Leukocyte Concentration Research Articles

Predictors of Acute Kidney Injury in Critically Ill Patient at Intensive Cardiac Care Unit

Background: Acute kidney injury (AKI) occurs frequently in the intensive cardiac care unit and recognized as a heterogeneous syndrome with variable etiology and clinical presentation that affects acute morbidity and mortality. AKI needs to be identified early and underlying causes must be treated
 Aim: The aim of this study is to identify the risk factors of AKI in critically ill patients at ICCU Sardjito General Hospital.
 Method: We performed a retrospective analysis of patient registry from Sardjito Cardiovascular Intensive Care (SCIENCE) between January 2021 and December 2021. This registry provided demographic data, risk factors, comorbidities, laboratory findings and survival outcomes. The KDIGO criteria were used to define AKI characterized by an increase in serum creatinine more or equal to 0.3 mg/dL in 48 hours, or an increase in serum creatinine more or equal to 1.5 times than previous value, or urine volume less than 0.5 mL/kg BW/hour for 6 hours. Univariate and multivariate data analyses were carried out.
 Results: This study included 428 patients with an incidence of AKI was 14,3 %. Univariate analysis showed that AKI was related to diabetes, acute heart failure, sepsis, APACHE score, SAPS, Sardjito score, MCARS, hemoglobin, leukocyte, and plasma albumin concentration. Furthermore, we did multivariate analysis and showed the independent predictor of AKI at ICCU admission is acute heart failure (OR 3.90; 95% CI 1.95–7.77; p <0.001), sepsis (OR 3.02; 95% CI 1.03-8.90; p 0.045) and high APACHE II score (OR 0.33; 95% CI 0.13-0.80; p 0.015).
 Conclusions: Acute heart failure, sepsis and high APACHE score at admission is independent predictors of AKI among critically ill in ICCU Sardjito General Hospital. The results of this study may contribute to the implementation of targeted therapies.

Delta Neutrophil Index as a New Marker of Purulent Inflammation in Men With Non-odontogenic Abscesses of the Neck.

Introduction If non-odontogenic abscesses and phlegmons (all purulent inflammations where the etiology is not a diseased tooth) of the neck are not promptly treated, they can lead to serious complications and even end in the death of the affected patient. Classical markers of inflammation such as plasma concentration of leukocytes (WBC), neutrophils (Neu), C-reactive protein (CRP), and erythrocyte sedimentation rate (ESR) are elevated in inflammatory neck diseases, but none of them has been proven as a definite marker in the prediction of this type of pathology. This necessitates the search and analysis of new indicators that could be used in the diagnosis, follow-up, and prognosis of patients with purulent neck infections. Potentially, such a marker could be the delta neutrophil index (DNI), which is increasingly entering clinical practice as a prognostic indicator in critically ill patients with life-threatening illnesses, including sepsis and systemic inflammatory response syndrome (SIRS). In the world literature, there are no data that have been studied in patients with purulent diseases of the neck of non-odontogenic origin, which is the aim of this original article. Materials and methods This retrospective study included 40 men with an average age of 46 (18-87) years with non-odontogenic abscesses and phlegmons of the neck who were hospitalized and operated on. In all of them, the concentration of leukocytes, eosinophils (Eo), neutrophils, and mature polymorphonuclear neutrophil leukocytes (PMN) was examined on an automatic 5-Diff hematology analyzer, ADVIA 2120i (SiemensMedical Solutions USA, Malvern, PA). Thus, it was calculated according to the formula for calculating DNI (DNI% = (Neu%+Eo%) - PMN%). Retrospectively, 30 healthy men with an average age of 42 (18-81) years were used as a control group, in which the same indicators were examined during a preventive examination. Results and discussion Comparing the mean values of WBC, Neu, and DNI between the studied patients with non-odontogenic purulent neck infections (n=40) and the healthy male controls (n=30) it was found that all three indicators of inflammation are significantly higher in the ill men, and these differences are statistically significant (p<0.05): 10.19 ±2.68x103/L versus 7.37 ±1.93x103/L for leukocytes, 7.68 ±2, 76x103/L versus 4.13 ±1.48x103/L for neutrophils, and 1.11±0.83% versus -1.07±1.22% for DNI. Therefore, the high mean numbers of measured WBC and Neu in men with non-odontogenic purulent neck infections were associated with an increase in their mean calculated DNI. This gives us reason to think that while WBC and Neualone are not sufficient for definitive diagnosis, treatment follow-up, and prediction of disease outcome, in combination with DNI they become reliable indicators in purulent neck infections. Conclusions The DNI correlates well with other well-known and established indicators of inflammation, such as the concentration of leukocytes and neutrophils in the peripheral blood of patients. Its calculation is fast as an implementation procedure and is economically beneficial. Its independent use in the diagnosis and treatment of these diseases is about to be investigated and analyzed.

