Human Plasma Research Articles

Inhibitory effect of flavonoids on multidrug and toxin extrusion protein 1 function: Implications for food/herb-drug interaction and drug-induced kidney injury.

Multidrug and toxin extrusion protein 1 (MATE1), an efflux transporter mainly expressed in renal proximal tubules, mediates the renal secretion of organic cationic drugs. The inhibition of MATE1 will impair the excretion of drugs into the tubular lumen, leading to the accumulation of nephrotoxic drugs in the kidney and consequently potentiating nephrotoxicity. Screening and identifying potent MATE1 inhibitors can predict or minimize the risk of drug-induced kidney injury. Flavonoids, a group of polyphenols commonly found in foodstuffs and herbal products, have been reported to cause transporter-mediated food/herb-drug interactions. Our objective was to investigate the inhibitory effects of flavonoids on MATE1 in vitro and in vivo and to assess the effects of flavonoids on cisplatin-induced kidney injury. Thirteen flavonoids exhibited significant transport activity inhibition (>50%) on MATE1 in MATE1-MDCK cells. Among them, the six strongest flavonoid inhibitors, including irisflorentin, silymarin, isosilybin, sinensetin, tangeretin, and nobiletin, markedly increased cisplatin cytotoxicity in these cells. In cisplatin-induced in vivo renal injury models, irisflorentin, isosilybin, and sinensetin also increased serum creatinine and blood urea nitrogen levels to different degrees, especially irisflorentin, which exhibited the most potent nephrotoxicity with cisplatin. The pharmacophore model indicated that the hydrogen bond acceptors at the 3, 5, and 7 positions may play a critical role in the inhibitory effect of flavonoids on MATE1. Our findings provide helpful information for predicting the potential risks of flavonoid-containing food/herb-drug interactions and avoiding the exacerbation of drug-induced kidney injury via MATE1 mediation.

The study of the influence of <i>Thlaspi arvense</i> L. on the characteristics of the reproductive system of male rats

Introduction. The development of new effective and safe herbal medicines capable of having a positive effect on reproductive status is an urgent task of domestic pharmacy. Medicinal plants are considered as alternative therapies aimed at increasing testosterone levels and fertility in men. The arsenal of medicinal plants used in traditional medicine for the treatment of diseases in men is mainly represented by phytoadaptogens, most of which have a small resource potential in Russia, therefore, it is relevant to search for plants with a sufficient raw material base and with the potential for cultivation. Thlaspi arvense L. is a promising medicinal plant, as it is widely used in folk medicine as a diuretic, anti-inflammatory, diaphoretic, antihistamine, hemostatic, astringent, has a positive effect on the processes of spermatogenesis and is widely distributed in our country. However, information on the chemical composition and biological activity of Thlaspi arvense L. they are insufficient, which shows the relevance of its further more detailed study in order to substantiate the possibility of application in practical medicine and solutions to the issues of standardization of medicinal plant raw materials.Aim. The study of the influence of Thlaspi arvense L. on the characteristics of the reproductive system of male rats.Materials and methods. The object of the study was an infusion of herbа Thlaspi arvense L. The features of the reproductive behavior of male rats after 21-day administration of the infusion of grass yarutka field were studied using tests that allow quantifying the severity of sexual motivation and sexual activity of males. The viability of spermatozoa in the ejaculate of rats was assessed, the total number of spermatozoa (ACS, million), degenerative and immobile forms (%) was calculated. Morphophysiological parameters of spermatogenesis of rats of control and experimental groups of animals were studied using classical histological methods. The concentration of testosterone in the blood serum of experimental groups of animals was determined by the enzyme immunoassay.Results and discussion. The results obtained allow us to conclude that the fertility of rats has increased against the background of the course administration of the infusion of herbа Thlaspi arvense L., as evidenced by an increase in the testosterone content in the blood, improvement of spermogram indicators and morphophysiological characteristics of spermatogenesis in the testicles of rats. The use of the infusion of herbа Thlaspi arvense L. has a protective effect on spermatogenesis. An increase in the thickness of the spermatogenic epithelium, the diameter of the cross-section of the convoluted seminal tubules and the index of spermatogenesis compared with the control group of animals was revealed. The positive effect of Thlaspi arvense L. on indicators of sexual motivation and sexual activity of male rats is shown.Conclusion. Thlaspi arvense L. is a promising medicinal plant, as it is able to have a positive effect on libido, the number and mobility of spermatozoa, the production of sex hormones, spermatogenesis, as well as on the pituitary-gonadal axis, which is associated with the content of a complex of biologically active substances in it.

