 132 and 36 to 936 nematodes/10g, respectively. Adult females were morphologically and molecularly characterized as (Oliveira et al. 2019; Subbotin et al. 2020). Female body length (n = 20) ranged from 653.3 to 806.3 μm (mean = 723 μm ± 52.7), stylet length from 11.0 to 12.3 μm (11.8 μm ± 0.3), body diameter from 14.8 to 17.9 μm (16.3 μm ± 1.1), post-uterine sac length from 38.7 to 51.9 μm (44.6 μm ± 5.1), vulva to anus from 145.5 to 223.2 μm (172.2 μm ± 22.4), and 26% of the vulva-anus distance. Genomic DNA was extracted (QIAGEN DNeasy® Blood & Tissue kit) from a pool of nematodes. The D2A/D3B (Tenente et al. 2004) primers were used to amplify and sequence the D2/D3 expansion region of the 28S rRNA gene. PCR product (~759 bp) was purified, sequenced, deposited in GenBank (OQ930285), and compared to previously deposited sequences (e.g., KX356756, KY510840, KY510839, KY510841, KT692694, KY510842, MH187565) by means of the BLAST algorithm. Similarly, 988F and 18SR-Burs (De Jesus et al. 2016) primers were used to amplify and sequence the near full-length 18S RNA gene (SSU). PCR product was purified, sequenced, deposited in GenBank (OQ954344), and compared to previously deposited sequences (e.g., KT454962, KT943534, KT943535, KY510835, KY510836, KY510837, KY510838, MH187565). Phylogenetic Bayesian analysis (Ronquist et al. 2012) of the of the D2/D3 and 18S regions placed this nematode from Colombia in the clade (PP = 100). To fulfill a modified Koch's postulates, the population described above was used in a greenhouse assay. In total, 120 soybean plants (cv. Flor Blanca) were infected with 200 (females + males)/plant. Briefly, at cotyledon stage (VC), 50 µl aliquot containing 50 was delivered onto each cotyledon and unifoliolate leaves (200 nematodes/plant). Sterile water was delivered to 80 plants which served as control. Plants were kept in the greenhouse at approximately 25°C and covered with clear plastic bag for 72 h to maintain over 90% relative humidity. After 15, 30, 45, and 60 days, soybean plants (n = 20) were processed, quantified, and the average reproduction factor (final population/initial population) was 0.1, 2.9, 14.0, and 1.8, respectively. Infected plants showed symptoms of blistering leaves with malformation (midrib vein twist), and was not observed in control plants. To our knowledge, this is the first report of parasitizing soybean buds and leaves in Colombia. Soybean is an important commodity for the Altillanura Region, and it is important to monitor the risk posed by this nematode. Furthermore, a better understanding of the nematode-host interaction and epidemiology in Colombia soybean producing regions is needed.
PubMed: 38085963
DOI: 10.1094/PDIS-06-23-1117-PDN -
Scientific Reports Dec 2023This in vitro study evaluated the potential hemostatic effect of fresh frozen plasma (FFP) ultrafiltration on clotting factors, coagulation parameters, and plasma...
This in vitro study evaluated the potential hemostatic effect of fresh frozen plasma (FFP) ultrafiltration on clotting factors, coagulation parameters, and plasma properties. ABO-specific units of FFP (n = 40) were prepared for the concentrated FFP and cryoprecipitate. Plasma water was removed from FFP by ultrafiltration using a dialyzer with a pump running at a 300 mL/min. The aliquot of each concentrated FFP after 50, 100, 200, and 250 mL of fluid removal were measured the standard coagulation assay, clotting activity, and plasma properties to compare those parameters of cryoprecipitate. Concentrated FFP contained 36.5% of fibrinogen in FFP with a mean concentration of 7.2 g/L, lower than the cryoprecipitate level. The levels of factor VIII (FVIII), von Willebrand factor (VWF):antigen (Ag), and VWF:ristocetin cofactor (RCo) were also lower in concentrated FFP, whereas the levels of factor V, factor IX, factor XIII, antithrombin and albumin was higher in concentrated FFP. Maximum clot firmness (MCF) in thromboelastometry was approximately one-half of that in cryoprecipitate. Although the levels of VWF:Ag, VWF:RCo, and FVIII differed depending on the ABO blood types, fibrinogen levels, and MCF were not significantly different among the ABO blood groups in FFP and concentrated FFP.
