Biochemical aspects of bupivacaine-induced acute muscle degradation

1986 ◽  
Vol 83 (1) ◽  
pp. 197-212
Author(s):  
S. Ishiura ◽  
I. Nonaka ◽  
H. Sugita
Keyword(s):  
Cathepsin B ◽  
Lysosomal Enzymes ◽  
Mononuclear Cells ◽  
Protein Pattern ◽  
Intracellular Localization ◽  
Adult Rat ◽  
Chronological Change ◽  
Rapid Dissolution ◽  
Biochemical Observation ◽  
And Control

A single injection of a local anaesthetic, bupivacaine, into the soleus muscle of adult rat has a severe mytoxic effect, i.e. rapid dissolution of myofilaments and degradation of myofibrillar proteins shortly after injection. Increased lysosomal enzymes were observed in homogenates of affected muscle. The activity of potent proteolytic enzyme, cathepsins B and L (assayed against a new synthetic substrate succinyl-Tyr-Met-naphthylamide), gradually increased and reached a plateau value that was 11-fold greater than the control 48 h after bupivacaine injection. The chronological change in the activity of cathepsins B and L was reflected in the myofibrillar protein pattern in bupivacaine-treated muscle. To determine whether the increase in lysosomal peptide hydrolases is due to activation of muscle lysosomes or not, mononuclear cells were separated from both injected and control muscles. The activity of cathepsins B and L in the lysate from injured muscle was 180-fold higher than the control. Affinity-purified antibody was used to study the intracellular localization of cathepsin B by immunohistochemical procedures. The results were consistent with the biochemical observation that the main source of cathepsin B in muscle homogenates was infiltrated mononuclear cells. Therefore, we conclude that the increased lysosomal enzymes may be derived mainly from mononuclear cells (macrophages), not from muscle lysosomes, in bupivacaine-induced acute muscle degeneration.

scholarly journals Evaluation of the phagocytic activity of peripheral blood monocytes of patients with Jorge Lobo's disease

2004 ◽  
Vol 37 (2) ◽  
pp. 165-168 ◽  
Author(s):  
Fátima Regina Vilani-Moreno ◽  
Luciana Moreira Silva ◽  
Diltor Vladimir Araújo Opromolla
Keyword(s):  
Incubation Time ◽  
Peripheral Blood ◽  
Phagocytic Activity ◽  
Healthy Subjects ◽  
Mononuclear Cells ◽  
Blood Monocytes ◽  
Peripheral Blood Monocytes ◽  
Significant Difference ◽  
Host Parasite ◽  
And Control

Studies on host-parasite interaction in Jorge Lobo's disease are scarce, with no report in the literature on the phagocytosis of Lacazia loboi by phagocytic mononuclear cells. Thus, the objective of the present study was to assess the phagocytic activity of blood monocytes in the presence of L. loboi in patients with the disease and in healthy subjects (controls) over 3 and 24 hours of incubation. Statistical analyses of the results showed no significant difference in percent phagocytosis of the fungus between patient and control monocytes. With respect to incubation time, however, there was a significant difference, in that percent phagocytosis was higher at 3 hours than at 24 hours (p <0.01). These results suggest that monocytes from patients with the mycosis are able to phagocyte the fungus, as also observed in control individuals.

scholarly journals Castration causes an increase in lysosomal size and upregulation of cathepsin D expression in principal cells along with increased secretion of procathepsin D and prosaposin oligomers in adult rat epididymis

PLoS ONE ◽  
2021 ◽  
Vol 16 (4) ◽  
pp. e0250454
Author(s):  
Lorena Carvelli ◽  
Andrea Carolina Aguilera ◽  
Leila Zyla ◽  
Laura Lucía Pereyra ◽  
Carlos R. Morales ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Complex Formation ◽  
Cathepsin D ◽  
Initial Segment ◽  
Lysosomal Enzymes ◽  
Sperm Maturation ◽  
Principal Cells ◽  
Adult Rat ◽  
Rat Epididymis ◽  
Lysosomal Proteins

In the epididymis, lysosomal proteins of the epithelial cells are normally targeted from the Golgi apparatus to lysosomes for degradation, although their secretion into the epididymal lumen has been documented and associated with sperm maturation. In this study, cathepsin D (CatD) and prosaposin (PSAP) were examined in adult epididymis of control, and 2-day castrated rats without (Ct) and with testosterone replacement (Ct+T) to evaluate their expression and regulation within epididymal epithelial cells. By light microscope-immunocytochemistry, a quantitative increase in size of lysosomes in principal cells of Ct animals was noted from the distal initial segment to the proximal cauda. Androgen replacement did not restore the size of lysosomes to control levels. Western blot analysis revealed a significant increase in CatD expression in the epididymis of Ct animals, which suggested an upregulation of its expression in principal cells; androgens restored levels of CatD to that of controls. In contrast, PSAP expression in Ct animals was not altered from controls. Additionally, an increase in procathepsin D levels was noted from samples of the epididymal fluid of Ct compared to control animals, accompanied by an increased complex formation with PSAP. Moreover, an increased oligomerization of prosaposin was observed in the epididymal lumen of Ct rats, with changes reverted to controls in Ct+T animals. Taken together these data suggest castration causes an increased uptake of substrates that are acted upon by CatD in lysosomes of principal cells and in the lumen by procathepsin D. These substrates may be derived from apoptotic cells noted in the lumen of proximal regions and possibly by degenerating sperm in distal regions of the epididymis of Ct animals. Exploring the mechanisms by which lysosomal enzymes are synthesized and secreted by the epididymis may help resolve some of the issues originating from epididymal dysfunctions with relevance to sperm maturation.