Biological therapy of bariatric surgery complications

In recent years, the number of performed bariatric interventions has increased significantly. Despite the high effectiveness of this method of treatment, the occurrence of complications is not uncommon. The most common complications are stapler line leaks, occurring in 1.5–2.4% of cases; anastomotic leakage after One Anastomosis Gastric Bypass – in 2.2–8% of cases. The problem with all methods is the increased duration of treatment and the need to go without food for significant periods of time, which means starting parenteral nutrition and risking malnutrition. The article demonstrates the effectiveness of biological therapy for complications of bariatric surgery in a series of clinical cases. Clinical cases of 2 patients with anastomotic leakage after One Anastomosis Gastric Bypass are described; also 2 patients with incompetent staple line after laparoscopic Sleeve Gastrectomy. PRP therapy, stromal vascular fraction (SVF) and fibrin glue were used. Platelet-rich plasma is an autologous blood serum containing high concentrations of platelets and growth factors. As a result, after 4 courses of PRP therapy, the patient with the failure of the upper third of the stapler line received an application of twocomponent fibrin glue “Kriofit” with a positive effect. In addition, positive dynamics was observed in the patient with the use of threecomponent therapy of anastomotic leakage. Based on the above facts, it can be concluded, that biological therapy demonstrates great potential value in the treatment of bariatric surgery complications such as stapler line leaks, and anastomotic leakage.

Pentacenequinone-Modulated 2D GdSn-PQ Nanosheets as a Fluorescent Probe for the Detection of Enrofloxacin in Biological and Environmental Samples.

The fate and effects of fluoroquinolone antibacterial (FQ) on the environment are important since there appears to be a surge in FQ resistance like enrofloxacin (ENR) in both environmental and clinical organisms. Numerous reports indicate that the sensing capabilities of these antibiotics need to be improved. Here, we have investigated the interaction of ENR with our synthesized pentacenequinone-modulated gadolinium-tin (GdSn-PQ) nanosheets and the formation of intermolecular interactions that caused the occurrence of aggregation-induced emission enhancement. The concept for designing hybrid metallic nanosheets comes from the unique features inherited from the parent organic precursor. Due to the distinct interaction between ENR and GdSn-PQ, the interstate conversion (ISC) between GdSn-PQ and ENR induces a significant wavelength shift in photoluminescence (PL), improving reliability, selectivity, and visibility compared to quenching- or AIEE-based methods without peak shifts, allowing for highly sensitive and visually detectable analyses. The fluorescence signal of GdSn-PQ exhibited a linear relationship (R2 = 0.9911), with the added ENR concentrations ranging from 5 to 90 nM, with a detection limit of 0.10 nM. We have demonstrated its potential and wide use in the detection of ENR in biological samples (human urine and blood serum) and environmental samples (tap water and seawater) with a recovery rate of 98- 108%. The current approach has demonstrated that the 2D GdSn-PQ nanosheet is a novel and powerful platform for future biological and environmental studies.

Single-Cell Dissection of the Immune Response After Acute Myocardial Infarction.

The immune system's role in ST-segment-elevated myocardial infarction (STEMI) remains poorly characterized but is an important driver of recurrent cardiovascular events. While anti-inflammatory drugs show promise in reducing recurrence risk, their broad immune system impairment may induce severe side effects. To overcome these challenges, a nuanced understanding of the immune response to STEMI is needed. For this, we compared peripheral blood mononuclear single-cell RNA-sequencing (scRNA-seq) and plasma protein expression over time (hospital admission, 24 hours, and 6-8 weeks post-STEMI) in 38 patients and 38 controls (95 995 diseased and 33 878 control peripheral blood mononuclear cells). Compared with controls, classical monocytes were increased and CD56dim natural killer cells were decreased in patients with STEMI at admission and persisted until 24 hours post-STEMI. The largest gene expression changes were observed in monocytes, associating with changes in toll-like receptor, interferon, and interleukin signaling activity. Finally, a targeted cardiovascular biomarker panel revealed expression changes in 33/92 plasma proteins post-STEMI. Interestingly, interleukin-6R, MMP9 (matrix metalloproteinase-9), and LDLR (low-density lipoprotein receptor) were affected by coronary artery disease-associated genetic risk variation, disease status, and time post-STEMI, indicating the importance of considering these aspects when defining potential future therapies. Our analyses revealed the immunologic pathways disturbed by STEMI, specifying affected cell types and disease stages. Additionally, we provide insights into patients expected to benefit most from anti-inflammatory treatments by identifying the genetic variants and disease stage at which these variants affect the outcome of these (drug-targeted) pathways. These findings advance our knowledge of the immune response post-STEMI and provide guidance for future therapeutic studies.

