High Variability of Mesenchymal Stem Cells Obtained via Bone Marrow Aspirate Concentrate Compared With Traditional Bone Marrow Aspiration Technique

Background: Bone marrow aspirate (BMA) is a common source for harvesting mesenchymal stem cells (MSCs), other progenitor cells, and associated cytokines and growth factors to be used in the biologic treatment of various orthopaedic pathologies. The aspirate is commonly centrifuged into a concentrated volume that can be immediately administered to a patient using commercially available kits. However, the handling and efficacy of BMA concentrate (BMAC) are still controversial. Purpose: To characterize BMA versus BMAC for MSC quantity, potency, and cytokine profile. Study Design: Controlled laboratory study. Methods: From 8 participants (age, 17-68 years), 30 mL of bone marrow was aspirated by a single surgeon from either the proximal humerus or distal femur and was separated into 2 equal samples. One sample was kept as BMA, and the other half was centrifuged into BMAC. The 2 samples then underwent flow cytometry for detection of MSCs, cell analysis for colony-forming units (CFUs), and cytokine profiling. A 2-tailed t test was used to detect differences between MSCs, CFUs, and cytokine density concentrations between BMA and BMAC. Results: The average concentration of MSCs in both BMA and BMAC was 0.001%. Average MSC events detected by flow cytometry were significantly higher in BMA versus BMAC (15.1 and 8.1, respectively; P < .045). Expanded MSCs demonstrated similar phenotypes, but CFUs were significantly increased in BMA compared with BMAC (104 vs 68 CFUs, respectively; P < .001). Total protein concentration and cytokine profiling demonstrated great variability between BMA and BMAC and between patients. Most importantly, BMAC failed to concentrate MSCs in 6 of 8 samples. Conclusion: There is great variability in MSC concentration, total protein concentration, and cytokine profile between BMA and BMAC. Clinical Relevance: When studying the clinical efficacy of BMAC, one must also evaluate the sample itself to determine the presence, concentration, and potency of MSCs if this is to be considered a cell-based therapy. Further standard operating procedures need to be investigated to ensure reproducible results and appropriate treatments.

Respiratory MUC5B Disproportion is Involved In Severe Community-Acquired Pneumonia

Background: Mucus production is a process involved in the pathogenesis of Community-acquired pneumonia (CAP). The study is to determine Mucin 5B (MUC5B) protein concentration and its proportion in the bronchoalveolar lavage fluid (BALF) of CAP patients and evaluate its value to help assess disease severity.Methods: A total of 118 patients were enrolled in this cross-sectional study, including 45 with severe CAP (SCAP) and 73 with non-severe CAP (NSCAP). MUC5B concentration in BALF were determined by immunoblotting analysis. Total protein concentration of BALF was detected by Pierce BCA kit. Cytokines IL6, IL10, IFNγ, IL13, and IL17 in BALF were measured using commercial enzyme-linked immunosorbent assay (ELISA). Spearman’s correlation analysis was applied to evaluate the relationships between MUC5B concentration or MUC5B/total protein ratio and the CURB-65 score, as well as cytokines. Logistic regression analysis was used to identify the independent factors associated with severe CAP. Receiver operating characteristic (ROC) curve was used to evaluate the assessment value of MUC5B/total protein ratio and other indexes for CAP severity.Results: MUC5B concentration in the BALF of NSCAP group was higher than that in SCAP group [NSCAP 13.56 µg/ml (IQR 5.92 – 25.79) vs. SCAP 8.20 µg/ml (IQR 4.97 – 14.03), p=0.011]. The total protein concentration in the BALF of NSCAP group was lower than that in SCAP group [NSCAP 0.38 mg/ml (IQR 0.15 - 1.10) vs. SCAP 0.68 mg/ml (IQR 0.46 - 1.69), p=0.002]. The MUC5B/total protein ratio was remarkably higher in NSCAP group than that in SCAP groups [NSCAP 3.66% (IQR 1.50% - 5.56%) vs. SCAP 1.38% (IQR 0.73% - 1.76%), p<0.001]. MUC5B/total protein ratio was negatively correlated with total protein concentration (rs= -0.576, p<0.001), IL6 (rs= -0.312, p= 0.001), IL10 (rs= -0.228, p=0.013) , IL17 (rs= -0.282, p=0.002) and CURB-65 score (rs= -0.239, p=0.009). Logistic regression identified that MUC5B/total protein ratio, IL6 level and CURB-65 score as independent variables related to CAP severity. ROC curve demonstrated best assessment value of MUC5B/total protein ratio for SCAP (AUC 0.803, p<0.001), with a sensitivity of 88.9% and a specificity of 64.4%.Conclusions: Respiratory MUC5B disproportion is related to CAP severity. MUC5B/total protein ratio may serve as an assessment marker and a potential therapeutic target for severe CAP.

