Advancements in the Management of Postoperative Air Leak following Thoracic Surgery: From Traditional Practices to Innovative Therapies.

Background: Postoperative air leak (PAL) is a frequent and potentially serious complication following thoracic surgery, characterized by the persistent escape of air from the lung into the pleural space. It is associated with extended hospitalizations, increased morbidity, and elevated healthcare costs. Understanding the mechanisms, risk factors, and effective management strategies for PAL is crucial in improving surgical outcomes. Aim: This review seeks to synthesize all known data concerning PAL, including its etiology, risk factors, diagnostic approaches, and the range of available treatments from conservative measures to surgical interventions, with a special focus on the use of autologous plasma. Materials and Methods: A comprehensive literature search of databases such as PubMed, Cochrane Library, and Google Scholar was conducted for studies and reviews published on PAL following thoracic surgery. The selection criteria aimed to include articles that provided insights into the incidence, mechanisms, risk assessment, diagnostic methods, and treatment options for PAL. Special attention was given to studies detailing the use of autologous plasma in managing this complication. Results: PAL is influenced by a variety of patient-related, surgical, and perioperative factors. Diagnosis primarily relies on clinical observation and imaging, with severity assessments guiding management decisions. Conservative treatments, including chest tube management and physiotherapy, serve as the initial approach, while persistent leaks may necessitate surgical intervention. Autologous plasma has emerged as a promising treatment, offering a novel mechanism for enhancing pleural healing and reducing air leak duration, although evidence is still evolving. Conclusions: Effective management of PAL requires a multifaceted approach tailored to the individual patient's needs and the specifics of their condition. Beyond the traditional treatment approaches, innovative treatment modalities offer the potential to improve outcomes for patients experiencing PAL after thoracic surgery. Further research is needed to optimize treatment protocols and integrate new therapies into clinical practice.

[Combined action of low-intensity laser radiation and autologous plasma with blood cells on the microcirculatory apparatus of the skin during involution changes.]

Involutional changes of the skin are associated with changes in its microcirculatory apparatus, among which spastic, atonic and mixed types are distinguished. The effect of low-intensity laser radiation in combination with autologous plasma with blood cells on the vascular component of the skin was studied. In the light of the restoration of the microcirculatory apparatus of the skin in different types, the clinical efficacy of the therapy was analyzed. The study of the effectiveness of low-intensity laser radiation in combination with autologous plasma with blood cells was conducted in 57 patients with clinical signs of involutive skin changes. Depending on the type of microcirculatory skin changes, all patients were divided into 3 comparable groups: the 1st - atonic type (n=24), the 2nd - spastic (n=12), the 3rd - mixed (n=21). The obtained data of a complex assessment of the dynamics of skin microcirculation, features of passive and active blood flow, indicate a significant pronounced positive effect of low-intensity laser radiation in combination with autologous plasma with blood cells in patients with spastic type of microcirculation.

A Histological and Clinical Evaluation of Plasma as a Graft Holding Solution and Its Efficacy in Terms of Hair Growth and Graft Survival.

Background There is a time lag between hair follicle harvesting and implantation, during this time hair follicles suffer ischemic injury. We need a holding medium or a solution to minimize or neutralize ischemic injury. Aims and Objective To evaluate plasma as a graft holding solution in terms of its efficacy in hair growth and hair graft survival. Method and Material A split scalp study was performed. The left side was designated as the control area (Group A) where grafts implanted were kept in Ringer's lactate, and the right side behaved as the test area (Group B) and received grafts preserved in autologous plasma. The p -value was calculated. Observations MTT staining for grafts stored in Ringer's lactate at 12 and 72 hours showed poor hair follicle cells' survival while grafts kept in plasma showed viable cells even after 72 hours. The hair count and density in plasma group were significantly higher than those in the Ringer's lactate group. There was an improvement in hair thickness in both groups from 6 months to 12 months. Conclusion Autologous plasma is an easily available graft holding solution. Platelets along with the plasma provide multiple growth factors promoting epithelialization, neovascularization, and action on hair follicle stem cells to improve growth. The fibrin coating around the graft makes it sticky and prevents dehydration. The split scalp controlled study certainly shows the advantages of using plasma over other extra cellular graft holding solutions.