Search in the Periphery for Potential Inflammatory Biomarkers of Dementia with Lewy Bodies and Alzheimer's Disease.

Neuroinflammation, with altered peripheral proinflammatory cytokine production, plays a major role in the pathogenesis of neurodegenerative diseases, such as Alzheimer's disease (AD), while the role of inflammation in dementia with Lewy bodies (DLB) is less known and the results of different studies are often in disagreement. The present study aimed to investigate the levels of TNFα and IL-6 in serum and supernatants, and the related DNA methylation in patients affected by DLB and AD compared to healthy controls (HCs), to clarify the role of epigenetic mechanisms of DNA promoter methylation on of pro-inflammatory cytokines overproduction. Twenty-one patients with DLB and fourteen with AD were frequency-matched for age and sex with eleven HCs. Clinical evaluation, TNFα and IL-6 gene methylation status, cytokine gene expression levels and production in serum and peripheral blood mononuclear cell (PBMC) supernatants were performed. In AD and DLB patients, higher serum levels of IL-6 and TNFα were detected than in HCs. Differences in LPS-stimulated versus spontaneous PBMCs were observed between DLB, AD, and HC in the levels of TNFα (p = 0.027) and IL-6 (p < 0.001). Higher levels were also revealed for sIL-6R in DLB (p < 0.001) and AD (p < 0.001) in comparison with HC.DNA hypomethylation in IL-6 and TNFα CpG promoter sites was detected for DLB and AD patients compared to the corresponding site in HCs. Our preliminary study documented increased levels of IL-6 and TNFα in DLB and AD patients to HCs. This overproduction can be due to epigenetic mechanisms regarding the hypomethylation of DNA promoters.

PECULIARITIES OF THE INFLUENCE OF TYLOZIN ON THE METABOLISM OF PROTEINS AND AMINO ACIDS IN CHICKEN

Over the last decade, poultry farming, both for meat and eggs, has been widely developed in our country. Poultry products are widely used in the diets of the population. In this problem, it is necessary to observe the careful application of antimicrobial drugs to productive animals, as they are used by humans and not to disturb the ecosystem and not cause the formation of resistant microflora. Taking into consideration that veterinary medicine should be more preventive than curative, the problems of poultry treatment are poorly described, it was decided that it is necessary to work with healthy organisms. Because the physiological reactivity of the body changes with pathologies even without medical interventions. Methods of using macrolides and other pharmacological agents in sick animals and poultry are described in the literature, but there are not enough works that would highlight the data on the effect of this agent on protein and amino acid metabolism in healthy chickens. In addition, it is known that metabolic processes in poultry take place with a fairly high intensity, and this is definitely reflected in the amount of doses of various drugs, and antibiotics in particular. Medicines in the bird's body, accordingly, are biotransformed more quickly and removed from the body faster with droppings and eggs, others accumulate in various organs and muscles. Therefore, it is important to know how the biosynthesis of the main proteins and the level of free amino acids changes under the action of the antimicrobial agent Tylozin on the days of administration and in the period after the administration of the antibiotic. The results of the research are informative. The work is devoted to the study of the influence of an antimicrobial drug from the pharmacological group of macrolides on the metabolism of amino acids and proteins in chickens in a comparative aspect. At the same time, egg production in laying hens aged 140-150 days was taken into account. The influence of different doses of Tylosin on the protein and amino acid composition of chickens during the period of use and after the end of drug administration was studied. It was established that changes in the metabolism of total protein, its fractions and amino acid nitrogen in the blood serum occur naturally, long-term, in different directions and, proportionally, depend on the dose of the drug. The effect of the drug was studied in minimum, average and maximum doses.

