Blood Chemistry Research Articles

Clinical Evaluation of a Novel, Patented Green Coffee Bean Extract (GCB70®), Enriched in 70% Chlorogenic Acid, in Overweight Individuals

Objective Obesity and overweight are challenging health problems of the millennium that lead to diabetes, hypertension, dyslipidemia, nonalcoholic fatty liver disease (NAFLD), and atherosclerosis. Green coffee bean exhibited significant promise in healthy weight management, potentiating glucose-insulin sensitization and supporting liver health. The safety and efficacy of a novel, patented water-soluble green coffee bean extract (GCB70® enriched in 70% total chlorogenic acid and <1% caffeine) was investigated in 105 participants for 12 consecutive weeks. An institutional review board and Drugs Controller General (India) (DCGI) approvals were obtained, and the study was registered at ClinicalTrials.gov. Method Body weight, body mass index (BMI), waist circumference, lipid profile, plasma leptin, glycosylated hemoglobin (HbA1c), and total blood chemistry were assessed over a period of 12 weeks of treatment. Safety was affirmed. Results GCB70 (500 mg BID) supplementation significantly reduced body weight (approximately 6%; p = 0.000**) in approximately 97% of the study population. About a 5.65% statistically significant reduction (p = 0.000**) in BMI was observed in 96% of the study volunteers. Waist circumference was significantly reduced by 6.77% and 6.62% in 98% of the male and female participants, respectively. Plasma leptin levels decreased by 13.6% in 99% of the study population as compared to the baseline value. Upon completion of 12 weeks’ treatment, fasting glucose levels decreased by 13.05% (p = 0.000**) in 79% of the study population. There was a statistically significant decrease in HbA1c levels in both male and female participants (p = 0.000**), while 86.7% of the study participants showed a statistically significant decrease in thyroid-stimulating hormone (TSH) levels (p = 0.000**). The mean decrease in TSH levels on completion of the treatment was 14.07% in the study population as compared to baseline levels. Total blood chemistry analysis exhibited broad-spectrum safety. Conclusions This investigation demonstrated that GCB70 is safe and efficacious in healthy weight management.

Low Phase Angle and Skeletal Muscle Index Increase Hospital-Acquired Infections During Stroke Rehabilitation

ObjectivesSarcopenia is common in patients with stroke and may increase the risk of medical complications such as infection. However, assessing sarcopenia in stroke patients with consciousness disturbance, aphasia or severe paralysis is challenging. This study aimed to investigate whether a combined assessment of phase angle (PhA) and skeletal muscle index (SMI), estimated using bioelectrical impedance analysis, was associated with 2 common nosocomial infections, hospital-acquired pneumonia (HAP) and urinary tract infection (UTI), during inpatient stroke rehabilitation. DesignSingle-center retrospective observational study. Settings and ParticipantsA total of 1068 patients with stroke admitted to a rehabilitation hospital between January 2016 and September 2019 were analyzed. MethodsThe study variables included demographic characteristics, comorbidities, stroke severity, blood chemistry and urine analysis, SMI, and PhA obtained using bioelectrical impedance analysis. Patients were classified as normal, low PhA only, low SMI only, and low PhA + SMI. Multivariate Cox proportional analysis was used to determine the variables associated with HAP and UTI. ResultsA combination of low PhA + SMI was observed in 429 (40.2%) patients. Over a median follow-up duration of 46 days, HAP occurred in 187 patients (17.5%) and UTI occurred in 155 patients (14.5%). The low PhA + SMI group showed a significantly higher incidence of HAP and UTI than the normal group (32.6% vs 4.6%, P < .001, for HAP; 20.7% vs 6.2%, P < .001, for UTI). In multivariate Cox analyses, low PhA and SMI were associated with significantly higher rates of HAP [hazard ratio (HR) 3.36, 95% CI 1.796-6.304, P < .001] and UTI (HR 1.71, 95% CI 1.002-2.947, P < .05) after adjusting for confounding variables. Conclusions and ImplicationsCombination of low PhA + SMI was independently associated with a higher risk of HAP and UTI in stroke patients who underwent inpatient rehabilitation. Measuring PhA and SMI using bioelectrical impedance analysis might be helpful in establishing care plans in these population.

Ramadan-specific nutrition education improves cardio-metabolic health and inflammation-a prospective nutrition intervention study from Pakistan.

There are recent reports that Ramadan fasting (RF) results in weight gain instead of weight loss. In addition, the data on the efficacy of brief nutrition education on healthy eating practices in Ramadan for better health are scarce. Therefore, a study was conducted to investigate the effects of brief nutrition education before the start of RF on healthy eating practices during RF. For this purpose, a prospective observational study focused on "Dietary Education and Awareness for Ramadan (DEAR)" as an intervention was carried out. The participants (n = 74) were recruited and divided into two groups, i.e., intervention and control groups (n = 37 each). As an intervention, nutrition education lessons were given before and during RF month. The control group did not attend these nutrition education lessons. Data on anthropometrics, dietary intake, and other parameters were collected at three time points: before, in the end, and 4 weeks after RF. Weight was measured in kg; height, waist circumference (WC), and hip circumference (HC) were measured in cm; and body mass index (BMI) was calculated. Waist-to-hip ratio (WHR) was calculated by dividing the waist value by the hip value. Body composition analysis was performed by the body composition analyzer (BF-907). Blood pressure (BP) was measured using a validated automated blood pressure. A 3-5 ml of venous blood was collected, and plasma and serum were separated. Serum and plasma samples were processed for general blood chemistry (blood lipid profile, glucose, and CRP) within 2 h. CRP was determined by the immunoturbidimetry method using an auto-analyzer. An enzyme-linked immunosorbent assay (ELISA) was used to determine cytokine/chemokines. Adherence to nutrition education (intervention) was assessed. The results show that nutrition education has positive effects on overall nutrition. Significant improvement in dietary adherence to dietary advice in the intervention group was noted. Significant BW loss (mean loss: 1.21 kg) in the intervention group was observed. The majority (63.3%) had lost BW ≥ 1.0 kg. Other changes observed as a result of the intervention included improvements in blood glucose, cholesterol, CRP levels, and systolic and diastolic BP. There was a notable shift in pro- and anti-inflammatory cytokine concentrations: IL-7, IL-4, and TGF-α decreased, while IL-2, TNF-α and resistin, IL-1 RA, IL-17 A, and sCD40 increased. In conclusion, RF resulted in a loss in mean BW and an improvement in related blood chemistry and cytokine profiles. Furthermore, nutrition education before RF resulted in better nutrition practices during RF and a desirable healthy BW, blood lipid, and cytokine profiles.