A clinical study on the usefulness of autologous plasma filler in the treatment of nasolabial fold wrinkles.

BACKGROUND: Recently, the efficacy of autologous plasma filler for the reduction of facial wrinkles has been demonstrated. OBJECTIVE: The aim of our study is to validate the efficacy and safety of autologous plasma filler in treating nasolabial fold wrinkles. MATERIAL AND METHODS: Twenty Korean patients with moderate-to-severe nasolabial fold wrinkles were enrolled. The patients were treated with one session of autologous plasma filler. The wrinkle improvement effects were evaluated at 1-week, 4-week, 8-week, and 12-week after the treatment. Three assessment methods were applied. First, two independent dermatologists assessed cosmetic results using a 5-point wrinkle assessment scale. Second, global aesthetic improvement score was used for assessment of the final cosmetic results. Third, patient satisfaction was surveyed. Also, the adverse effects associated to treatment were observed. RESULTS: Mean age of the patients was 44.5 years. The average 5-point wrinkle assessment scale score was significantly improved at 1, 4, 8, and 12 weeks after treatment, comparing to before treatment (p < 0.01). The patients' average global aesthetic improvement score also indicated better cosmetic outcomes. CONCLUSION: The clinical improvement with sufficient patients' satisfaction and no significant adverse events demonstrated that novel autologous plasma filler could be considered as efficient and safety treatment option for nasolabial fold wrinkles.

Histamine release positive test associates with disease remission in chronic spontaneous urticaria: a proof-of-concept study.

SUMMARY: Background. Histamine release (HR) test has previously been shown to predict the presence of endogenous histamine-releasing factors in chronic spontaneous urticaria (CSU). Objectives and methods. Twenty CSU patients unresponsive to antihistamine treatment were enrolled in order to evaluate the correlations between HR test results and demographic features, quality of life, disease activity, clinical course, and autologous serum and plasma skin tests (ASST and APST). Results. All patients with positive HR test (9/9, 100%) had a more severe disease activity at onset (urticaria activity score, UAS > 2) when compared to negative HR test patients (5/11; p = 0.04). Quality of life questionnaire's results were not substantially different between HR positive and negative subgroups at baseline (p > 0.05), and results of HR test and ASST/APST did not co-segregate (p > 0.05). After 12 months, patients with a positive HR test had a significant reduction of disease activity (p = 0.003) whereas patients with a negative HR test did not (p > 0.05), leading to disease remission and antihistamine treatment withdrawal in 67% (6/9) of positive HR test patients versus 18% (2/11) of negative HR test patients (p = 0.027). Conclusions. Positive HR test may predict spontaneous CSU remission at 12 months.

Monocytes of Patients with Type 1 Diabetes Harbour Enterovirus RNA.

BACKGROUND: Intracellular enterovirus (EV) RNA was detected in blood of patients with type 1 diabetes (T1D). The presence of EV RNA in subsets of peripheral blood mononuclear cells (PBMCs) of patients, and the in vitro infection of these cells with an EV, was investigated. MATERIALS AND METHODS: Blood was collected from 42 patients with T1D, PBMCs were isolated and monocytes were purified. Interferon alpha (IFNα) mRNA and EV RNA were investigated using RT-PCR. Levels of IFNα in plasma were measured using an immunoassay. Cells were inoculated with Coxsackievirus B4 (CBV4) in vitro, and infection was assessed by indirect immunofluorescence (IFI). RESULTS: Interferon alpha mRNA was detected in blood and in monocytes of 12 of 42 patients with T1D, but not in monocyte-depleted PBMCs of the same individuals. Significant plasma levels of IFNα (≥ 5 IU/mL) were found in six patients. EV RNA was detected in whole blood and in monocytes of seven patients and negative-strand EV RNA was found in monocytes of 6 of them. When monocytes of patients with IFNα and/or EV RNA in their blood were inoculated with CVB4, the proportion of cells stained by an anti-VP1 antibody was 8.8 ± 1%, whereas no VP1 was detected in the monocytes of IFNα, EV RNA negative patients. Nevertheless, when CBV4 was mixed with plasma, VP1 was detected in monocytes of all patients with T1D (staining ranging from 12 to 36%). CONCLUSIONS: Our data indicate that monocytes of patients with T1D can harbor EV RNA and IFNα mRNA and can be infected with an EV in vitro.