INVESTIGATION OF THE EFFICIENCY OF NON-STEROIDAL ANTI-INFLAMMATORY VETERINARY MEDICINAL PRODUCT IN THE COMPLEX TREATMENT OF DISEASES OF THE MUSCULOSKELETAL SYSTEM OF PIGS

The article presents the results of clinical trials of a non-steroidal anti-inflammatory veterinary drug when administered to fattening piglets with clinical signs of musculoskeletal diseases. For the treatment of the diseases, piglets of the experimental (I) and control (II) groups received an antimicrobial drug based on the tetracycline antibiotic. The animals of the experimental group were additionally administered orally with drinking water the test non-steroidal anti-inflammatory drug based on acetylsalicylic, ascorbic and citric acids, in the recommended dosage by the manufacturer. Blood for laboratory tests was selected from piglets before and after the use of drugs. Studies of hematological and biochemical parameters were conducted according to generally accepted methods. The therapeutic efficiecy of the drug and its effect on the morphofunctional state of the body was evaluated by comparing the statistically processed data obtained before and after treatment, as well as in comparison with the indicators of animals of the control group. The analysis of hematological parameters of piglets during the experiment showed a decrease in the number of leukocytes in the animal’s blood after the use of drugs. The normalization of the leukogram was also noted by reducing the relative number of rod-nuclear neutrophils. In particular, this indicator decreased by 17.4% in group I piglets, and by 12% in group II piglets. It is noted that the clinical response to therapy was faster and more pronounced in animals, which used a non-steroidal anti-inflammatory drug in combination with an antimicrobial drug than in piglets, which were treated only by antibiotic. According to the results of biochemical studies, it was found that the content of urea in the serum of animals of both groups before the use of drugs exceeded more than 3 times the upper limit of physiological values for a given age group of piglets. After therapy with the drugs a significant decrease in the urea content in the blood serum of groups 1 and 2 animals was noted by 3.42 and 2.98 times, respectively. It is established that the activity of marker enzymes of the liver physiological state (ALT, AST) in the serum of group I piglets after complex treatment with the studied drugs was slightly reduced and was within the physiological values. In the serum of group II animals, which were treated only with antimicrobial drug, ALT activity increased by 12,0 %, and AST – by 38,6 %, compared to the initial data, and exceeded the limit of physiological values. An increase in the content of iron by 37.4 % in the blood of group I piglets, compared to the period before drug administration, was noted. No adverse reactions and negative phenomena were detected during the use of the test drug, and the drug was well tolerated. According to the clinical condition of the experimental group animals and the results of laboratory tests it was established that the use of non-steroidal anti-inflammatory drugs based on acetylsalicylic, ascorbic and citric acids simultaneously with antimicrobial therapy led to a faster improvement in the state of the musculoskeletal system. Observation during the month of the piglet’s clinical conditions confirmed the safety of the drug in the recommended dosage and method of administration in the combined treatment of inflammatory processes of the musculoskeletal system.

Polydimethylsiloxane Surface Modification of Microfluidic Devices for Blood Plasma Separation.

Over the last decade, researchers have developed a variety of new analytical and clinical diagnostic devices. These devices are predominantly based on microfluidic technologies, where biological samples can be processed and manipulated for the collection and detection of important biomolecules. Polydimethylsiloxane (PDMS) is the most commonly used material in the fabrication of these microfluidic devices. However, it has a hydrophobic nature (contact angle with water of 110°), leading to poor wetting behavior and issues related to the mixing of fluids, difficulties in obtaining uniform coatings, and reduced efficiency in processes such as plasma separation and molecule detection (protein adsorption). This work aimed to consider the fabrication aspects of PDMS microfluidic devices for biological applications, such as surface modification methods. Therefore, we studied and characterized two methods for obtaining hydrophilic PDMS surfaces: surface modification by bulk mixture and the surface immersion method. To modify the PDMS surface properties, three different surfactants were used in both methods (Pluronic® F127, polyethylene glycol (PEG), and polyethylene oxide (PEO)) at different percentages. Water contact angle (WCA) measurements were performed to evaluate the surface wettability. Additionally, capillary flow studies were performed with microchannel molds, which were produced using stereolithography combined with PDMS double casting and replica molding procedures. A PDMS microfluidic device for blood plasma separation was also fabricated by soft lithography with PDMS modified by PEO surfactant at 2.5% (v/v), which proved to be the best method for making the PDMS hydrophilic, as the WCA was lower than 50° for several days without compromising the PDMS's optical properties. Thus, this study indicates that PDMS surface modification shows great potential for enhancing blood plasma separation efficiency in microfluidic devices, as it facilitates fluid flow, reduces cell aggregations and the trapping of air bubbles, and achieves higher levels of sample purity.