The effect of the combined use of tulatromycin and thymalin in genital mycoplasmosis in cows on cellular immunity indicators

Modern technologies of industrial animal husbandry imply the most effective use of the reproductive potential of females and obtaining healthy young from them. The limiting factor in achieving this goal are diseases characterized by smoothed symptoms, which complicates their timely diagnosis and therapy, one of which is genital mycoplasmosis of cattle. The presence of a long latent period and unclear symptoms allows this disease to remain unnoticed and spread in the herd. During the latent period, morphofunctional changes develop in the body of females, leading to partial or complete loss of reproductive ability. Tetracycline, fluoroquinolone and macrolide antibiotics have been used with varying degrees of effectiveness for the treatment of cows with genital mycoplasmosis, however, antibiotic therapy alone does not restore all functions disrupted during the disease, and in particular does not eliminate immunodeficiency. In this regard, the purpose of our study was to study the inclusion of the immunomodulator thymalin in the treatment regimen of cows with genital mycoplasmosis and its effect on the content of leukocytes, lymphocytes and their subpopulations in the blood. Three groups of animals were formed for the experiment: the first group - pregnant cows with genital mycoplasmosis for the treatment of which the antibiotic traksovet 100 (tulatromycin) was used at a dose of 2.5 mg per 1 kg of animal body weight, subcutaneously, once 40 days before the expected delivery; the second group - pregnant cows with genital mycoplasmosis for the treatment of which the antibiotic traksovet 100 was used at the same dose and the immunomodulator timalin at a dose of 0.1 mg per 1 kg of animal body weight, intramuscularly, twice with an interval of 72 hours; the third group – clinically healthy pregnant cows. In all groups of cows, the number of leukocytes, the percentage of lymphocytes, the absolute and relative content of T-, B- and 0-lymphocytes were determined. It has been established that the use of tulatromycin for the treatment of cows with genital mycoplasmosis gives a good therapeutic effect, but provides only partial restoration of cellular immunity. The combined use of tulatromycin and thymalin has the maximum therapeutic effect and leads to the complete restoration of cellular immunity, and some indicators even slightly exceed their value in clinically healthy cows.

Leukocyte Concentrations Are Isometric in Reptiles Unlike in Endotherms.

AbstractHow do large and small reptiles defend against infections, given the consequences of body mass for physiology and disease transmission? Functionally equivalent mammalian and avian granulocytes increased disproportionately with body mass (i.e., scaled hypermetrically), such that large organisms had higher concentrations than expected by a prediction of proportional protection across sizes. However, as these scaling relationships were derived from endothermic animals, they do not necessarily inform the scaling of leukocyte concentration for ectothermic reptiles that have a different physiology and evolutionary history. Here, we asked whether and how lymphocyte and heterophil concentrations relate to body mass among more than 120 reptile species. We compared these relationships to those found in birds and mammals and to existing scaling frameworks (i.e., protecton, complexity, rate of metabolism, or safety factor hypotheses). Both lymphocyte and heterophil concentrations scaled almost isometrically among reptiles. In contrast, functionally equivalent granulocytes scaled hypermetrically and lymphocytes scaled isometrically in birds and mammals. Life history traits were also poor predictors of variation in reptilian heterophil and lymphocyte concentrations. Our results provide insight into differences in immune protection in birds and mammals relative to that in reptiles through a comparative lens. The shape of scaling relationships differs, which should be considered when modeling disease dynamics among these groups.

Assessment of the clinical and hematological status in acute radiation damage of Leningrad calico chickens