The relation of autologous serum and plasma skin test results with urticarial activity score, sex and age in patients with chronic urticaria.

INTRODUCTION: Some previous studies reported autoimmunity as an etiologic factor in chronic urticaria (CU), but the results of some autoimmunity tests in these studies are conflicting. AIM: To concretize whether there was any relation of autologous serum skin test (ASST) and autologous plasma skin test (APST) results with sex, age and urticarial activity score (UAS) in patients with CU. MATERIAL AND METHODS: Fifty patients with CU and twenty healthy subjects admitted to our dermatology clinic were included in the present study. The ASST and APST were applied to all individuals. RESULTS: The positiveness rates of ASST and APST were significantly higher in the patient group than controls (p = 0.027, p = 0.001, respectively). Among patients, the APST positiveness rate (72%) was significantly (p < 0.05) higher than ASST (46%). It was seen that 48% of patients with negative ASST results had positive APST. However, no patient with negative APST results had positive ASST. There were significant (p < 0.05) relations of the tests' positiveness rates with sex and old age but with UAS. The diameter of the erythematous papule was remarkably (p < 0.05) larger in APST than ASST and also significantly (p < 0.05) larger in females compared to males in both tests (p < 0.05). It was positively increased with old age (p < 0.05). CONCLUSIONS: We can suggest that APST is more sensitive than ASST in the assessment of autoimmunity in CU. A high positiveness rate of APST results may be attributed to high numbers of autoantibodies and coagulation factors present in plasma that might probably play a role in etiopathogenesis of CU.

Autologous exosome facilitates load and target delivery of bioactive peptides to repair spinal cord injury.

Spinal cord injury (SCI) causes motor, sensory and automatic impairment due to rarely axon regeneration. Developing effective treatment for SCI in the clinic is extremely challenging because of the restrictive axonal regenerative ability and disconnection of neural elements after injury, as well as the limited systemic drug delivery efficiency caused by blood spinal cord barrier. To develop an effective non-invasive treatment strategy for SCI in clinic, we generated an autologous plasma exosome (AP-EXO) based biological scaffold where AP-EXO was loaded with neuron targeting peptide (RVG) and growth-facilitating peptides (ILP and ISP). This scaffold can be targeted delivered to neurons in the injured area and elicit robust axon regrowth across the lesion core to the levels over 30-fold greater than naïve treatment, thus reestablish the intraspinal circuits and promote motor functional recovery after spinal cord injury in mice. More importantly, in ex vivo, human plasma exosomes (HP-EXO) loaded with combinatory peptides of RVG, ILP and ISP showed safety and no liver and kidney toxicity in the application to nude SCI mice. Combining the efficacy and safety, the AP-EXO-based personalized treatment confers functional recovery after SCI and showed immense promising in biomedical applications in treating SCI. It is helpful to expand the application of combinatory peptides and human plasma derived autologous exosomes in promoting regeneration and recovery upon SCI treatment.

How CD4+ T Cells Transcriptional Profile Is Affected by Culture Conditions: Towards the Design of Optimal In Vitro HIV Reactivation Assays.

Most of the current assays directed at the investigation of HIV reactivation are based on cultures of infected cells such as Peripheral Blood Mononuclear Cells (PBMCs) or isolated CD4+ T cells, stimulated in vitro with different activator molecules. The culture media in these in vitro tests lack many age- and donor-specific immunomodulatory components normally found within the autologous plasma. This triggered our interest in understanding the impact that different matrices and cell types have on T cell transcriptional profiles following in vitro culture and stimulation. METHODS: Unstimulated or stimulated CD4+ T cells of three young adults with perinatal HIV-infection were isolated from PBMCs before or after culture in RPMI medium or autologous plasma. Transcriptomes were sequenced using Oxford Nanopore technologies. RESULTS: Transcriptional profiles revealed the activation of similar pathways upon stimulation in both media with a higher magnitude of TCR cascade activation in CD4+ lymphocytes cultured in RPMI. CONCLUSIONS: These results suggest that for studies aiming at quantifying the magnitude of biological mechanisms under T cell activation, the autologous plasma could better approximate the in vivo environment. Conversely, if the study aims at defining qualitative aspects, then RPMI culture could provide more evident results.