A tick bite patient with fever and meningitis co-infected with Rickettsia raoultii and Tacheng tick virus 1: a case report

Background Increasing numbers of tick-borne pathogens are being discovered, including those that infect humans. However, reports on co-infections caused by two or more tick-borne pathogens are scarce. Case presentation A 38-year-old male farmer was bitten by a hard tick, presented with fever (37.7 °C), severe headache and ejection vomiting. Lumbar puncture was performed in the lateral decubitus. The cerebrospinal fluid (CSF) was clear, and analysis showed severe increased pressure (320 mm H2O), mild leukocytosis (126.0 × 106/L, mononuclear cells accounting for 73%) and elevated total protein concentration (0.92 g/L). Bacterial cultures of CSF and blood were negative. The diagnosis of Rickettsia raoultii and Tacheng tick virus 1 (TcTV-1) co-infection was confirmed by amplifying four rickettsial genetic markers and the partial small (S) RNA segment of TcTV-1 from the patient’s blood. The patient gradually recovered after treatment with levofloxacin and ribavirin. Conclusions This is the first reported co-infection case with fever and meningitis caused by R. raoultii and TcTV-1. It is vital to screen for multiple pathogens in tick-bitten patients, especially in those with severe complex symptoms.

Effect of Adding Water-Soluble Chitosan on Some Physiological Traits of Quail Males

The study was conducted to determine the effect of adding water-soluble chitosan on some hematological and biochemical traits in the quail during rearing for meat production, twenty-seven male quail at nine weeks of age were randomly assigned to three treatments with three replicates, it was treated with chitosan for 28 days, according to the following concentrations, the first treatment (control treatment), the second and third treatments, water-soluble chitosan was added at a concentration of 0.2 and 0.3 g/L, respectively. The results showed that there were no significant differences among treatments in RBC, PCV, Hb, MCV, MCH, glucose concentration and AST and ALT enzymatic activity, while there were significantly increased WBC in both experimentsin addition treatments compared to the control treatment. The adding of the water-soluble chitosan at a concentration of 0.2 and 0.3 g/L resulted in a significant decrease in the concentration of cholesterol and triglycerides, while the adding 0.3 g/Lled to a significant increase (P≤ 0.05) in the total protein concentration, albumin and Globulin compared with the control group. We conclude from this research that the adding of water-soluble chitosan at a concentration of 0.3 g/L has the potential to improve productive performance and enhance bird health.

Therapeutic Artemether-Lumefantrine Modulated Monosodium Glutamate-Related Adversity on Rats’ Kidney Histology and Antioxidant Response Bio-Indicators