The problem of radiation damage to the human body, animals, birds and its individual systems and organs against the background of the use of ionizing radiation sources has acquired relevance throughout the world over the past decades. In the event of an emergency, acute radiation damage in poultry will be massive and increase the risks of developing infectious and noninfectious diseases, due to a decrease in the level of specific resistance, low effectiveness of therapeutic and preventive measures and, as a result, will lead to a decrease in the quality of the resulting product. In view of this, studies aimed at studying the features of the course of radiation pathology in chickens are very relevant and will be taken into account when developing and applying radioprotective formulations.The study of the features of the course of acute radiation damage caused by uniform single irradiation at doses of 7 Gy; 9 Gy; 11 Gy and 13 Gy, at a power of 0.99 Gy/min, was carried out on Leningrad calico chickens. Resistance to ionizing radiation was assessed by generally accepted clinical and hematological methods. Pronounced changes in clinical manifestations and shifts in morphological parameters of peripheral blood were noted under the influence of gamma radiation. Thus, irradiation of chickens with doses of 11 Gy and 13 Gy caused the development of acute radiation damage of an extremely severe degree, which led to severe depression, refusal of feed, shortening the duration or absence of a latent period, against the background of severe leuko- and erythropenia. Irradiation with a dose of 9 Gy caused severe acute radiation damage characterized by adynamia, reduced feed intake, diarrhea with an admixture of blood. Leukopenia was noted in the blood, which by day 12 values were lower than before irradiation by 41.9% and erythropenia. During acute radiation damage in chickens, when irradiated at a dose of 7 Gy, the development of the initial, latent, peak and recovery period in the surviving individuals was clearly traced. The clinical manifestation was characterized by a complex of dyspeptic and neuromotor disorders, moderate leukopenia was noted in the blood. In all experimental chickens, a violation of egg production was noted, up to complete cessation against the background of irradiation at doses of 11 Gy and 13 Gy. It is concluded that uniform single irradiation of chickens in doses from 7 Gy to 13 Gy leads to acute radiation damage and negatively affects the productive qualities of poultry. In order to conduct the experiment on the principle of pairs of analogues, 5 groups of animals were formed, 10 chickens in each. Determination of the content of erythrocytes, leukocytes and platelets in the blood was determined by counting in the Goryaev chamber. External total single exposure to gamma radiation at doses of 7 Gy, 9 Gy, 11 Gy and 13 Gy led to the development of acute radiation exposure of medium, severe and extremely severe degrees, respectively.

Лейкоцити крові щурів різного віку при ініціації десинхронозу на тлі введення кріоконсервованої кордової крові

It has been suggested that the preventive administration of cryopreserved cord blood nucleated cells (CBNCs) prior to the onset of desynchronosis may help to correct its negative effects on the body. The efficacy of CBNCs on blood leukocyte indicators in young and old rats (6 and 18 months old) was investigated. Desynchronosis was modelled by shifting the light regime: the duration of the light period was increased by 12 h, resulting in a light period of 24 h. Leukocyte types were determined in blood smears. Integral leukocyte indices were used to assess the state of the immune system. Desynchronosis caused leukocytosis in young rats and leukopenia in aged rats. The number of banded neutrophils increased in both, segmented neutrophils decreased in young rats and increased in aged rats; lymphocytes changed on the contrary; eosinophils decreased in young rats but did not change in aged rats. In young rats with desynchronosis, the predominance of young cells, macrophages, activation of the humoral link of the immune system, auto- and endogenous intoxication, acceleration of hypersensitivity of the immediate type processes, and increased body adaptation were noted. In aged rats, infectious intoxication, cellular link of immune system predominance, and a decrease in allergy and adaptation were observed. After CBNCs injection before desynchronosis, leukocytosis remained, segmented neutrophils and eosinophils recovered, banded neutrophils increased and lymphocytes decreased in young rats. In old rats, the total number of leukocytes, monocytes and eosinophils recovered, but lymphopenia increased. Regardless of age, the cells of non-specific protection predominated; the cellular link of the immune system activated, adaptation decreased and autointoxication was manifested. In young rats, young forms of neutrophils increased, allergy and immunoreactivity decreased, and the affective link of the immune system and macrophage in aged rats was activated. Therefore, after the preventive use of human CBNCs before desynchronosis initiation in aged rats, the total leukocyte content and the number of eosinophils and monocytes were restored. In 6-month-old rats, the content of young forms of neutrophils increased, indicating stimulation of leukopoiesis. The allergy index decreased in both groups.

Воздействие бовгиалуронидазы азоксимера на бактериальные биопленки в эякуляте пациентов с хроническим простатитом.

Introduction. The use of molecular biological methods in the analysis of bacterial diversity (metataxonomics) of prostate secretion and ejaculate to assess the male microbiota made it possible to identify the bacterial diversity of the microbiota in chronic prostatitis. At the same time, many strains of uropathogens identified in chronic prostatitis show the ability to form biofilms, while there are no generally accepted methods for treating infections caused by bacterial biofilms. Purpose of the study. To study the effect of bovhyaluronidase azoximer in chronic prostatitis, which may be due to the enzymatic action of hyaluronidase on the matrix of bacterial biofilms, which increases the bioavailability of antibiotics, leading to the transition of bacteria into a planktonic state. Material and methods. An electron microscopic study of the ejaculate of 42 patients with chronic prostatitis was performed, of which 32 randomly selected patients received combined antibiotic treatment (4-6 weeks) together with bovhyaluronidase azoxymer (12 weeks) (group I), 10 patients received only antibiotic therapy according to clinical guidelines (group II). On ultrathin sections, the number of bacterial microcolonies and neutrophilic leukocytes per 100 spermatozoa was counted. Results. In the semen of 30 patients, bacterial biofilms with a characteristic morphology were detected: the presence of heterogeneous microorganisms in one microcolony and the presence of a fibrous matrix in which bacteria are immersed. After therapy, in group I, a decrease in the number of bacterial microcolonies, the disappearance of the extracellular matrix, and isolated single bacteria (planktonic state) were detected, in some cases, damage to the bacterial cell wall was detected. In group II quantitative changes in the content of microcolonies and neutrophilic leukocytes were also revealed, however, the morphology of microcolonies in group II did not change. Conclusions. electron microscopy in the study of the ejaculate of patients with chronic prostatitis allows us to reveal the features of the structure of bacterial biofilms. In combined antibiotic therapy together with bovhyaluronidase azoxymer, the biofilm matrix changes and the bacteria transform into a planktonic form.