Optimization of human umbilical cord blood-derived mesenchymal stem cell isolation and culture methods in serum- and xeno-free conditions.

BACKGROUND: Although umbilical cord blood (UCB) is identified as a source of mesenchymal stem cells (MSCs) with various advantages, the success in cell isolation is volatile. Therefore, it is necessary to optimize methods of cord blood-derived MSC (UCB-MSC) isolation and culture. In this study, we evaluated the efficiency of UCB-MSC isolation and expansion using different commercially available serum- and xeno-free media and investigated the capacity of autologous serum and plasma as a supplement to support cell proliferation. Additionally, we defined the presence of multilineage-differentiating stress-enduring (Muse) cells in the UCB-MSC population. Functions of UCB-MSC in in vitro angiogenesis processes and anti-cancer were also verified. METHODS: Mononuclear cells were isolated using density gradient separation and cultured in four commercial media kits, as well as four surface coating solutions. UCB-MSCs were characterized and tested on tube formation assay, and co-cultured with SK-MEL cells in a transwell system. RESULTS: The results showed that only StemMACS™ MSC Expansion Media is more appropriate to isolate and culture UCB-MSCs. The cells exhibited a high cell proliferation rate, CFU forming capability, MSC surface marker expression, trilineage differentiate potential, and chromosome stability. In addition, the culture conditions with autologous serum coating and autologous plasma supplement enhanced cell growth and colony forming. This cell population contained Muse cells at rate of 0.3%. Moreover, UCB-MSCs could induce the tube formation of human umbilical vein endothelial cells and inhibit more than 50% of SK-MEL cell growth. CONCLUSIONS: UCB-MSCs could be high-yield isolated and expanded under serum- and xeno-free conditions by using the StemMACS™ MSC Expansion Media kit. Autologous serum coating and plasma supplement enhanced cell proliferation. These UCB-MSCs had effected the tube formation process and an anti-cancer impact.

New Translational Trends in Personalized Medicine: Autologous Peripheral Blood Stem Cells and Plasma for COVID-19 Patient.

The COVID-19 pandemic, caused by Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), still remains a severe threat. At the time of writing this paper, the second infectious wave has caused more than 280,000 deaths all over the world. Italy was one of the first countries involved, with more than 200,000 people reported as infected and 30,000 deaths. There are no specific treatments for COVID-19 and the vaccine still remains somehow inconclusive. The world health community is trying to define and share therapeutic protocols in early and advanced clinical stages. However, numbers remain critical with a serious disease rate of 14%, ending with sepsis, acute respiratory distress syndrome (ARDS), multiple organ failure (MOF) and vascular and thromboembolic findings. The mortality rate was estimated within 2-3%, and more than double that for individuals over 65 years old; almost one patient in three dies in the Intensive Care Unit (ICU). Efforts for effective solutions are underway with multiple lines of investigations, and health authorities have reported success treating infected patients with donated plasma from survivors of the illness, the proposed benefit being protective antibodies formed by the survivors. Plasma transfusion, blood and stem cells, either autologous or allograft transplantation, are not novel therapies, and in this short paper, we propose therapeutic autologous plasma and peripheral blood stem cells as a possible treatment for fulminant COVID-19 infection.

Autologous plasma versus fetal calf serum as a supplement for the culture of neutrophils.

OBJECTIVE: Currently, the replacement of fetal calf serum (FCS) by a more suitable alternative is a sought aim in the field of tissue and cell culture research. Autologous plasma (AP) and especially autologous serum (AS) have been shown to be effective substitutes of FCS in culture media for some of the cell types. Nevertheless, there is no comparative data on the most appropriate supplement for cell media in neutrophil studies, it is now unclear whether AP have a relatively equal, superior or inferior performance to FCS in neutrophil cell culture. In the present study, human blood neutrophils were isolated and cultured in FCS- or AP-supplemented medium. After 12, 36 and 60 h of incubation, cell viability, oxidative burst and CD11b expression were determined by flow cytometry. RESULTS: Compared to the culture of neutrophils in FCS 10% medium, the culture of neutrophils in a medium with AP 10% could prolong their life span without affecting their function. The findings introduce AP as a better supplement for human neutrophil cell culture than FCS and propose a simple and economical procedure for neutrophil isolation and culture.