Wearable Fabric Sensor Using Functionalized MWCNT Nanocomposite Material

Despite the recent advancements in sensor detection of biomarkers in sweat, there is no sensor capable of long-term detection in constricted or load-bearing applications where other flexible plastic sensors might cause discomfort. In this work, a carbon nanotube-based fabric sensor capable of real-time detection of sodium in human sweat is presented. The sensor consists of an electrospun nylon-6 base functionalized with multi-walled carbon nanotubes for enhanced conductivity and p-tert-calix[4]arene for enhanced sodium selectivity. Monitoring sweat is a compelling choice to gain insight into a person’s hydration at a molecular level, as sweat is rich in physiological and metabolic information that can be obtained non-invasively. Body surface sweat is approximately 99% water but also contains over 40 other compounds, most notably the presence of extracellular ions: sodium and chloride. Sodium has been shown to have a notable correlation to body fluid balance and is lost in ample quantities during exercise, making it a promising candidate for the detection of dehydration (i.e., hypernatremia) and overhydration (i.e., hyponatremia). Other compounds commonly found in surface sweat include intracellular ions, metabolites, hormones, small proteins, and peptides. This rich composition and its correspondence to blood chemistry can be the key to accessing the body’s biomolecular (health) state through non-invasive monitoring and diagnostics.When monitoring biomarkers in sweat, the sampling step has the greatest impact on accuracy. Traditional methods for monitoring sweat sodium concentration involve collecting a sweat sample and performing chemical analysis in two separate steps. Typical sweat collection procedures include whole body washdown, sweat collection patches, arm bags, and Macroducts® which require some combination of tedious procedures, trained professionals, and large equipment for analysis. These methods are often limited by insufficient sample volumes, non-negligible sample evaporation, possible analyte degradation between sampling and analysis all of which strongly impacts the reliability and sensitivity of the measurement.To circumvent the problems caused by separating sampling and analysis, sweat sampling can also be completed using wearable sensors. Through in-situ sampling and analysis, wearable sweat sensors can perform the necessary real-time measurements with freshly generated sweat rather than a mixture of new and old sweat. Functional absorbent materials (e.g., paper, nonwoven, cellulosic, or hydrogels) are typically introduced between the skin and the sensing component; although as shown in this work, they can also be introduced as sensing components during sampling. The advantages of functional absorbent sensing materials are their low cost, multiple functions, real-time and continuous sensing, and most importantly the enhanced breathability of sweat glands. As such, these wearable sensors avoid the issue of altering the sweat composition underneath because they remove the old sweat accumulation as new sweat wicks into the material and washes away old sweat. This also allows physiological monitoring for longer periods of time than traditional epidermal microfluidic devices. Additionally, functional absorbent materials can also be integrated into fabrics with inherent moisture wicking properties to bring sweat to a sensing area.This work presents the characterization and on-body testing of a previously developed functional absorbent material sensor made of a multi-walled carbon nanotube (MWCNT) functionalized nylon-6 nanocomposite. The wearable-fabric sensor works by measuring resistance of the nanocomposite material as sodium binds to the sensor fabric. The functionalization of the nanocomposite material was verified by FTIR; while XRD spectra show that the electrospun nylon-6 is made of -form crystals and the resulting nanocomposite is intercalated/exfoliated with an increase in crystalline size. The optimized MWCNT/nylon-6 nanocomposite sensor has a MWCNT loading close to the percolation threshold at approximately 1-2 wt%. Experimental results demonstrated that the sensor is selective to sodium in sweat at a sensitivity of 10 mM sodium. When worn on the body (attached to human skin), the sensor is capable of real-time monitoring of sodium concentration in sweat in the physiologically relevant range of 10-110 mM with up to 85% accuracy. Additionally, the obtained signal can be sent out via Bluetooth so that changes in hydration status can be detected in real-time.Overall, this carbon nanotube-based fabric sensor can be integrated into “smarter” clothing to read an array of biomarkers in sweat. This could lead to better understanding of health and disease processes (e.g., analysis of common diabetic neurological complications, diagnosis of cystic fibrosis (CF), sweat monitoring for advanced prosthetic limb applications and bedridden patients, and athlete performance tracking) resulting in better treatments and health outcomes for all patients.

Experience of inulin use for correcting intestinal microbiota in patients suffering from recurrent vulvovaginal candidiasis: A prospective cohort comparative study

Aim. To evaluate the effectiveness of inulin for the adjustment of intestinal microflora in patients with recurrent vulvovaginal candidiasis (VVC).&#x0D; Materials and methods. A prospective cohort comparative study included 79 women aged 18 to 50 years. They were divided into three groups: the main group included 32 patients receiving complex treatment with a dietary supplement containing inulin derived from the "Extra" variety of Jerusalem artichoke root in combination with fluconazole; the comparison group included 27 patients receiving fluconazole only; and the control group included 20 healthy women. The patients were followed up for 12 months. Examination included stool culture for dysbiosis, complete blood count, urinalysis, blood chemistry (glucose, bilirubin, alanine aminotransferase, aspartate aminotransferase, total protein, urea, creatinine), examination of the vulva and vaginal mucosa, vaginal smear microscopy for microflora, Gram staining, real-time polymerase chain reaction for Mycoplasma hominis, Ureaplasma parvum, Trihomonas vaginalis, Chlamydia traсhomatis, Candida albicans. Subsequently, all subjects every three months underwent a bimanual examination, speculum examination of the cervical mucosa and vagina, Gram staining of vaginal mucosa and cervix swabs, a thorough interview, and history taking. At 12 months, stool was cultured for dysbiosis again, and the data were analyzed.&#x0D; Results. The VVC therapy in the group of inulin-containing dietary supplement was more effective: the recurrence rate was 3 times lower than with the standard treatment regimen.&#x0D; Conclusion. The use of dietary supplement containing inulin derived from the "Extra" variety of Jerusalem artichoke root in complex therapy for recurrent VVC can significantly improve the gut and vagina microflora condition and prevent the recurrence and normalization of stool in patients.