Modulation of the photobehavior of gefitinib and its phenolic metabolites by human transport proteins

The photobiological damage that certain drugs or their metabolites can photosensitize in proteins is generally associated with the nature of the excited species that are generated upon interaction with UVA light. In this regard, the photoinduced damage of the anticancer drug gefitinib (GFT) and its two main photoactive metabolites GFT-M1 and GFT-M2 in cellular milieu was recently investigated. With this background, the photophysical properties of both the drug and its metabolites have now been studied in the presence of the two main transport proteins of human plasma, i.e., serum albumin (HSA) and α1-acid glycoprotein (HAG) upon UVA light excitation. In general, the observed photobehavior was strongly affected by the confined environment provided by the protein. Thus, GFT-M1 (which exhibits the highest phototoxicity) showed the highest fluorescence yield arising from long-lived HSA-bound phenolate-like excited species. Conversely, locally excited (LE) states were formed within HAG, resulting in lower fluorescence yields. The reserve was true for GFT-M2, which despite being also a phenol, led mainly to formation of LE states within HSA, and phenolate-like species (with a minor contribution of LE) inside HAG. Finally, the parent drug GFT, which is known to form LE states within HSA, exhibited a parallel behavior in the two proteins. In addition, determination of the association constants by both absorption and emission spectroscopy revealed that the two metabolites bind stronger to HSA than the parent drug, whereas smaller differences were observed for HAG. This was further confirmed by studying the competing interactions between GFT or its metabolites with the two proteins using fluorescence measurements. These above experimental findings were satisfactorily correlated with the results obtained by means of molecular dynamics (MD) simulations, which revealed the high affinity binding sites, the strength of interactions and the involved amino acid residues. In general, the differences observed in the photobehavior of the drug and its two photoactive metabolites in protein media are consistent with their relative photosensitizing potentials.

Synthesis of highly luminescent core-shell nanoprobes in a single pot for ofloxacin detection in blood serum and water.

Antibiotics are commonly used as antibacterial medications due to their extensive and potent therapeutic properties. However, the overconsumption of these chemicals leads to their accumulation in the human body via the food chain, amplifying drug resistance and compromising immunity, thus presenting a significant hazard to human health. Antibiotics are classified as organic pollutants. Therefore, it is crucial to conduct research on precise methodologies for detecting antibiotics in many substances, including food, pharmaceutical waste, and biological samples like serum and urine. The methodology described in this research paper introduces an innovative technique for producing nanoparticles using silica as the shell material, iron oxide as the core material, and carbon as the shell dopant. By integrating a carbon-doped silica shell, this substance acquires exceptional fluorescence characteristics and a substantial quantum yield value of 80%. By capitalising on this characteristic of the substance, we have effectively constructed a fluorescent sensor that enables accurate ofloxacin analysis, with a detection limit of 1.3 × 10-6 M and a linear range of concentrations from 0 to 120 × 10-6 M. We also evaluated the potential of CSIONPs for OLF detection in blood serum and tap water analysis. The obtained relative standard deviation values were below 3.5%. The percentage of ofloxacin recovery from blood serum ranged from 95.52% to 103.28%, and from 89.9% to 96.0% from tap water.

Radiolabeling and preclinical animal model evaluation of DTPA coupled 99mTc-labelled flutamide complex ([99mTc]DTPA-FLUT) as a potential radiotracer for cancer imaging.