The Relation of Autologous Serum Skin Test and Autologous Plasma Skin Test Result with Various Clinical and Laboratory Findings in Patients with Chronic Spontaneous Urticaria.

BACKGROUND: Despite the autologous serum skin test (ASST) and autologous plasma skin test (APST) is widely used test accessing whether a patient with chronic spontaneous urticaria (CSU) has autoreactivity or not, the clinician often encounter difficulty making correlation between the test result and clinical implications. OBJECTIVE: This study was aimed to find any clinical and laboratory findings related to the ASST and APST response. Agreement and correlation between the two tests was also analyzed. METHODS: A retrospective study was conducted on 300 CSU patients who underwent ASST, APST. The subjects were divided into four groups according to the skin test result. Also, the degree of serum and plasma response was recorded. RESULTS: Both ASST and APST positive group had shorter duration of the disease, higher incidence of at least one episode of angioedema than negative group. There were no significant differences in the positivity for autoantibodies including antinuclear, ds-DNA, and thyroid-related between the two groups. The predicted positive rate of ASST and APST according to age showed bimodal peak and decreasing pattern according to disease duration. Predicted positivity of both tests declined with increase in total immunoglobuline E (IgE) level. In the correlation study, the two tests showed high correlation coefficients. CONCLUSION: ASST and APST positivity may be related to disease duration and severity of CSU. The two tests showed a generally consistent result. Autoreactivity may be gradually lost as disease continues. We suggest the autoreactivity in CSU could arise independently from IgE mediated immune process.

A Histological and Clinical Evaluation of Plasma as a Graft-holding Solution and Its Efficacy in Terms of Hair Growth and Graft Survival.

Background: There is a time lag between hair follicle harvesting and implantation; during this time, hair follicles have ischemic injury. We need a holding media or a solution to minimize or neutralize ischemic injury. Aim and Objectives: The aim of this study was to evaluate plasma as a graft-holding solution in terms of its efficacy in hair growth and hair graft survival. Materials and Methods: A split-scalp study was carried out. The left side was designated as the control area (Group A), where graft implanted was kept in Ringer's lactate (RL), and the right side behaved as the test area (Group B), which received grafts preserved in autologous plasma. The P value was calculated. Observations: MTT [3-(4,5-Dimethylthiazol-2-Yl)-2,5-Diphenyltetrazolium Bromide] staining for graft stored in RL at 12-72h showed poor hair follicles cells survival, whereas graft kept in plasma showed viable cells even after 72h. The hair count and density in plasma group were significantly higher than the RL group. There was an improvement in hair thickness in both groups from 6 to 12 months. Conclusion: Autologous plasma is an easily available graft-holding solution. Platelets along with the plasma provide multiple growth factors promoting epithelialization, neovascularization, and action on hair follicle stem cells to improve growth. The fibrin coating around graft makes it sticky and prevents dehydration. The split-scalp controlled study certainly shows the advantages of using plasma over other extracellular graft-holding solutions.

Ovarian Rejuvenation Through Platelet-Rich Autologous Plasma (PRP)-a Chance to Have a Baby Without Donor Eggs, Improving the Life Quality of Women Suffering from Early Menopause Without Synthetic Hormonal Treatment.

Due to the fact that modern American and European women postpone childbirth until later in life, they are more likely to face the problem of ovarian insufficiency by the time they are ready to have children. So, the ability to restore the ovarian function safely is crucially important. Our study involved 38 women 31-45 years of age with low ovarian reserves and at least two unsuccessful attempts to receive their oocytes through IVF. The blood from the patients was collected into two BD vacutainers for PRP preparation. The platelet concentration in the PRP was 1 × 106 µl. PRP injections into the ovaries were performed by a gynecologist with a special 25G needle, 20 cm in length, as an ultrasound-guided procedure or a laparoscopic-assisted approach. After PRP treatment, women were tested with several criteria for 12 months. We saw a significant improvement in hormone levels; six healthy babies were born, ten pregnancies were achieved, and four out of the ten were from natural conception. The PRP injections into the ovaries are safe, productive, and a natural treatment that may help women with premature ovarian insufficiency to give birth to their own child. The difference and novelty with our method of ovarian rejuvenation is in obtaining a higher platelet concentration (about 1 × 106 µl), which allows us to achieve long-lasting results, within 12 months, after a single procedure. Additionally, for the first time, we proposed and successfully performed a laparoscopically assisted technique for administering PRP into the ovary.