Toxoplasmosis in jungle dog (speothos venaticus): case study

This clinical case describes a bush dog (Speothos venaticus), a vulnerable species in Ecuador. The objective was to document a case of toxoplasmosis in a captive specimen at the Yanacocha Animal Rescue Center in the Amazon region of Ecuador. A 2-year-old, 6.7 kg specimen was brought to the VitalPet specialty clinic in Ambato with symptoms of ataxia, lethargy, and anorexia of 24 hours’ duration. Clinical examinations and blood tests, including complete blood count, comprehensive blood chemistry, hemoparasite tests, and immunoglobulin analysis for toxoplasmosis, were performed. Results showed neutrophilic leukocytosis, a hematocrit of 50%, and slight hyperglycemia. Infections by Ehrlichia, Anaplasma, Lyme, and Dirofilaria were ruled out. Clinical signs and high IgG and IgM antibody titers indicated that the patient was affected by toxoplasmosis

Synthetic and phytogenic antioxidants improve productive performance, antioxidant activity, gene expression, and offspring quality in breeder hens subjected to heat stress

This study aimed to investigate the efficacy of a synthetic source (a combination of vitamin E, vitamin C, selenium, and L-carnitine) and phytogenic sources (a combination of clove, green tea pomace, and Vietnamese coriander) in overcoming heat stress (HS) damage in female breeder hens on production, blood chemistry, sperm survival in the oviduct, antioxidant properties, gene expression, and quality of offspring. One hundred SUT female breeder hens were housed in individual cages and divided into 4 treatment groups: T1) basal diets in the thermoneutral (TN) zone; T2) basal diets under HS; 3) basal diets with synthetic antioxidants under HS; and T4) basal diets with phytochemical antioxidants under HS. The result revealed that HS condition had a negative effect on reducing final body weight, egg weight, and 1-day-old chick weight while increasing water intake and FCR and altered blood chemicals in breeder hens compared to TN breeder hens (P < 0.05). However, either synthetic or phytogenic antioxidants resulted in increased egg production and hatchability, while decreasing the number of late stages of embryo death during the incubation (P < 0.05). Furthermore, the synthetic antioxidants also improved the uniformity of chicks and reduced late-stage embryo death compared with phytogenic antioxidants (P < 0.05). HS breeder hens fed with either of the antioxidant sources exhibited higher antioxidant capacity in terms of DPPH and ABTS radical scavenging (in yolk, liver, and breast meat) and FRAP radical scavenging (in yolk and liver) and lower liver malondialdehyde than HS breeder hens fed with the control diet (P < 0.05). Additionally, the gene expression of antioxidant enzymes (superoxide dismutase, catalase, and glutathione peroxidase) in the liver was upregulated, whereas the expression of proinflammatory cytokines (nuclear factor-κB) and heat shock proteins (HSP70 and HSP90) was downregulated in breeder hens that received both antioxidant sources (P < 0.05). Future investigations should focus on the potential for combinations of synthetic and phytogenic antioxidants in diets for HS breeder hens.

Biological Selenium Nanoparticles in Quail Nutrition: Biosynthesis and its Impact on Performance, Carcass, Blood Chemistry, and Cecal Microbiota.

This study was conducted to examine the influence of dietary supplementation of biological nano-selenium (BNSe) on productive performance, hematology, blood chemistry, antioxidant status, immune response, cecal microbiota, and carcass traits of quails. In total, 180 Japanese quails (1week old) were randomly allocated into four groups, with five replicates of nine chicks each in a complete randomized design. The 1st group was fed a control diet without BNSe, and the 2nd, 3rd, and 4th treatments were fed diets supplemented with BNSe (0.2, 0.4, and 0.6g /kg feed, respectively). The best level of BNSe in body weight (BW) and body weight gain (BWG) parameters was 0.4g/kg diet. Feed conversion was improved (P < 0.01) by adding BNSe in quail feed compared with the basal diet without any supplementation. The inclusion of different BNSe levels (0.2, 0.4, 0.6g/kg) exhibited an insignificant influence on all carcass traits. The dietary addition of BNSe (0.4 and 0.6g/kg) significantly augmented aspartate aminotransferase (AST) activity (P = 0.0127), total protein and globulin (P < 0.05), white blood cells (WBCs) (P = 0.031), and red blood cells (RBCs) (P = 0.0414) compared with the control. The dietary BNSe supplementation significantly improved lipid parameters, antioxidant and immunological indices, and increased selenium level in the blood (P < 0.05). BNSe significantly increased (P = 0.0003) lactic acid bacteria population number and lowered the total number of yeasts, molds, total bacterial count, E. coli, Coliform, Salmonella, and Enterobacter (P < 0.0001). In conclusion, adding BNSe up to 0.4 and 0.6g/kg can boost the growth, lactic acid bacteria population number, hematology, immunological indices, antioxidant capacity, and lipid profile, as well as decline intestinal pathogens in growing quail.

The Effects of Geranylgeraniol on Blood Safety and Sex Hormone Profiles in Healthy Adults: A Dose-Escalation, Randomized, Placebo-Controlled Trial

Geranylgeraniol (GG), an ingredient extracted from the South American annatto plant, has been shown to benefit bone and muscle health, is crucial in the biosynthesis of menaquinone-4 and coenzyme Q10, and has pain and inflammation reduction activities. However, no known studies to date have demonstrated the safety and impact of GG supplementation in humans. This 8-week, randomized, placebo-controlled, dose-escalated trial was conducted to determine the effect of GG on blood safety and hormone markers in healthy adults. Sixty-six males and females between 30 and 49 years of age were supplemented with either GG or a placebo (PLA) for 8 weeks, with dose escalation from 150 mg to 300 mg occurring after 4 weeks in the treatment group. Changes in complete blood count and comprehensive metabolic panels were analyzed for whole study groups (males and females) while changes in sex hormone panels were analyzed for males and females independently. There were no significant changes in complete blood count, comprehensive metabolic panel, progesterone, estradiol, sex–hormone binding globulin, or dihydrotestosterone (p &gt; 0.05). An exploratory analysis of testosterone levels in a subgroup of males with baseline (Pre) total testosterone &lt; 700 ng/dL (GG = 15; PLA = 13) demonstrated a significant increase (p &lt; 0.05) from Pre to Week 8 in total-, free-, and bioavailable testosterone (+7.5%, +15.0%, and +14.8%, respectively). This study demonstrates that GG does not significantly change the composition of blood chemistry, hematology, or sex hormone profiles in adult males or females. Given the effects observed in the exploratory analysis in a subgroup of males, GG supplementation may be beneficial for testosterone enhancement in male populations subject to low testosterone (i.e., aging males and those with late-onset hypogonadism), but further research is needed.