Advances in molecular imaging strategies have had an effect on precise diagnosis and treatment. Research has been intensified to develop more effective and versatile radiopharmaceuticals to uplift diagnostic efficiency and, consequently, the treatment. To label the flutamide (FLUT) coupled with diethylenetriamine pentaacetate (DTPA) with technetium-99 m (99mTc) and to evaluate its binding efficiency with rhabdomyosarcoma (RMS) cancer cells. Radiolabeling of FLUT with 185 MBq freshly eluted 99mTcO4-1 was carried out via DTPA bifunctional chelating agent using stannous chloride reducing agent at pH 5. The labeled compound was assessed for its purity using chromatography analysis, stability in saline and blood serum, AND charge using paper electrophoresis. Normal biodistribution was studied using a mouse model, while binding affinity with RMS cancer cells was studied using an internalization assay. The in vivo accumulation of RMS cancer cells in a rabbit model was monitored using a SPECT gamma camera. Radiolabeling reaction displayed a pharmaceutical yield of 97% and a stability assay showed >95% intact radiopharmaceutical up to 6 h in saline and blood serum. In vitro internalization studies showed the potential of [99mTc]DTPA-FLUT to enter into cancer cells. This biodistribution study showed rapid blood clearance and minimum uptake by body organs, and scintigraphy displayed the [99mTc]DTPA-FLUT uptake by lesion, induced by RMS cancer cell lines in rabbit. Stable, newly developed [99mTc]DTPA-FLUT seeks its way to internalize into RMS cancer cells, indicating it could be a potential candidate for the diagnosis of RMS cancer.

Wall Shear Stress Measured with 4D Flow CMR Correlates with Biomarkers of Inflammation and Collagen Synthesis in Mild-to-Moderate Ascending Aortic Dilation and Tricuspid Aortic Valves.

Understanding the mechanisms underlying ascending aortic dilation is imperative for refined risk stratification of these patients, particularly among incidentally identified patients, most commonly presenting with tricuspid valves. The aim of this study was to explore associations between ascending aortic hemodynamics, assessed using four-dimensional flow cardiovascular magnetic resonance imaging (4D Flow CMR), and circulating biomarkers in aortic dilation. Forty-seven cases with aortic dilation (diameter ≥40 mm) and 50 sex-and age-matched controls (diameter <40 mm), all with tricuspid aortic valves, underwent 4D flow CMR and venous blood sampling. Associations between flow displacement, wall shear stress (WSS), and oscillatory shear index in the ascending aorta derived from 4D Flow CMR, and biomarkers including interleukin-6, collagen type I α1 chain, metalloproteinases (MMPs), and inhibitors of MMPs derived from blood plasma, were investigated. Cases with dilation exhibited lower peak systolic WSS, higher flow displacement, and higher mean oscillatory shear index compared to controls without dilation. No significant differences in biomarkers were observed between the groups. Correlations between hemodynamics and biomarkers were observed, particularly between maximum time-averaged WSS and interleukin-6 (r = 0.539, p < 0.001), and maximum oscillatory shear index and collagen type I α1 chain (r = -0.575, p < 0.001 in cases). Significant associations were discovered between 4D flow CMR derived whole-cardiac cycle WSS and circulating biomarkers representing inflammation and collagen synthesis, suggesting an intricate interplay between hemodynamics and the processes of inflammation and collagen synthesis in patients with early aortic dilation and tricuspid aortic valves.

Bioanalytical Method Development and Validation and forced degradation of Sitagliptin and determination of Pharmacokinetic application study in Human Plasma by RP-HPLC method

In the present investigation, an attempt was made for the development, validation, forced degradation and pharmacokinetic application of sitagliptin in the human plasma spiking studies by UV - HPLC method. The experimentation was developed based on the extensive literature survey and ascertained by the statistical parameters of the sampling. A simplified, accurate method was created by the liquid chromatographic system from Shimadazu LC 20AD consisting of manual injection. The optimized chromatogram was obtained with acetonitrile in the isocratic mobile phase method at a 1.0 mL/min flow rate. A thermo C-8 column (4.6X250mm,5μm) was used as a stationary phase, and 265.0nm was selected as the detection wavelength with the aid of a UV-Vis detector. The proposed method was validated as per ICH guidelines. The technique was linear in the range of 10-50μg/mL with correlation coefficient R2 = 0.9746, respectively. Recovery studies postulated that % RSD 19.14, 3 &amp; 9.95 respectively. Injection repeatability values were found to be % RSD 17 &amp; 10.63 for intraday and interday, respectively. Stress degradation studies revealed that sitagliptin degrades more rapidly when subjected to 0.1 NaoH. Human plasma spiking studies reported 3.02 ng/mL at 3.02+/-60 min of C and T max, respectively. Keywords: Sitagliptin, Method development, HPLC, Validation, Stress degradation studies, Human plasma spiking studies