Topics: Hemostatics; von Willebrand Factor; Ultrafiltration; Renal Dialysis; Factor VIII; Fibrinogen; Plasma; Factor V
PubMed: 38062086
DOI: 10.1038/s41598-023-48759-1 -
The International Journal of... Oct 2023With an increased demand for rapid, diagnostic tools for TB and drug resistance detection, Truenat MTB-RIF assay has proven to be a rapid point of care molecular test....
With an increased demand for rapid, diagnostic tools for TB and drug resistance detection, Truenat MTB-RIF assay has proven to be a rapid point of care molecular test. The present study aimed to establish a proof of concept of using Trueprep-extracted DNA for line-probe assay (LPA) testing. A total of 150 sputum samples (MTB-positive at Truenat sites) were divided into two aliquots. One aliquot was used for DNA extraction using the Trueprep device and MTB testing. The second aliquot of the sample was subjected to GenoLyse DNA extraction. DNA from both the Trueprep and GenoLyse methods was subjected to first-line (FL) and second-line (SL) LPA testing. Of 139 Trueprep-extracted DNA, respectively 135 (97%) and 105 (75%) had interpretable results by FL and SL-LPA testing. Of 128 GenoLyse-extracted DNA, all 128 (100%) had interpretable FL-LPA results and 114 (89%) had interpretable SL-LPA results. The results obtained in this study indicate that Trueprep-extracted DNA can be used in obtaining valid LPA results. However, the study needs to be conducted on a larger sample size before our recommendations can be used for policy-making decisions.
Topics: Humans; Rifampin; Mycobacterium tuberculosis; Tuberculosis, Multidrug-Resistant; Point-of-Care Testing; Sputum; Sensitivity and Specificity
PubMed: 37749831
DOI: 10.5588/ijtld.23.0003 -
Animals : An Open Access Journal From... Dec 2023Cryopreservation deteriorates boar sperm quality and lifespan, which restricts the use of artificial insemination with frozen-thawed boar semen in field conditions. The...
Cryopreservation deteriorates boar sperm quality and lifespan, which restricts the use of artificial insemination with frozen-thawed boar semen in field conditions. The objective of this study was to test the effects of post-thaw storage time and temperature on boar sperm survival. Semen ejaculates from five Landrace boars (one ejaculate per boar) were collected and frozen following a 0.5 mL-straw protocol. Straws from the five boars were thawed and diluted 1:1 (v:v) in BTS. The frozen-thawed semen samples were aliquoted into three parts and respectively stored at 5 °C, 17 °C, and 37 °C for up to 6 h. At 0.5, 2, and 6 h of storage, sperm motility, viability, mitochondrial membrane potential, and intracellular reactive oxygen species (ROS) levels and apoptotic changes were measured. Antioxidant and oxidant levels were tested in boar sperm (SPZ) and their surrounding environment (SN) at each timepoint. The results showed significant effects of post-thaw storage time and temperature and an impact on boar sperm quality (total and progressive motility, VCL, viability, acrosome integrity), early and late sperm apoptotic changes, and changes in MDA levels in SPZ and SN. Compared to storage at 5 °C and 37 °C, frozen-thawed semen samples stored at 17 °C displayed better sperm quality, less apoptotic levels, and lower levels of SPZ MDA and SN MDA. Notably, post-thaw storage at 17 °C extended boar sperm lifespan up to 6 h without obvious reduction in sperm quality. In conclusion, storage of frozen-thawed boar semen at 17 °C preserves sperm quality for up to 6 h, which facilitates the use of cryopreserved boar semen for field artificial insemination.