Multiple cryosauna sessions for post-exercise recovery of delayed onset muscle soreness (DOMS): a randomized control trial.

The main goal was to investigate the effectiveness of cryosauna in preventing the development of delayed onset muscle soreness and to analyze the regenerative changes within muscles after acute fatigue-induced exercises. Thirty-one volunteers were assigned into two groups: 1) an intervention group that participated in cryostimulation after fatigue-induced exercise protocol (CRYO, n = 16) and a control group that performed fatigue-induced exercise protocol, but without any intervention (CONT, n = 15). Main outcome measures include at baseline: blood sample testing (leukocyte content, myoglobin concentration, and creatine kinase activity) and muscle stiffness of lower extremity; immediately after (stiffness), and 24-48-72-96h post-exercise (blood samples and stiffness). Both groups performed an exercise-induced muscle damage protocol based on repeated countermovement jumps (10 sets, 10 repetitions). The CRYO group underwent a cryosauna (temperature: -110°C, time: 1.5min per session) intervention during four sessions (i.e., immediately after, 24-48-72h post-exercise). Leukocyte content was significantly greater 24-48-72h after exercise in CONT, compared with the CRYO group (p ≤ 0.05 for all), while creatine kinase activity was greater 24-48-96h in CONT, compared with the CRYO group (p ≤ 0.05 for all). Muscle stiffness increased significantly in rectus femoris, tibialis anterior, and fibula muscle after 48h post-exercise (p ≤ 0.05 for all), as well as in tibialis anterior and fibula after 72h post-exercise (p ≤ 0.05 for all) in the CRYO group. Multiple cryosauna was an effective recovery strategy that reduced blood biomarkers and muscle stiffness after exercise-induced muscle damage. Moreover, the development of delayed onset muscle soreness, expressed by a greater muscle stiffness post-exercise, was attenuated to the first 48h.

Seminal biomarkers and their correlations to semen parameters in subfertile men.

To assess the levels of seminal biomarkers fructose, zinc and citrate and their correlations to semen parameters in infertile men. 200 infertile male participants undergoing fertility assessment at Singapore General Hospital (SGH), Singapore were recruited prospectively, from June 2020 to August 2021. Their semen samples were assessed for seminal parameters, biomarker levels of fructose, citrate and zinc, leukocyte concentrations and aerobic cultures. They were also assessed for their smoking habits. Sperm concentrations were negatively correlated to seminal fructose levels, r=-0.262, P<0.001. Progressive motility were positively correlated to seminal citrate levels, r=0.181, P=0.014. Sperm morphology and total motile sperm count (TMSC) were positively correlated to seminal zinc and citrate levels, P<0.05. Zinc and citrate levels were significantly reduced in teratozoospermia, asthenoteratozoospermia and oligoasthenoteratozoospermia groups compared to normozoospermia, P<0.05. The presence of infection was associated with elevated leukocyte concentrations, lower sperm concentration (12.5 vs 55.8 million/mL, P=0.024) and fructose levels (35.5 vs 49.2µmol/ejaculate). Heavy smokers compared to light smokers, had lower sperm concentrations (35.3 vs 49.4 million/mL), TMSC (30.9 vs 47.5 million) and zinc levels (4.9 vs 6.7µmol/ejaculate) and significantly lower citrate levels (52.6 vs 79.2µmol/ejaculate, P=0.029). Higher zinc and citrate levels correlated with better progressive motility, sperm morphology and TMSC. Smoking negatively impacted zinc and citrate levels, thereby affecting sperm quality. In conclusion, the inclusion of biomarkers in basic male work-up assessment would assist in identifying common deficiencies and aid in adequate replacement therapy.

Umbilical Cord Stem Cell Lysate: A New Biologic Injectate for the Putative Treatment of Acute Temporomandibular Joint Inflammation.