Commentaries on “Reconsidering the path for neural and physiological methods in consumer psychology”

AbstractThe initial version of the article by Clithero, Karmarkar, Nave, and Plassmann (Journal of Consumer Psychology, 2024) was critiqued by open comments from a small group of scholars. Their suggestions encouraged the authors to clarify challenging relationships between brain processes and emotions, beliefs, and actions. The revision expanded fMRI and EEG to include measures of vision, facial expression, breathing, heart rhythms, and blood chemistry. The paper provides multiple avenues of joint work between neurological and psychological scholars. The comments below reflect different reactions to the final article. Wes Hutchinson acknowledges that neuroscience insights complement cognitive measures that generate explicit measures of thought, emotion, or preferences, but he warns that repeated measures over time are problematic for both types of measurement, and the inherent complexity of brain–behavior relationships is often underestimated. With both orientations, understanding the functioning of human behavior is akin to making sense of an orchestra, where the interactive blending of different instruments and musicians reflects a complex activity that generates sounds, emotions, and stories. Both consumer neuroscientists and psychologists need to broaden their paradigmatic approaches with bodily measures and advanced psychological procedures to overcome challenges to joint progress. Martin Reiman asserts that despite difficulties with measures that have different levels of abstraction or velocity, research has provided remarkable associations between brain activity and consumer behavior. Effective studies merging brain and behavior can effectively proceed with studies that differ in two dimensions: first, by altering the number of variables, and second, by shifting whether the scientific paradigm is inductive or deductive. In its simple form, the Excavation path explores brain activity when a person is exposed to specific statements or emotions. In its most challenging form, Integrative Studies generate predictions from theories that test the convergent validity of divergent measures and leverage skills from different researchers. Studies reflecting high levels on one dimension but low levels on the other can also provide fruitful research opportunities. Brian Knutson, like Reimann, counters the idea that consumer psychology has not lived up to its promises. He references studies showing that activity from very specific areas of the brain reliably predicts choices better than explicit ratings or choices. Such research generates deductions from increasingly precise neural maps that enable confirmation of theory. That said, he acknowledges that consumer neuroscience is not able to identify a brain button that would alter choice through manipulated neurostimulation. However, since human brains are similar across people, the depth of neural insights that are consistent across a small sample of 40 respondents may generate greater insights than conventional marketing research with 2000 respondents. The cost of neuroscience will further decrease with gains in reliability, validity, and generalizability, particularly if augmented with bodily measures. He acknowledges that the theoretical side has developed more slowly than applications, particularly applications that are supported by sponsoring organizations more satisfied with local insights than general models.

Therapeutic effects of in vivo administration of an inhibitor of tryptophan 2,3-dioxygenase (680c91) for the treatment of fibroids: a preclinical study

ObjectiveFibroids are characterized by marked overexpression of Tryptophan 2,3 dioxygenase (TDO2). The objective of this study was to determine the effectiveness of in vivo administration of an inhibitor of TDO2 (680C91) on fibroid size and gene expression. DesignAnimal and ex vivo human study. SubjectsSCID mice bearing human fibroid xenografts treated with vehicle and TDO2 inhibitor. InterventionDaily intraperitoneal administration of 680C91 or vehicle for two months and in vitro studies with fibroid explants. Main Outcome MeasuresTumor weight and gene expression profile of xenografts and in vitro mechanistic experiments using fibroid explants. ResultsCompound 680C91 was well-tolerated with no effects on blood chemistry and body weight. Treatment of mice with 680C91 resulted in 30% reduction in the weight of fibroid xenografts after two months of treatment and as expected lower levels of kynurenine, the byproduct of tryptophan degradation and an endogenous ligand of AhR (aryl hydrocarbon receptor) in the xenografts. The expression of CYP1B1 (cytochrome P450 family 1 subfamily B member 1), TGF-β3 (transforming growth factor beta 3), FN1 (fibronectin), CDK2 (cyclin-dependent kinase 2), E2F1 (E2F transcription factor 1), IL-8 (interleukin 8) and SPARC (secreted protein acidic and cysteine rich) mRNA, were lower in the xenografts of mice treated with 680C91 as compared with vehicle controls. Similarly, the protein abundance of collagen, FN1, CYP1B1 and SPARC were lower in the xenografts of 680C91 treated mice as compared with vehicle controls. Immunohistochemical analysis of xenografts indicated decreased expression of collagen, Ki67 and E2F1 but no significant changes in cleaved caspase 3 expression in mice treated with 680C91. The levels of kynurenine in the xenografts showed a direct correlation with the tumor weight and FN1 levels. In vitro studies with fibroid explants showed a significant induction of CYP1B1, TGF-β3, FN1, CDK2, E2F1, IL8 and SPARC mRNA by tryptophan which could be blocked by co-treatment with 680C91 and the AhR antagonist CH-223191. ConclusionThe results indicate that correction of aberrant tryptophan catabolism in fibroids could be an effective treatment through its effect to reduce cell proliferation and ECM accumulation.

Physiological response of roe deer (Capreolus capreolus) during stalking hunts depending on age

BackgroundThe European roe deer (Capreolus capreolus) is a species particularly reactive to all kinds of negative stimuli. Hunting activity is one of the most potent stressors that disturbs the welfare of wild animals. During stress, various endocrine responses are elicited to improve the physical performance of the affected individual. A commonly assessed hormone for overcoming stressful situations is cortisol (CORT). In this study, plasma CORT levels in roe deer were assessed during the season of the most intense stalking hunts in Poland (summer vs. late autumn), the sex of the harvested animals (males vs. females), and age of harvest animal. In addition, the health status of the roe doe was evaluated on the basis of selected indices of blood chemistry, which could be associated with circulating cortisol levels.ResultsThe mean cortisol levels were 58.066 ng/ml in the male group (summer) and 27.694 ng/ml in the female group (late autumn). Higher CORT levels were associated with a significantly lower of total cholesterol, lactate dehydrogenase, and uric acid (p < 0.05). Moreover, the mean concentration of uric acid was negatively correlated with the level of CORT in the male and female groups (p < 0.05). Together with the increase in mean CORT level, the HDL cholesterol of all the tested animals increased significantly (p < 0.05).ConclusionsHigher CORT in males during the summer than in females during the late autumn were most likely due to the arousal with the mating season. The level of CORT increased with the animals’ age. Uric acid and age are both predictors of roe deer’s serum CORT level.