Xenogen-free isolation and culture of human adipose mesenchymal stem cells.

BACKGROUND: Adipose-derived Stem Cells (ASCs) present great potential for reconstructive procedures. Currently, isolation by enzyme digestion and culturing using xenogenic substances remain the gold standard, impairing clinical use. METHODS: Abdominal lipo-aspirate and blood samples were obtained from healthy patients. A novel mechanical isolation method for ASCs was compared to (the standard) collagenase digestion. ASCs are examined by flowcytometry and multilineage differentiation assays. Cell cultures were performed without xenogenic or toxic substances, using autologous plasma extracted from peripheral blood. After eGFP-transfection, an in vivo differentiation assay was performed. RESULTS: Mechanical isolation is more successful in isolating CD34+/CD31-/CD45-/CD13+/CD73+/CD146- ASCs from lipo-aspirate than isolation via collagenase digestion (p <0 .05). ASCs display multilineage differentiation potential in vitro. Autologous plasma is a valid additive for ASCs culturing. eGFP-ASCs, retrieved after 3 months in vivo, differentiated in adipocytes and endothelial cells. CONCLUSION: A practical method for human ASC isolation and culturing from abdominal lipo-aspirate, without the addition of xenogenic substances, is described. The mechanical protocol is more successful than the current gold standard protocol of enzyme digestion. These results are important in the translation of laboratory-based cell cultures to clinical reconstructive and aesthetic applications.

Platelet rich plasma hydrogels promote in vitro and in vivo angiogenic potential of adipose-derived stem cells.

Despite great advances in skin wound care utilizing grafting techniques, the resulting severe scarring, deformity and ineffective vascularization remains a challenge. Alternatively, tissue engineering of new skin using patient-derived stem cells and scaffolding materials promises to greatly increase the functional and aesthetic outcome of skin wound healing. This work focused on the optimization of a polyethylene glycol modified (PEGylated) platelet-rich plasma (PRP) hydrogel for the protracted release of cytokines, growth factors, and signaling molecules and also the delivery of a provisional physical framework for stem cell angiogenesis. Freshly collected whole blood was utilized to synthesize PEGylated PRP hydrogels containing platelet concentrations ranging from 0 to 200,000 platelets/µl. Hydrogels were characterized using thromboelastography and impedance aggregometry for platelet function and were visualized using scanning electron microscopy. To assess the effects of PEGylated PRP hydrogels on cells, PRP solutions were seeded with human adipose-derived stem cells (ASCs) prior to gelation. Following 14 days of incubation in vitro, increased platelet concentrations resulted in higher ASC proliferation and vascular gene and protein expression (assessed via RT-PCR, ELISA, and immunochemistry). Using a rat skin excision model, wounds treated with PRP + ASC hydrogels increased the number of vessels in the wound by day 8 (80.2 vs. 62.6 vessels/mm2) compared to controls. In conclusion, the proposed PEGylated PRP hydrogel promoted both in vitro and transient in vivo angiogenesis of ASCs for improved wound healing. STATEMENT OF SIGNIFICANCE: Our findings support an innovative means of cellular therapy intervention to improve surgical wound healing in a normal wound model. ASCs seeded within PEGylated PRP could be an efficacious and completely autologous therapy for treating patients who have poorly healing wounds caused by vascular insufficiency, previous irradiation, or full-thickness burns. Because wound healing is a dynamic and complex process, the application of more than one growth factor with ASCs demonstrates an advantageous way of improving healing.

Red blood cell aggregation, disaggregation and aggregate morphology in autologous plasma and serum in diabetic foot disease.