scholarly journals 30USE OF ADULT STEM/PROGENITOR CELLS AS NUCLEAR DONORS TO PRODUCE CLONED PORCINE EMBRYOS

2004 ◽  
Vol 16 (2) ◽  
pp. 137
Author(s):  
P. Bosch ◽  
S.L. Pratt ◽  
E. Sherrer ◽  
C.A. Hodges ◽  
E. Ivy Hill ◽  
...  
Keyword(s):  
Alcian Blue ◽  
Mononuclear Cells ◽  
Electric Pulse ◽  
Adipogenic Differentiation ◽  
Blastocyst Stage ◽  
Calcium Deposition ◽  
Nuclear Reprogramming ◽  
Live Animal ◽  
And Control ◽  
Oil Red O

Incomplete or defective nuclear reprogramming may be responsible for low cloning efficiencies. Less differentiated stem cells are thought to be more easily reprogrammed, resulting in improved survival of cloned mice (Rideout WM III et al., 2000 Nat. Genet. 24, 109–110). Our objective was to establish porcine mesenchymal stem cell (MSC) cultures and use these as donor cells in nuclear transfer (NT). A bone marrow (BM) aspirate was collected from an anesthetized gilt. BM mononuclear cells were isolated by centrifugation over a density gradient (Histopaque-1077; Sigma, St. Louis, MO, USA), resuspended in low glucose DMEM (Gibco) plus 10% FBS and plated on flasks; fibroblast-like MSCs were later passaged. Ear skin fibroblast (SF) cultures from the same BM donor gilt were established. Cultures of MSC and SF were exposed to lipogenic, osteogenic or chondrogenic differentiation media (Pittenger MF et al., 1999 Science 284, 143–147) for 14 days. Cells cultured in DMEM with 10% FBS served as controls. Differentiation was assessed by histochemical methods. Calcium deposits and alkaline phosphatase (AP) activity (Vector Red AP Substrate Kit, Vector Labs) were indicative of osteogenic differentiation. MSCs cultured under osteogenic conditions were positive for AP activity and developed a black color after von Kossa staining, indicative of calcium deposition. Oil red O stain identified cellular lipid accumulation. When exposed to adipogenic differentiation media, 10–15% of MSCs developed an adipocyte phenotype with lipid droplet accumulation and oil red O staining. Lipogenic differentiation was not observed in SF and control cultures. Presence of acidic mucopolysaccharides associated with cartilage formation was determined by alcian blue stain. MSCs exposed to chondrogenic conditions were alcian blue-positive, and SF and control cultures were alcian blue negative. For NT, confluent (passage 2) MSC and SF cultures were exposed to roscovitine (15μM; Sigma) for 24h. In vitro-matured oocytes were enucleated and a single cell (MSC or SF) was transferred into the periviteline space. Cell-oocyte couplets were fused in Zimmerman’s medium with a single electric pulse (250V/mm for 20μs) delivered through a needle-type electrode. NT units were electrically activated (2 pulses of 100V/mm for 60μs separated by 5s) in a chamber 1h after fusion and transferred to NCSU-23 medium. Embryos were examined for cleavage and blastocyst formation at 2 and 7 days after NT, respectively. Cleavage rates were 53.3% (40/75) for MSC and 59.7% (46/77) for SF NT embryos. Development to blastocyst stage was 6.6% (5/75) in the MSC group and 1.2% (1/77) in SF group. In conclusion, we established an adult MSC line from a live animal using a minimally invasive BM aspiration technique. Additionally, MSC donor-derived NTs developed to the blastocyst stage. Further experiments will determine nuclear reprogramming in MSC-derived NT embryos.

scholarly journals Association of Strong Immune Responses to PPE Protein Rv1168c with Active Tuberculosis

2008 ◽  
Vol 15 (6) ◽  
pp. 974-980 ◽  
Author(s):  
Nooruddin Khan ◽  
Kaiser Alam ◽  
Shiny Nair ◽  
Vijaya Lakshmi Valluri ◽  
Kolluri J. R. Murthy ◽  
...  
Keyword(s):  
Mononuclear Cells ◽  
Tuberculin Test ◽  
Interferon Response ◽  
Peripheral Blood Mononuclear ◽  
Purified Protein Derivative ◽  
Conventional Tests ◽  
Immunodominant Antigens ◽  
Smear Negative ◽  
And Control ◽  
Immunogenic Properties