Description of box trapping, immobilisation, anaesthesia monitoring and blood chemistry and serology in free-ranging European wildcats (Felis silvestris) in Southwest Germany

Specific questions in wildlife research and surveillance require safe and efficient capture, handling and anaesthesia protocols to enable sampling and transmitter placement in free-ranging individuals. For wild felids, various protocols are available, but detailed reports for European wildcats (Felis silvestris) are scarce. In particular, tools for anaesthesia monitoring under field conditions and reference values for heart rate, respiratory rate, oxygen saturation and body temperature are missing. In the present study, European wildcats were caught in box traps before being released into catch bags for manual restraint. Inside the bags, ketamine-xylazine anaesthesia was applied via intramuscular injection, adjusted to the animal’s body weight. During anaesthesia, samples were taken, and vital variables were monitored continuously. Haematology and blood chemistry parameters were obtained, along with serological markers for antibodies against feline immunodeficiency virus (FIV), feline coronavirus and antigens of feline leukaemia virus (FeLV). In total, 29 wildcats were captured, of which 21 were examined and marked with passive integrated transponders. Twelve wildcats were collared with GPS transmitters. Handling time under anaesthesia averaged 30 min (range 26–35 min). Heart rate ranged between 76 and 170 beats/min and respiratory rate between 20 and 52 breaths/min. Relative arterial oxygen saturation stayed mainly between 93 and 99%, and rectal temperature ranged between 36.2 and 40.2 °C. Further, FeLV antibodies were detected in 2/21 samples. The applied protocol facilitated safe and sufficient examination, sampling and transmitter placement, as well as the establishment of haematological and blood chemical values in free-ranging European wildcats for the first time.

Identifying human‐relevant genetic interactors of APOE using genetically diverse mouse strains

AbstractBackgroundAPOEe4 is the greatest risk factor for late‐onset Alzheimer’s disease (AD), accounting for ∼30% of genetic risk and significantly decreasing age of onset. APOE is implicated to play central roles across multiple pathways such as neuroinflammation, lipid metabolism and vascular health. Current mechanistic work has introduced humanized APOE alleles in mouse models of AD on a single laboratory mouse strain, C57BL/6J (B6). Recent work from our lab has taken advantage of genetically diverse wild‐derived strains which were captured from different geographical regions and then inbred, thus, their genomes are closer to recapitulating combinations that would exist in the natural world. Here, we seek to identify and validate human‐relevant genetic interactors of APOE within these contexts and in the presence of amyloid.MethodNovel wild‐derived mice were developed by backcrossing the respective strain (PWK/PhJ, WSB/EiJ) to B6.APOEe4 (N5). These were crossed with congenic APP/PS1 carriers, and offspring intercrossed to generate cohorts of 4‐ and 8‐month males and females homozygous for APOEe4, with and without APP/PS1. A cohort of B6.APOEe4 carrying APP/PS1 were also generated for comparison. Neuropathology was assessed along with blood chemistry, and transcriptional profiling was performed on brain hemispheres.ResultBrain transcriptional profiling revealed that APOEe4 had the greatest impact on differential gene expression (dE) in the PWK genetic context at both timepoints, with the strongest effect in females. Comparison with AMP‐AD human‐derived consensus clusters also indicated the broadest overlap when variants were present in the PWK context. Furthermore, APOEe4 genotype was sufficient to cause neuronal damage at 8 months, despite other studies indicating neuronal resilience to amyloid and aging in PWK. In both WSB and B6 carrying APP/PS1, APOEe4 increased the incidence and severity of cerebral amyloid angiopathy (CAA). Further neuropathological assessments are ongoing, and will be correlated with dE.ConclusionThis work highlights the importance of assessing risk alleles for AD in a variety of genetic contexts. We have also created genetically diverse panels of mice that carry humanized amyloid beta and humanized tau in that hope that these strains will provide an improved preclinical platform for therapeutic discovery and testing.

Pharmacology of a CpG-Linked Anti-Mir-126 Oligonucleotide Targeting Acute Myeloid Leukemia (AML) in Rats and Non-Human Primates (NHP)