To compare in vivo, the acute anti-inflammatory effects of a lysate derived from human umbilical perivascular mesenchymal cells with the cells themselves in both an established hind-paw model of carrageenan-induced inflammation and also in the inflamed temporomandibular joint. Human umbilical cord perivascular cells were harvested and cultured in xeno- and serum-free conditions to P3. In addition, P3 cells were used to prepare a proprietary 0.22 micron filtered lysate. First, CD1 immunocompetent mice underwent unilateral hind-paw injections of carrageenan for induction of inflammation, followed immediately by treatment with saline (negative control), 1% cell lysate, or viable cells. The contralateral paw remained un-injected with carrageenan. Paw circumference was measured prior to injections and 48 hr later and myeloperoxidase and TNF-alpha concentrations were measured post-sacrifice in excised tissue. Second, immunocompetent Male Wistar rats underwent unilateral intra-articular temporomandibular (TMJ) injections from the same treatment groups and were sacrificed at 4 and 48 hr post-injection. The contralateral TMJ remained un-injected with carrageenan. Articular tissue and synovial aspirates, from the treated TMJ were obtained for histologic and leukocyte infiltration analyses. The lysate and cell-treated hind-paw demonstrated reduced tissue edema, and significantly lower concentrations of myeloperoxidase and TNF-alpha at 48 hr compared to untreated controls. Treated TMJs demonstrated lower concentrations of leukocytes in the synovium compared to controls and histologic evidence, in the peri-articular tissue, of reduced inflammation. In this preliminary study, both the human umbilical perivascular cells and a highly diluted lysate produced therefrom were anti-inflammatory.

Hematological changes in descendants of animals irradiated in different doses

An urgent problem today is not only the study of changes in the stability of the genome of somatic cells, the clarification of the mechanisms of its destabilization under the influence of a complex of environmental factors, including radiation, but also the study of the possibility of hereditary transmission of these changes. In the modern experimental and clinical medicine, the most important problem is the harmful effect of ionizing radiation on the descendants of irradiated parents and the specifics of the effect of radiation on the developing organism. An important aspect of the problem of long-term post-radiation effects is the state of reproductive function and health of the descendants of the irradiated population, due to the physiological characteristics inherent to the mother and child - high sensitivity to the action of ionizing radiation, the consequences of which may manifest after a long period of imaginary well-being. The purpose of the work is to investigate the dynamics of body weight and hematological indiexes in the descendants of intact sexually mature animals and descendants born to animals irradiated at different doses, which were exposed to radiation at a dose of 1.0 Gy. The authors revealed that in descendants born to animals irradiated at different doses, which were exposed to radiation at a dose of 1.0 Gy, it was established that by the 30th day after irradiation at a dose of 1.0 Gy, the general condition of pup rats born from animals irradiated at a dose of 0.5 Gy and exposed to irradiation at a dose of 1.0 Gy is satisfactory against the background of an insignificant decrease in the body weight. This also applies to pup rats born to animals irradiated at a dose of 1.0 Gy, whose general condition slightly improves compared to the previous period of the study. The data obtained showed in the peripheral blood after 1 day in the descendants born to animals irradiated at a dose of 0.5 Gy, after irradiation at a dose of 1.0 Gy, a reduced content of hemoglobin and erythrocytes against the background of an increase in the content of platelets, leukocytes, lymphocytes and reticulocytes in comparison with non-irradiated animals, which was maintained throughout the experiment. Pronounced changes in hematological parameters were observed in the descendants born to animals irradiated at a dose of 1.0 Gy, which were exposed to total γ-irradiation at a dose of 1.0 Gy. The authors suppose their data are in favour of the pronounced adaptationof organisms’ regulatiory systems to ionizing irradiation effect that, firstly, outline the adaptative physiologcal mechanisms and, secondly, show the main launches of pathophysiologcial mechamisns in case of irradiation-induced hematological changes.

Применение кормовой добавки на основе растительного жмыха и лактулозы в рационах кроликов на откорме

Purpose. Study the effectiveness of a new feed additive based on vegetable cake and lactulose introduced into the diet of young rabbits in order to increase productivity. Materials and Methods. During the research, the objects were rabbits of the Californian breed, which were divided into 2 groups (control and experimental), in each group of 15 heads, according to the principle of analogues, as well as a feed additive based on milk thistle cake and lactulose. Experimental studies on the determination of growth dynamics, determination of slaughter indicators, as well as biochemical and morphological indicators of blood were carried out in a complex analytical laboratory by using standardized research methods. Results. During the experiment, it was found that the introduction of "Lactu-Super" in the amount of 5% by weight of feed contributed to an increase in the concentration of erythrocytes, leukocytes and hemoglobin, which corresponded to a higher level of metabolism, as well as the pre-slaughter weight of animals by 3.7% and slaughter weight, slaughter yield of 141.6 g and 2.0% relative to the control group that did not receive the supplement. Conclusion. The results of scientific and economic experience have demonstrated the effectiveness of using a new feed additive based on vegetable cake and lactulose for fattening hybrid rabbits under intensive technology from the age of 45 days.