BACKGROUND: Diabetes mellitus is frequently associated with microcirculation pathology and hemorheological disorders. METHODS: 24 patients with diabetic foot and 22 healthy subjects were recruited. RBC aggregation, disaggregation and morphology of aggregates were determined in autologous plasma and serum. RESULTS: The RBC aggregation in patients with diabetic foot increased in autologous plasma and serum. Increased red blood cell aggregate strength in these patients was observed only in autologous plasma. Microscopic images of RBC aggregates of patients with diabetic foot show the formation of pathologic globular structures of aggregates in autologous plasma and serum. CONCLUSION: The RBC aggregation in autologous plasma and autologous serum in patients with diabetic foot is significantly higher than in healthy subjects. Increase in strength of RBC aggregates in diabetic foot patients was observed only in autologous plasma. The microscopic images of RBC aggregates in patients with diabetic foot indicate the formation of globular (pathologic) structures of aggregates in autologous plasma and serum. The differences in the morphology of RBC aggregates in autologous plasma and serum between healthy subjects and diabetic foot patients, obtained by microscopic image analysis with high magnification light microscope, can be used as an additional diagnostic tool in medical practice.

Peripheral Blood Plasma Clot as a Local Antimicrobial Drug Delivery Matrix.

Platelet-free blood plasma clots were loaded either with antibiotics (vancomycin, gentamicin, or linezolid) at concentrations of 5-300 µg/mL or with silver ions (silver acetate) at concentrations of 3.3-129 µg/mL. The release of antibiotics or silver from the clot matrix was analyzed after repeated immersion of the plasma clots using reversed-phase high-performance liquid chromatography (RP-HPLC) or atomic absorption spectroscopy (AAS). The antimicrobial activity was tested against Staphylococcus aureus; tissue cell compatibility was analyzed using human mesenchymal stem cells (hMSC). Fibrin fiber thickness of the clots was analyzed by scanning electron microscopy. While addition of linezolid and vancomycin did not significantly change the fibrin fiber thickness, gentamicin and silver ions led to an increase in fiber thickness. All antibiotics showed a concentration-dependent burst-like release from the plasma clots within 1 h followed by a general decay in elution. The release of vancomycin and gentamicin, or silver lasted up to 7 days (depending on initial concentrations), but lasted only up to 4 h for linezolid. A correlation (p < 0.0001) was noted between the concentration of released antibiotics analyzed by HPLC and antimicrobial activity (agar diffusion test). A decrease in antibacterial activity of gentamicin- and vancomycin-containing clots occurred within 4 or 5 days. In contrast, the corresponding antibacterial activity of plasma clots containing linezolid was limited to 3 h. Antibacterial activity of plasma clots containing silver at the highest concentrations decreased after day 3, but clots with lower concentrations induced incomplete bacterial lysis or displayed no antibacterial activity. The antibiotic-containing clots did not induce cytotoxic effects on the embedded hMSC in contrast to all clots containing silver. Our results indicate that an autologous plasma clot can be used to deliver antibiotics such as vancomycin and gentamicin in combination with hMSC and the antibacterial effects persist for days without inducing cytotoxic effects on the embedded stem cells.

Autologous Serum and Plasma Skin Tests in Chronic Spontaneous Urticaria: A Reappraisal.

AIM: The objective of this study was to assess autologous serum skin test (ASST) vs autologous plasma skin test (APST) response in chronic spontaneous urticaria (CSU) patients and study the significance of intensity of positive responses in relation to clinicoepidemiological parameters. MATERIALS AND METHODS: One hundred CSU patients and 100 age and sex-matched controls were recruited. The demographic and clinical features were recorded in all patients and routine investigations were performed. ASST and APST tests were performed as per the standard guidelines. RESULTS: The mean duration of illness was 4.85 ± 5.07 years, 90% patients were APST (+), 68% ASST (+), and 22 patients were only APST (+). Positive predictive value (PPV) of ASST and APST was 90.7% and 95.7%, respectively. A significant inverse association was seen between thyroid status and serum IgE levels with APST and ASST positivity. CONCLUSION: APST appears to have better PPV and high intensity of positive response on autologous tests, and correlates with ANA positivity and angioedema.