Co-intake-related interactive-synergistic influence of artemether-lumefantrine, AL and monosodium glutamate, MSG that separately mediated oxidative stress could be significant on the kidney actively involved in xenobiotic detoxification and elimination. Thus, influence of AL on rats’ kidney histomorphology and antioxidant bio-indicators following MSG-challenge was assessed. For 7 days, thirty rats (n = 5) were respectively exposed to vehicle (distilled water), therapeutic AL (TAL), high AL (HAL), MSG, MSG plus TAL or MSG plus HAL. Significant (P<0.05) results comparison showed highest and least (P<0.05) albumin concentration (Mg/dl) in TAL-fed (3.76±0.33) and MSG-fed (1.88±0.70), rats. Total protein concentration (Mg/dl) in MSG-fed (4.04±2.04) and HAL-fed (4.76±1.92), rats lowered markedly. Highest glutathione peroxidase activity (IU/L) in TAL-fed (30.74±12.46) lowered in MSG plus HAL-fed (20.11±6.08) and MSG-fed (20.33±4.85), rats. Catalase activity (IU/L) in control was highest (4.89 ± 0.26) but least (2.58 ± 1.06) in MSG-fed rats. Zinc and Magnesium concentration (Mg/dl) was respectively highest (58.99±5.10) and least (3.48±0.31) in MSG plus HAL-fed but least (18.80±7.77) and highest (4.38±1.67) in MSG-fed, rats. Malondialdehyde concentration (µmol/ml) in MSG plus HAL-fed rats (4.04±0.67) was highest (P<0.05) and least (P<0.05) in HAL-fed rats (1.18±0.11). Differences in superoxide dismutase activity (IU/L) were, however, non-significant (P>0.05).Rats’ kidney photomicrographs (H&E × 400) revealed normal histo-architecture in control but varied degree of fibroplasias (TAL- ,HAL- and MSG plus TAL-fed) and necrosis with infiltrations (MSG plus HAL-and MSG-fed), rats. These demonstrated MSG-related adversity and significant modulation response of TAL, unlike HAL, on the rats’ kidney histology and studied antioxidant response bio-indicators.

Stimulated saliva composition in patients with cancer of the head and neck region

Background To analyse over time changes in stimulated whole saliva regarding total protein, Immunoglobulin A (IgA), and mucin type O-glycans (mostly MUC5B and MUC7) in head and neck cancer patients. Methods 29 dentate patients (20 men and 9 women, 59 ± 8 years) treated with curative radiation therapy and chemotherapy for cancer of the head and neck region were included. The stimulated whole salivary secretion rate was determined and saliva collected at four time-points: at pretreatment, and at 6 months, 1 and 2 years post treatment. The total protein concentration was determined spectrophotometrically by using Bicinchoninic Acid assay and Immunoglobulin A (IgA) by using ELISA technique. Glycosylation pattern of salivary mucins was determined in samples collected pre- and post treatment by using LC/MS electrospray and mucin content quantified using SDS-AgPAGE gels and PAS staining. Results Compared with pretreatment, the total protein concentration was increased already at 6 months post treatment (p < 0.01), and continued to increase up to 2 years post treatment (p < 0.001). During that period no significant changes in IgA concentration was detected. At pretreatment, the output/min of both total protein and IgA was significantly higher than at all time-points post treatment. Saliva from the cancer patients showed a low abundance/no detectable MUC7, while the MUC5B level remained, compared to saliva from a healthy control. The glycomic analysis showed that the percentage of core 2 O-glycans was increased as core 1, 3 and 4 O-glycans were decreased. The level of sialylation was higher at 6 months post treatment, while sulfation was lower. Conclusion A decreased output per minute of proteins at decreased salivary secretion rate, as well as reduced sulfation of MUC5B at 6 months post treatment tended to correlate with the patients’ experience of sticky saliva and oral dryness. At 2 years post treatment, the decreased amount of IgA combined with a lowered salivary secretion rate indicate a reduced oral defense with increased risk of oral infections.

Silver nanoparticle enhances secretion of exosomes in SH-SY5Y cells: Potential therapeutic strategy for human neuroblastoma cancer