Leukocyte-poor platelet-rich plasma and leukocyte-rich platelet-rich plasma promote myoblast proliferation through the upregulation of cyclin A, cdk1, and cdk2.

Muscle injuries are common among athletes and often treated with platelet-rich plasma (PRP). However, whether the leukocyte concentration affects the efficacy of PRP in treating muscle injuries remains unclear. This study investigated the effects of leukocyte-poor platelet-rich plasma (LP-PRP) and leukocyte-rich platelet-rich plasma (LR-PRP) on myoblast proliferation and the molecular mechanisms underlying these effects. Myoblasts were treated with 0.5% LP-PRP, 0.5% LR-PRP, 1% LP-PRP, or 1% LR-PRP for 24 h. The gene expression of the LP-PRP- and LR-PRP-treated myoblasts was determined using RNA sequencing analysis. Cell proliferation was evaluated using an bromodeoxyuridine (BrdU) assay, and cell cycle progression was assessed through flow cytometry. The expression of cyclin A, cyclin-dependent kinase 1 (cdk1), and cdk2 was examined using Western blotting. The expression of myoblast determination protein 1 (MyoD1) was examined through Western blotting and immunofluorescence staining. The LP-PRP and LR-PRP both promoted the proliferation of myoblasts and increased differential gene expression of myoblasts. Moreover, the LP-PRP and LR-PRP substantially upregulated the expression of cyclin A, cdk1, and cdk2. MyoD1 expression was induced in the LP-PRP and LR-PRP-treated myoblasts. Our results corroborate the finding that LP-PRP and LR-PRP have similar positive effects on myoblast proliferation and MyoD1 expression.

Review of Dohan Eherenfest et al. (2009) on “Classification of platelet concentrates: From pure platelet-rich plasma (P-PRP) to leucocyte- and platelet-rich fibrin (L-PRF)”

This classic discusses the original publication of Dohan Eherenfest etal. on "Classification of platelet concentrates: from pure platelet-rich plasma (P-PRP) to leucocyte- and platelet-rich fibrin (L-PRF)", in which the authors propose four categories of platelet concentrates depending on their leucocyte and fibrin content (P-PRP, leucocyte- and platelet-rich plasma (L-PRP), pure platelet-rich fibrin (P-PRF), and L-PRF) to group a "jungle" of products in which the term platelet-rich plasma (PRP) was used indistinctly. They were able to identify common factors such as: (1) the use of anticoagulants and immediate centrifugation of the blood after its collection; (2) most preparation techniques allowed platelet concentrate preparation within an hour; (3) the centrifugation aimed to separate the blood in layers that would allow the extraction of specific fractions; and (4) the product was activated with thrombin or calcium chloride. The reviewed manuscript has been listed among the most cited PRP articles in regenerative medicine, with more than 800 citations, driving current scientific research and clinical practise by categorising L-PRP and P-PRP (now, leucocyte-poor PRP). The classification has also opened the door to understanding intrinsic biological mechanisms between platelets, leukocytes, fibrin, and growth factors, which will later be considered for studying the proliferation and differentiation of cells in different tissues affected by PRP. Since the initial classification of platelet concentrates, several other classification systems have been proposed and published in the current literature such as platelet, activation, white blood cell (PAW), Mishra, platelet, leucocyte, red blood cells, and activation (PLRA), dose of platelet, efficiency, purity, and activation (DEPA), method, activation, red blood cells, spin, platelets, image guidance, leukocytes, and light activation (MARSPILL), etc. These classifications have identified important aspects of PRP that affect the biological composition and, ultimately, the indications and outcomes. To date, there is still a lack of standardisation in sample preparation, cohort heterogeneity, and incomplete reporting of sample preparation utilised, leading to a lack of clarity and challenging researchers and clinicians.