Preclinical pharmacokinetic studies of villocarine A, an active Uncaria alkaloid.

Villocarine A is a bioactive indole alkaloid isolated from the Uncaria genus. It has demonstrated vasorelaxation activity and potential to protect the central nervous system. To identify the pharmacokinetic properties of villocarine A, a series of in vitro and in vivo studies have been performed. Villocarine A was found to be highly permeable (15.6 ± 1.6*10-6cm/s) across human colorectal adenocarcinoma cell monolayer with high protein binding (>91%) in both rat and human plasma. Hepatic extraction ratio of villocarine A was 0.1 in pooled rat liver and 0.2 in human liver microsomes and was found stable in rat plasma at 37°C. Due to the high permeability and low rate of metabolism properties, villocarine A was initially considered suitable for preclinical development and an ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method for quantification (linearity: 1-150 ng/ml) in rat plasma was developed and validated for in vivo studies. Essential pharmacokinetic parameters included the volume of distribution and clearance of villocarine A, which were found to be 100.3 ± 15.6L/kg and 8.2 ± 1.1L/h/kg, respectively, after intravenous administration in rats. Following oral dosing, villocarine A exhibited rapid absorption as the maximum plasma concentration (53.2 ± 10.4ng/ml) occurred at 0.3 ± 0.1h, post-dose. The absolute oral bioavailability of villocarine A was 16.8 ± 0.1%. To our knowledge, this was the first pharmacokinetic study of villocarine A, which demonstrated the essential pharmacokinetic properties of villocarine A: large volume distribution, high clearance, and low oral bioavailability in rats.

The Role of Fibrinolytic Activity of Blood Plasma in the Development of Obstetric Complications in Overweight Pregnant Women with Metabolic Dysfunction-Associated Steatohepatitis: A Prospective Cohort Study

The Role of Fibrinolytic Activity of Blood Plasma in the Development of Obstetric Complications in Overweight Pregnant Women with Metabolic Dysfunction-Associated Steatohepatitis: A Prospective Cohort Study

A Validated Chiral Chromatography Method for Enantiomeric Separation of Pomalidomide in Human Plasma.

In the present work, new chiral stationary phase high-performance liquid chromatography (CSP-HPLC) method was established and validated for the quantification of pomalidomide (PMD) enantiomers in human plasma. The chromatographic enantiomeric separation was achieved on a Daicel-CSP, Chiralpack IA 4.6 × 250mm, 5μm; because of its advantages of high degree of retention, high resolution capacity, better reproducibility, ability to produce lower back pressure and low degree of tailing. The mobile phase was maintained as methanol: glacial acetic acid (499.50ml:50μL). Ultraviolet wavelength for detection was 220nm. PMD enantiomer-I and enantiomer-II were separated at 8.83 and 15.34min, respectively. Limit of detection and limit of quantification for each enantiomer and the calibration curve of standard PMD was linear in range between 10-5,000ngmL-1. The method was validated according to The International Council for Harmonisation of Technical Requirements of Pharmaceuticals for Human Use (ICH(Q2R1)) specific guidelines. We found no interference peak with PMD chromatogram obtained. This is a simple, reliable and specific method for detection and quantification of enantiomer of PMD in human plasma sample.