Background: Exosomes—a subset of extracellular vesicles (EVs)—are secreted by virtually all cells, including human neuroblastoma cancer (SH-SY5Y) cells, into bodily fluids. Oxidative stress is critically involved in exosome biogenesis and release. Silver nanoparticles (AgNPs) induce cytotoxicity, oxidative stress, and apoptosis in cancer and non-cancer cells. Methods: Here, we studied whether AgNPs-induced oxidative stress could enhance exosome biogenesis and release under low serum conditions in the presence of AgNPs. Although several studies have reported various mechanisms that contribute to EV biogenesis and release from cells, none exists on the involvement of external stimuli by controlling acetylcholinesterase (AChE) and neutral-sphingomyelinase (n-SMase) activities, total protein concentration of exosomes, and exosome count. Results: Owing to cytotoxic and oxidative stresses, AgNPs-treated cells and exosome release were significantly facilitated, which strongly correlated with the AgNPs-induced oxidative stress. Moreover, the expression levels of some important exosome biomarkers were found to be significant under oxidative stress conditions. N-acetylcysteine prevented oxidative stress-induced exosome biogenesis and release. Furthermore, we identified the involvement of the ceramide pathway in exosome functions by inhibiting AChE and n-SMase activities, and exosome protein/counts. These data contribute to the understanding of how AgNPs and intracellular molecular pathways affect exosome biogenesis and release in SH-SY5Y cells. Conclusion: To the best of our knowledge, this is the first study showing that AgNPs stimulate exosome biogenesis and release by inducing oxidative stress and ceramide pathways.

Effect of Encapsulated Beet Extracts (Beta vulgaris) Added to Yogurt on the Physicochemical Characteristics and Antioxidant Activity

Beet has been used as an ingredient for functional foods due to its high antioxidant activity, thanks to the betalains it contains. The effects of the addition of beet extract (liquid and lyophilized) on the physicochemical characteristics, color, antioxidant activity (AA), total betalains (TB), total polyphenols (TP), and total protein concentration (TPC) were evaluated on stirred yogurt. The treatments (T1-yogurt natural, T2-yogurt added with beet juice, T3-added extract of beet encapsulated with maltodextrin, and T4-yogurt added with extract of beet encapsulated with inulin) exhibited results with significant differences (p < 0.05). The highest TB content was observed in T2 (209.49 ± 14.91), followed by T3 (18.65 ± 1.01) and later T4 (12.96 ± 0.55). The highest AA was observed on T2 after 14 days (ABTS˙ 0.819 mM TE/100 g and DPPH˙ 0.343 mM TE/100 g), and the lowest was found on T1 at day 14 (ABTS˙ 0.526 mM TE/100 g and DPPH˙ 0.094 mM TE/100 g). A high content of TP was observed (7.13 to 9.79 mg GAE/g). The TPC varied between 11.38 to 12.56 µg/mL. The addition of beet extract significantly increased AA in yogurt, betalains being the main compounds responsible for that bioactivity.

Chronic Inflammatory Pain Management by BDNF Modulation: A Comparative Study of Gabapentin, Indomethacin and Their Combination on Arthritis Rat Model

Brain derived neurotropic factors (BDNF) can be secreted either as a pro-BDNF or a mature form (mBDNF) through γ-amino-butyric acid (GABAA) receptors activation. Depolarization of GABA neurons in spinal cord can be mediated through N-methyl-D-aspartate (NMDA) receptor due to the exogenous secretion of over expressed BDNF. This over expressed BDNF further modulate the excitation and sensitization of nociceptors. We investigated the modulation of BDNF by GABAA agonist i.e., gabapentin, indomethacin (non-steroidal anti-inflammatory) and their low-dose combination on adjuvant-induced inflammatory arthritis. Mycobacterium tuberculosis H37Ra strain, as adjuvant, was injected in the tail base of female Sprague-Dawley rats. Gabapentin (5 mg/kg), indomethacin (5 mg/kg) and low dose combination of gabapentin (1.5 mg/kg) + indomethacin (2.5 mg/kg) were used. Paw edema was measure by plethysmometer and chronic pain was measured by plantar apparatus. Nitric oxide, peroxide and superoxide dismutase levels were also measured to analyze the free radical and antioxidant balance in different treatment groups along with the total protein concentration. Significant reduction of nitric oxide, peroxide as well as total protein concentration was found in low dose combination. Immunohistochemistry data also showed that the low dose combination has more potential in lowering the BDNF expression in cortex as well as in hippocampus region of brain as compared to their mono therapies. Our study suggested that low-dose combination of gabapentin and indomethacin has a significant impact on lowering the chronic and its associated markers as compare to their monotherapy. Thus indicated the additive effects of the combination therapy on neuropathic pain.