Platelet-Rich Plasma Injections for Chronic Tendinopathies in the Upper Extremities

&#x0D; Findings from evidence syntheses describing the comparative clinical effectiveness of platelet-rich plasma injection (PRPi) in the treatment of chronic tendinopathies of the upper extremities were variable.&#x0D; Whereas findings describing function showed no clear demonstration of effect, some findings describing pain suggested there may be potential for a benefit of PRPi with higher concentrations of leukocytes or longer durations of follow-up.&#x0D; Lack of a clear demonstration of the comparative clinical effectiveness of PRPi in chronic tendinopathies of the upper extremities does not currently support decision-making in favour of its use.&#x0D;

Hematological changes and structural morphological changes in the liver and spleen in mice against the background of acute toxic effects of cyclophosphan

Background. Disturbances in the body associated with bone marrow failure caused by the administration of lethal doses of cyclophosphan to animals make it possible to use this experimental model to solve important pathophysiological problems.&#x0D; Aim. To reveal hematological changes and structural morphological changes in the liver and spleen in mice against the background of acute toxic effects of lethal doses of.cyclophosphan.&#x0D; Material and methods. The studies were performed on 200 outbred male mice weighing 2729 g. Acute cytotoxic syndrome was modeled by a single intraperitoneal injection of a solution of cyclophosphan at doses of 500 and 750 mg/kg of body weight. Blood sampling for hematological studies was carried out on the 3rd, 6th, 10th and 15th days of intoxication, and the removal of the liver and spleen for morphological studies was performed on the 6th and 15th days. Statistical analysis was performed using Statistica 10 and MS Excel. Results were presented as mean (M) standard error of the mean (mx). To determine the significance of differences between groups with a significance level of p 0.05, Fisher's test and MannWhitney U-test were used.&#x0D; Results. Mortality against the background of acute cytotoxic action of cyclophosphan at doses of 500 and 750 mg/kg reached 83.3 and 100%. The initial content of leukocytes in the peripheral blood of mice was (8.62.3)109/l, and on the 6th day of intoxication it decreased to (1.00.3)109/l and (0.60.1)109/l (p=0.01). Blast cells of all hematopoietic lineages were present in blood smears. In the morphological picture of the spleen, the connective tissue stroma was sharply distinguished, and the white and red pulps were almost completely absent. Signs of acute toxic effects of cyclophosphan were found in the liver tissue.&#x0D; Conclusion. The course of acute cytotoxic syndrome caused by cyclophosphan is accompanied by high mortality, the development of pancytopenia, pronounced destructive changes in the spleen tissue and toxic changes in the liver tissue.

The effect of two platelet-rich plasma aspiration techniques on plasma cellular concentrations using a double syringe gravitational centrifugation system.

To identify which aspiration technique increased plasma platelet concentration and which technique minimized plasma leukocyte and erythrocyte concentrations using a gravitational double-syringe platelet rich plasma (PRP) system. Controlled laboratory study. Thirty adult dogs. Whole blood was collected into two autologous conditioned plasma (ACP) syringes and an ethylenediaminetetraacetic acid (EDTA tube) (control samples). The ACP syringes were centrifuged for 5 min at 1500 rpm. The proximal 2 mL of plasma from one ACP syringe was deposited in an EDTA tube (preflash samples). Plasma from the second ACP syringe was withdrawn until the buffy coat was pierced, producing a "flash" of red blood cells, agitated and deposited into an EDTA tube (flash samples). Complete blood counts were performed. Mean plasma platelet concentrations of the control, preflash, and flash samples were 2.4 × 105/dL, 3.3 × 105/dL and 4.1 × 105/dL, respectively. The mean platelet concentration of the flash samples was 7.9 × 104/dL higher than the preflash samples (p = .005). The mean platelet concentration was lower in the control samples than the preflash (p = .002) and flash (p < .0001) samples. The median plasma leukocyte concentration of the preflash samples (0/dL) was lower than in the flash samples (2.4 × 103/dL) (p = .001). The median plasma hematocrit value of the preflash samples (0%) was lower than in the flash samples (1.0%) (p = .002). The flash method is not necessary to produce a PRP sample. Both methods produced PRP. However, clinicians should avoid aspirating the buffy coat when processing PRP for therapies where leukocytes and erythrocytes are contraindicated.

Combined effects of green tea supplementation and eccentric exercise on nuclear factor erythroid 2-related factor 2 activity

PurposeThis study investigated whether combining eccentric exercise and green tea supplementation synergistically increased nuclear factor erythroid 2-related factor 2 (NRF2) activity, a transcription factor responsible for coordinating endogenous antioxidant expression.MethodsIn a double-blinded, randomized, between-subjects design, 24 males (mean [SD]; 23 [3] years, 179.6 [6.1] cm, 78.8 [10.6] kg) performed 100 drop jumps following a 6 days supplementation period with either green tea (poly)phenols (n = 12; 500 mg·d−1) or a placebo (n = 12; inulin). NRF2/antioxidant response element (ARE) binding in peripheral blood mononuclear cells (PBMCs), catalase (CAT) and glutathione reductase (GR) activity, 8-hydroxy-2′-deoxyguanosine (8-OHdG) excretion, and differential leukocyte counts were measured pre-, post-, 1 h and 24 h post-exercise.ResultsExercise did not increase NRF2/ARE binding (p = 0.12) (fold change vs rest: green tea = [post] 0.78 ± 0.45, [1 h] 1.17 ± 0.54, [24 h] 1.06 ± 0.56; placebo = [post] 1.40 ± 1.50, [1 h] 2.98 ± 3.70, [24 h] 1.04 ± 0.45). Furthermore, CAT activity (p = 0.12) and 8-OHdG excretion (p = 0.42) were unchanged in response to exercise and were not augmented by green tea supplementation (p > 0.05 for all). Exercise increased GR activity by 30% (p = 0.01), however no differences were found between supplement groups (p = 0.51). Leukocyte and neutrophil concentrations were only elevated post-exercise (p < 0.001 for all).ConclusionEccentric exercise, either performed alone or in conjunction with green tea supplementation, did not significantly increase NRF2 activity in PBMCs.Trial registration numberosf.io/kz37g (registered: 15/09/21).Graphical abstract