The therapeutic effect of the oily extract of Cannabis sp. in Aluminum Chloride-Induced Alzheimer's Disease in Rats

Objective: To evaluate the therapeutic potential of Cannabis sp extract in a model of Alzheimer's disease induced by aluminum chloride in rats. Methods: Thirty Wistar rats were divided into five groups (n=6) where one group received no treatment, another group received only aluminum chloride and the others, in addition to aluminum chloride, received different doses (50, 100 and 150µl) of oily extract of Cannabis sp. The animals were euthanized one day after treatment, where the brain was collected, fixed, processed and embedded in paraffin. Histological sections of the hippocampus were used for qualitative and quantitative assessment of morphological changes. Blood serum was also obtained to determine indicators of oxidative status. Results: Reduction of more than 80% in cell death in the dentate gyrus region of the hippocampus in animals treated with cannabis extract. Furthermore, a reduction in total protein levels by approximately 20% and an increase (15%) in antioxidant enzymes in relation to the positive control were also observed in the serum. Conclusion: The use of the oily extract of Cannabis sp. in a 2:1 ratio of delta-9-tetrahydrocannabinol (THC): Cannabidiol (CBD) effectively reduced hippocampal neuron deaths from amyloidosis induced in chronic aluminum chloride intoxication.

Apolipoprotein-CIII O-Glycosylation Is Associated with Micro- and Macrovascular Complications of Type 2 Diabetes.

Apolipoprotein-CIII (apo-CIII) inhibits the clearance of triglycerides from circulation and is associated with an increased risk of diabetes complications. It exists in four main proteoforms: O-glycosylated variants containing either zero, one, or two sialic acids and a non-glycosylated variant. O-glycosylation may affect the metabolic functions of apo-CIII. We investigated the associations of apo-CIII glycosylation in blood plasma, measured by mass spectrometry of the intact protein, and genetic variants with micro- and macrovascular complications (retinopathy, nephropathy, neuropathy, cardiovascular disease) of type 2 diabetes in a DiaGene study (n = 1571) and the Hoorn DCS cohort (n = 5409). Mono-sialylated apolipoprotein-CIII (apo-CIII1) was associated with a reduced risk of retinopathy (β = -7.215, 95% CI -11.137 to -3.294) whereas disialylated apolipoprotein-CIII (apo-CIII2) was associated with an increased risk (β = 5.309, 95% CI 2.279 to 8.339). A variant of the GALNT2-gene (rs4846913), previously linked to lower apo-CIII0a, was associated with a decreased prevalence of retinopathy (OR = 0.739, 95% CI 0.575 to 0.951). Higher apo-CIII1 levels were associated with neuropathy (β = 7.706, 95% CI 2.317 to 13.095) and lower apo-CIII0a with macrovascular complications (β = -9.195, 95% CI -15.847 to -2.543). In conclusion, apo-CIII glycosylation was associated with the prevalence of micro- and macrovascular complications of diabetes. Moreover, a variant in the GALNT2-gene was associated with apo-CIII glycosylation and retinopathy, suggesting a causal effect. The findings facilitate a molecular understanding of the pathophysiology of diabetes complications and warrant consideration of apo-CIII glycosylation as a potential target in the prevention of diabetes complications.

Strategy to Empower Nontargeted Metabolomics by Triple-Dimensional Combinatorial Derivatization with MS-TDF Software.

Chemical derivatization is a widely employed strategy in metabolomics to enhance metabolite coverage by improving chromatographic behavior and increasing the ionization rates in mass spectroscopy (MS). However, derivatization might complicate MS data, posing challenges for data mining due to the lack of a corresponding benchmark database. To address this issue, we developed a triple-dimensional combinatorial derivatization strategy for nontargeted metabolomics. This strategy utilizes three structurally similar derivatization reagents and is supported by MS-TDF software for accelerated data processing. Notably, simultaneous derivatization of specific metabolite functional groups in biological samples produced compounds with stable but distinct chromatographic retention times and mass numbers, facilitating discrimination by MS-TDF, an in-house MS data processing software. In this study, carbonyl analogues in human plasma were derivatized using a combination of three hydrazide-based derivatization reagents: 2-hydrazinopyridine, 2-hydrazino-5-methylpyridine, and 2-hydrazino-5-cyanopyridine (6-hydrazinonicotinonitrile). This approach was applied to identify potential carbonyl biomarkers in lung cancer. Analysis and validation of human plasma samples demonstrated that our strategy improved the recognition accuracy of metabolites and reduced the risk of false positives, providing a useful method for nontargeted metabolomics studies. The MATLAB code for MS-TDF is available on GitHub at https://github.com/CaixiaYuan/MS-TDF.