Introduction: Acute myeloid leukemia (AML) is a lethal hematopoietic malignancy, characterized by the accumulation of clonal myeloid progenitor cells arrested in development. AML patients have dismal outcomes with a 5-year overall survival (2011 - 2017) of 29.5% (SEER). The persistence of leukemia stem cells (LSCs), a treatment-resistant primitive cell population able to inititiate and maintain AML, is thought to underlie relapse. Even newly approved drugs do not target LSCs efficiently. MicroRNA (miR)-126 is a non-coding regulatory RNA that contributes to LSC homeostasis in the bone marrow niche. Several lines of evidence validate miR-126 as a target in AML. Thus, we designed and made a novel anti-miR-126 oligonucleotide (oligo) linked to a CpG moiety, termed miRisten. miRisten incorporates modifications such as 2'-fluoro and phosphorothioate substituting phosphodiester in the sugar moiety of the nucleotide to enhance in vivo stability. miRisten eradicates LSCs in mouse models of AML and synergizes with FDA-approved AML drugs (PMID: 29505034, 34372909). Here, the results of IND-enabling toxicokinetic (TK) studies of miRisten in rats and NHP are presented. Objective: To investigate the pharmacological properties of miRisten to establish a safe dosing strategy to achieve prolonged knockdown of miR-126 for a first-in-human trial. Experimental Plan and Results: Good laboratory practice (GLP) TK studies were carried out in Sprague-Dawley rats. miRisten was given IV b.i.d. for 19 days at human equivalent doses (HED) of 1.6 and 4 mg/kg. miRisten was measured in plasma after the first dose. Plasma miRisten C max average was 3622 ± 335 and 6010 ± 175 nM post first doses of 1.6 and 4mg/kg, respectively. Plasma t 1/2 averaged 20.0 ± 3.8 minutes across both doses. The area under the curve (AUC 0 - ¥) were 638.9 ± 32 nMxhr (1.6 mg/kg) and 1313.5 ± 439 (4 mg/kg). A parallel toxicology study was undertaken documenting cage-side observations and body weight; complement activation, blood chemistries, urine and hematology were also evaluated. There were no unscheduled deaths in the toxicology segment. Four single NHP exploratory studies were also carried out (Fig. 1: Treatment schema). In each study, miRisten was given IV as a slow bolus (~5') b.i.d. and PK parameters evaluated in plasma. Study #1 &amp; #2: 1 wk at 4mg/kg; #3: 1 wk at 5 mg/kg; #4: 18 days at 4 mg/kg. In all studies, miRisten dosage was ramped up (1, 2, 4 mg/kg for all and 5 mg/kg at dose 4 for Study #3). Solumedrol was given as premedication for the first 3 doses in all studies to prevent known phosphorothioate-mediated complement activation and cytokine release syndrome. Blood draws for PK were carried out immediately following doses 3 and 12 (Studies 1 - 3) and after the last dose on Day 18 (Study 4). The average area under the curve AUC was 3860 ± 1189 (4 mg/kg) and 3954 ± 1569 nMxhr (5 mg/kg) which was greater than observed in mice given the equivalent HED in bolus doses (~6x, 465.0). The t 1/2 of plasma miRisten was 11.7 min on Day 3 and 12.0 min on Day 7, comparable to mouse (9.6 min) and rat (20 min). Cage-side observations, body weight, complement activation, blood chemistries, urine and hematology were also evaluated. There was no terminal sacrifice of NHP however, all animals tolerated the treatment without observable adverse events. Conclusion: The dose ramp-up in NHP combined with pre-medication with solumedrol were undertaken to prevent possible acute inflammatory reactivity leading potentially to death. There were no unscheduled deaths in any species studied. These data are consistent with the safety of miRisten given IV with a dose ramping regimen combined with solumedrol. The greater AUC in NHP may predict better potency in humans. In mice, miRisten at 1.6 mg/kg HED q.d. effectively eradicated LSCs [PMID 29505034].

Qpctl Inhibition, By Targeting the Inflammatory Tumor Microenvironment, Constitutes a Novel Therapeutic Approach for Diffuse Large B-Cell Lymphoma

We have previously described that diffuse large B-cell lymphomas (DLBCL) harboring an inflammatory and immunosuppressive microenvironment (i.e., IN-LME) carry independent poor prognosis while proof-of-principle experiments modifying the composition of the LME were sufficient to induce lymphoma regression in mice models ( Kotlov, Cancer Discovery 2021). However, identification of pharmaceutical approaches that by modifying the LME will induce an anti-lymphoma effect while being suitable for short-term human translation remains challenging. The IN-LME is characterized by the presence of pro-inflammatory and immunosuppressive chemokines and cytokines that frequently undergo post-translational modifications. QPCTL (glutaminyl-peptide cyclotransferase-like) is an intracellular enzyme that mediates a N-terminal pyroglutamylation of CD47 and several chemokines, including CCL2 and CCL7, ultimately resulting in their sustained expression and/or secretion. SC-2882 is a first-in-class specific QPCTL inhibitor that induces secondary proteolytic degradation of the monocyte chemoattractants CCL2 and CCL7 ( da Silva, Nature Immunology 2022) and inactivation of the “do-not-eat-me” signal CD47 ( LogtenbergNature Medicine 2019). In a dataset of 470 DLBCL patients we found that the expression of QPCTL correlates negatively with overall survival and progression free survival (0.019 and &amp;lt;0.001, respectively). To determine whether QPCTL could be a potential target in DLBCL, we investigated its expression along with its targets CCL2, CCL7 and CD47 in additional 216 DLBCL cases categorized by LME. We found that while QPCTL and CD47 are expressed at similar levels in all DLBCL cases regardless their COO class and LME category, CCL2 was highly expressed in DLBCL harboring a IN-LME (p &amp;lt;0.05 vs. other LME categories), that also correlate with higher presence of tumor-associated macrophages (TAMs) and exhausted CD8+PD1 HIGH T cells (p &amp;lt;0.01 and p&amp;lt;0.001, vs. other LME categories, respectively). CCL7 was not expressed at detectable levels in majority of LMEs in DLBCL. In preclinical models, murine and human DLBCL cell lines expressed baseline QPCTL and CD47 while CCL2 and CCL7 expression (qPCR) and secretion (ELISA) increased at higher levels upon culturing these cells for 24 h embedded in splenic extracellular matrix 3D cultures. Exposure of a panel of 3 murine and 5 human DLBCL cells to increasing concentrations of SC-2882 (vs. vehicle) for up to 72 h and up to a 10 micromolar dose had no noticeable decrease in cell viability or proliferation. However, SC-2882 (vs. vehicle) at nanomolar concentrations increased anti-Cd20-dependent lymphoma cell phagocytosis by in vitro differentiated macrophages (from splenic monocytes in mice and THP1 cells in humans) in 3D co-cultures (p &amp;lt; 0.05, for all conditions). To further test the anti-lymphoma effect of SC-2882 we used a murine GCB-DLBCL model by implanting GEMM-derived aggressive Ezh2 mutant (EZH2 Y646equivalent) + BCL2 amplified lymphoma cells in the spleen of immunocompetent C57BL/mice (n=14). After implantation, mice were randomized to receive SC-2882 by oral gavage (n=7) or vehicle (n=14) for 14 days. Tumor burden was followed by luciferase imaging (Caliper), and spleens were analyzed at sacrifice by multiparametric imaging, flow-cytometry, and spatial transcriptomics. Oral SC-2882 (vs. vehicle) induced a significant decrease in tumor growth (p=0.0079, Figure 1) without evidence of systemic toxicity by body weight, blood cell count and chemistry panel. Analysis of the LME in remnant splenic tumors showed in SC-2882 (vs. vehicle) treated mice a significant decrease in lymphoma cells and Ki67+ cells (p&amp;lt;0.001) as well as F480+ TAMs (p&amp;lt;0.01) while there was an increase of CD3+ T cell infiltration (p&amp;lt;0.001). We also found a decrease in Cd4+Foxp3+ Tregs and increase in Cd8+Gzb+ cells in SC-2882 treated mice by flow-cytometry. These changes mirrored the findings in the spatial transcriptomics. Additional analysis of LME cell subpopulations by SC-RNA-seq is ongoing. Overall, these data indicate that QPCTL is a potential target in DLBCL, and that QPCTL inhibition exhibits potent anti-lymphoma effects primarily by targeting the IN-LME.