PubMed: 38200818
DOI: 10.3390/ani14010087 -
The Science of the Total Environment Dec 2023Soil is the basis for almost all global agriculture and the medium in which most terrestrial biological activity occurs. Viticulture represents an important agricultural... (Review)
Review
Soil is the basis for almost all global agriculture and the medium in which most terrestrial biological activity occurs. Viticulture represents an important agricultural practice in the Castilla-La Mancha (CLM) community. In this region, there are several protected denominations of origin (PDO), the largest being Valdepeñas. This paper describes the accumulation pattern of sulphur (S) in the vineyard soils of this PDO. Samples were collected from 90 vineyard soil profiles. Sulphur content was determined using an X-ray Fluorescence spectrometer in the solid mode on a powdered aliquot of each sample. The results indicated that the total S in soils varied from 0.54 to 6.90 (g·kg) in surface soil (0-30 cm) and from 0.39 to 2.80 (g·kg) on the subsurface layer (30-80 cm). When comparing the mean values of surface horizons to the subsurface horizons, S content lowered as soil depth increased. Kurtosis exceeded 45 % in all cases, which indicates a wide variability of concentrations. These findings can be explained by the continuous fertiliser and fungicide applications (and therefore S) in these production systems. Using the geoaccumulation index (Igeo), most soils were included in Class 0 (Igeo <0) and were, thus, S uncontaminated; only a few points can be considered pollutants. The obtained results should contribute to extend the scarce existing database on S in Mediterranean regions like that herein studied.
PubMed: 37647963
DOI: 10.1016/j.scitotenv.2023.166642 -
International Journal of Molecular... Nov 2023Oxidative stress is involved in the development, progression, and complications of diabetes mellitus (DM). Oxidative modification of human serum albumin's cysteine-34 is...
Oxidative stress is involved in the development, progression, and complications of diabetes mellitus (DM). Oxidative modification of human serum albumin's cysteine-34 is a marker for oxidative stress-related pathological conditions. We aimed to evaluate the redox state of albumin in patients with DM to investigate possible correlations with age, diabetes duration, and disease control status. Plasma aliquots were collected from 52 participants (26 type 1 and 26 type 2 DM). Patients were divided into two groups according to their glycated hemoglobin levels less than or equal to and greater than 58 mmol/L. Albumin redox state was assessed with high-performance liquid chromatography by fractionating it into human mercaptalbumin (HMA) and human nonmercaptalbumin 1 and 2 (HNA1 and HNA2). Albumin redox fractions were differently related to the age of study participants. In age-matched T1DM and T2DM groups, the albumin redox state was essentially the same. Irreversibly oxidized HNA2 was positively correlated with diabetes duration, especially in the T1DM group. HNA was increased in people with an increased HbA1c (>58 mmol/mol). Our results support the hypothesis that oxidative stress plays a crucial role in DM pathogenesis and emphasize the importance of diabetes control on systemic oxidative burden.
Topics: Humans; Glycated Hemoglobin; Diabetes Mellitus, Type 1; Serum Albumin, Human; Diabetes Mellitus, Type 2; Serum; Oxidative Stress; Oxidation-Reduction
PubMed: 38003446
DOI: 10.3390/ijms242216256 -
Clinical Chemistry and Laboratory... Nov 2023C-peptide and insulin are peptide hormones and their stability is affected by a number of pre-analytical factors. The study aimed to investigate the impact of sample...
OBJECTIVES
C-peptide and insulin are peptide hormones and their stability is affected by a number of pre-analytical factors. The study aimed to investigate the impact of sample type, storage temperature and time delays before centrifugation and analysis on the stability of C-peptide and insulin.