Viability of mononuclear cells in leukocyte concentrates at the stages of their preparation, freezing, and thawing

Aim. To evaluate the viability of mononuclear cells (MNCs) in leukocyte concentrates (LCs) at the stages of their preparation, freezing, and thawing.Materials and methods. The study material included 44 LCs from donors of allogeneic hematopoietic stem cells (HSCs) and 189 autologous LCs from patients with oncohematological disorders. LCs were obtained from donors and patients by leukocytapheresis after mobilization of HSCs. LCs from patients were frozen with dimethyl sulfoxide (DMSO) used as a cryoprotectant at a final concentration of 5% and stored in liquid nitrogen. LCs were thawed before transplantation. A total of 161 LCs were immediately transfused to the recipient after thawing, and 28 LCs were washed from DMSO before transfusion. Flow cytofluorometry was used to determine the percentage of MNC populations that excluded 7-aminoactinomycin D (7-AAD).Results. The viability of peripheral blood MNCs in donors and patients was close to 100%. It was found that leukocytapheresis and cryopreservation with DMSO did not affect the viability of MNCs. The freezing of LCs with DMSO, storage in liquid nitrogen, and thawing resulted in a significant decrease in the content of viable MNCs (p = 0.0025), while no effect of LC storage duration on the viability of MNCs was revealed. Following DMSO removal from LCs, significantly more HSCs remained in a viable state than without washing (94.4 [94.5; 95.2] % vs. 86.7 [67.6; 92.9] %, (p = 0.0051); for other MNC populations, except monocytes, the differences in the viability index were also statistically significant.Conclusion. The viability of MNCs in LCs is recommended to be used as an independent characteristic of the transplant quality. In obtaining LCs and mixing them with the cryoprotectant DMSO, the viability of MNCs does not decrease, while in thawed LCs, it decreases significantly. Thawing of LCs with removal of DMSO allows to achieve the best viability of HSCs and most MNC populations.

Safety of troclosene sodium solution in decontamination of wounds: A prospective clinical and laboratory study.

Debate persists regarding the safety of hypochlorite-containing solutions in the decontamination of infected wounds. In 2006, the Israeli Ministry of Health withdrew licensing approval for troclosene sodium as a wound irrigation solution. The aim of this prospective clinical and laboratory study was to investigate the safety of troclosene sodium solution for decontamination of infected wounds. Troclosene sodium solution was used to treat 30 patients with 35 infected skin wounds of various etiologies and body areas, over a treatment period of 8 days. Data were gathered according to a prospectively designed protocol including general findings, wound-specific observations on Day 1 and Day 8 and laboratory parameters on Day 1 and Day 8. Wound swabs and tissue biopsy for culture were taken on Day 1 and Day 8. Statistical analysis was executed. Tests were 2-sided and p values of <0.05 were considered statistically significant. Eighteen males and 12 females, with 35 infected skin wounds were enrolled. There were no adverse clinical events. No significant changes were observed in general clinical observations. Statistically significant improvements were observed in: pain (p < 0.0001); edema (p < 0.0001); area of wound covered by granulation tissue (p < 0.0001); exudate (p < 0.0001); and erythema (p = 0.002). Prior to treatment, bacteria were demonstrated on microscopy or on culture in 90% of wound samples. On Day 8, this frequency reduced to 40%. There were no abnormal laboratory tests. Serum sodium concentration increased significantly between Day 1 and Day 8, whilst serum concentration of urea and concentrations of thrombocytes, leucocytes and neutrophils showed statistically significant reductions, but all values remained within normal laboratory ranges throughout the study period. Troclosene sodium solution is clinically safe in the management of infected wounds. These findings were presented to the Israel Ministry of Health and as a result, troclosene sodium was re-approved and licensed for decontamination of infected wounds in Israel.