Development of Thioredoxin Monothiol Derivatives for Mitigating Radiation Induced Hematopoietic Injury

Hematopoietic stem cells (HSCs) are among the most sensitive cells to radiation injury and contribute to many of the manifestations of acute radiation syndrome. There is an unmet need for effective agents that can be used to rescue radiation injury and enhance HSC recovery especially when administered post-irradiation. We previously identified the stress-protective human thioredoxin-1 protein (TRX) as a novel molecule for enhancing HSC recovery in mouse models of radiation injury. Treatment of lethally irradiated mice with recombinant TRX 24 hours after irradiation increased survival by 45%. However, native or recombinant TRX has a very short half-life (0.5-1 hr), which limits its clinical application, and the ability of exogenous TRX to internalize to the cytosol and support cellular proliferation is a potential concern. To overcome these limitations, an engineered monocysteinic, chemically pre-reduced TRX variantin which the second active-site Cys at position 35 has been replaced by Ser was developed (ORP100S). This modification stabilizes the mixed-disulfide transition state intermediate formed during docking and reduction of TRX-specific target protein Cys disulfide bonds. The resulting ORP100S Cys 32-target Cys linkage shifts the thiol-disulfide equilibrium and confers a longer duration and more potent activity compared to native TRX. Compared to TRX, ORP100S does not increase multiplication of EML hematopoietic cells in vitro, consistent with enhanced extracellular regulatory/stress-protective activity and attenuated intracellular proliferative activity. We first tested the effectiveness of ORP100S in mitigating lethal total body irradiation (TBI) in C57Bl/6 mice. Animals were irradiated (9.5 Gy) and 24 hours later were administered (IV, tail-vein injection) either PBS control buffer or ORP100S in PBS at 32 μg/mouse, every other day for a total of five doses. While all lethally-irradiated mice receiving PBS alone died within 2 weeks, half of the mice receiving ORP100S survived through termination of the study at 40 days. We further performed a dose-ranging efficacy study. At 24 hrs after TBI (8.45Gy), C57Bl/6 mice were given PBS or ORP100S in PBS (32 μg, 64 μg, 128 μg, 320 μg IV; 128μg SQ) every other day for a total of five doses. Nine of ten 128 μg SQ ORP100S-treated mice and all treatment groups given ORP100S at greater than 64 μg IV survived to the end of study on Day 45. Because subcutaneous injection is a preferred route of administration in the event of radiation exposure incidents where large populations may require treatment in the field, we performed a pharmacokinetic (PK) study to establish optimal SQ dosing of ORP100S, the timing of initial administration (24 hr or 48 hr after radiation), and the duration of treatment (one injection or five injections). PK and pharmacodynamic (PD) studies were also performed to compare IV and SQ administration of 64 μg or 128 μg ORP100S. Plasma samples were collected before injection and at 15 min, 30 min, 60 min, 2 hr, 4 hr, 8 hr and 16 hr. ORP100S levels in plasma were analyzed using a custom hybrid immunocapture LC/MS-MS assay with selective quantification of the ORP100S tryptic peptide SMPTFQFFK following immunoprecipitation. We found that at two hr post injection, SQ administration of ORP100S achieved similar plasma level of ORP100S compared to IV injection. We also performed a repeated-dose toxicology study of ORP100S in C57Bl/6 mice. Animals were injected with 0, 128 μg, 1,280 μg, 2,560 μg, 5,120 μg IV or SQ in a volume of 200 μl once daily for five days. The mice were weighed daily and monitored for any changes in activities and behaviors. Terminal blood samples were collected at euthanasia the day following the final dose. Blood cell counts, chemistry panels, and biomarkers relevant to radiation effect were measured, and organs (brain, lung, heart, kidney, liver, spleen, bone marrow) were harvested for histological examination. A confirmatory study utilizing non-human primates (cynomolgous macaques) is in progress utilizing a sub-lethal 4 Gy radiation dose. In summary, we demonstrate that ORP100S, a monocysteinic, thioredoxin derivative, is a potent mitigator of radiation-induced hematopoietic injury even when delivered many hours post-exposure. Our preclinical study results support safety and efficacy with clear potential for this novel agent to advance to clinical studies for the treatment of acute radiation syndrome.

Assessment of Circulating Retinol Binding Protein-4 Levels in Patients with Compound Heterozygosity for Hb S and β-Thalassemia