METHODS
Ten healthy non-diabetic adults in fasting and non-fasting state were enrolled. 40 mL of blood was collected from each participant into SST and dipotassium EDTA tubes. Samples were centrifuged immediately or at timed intervals (8, 12, 48 and 72 h). After baseline measurements on the Roche Cobas e602 analyzer using electrochemiluminescence immunoassays, aliquots were stored at room temperature (RT), 2-8 and -20 °C for 4 h to 30 days. The percentage deviation (PD) from baseline was calculated and a change greater than desirable biological variation total error was considered clinically significant.
RESULTS
C-peptide was more stable in separated serum than plasma (PD of -5 vs. -13 %) samples stored at 2-8 °C for 7 days and was most unstable at RT when centrifugation was delayed (PD -46 % in plasma and -74 % in serum after 48 h). Insulin was more stable in plasma than in serum under the different storage conditions with a minimum PD of -1% when stored at -20 °C for 30 days. When samples were kept unspun at RT for 72 h, PD was -23 and -80 % in plasma and serum, respectively.
CONCLUSIONS
C-peptide was more stable in serum provided the sample was centrifuged immediately and stored in the fridge or freezer while insulin was found to be more stable in EDTA plasma.
Topics: Adult; Humans; Insulin; C-Peptide; Edetic Acid; Plasma; Serum; Blood Specimen Collection; Temperature
PubMed: 37409980
DOI: 10.1515/cclm-2023-0339 -
Veterinary Clinical Pathology Dec 2023The Platelet function analyzer-200 (PFA-200) can determine the effect of clopidogrel in cats, but analysis traditionally must be performed at point-of-care (POC). The...
BACKGROUND
The Platelet function analyzer-200 (PFA-200) can determine the effect of clopidogrel in cats, but analysis traditionally must be performed at point-of-care (POC). The ability to ship samples of blood to a laboratory would allow widespread access.
OBJECTIVES
We aimed to validate the shipping of blood samples for PFA-200 analysis in cats to determine the effect of clopidogrel.
METHODS
Twenty healthy cats and 10 cats receiving clopidogrel were recruited. Blood was collected from cats and aliquoted into two samples, one was analyzed at POC within 2 hours using the PFA-200, and the other was packaged and transported to a location 4 km away, stored, and transported back to the lab for analysis the following day.
RESULTS
Median closure times (CTs) with the collagen/adenosine diphosphate (COL/ADP) cartridge in healthy cats were 51.5 seconds (POC) and 78.8 seconds (shipped), which were significantly different (P < 0.001), and for cats on clopidogrel, median CTs were 147.5 seconds (POC) and 190 seconds (shipped), which were not significantly different (P = 0.131). Median CTs with the P2Y cartridge in healthy cats were 50.5 seconds (POC) and 64.9 seconds (shipped), which were significantly different (P = 0.03), and in cats receiving clopidogrel, median CTs were 300 seconds (POC) and 300 seconds (shipped) which were not significantly different (P = 1.000). Reference intervals for CTs differed for COL/ADP at POC (19.8-89.7 seconds) and shipped (50.9-161.6 seconds) and for P2Y at POC (35.5-118.8 seconds) and shipped (35.1-108.9 seconds). Receiver operating characteristics showed similar areas under the curve (AUCROCs) regarding the effect of clopidogrel for COL/ADP at POC (0.994 seconds) and shipped (0.932) and for P2Y at POC (0.904 seconds) and shipped (0.975 seconds). When classifying for the presence of clopidogrel effects, Cohen's Kappa was 0.62 for COL/ADP and 1.00 for P2Y.
CONCLUSIONS
Shipping blood samples for PFA analysis are feasible with similar performance to POC analyses for determining the effect of clopidogrel in cats.
Topics: Animals; Cats; Adenosine Diphosphate; Blood Platelets; Clopidogrel; Platelet Aggregation; Platelet Aggregation Inhibitors; Platelet Function Tests; Specimen Handling
PubMed: 37488077
DOI: 10.1111/vcp.13265