Background: Retinol Binding Protein-4 (RBP-4) is a member of the lipocalin family of proteins and the major transport protein of the hydrophobic molecule retinol, also known as vitamin A, in the circulation. Expression of RBP4 is highest in the liver, where most of the body's vitamin A reserves are stored as retinyl esters. For the mobilization of vitamin A from the liver, retinyl esters are hydrolyzed to retinol, which then binds to RBP-4 in the hepatocyte. After associating with transthyretin (TTR), the retinol/RBP-4/TTR complex is released into the bloodstream and delivers retinol to tissues via binding to specific membrane receptors. While, after its dissociation from TTR and retinol release, retinol-free apo-RBP4 in the circulation is filtrated by the kidney. More than 99% of that is reabsorbed by the proximal renal tubule, which renders urinary RBP-4 a highly sensitive marker for tubular dysfunction. We and other have shown previously that the patients with Sickle Cell Disease (SCD) have deficiencies or suboptimal levels of vitamin A, while recently (Br J Haematol. 2023;doi:10.1111/bjh.18862) higher urine RBP-4 in patients with SCD especially in those with persistent albuminuria but without an association of urine RBP-4 levels with estimated glomerular filtration rate (eGFR). In this context, we aimed to investigate the potential clinical significance of circulating plasma RBP-4 levels and its association with disease features in Caucasian patients with compound heterozygosity for HbS and β-thalassemia (HbS/β thal). Patients and Methods: Ninety Caucasian adult patients with HbS/β thal at steady phase were included in the study, while 22 apparently healthy individuals of similar age and gender served as controls. None of the patients was diabetic and has received any transfusions at least 6-monthes before enrollment in the study. RBP-4 concentration was measured with an immunoturbidimetric assay applied in an automated chemistry analyzer, along with hematologic, and series of blood chemistry parameters reflecting renal, cardiac and endothelial dysfunction. eGFR values were calculated using the CKD-EPI (Cystatin C and/or Creatinine equations). Results: We found that: a) Plasma RBP-4 levels were significantly lower in patients with HbS/β thal compared to controls, 30.5±2.7mg/L (Confidence Intervals (CI): 28.8;32.2mg/mL) vs 36.3±3.6mg/L (CI: 32.7;39.9mg/mL), p&amp;lt;0.003. The patients with β + genotypehave higher RBP-4 levels than those with β 0 genotype (p&amp;lt;0.05), whereas patients treated with hydroxycarbamide(50/90) are more likely to express higher RBP-4 levels, p-trend=0.06; b) RBP-4 levels in patients with HbS/β thal correlated positively with their age, body mass index (BMI) and Hb levels (r=0.264, p=0.012; r=0.334, p=0.001 and r=0.209, p&amp;lt;0.05, respectively; c) RBP-4 levels in patients with HbS/β thal correlated with total bilirubin, total cholesterol, triglycerides and uric acid (r=-0.452, p&amp;lt;0.001; r=0.395, p=0.004; r=0.292, p=0.015 and r=0.435, p&amp;lt;0.001, respectively); d) RBP-4 levels in patients with HbS/β thal correlated negatively with eGFR values rho=-0.360, p=0.003 and hs-CRP concentrations r=-0.309, p=0.004 and e) No correlations were found between RBP-4 levels with markers of cardiac and endothelial dysfunction such as High Sensitivity Troponin T, Growth Differentiation Factor-15, von Willebrand factor antigen, Α Disintegrin and Metalloproteinase with Thrombospondin Type 1 Motif 13 antigen and P-Selectin, as well as with number of vaso-occlusive crisis and mean pulmonary arterial pressure in the patients. Conclusions: These findings demonstrate a multifactorial role of RBP-4 in patients with HbS/β thal as the circulating levels of the protein correlate significantly with markers of erythropoiesis, inflammation and renal function. It is unclear of this study if RBP-4 has any specific functions in patients with HbS/β thal. However, RBP-4 should be more investigated in order to determine its clinical significance and in the case of sickle cell nephropathy both plasma and urine measurements should be considered.

UROLITHIASIS IN DOMESTIC MALE CAT

Urolithiasis is a disease caused by the presence of uroliths, calculi, crystals, or excessive sediment in the urinary tract. The aim is to find out about the methods of diagnosis, as well as control and treatment that are permanent and efficient and in accordance with the causes of cases of urolithiasis that occur in cats. A local male cat named Benji, 4 years old and weighing 5.42 kg, has complaints of decreased appetite, weakness, stranguria, hematuria, and difficulty urinating which has been going on for three days. Previously case animals also show pain responses when they want to urinate. The physical examination results found abdominal distension, pain on palpation, and an enlargement of the urinary bladder. Urinalysis results showed leukocyt +5, protein +3, and blood cells +3, blood chemistry examination showed an increase in glucose (191 mg/dL), globulin (5.3 mg/dL), and a decrease in alkaline phosphate (&lt;10 U/L), ultrasonographic examination showed a hyperechoic mass formation located in the lumen of the bladder and the presence of sediment in the bladder, radiographic examination showed enlargement of the bladder, urine sedimentation examination showed the presence of struvite. The therapy was Pentosan polysulphate 3 mg/kg, 0.16 ml/kg given only once, Prazosin 0.5 mg/kg BW twice a day for five days, Meloxicam 0.18 ml once a day for three days, and Amoxiclav twice a day for 10 days. It can be concluded that based on anamnesis, clinical examination, and supporting examinations, the case animal was diagnosed with urolithiasis. After giving therapy for four days, the case cat can urinate normally and the urine looks clear. Suggestions in order to prevent the recurrence of urolithiasis infections, it is necessary to educate clients about cat care, feed and drink. This indicates that the case cat's condition is getting better.

Glutathione and bicarbonate nanoparticles improve mucociliary transport in cystic fibrosis epithelia.

Cystic fibrosis (CF) airway disease is characterized by thick mucus and impaired mucociliary transport (MCT). Loss of functional cystic fibrosis transmembrane receptor (CFTR) leads to acidification and oxidation of airway surface mucus. Replacing bicarbonate (HCO3 -) topically fails due to rapid reabsorption and neutralization, while the scavenging antioxidant, glutathione sulfhydryl (GSH), is also rapidly degraded. The objective of this study is to investigate GSH/NaHCO3 nanoparticles as novel strategy for CF airway disease. GSH/NaHCO3 poly (lactic-co-glycolic acid) nanoparticles were tested on primary CF (F508del/F508del) epithelial cultures to evaluate dose-release curves, surface pH, toxicity, and MCT indices using micro-optical coherence tomography. In vivo tests were performed in three rabbits to assess safety and toxicity. After 1week of daily injections, histopathology, computed tomography (CT), and blood chemistries were performed and compared to three controls. Fluorescent nanoparticles were injected into a rabbit with maxillary sinusitis and explants visualized with confocal microscopy. Sustained release of GSH and HCO3 - with no cellular toxicity was observed over 2weeks. Apical surface pH gradually increased from 6.54±0.13 (baseline) to 7.07±0.10 (24h) (p<0.001) and 6.87 ± 0.05 at 14 days (p<0.001). MCT, ciliary beat frequency, and periciliary liquid were significantly increased. When injected into the maxillary sinuses of rabbits, there were no changes to histology, CT, or blood chemistries. Nanoparticles penetrated rabbit sinusitis mucus on confocal microscopy. Findings suggest that GSH/NaHCO3 - nanoparticles are a promising treatment option for viscous mucus in CF and other respiratory diseases of mucus obstruction such as chronic rhinosinusitis.