ABSTRACT Accurate diagnosis of tuberculosis (TB) infection is critical for the treatment, prevention, and control of TB. Conventional diagnostic tests based on purified protein derivative (PPD) do not achieve the required diagnostic sensitivity. Therefore, in this study, we have evaluated the immunogenic properties of Rv1168c, a member of the PPE family, in comparison with PPD, which is routinely used in the tuberculin test, and Hsp60 and ESAT-6, well-known immunodominant antigens of Mycobacterium tuberculosis. In a conventional enzyme immunoassay, the recombinant Rv1168c protein displayed stronger immunoreactivity against the sera obtained from patients with clinically active TB than did PPD, Hsp60, or ESAT-6 and could distinguish TB patients from Mycobacterium bovis BCG-vaccinated controls. Interestingly, Rv1168c antigen permits diagnosis of smear-negative pulmonary TB as well as extrapulmonary TB cases, which are often difficult to diagnose by conventional tests. The immunodominant nature of Rv1168c makes it a promising candidate to use in serodiagnosis of TB. In addition, our studies also show that Rv1168c is a potent T-cell antigen which elicits a strong gamma interferon response in sensitized peripheral blood mononuclear cells obtained from TB patients.

scholarly journals Depression, GABA, and Age Correlate with Plasma Levels of Inflammatory Markers

2019 ◽  
Vol 20 (24) ◽  
pp. 6172
Author(s):  
Amol K. Bhandage ◽  
Janet L. Cunningham ◽  
Zhe Jin ◽  
Qiujin Shen ◽  
Santiago Bongiovanni ◽  
...  
Keyword(s):  
Inflammatory Markers ◽  
Major Depressive Episode ◽  
Mononuclear Cells ◽  
Gamma Aminobutyric Acid ◽  
Protein Markers ◽  
Major Depressive ◽  
Peripheral Blood Mononuclear ◽  
Inhibitory Neurotransmitter ◽  
Mental Diseases ◽  
And Control

Immunomodulation is increasingly being recognised as a part of mental diseases. Here, we examined whether levels of immunological protein markers changed with depression, age, or the inhibitory neurotransmitter gamma-aminobutyric acid (GABA). An analysis of plasma samples from patients with a major depressive episode and control blood donors (CBD) revealed the expression of 67 inflammatory markers. Thirteen of these markers displayed augmented levels in patients compared to CBD. Twenty-one markers correlated with the age of the patients, whereas 10 markers correlated with the age of CBD. Interestingly, CST5 and CDCP1 showed the strongest correlation with age in the patients and CBD, respectively. IL-18 was the only marker that correlated with the MADRS-S scores of the patients. Neuronal growth factors (NGFs) were significantly enhanced in plasma from the patients, as was the average plasma GABA concentration. GABA modulated the release of seven cytokines in anti-CD3-stimulated peripheral blood mononuclear cells (PBMCs) from the patients. The study reveals significant changes in the plasma composition of small molecules during depression and identifies potential peripheral biomarkers of the disease.

Bone Marrow Mononuclear Cells Increase Retinal Ganglion Cell Survival and Axon Regeneration in the Adult Rat

2011 ◽  
Vol 20 (3) ◽  
pp. 391-406 ◽  
Author(s):  
Camila Zaverucha-Do-Valle ◽  
Fernanda Gubert ◽  
Michelle Bargas-Rega ◽  
Juliana L. L. Coronel ◽  
Louise A. Mesentier-Louro ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Ganglion Cell ◽  
Cell Survival ◽  
Retinal Ganglion Cell ◽  
Mononuclear Cells ◽  
Axon Regeneration ◽  
Bone Marrow Mononuclear Cells ◽  
Retinal Ganglion ◽  
Adult Rat ◽  
Retinal Ganglion Cell Survival

scholarly journals Changes of Regulatory T and B Cells in Patients with Papillary Thyroid Carcinoma after131I Radioablation: A Preliminary Study

2013 ◽  
Vol 2013 ◽  
pp. 1-8
Author(s):  
Lei Jiang ◽  
Yanxia Zhan ◽  
Yusen Gu ◽  
Yi Ye ◽  
Yunfeng Cheng ◽  
...  
Keyword(s):  
B Cells ◽  
Papillary Thyroid Carcinoma ◽  
Thyroid Carcinoma ◽  
B Cell ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Papillary Thyroid ◽  
T And B Cells ◽  
Significant Difference ◽  
And Control

Introduction. Lymphocytic infiltration and specific lymphocytes subsets may play important roles in papillary thyroid carcinoma (PTC) progression and prognosis. In this study, we try to understand the influence of131I radioablation on the important lymphocytes subtypes of regulatory T and B cells (Tregs and Bregs).Methods. Peripheral blood mononuclear cells from 30 PTC patients before and after131I therapy, and 20 healthy donors were collected. The expression of Tregs (CD4+CD25+CD127-/low) and B cell (CD5+CD19+) and production and secretion of interleukin 10 (IL-10) were analyzed by FACS and ELISA assay, respectively.Results. For Tregs percentage in peripheral blood lymphocytes, there was no difference between pretreatment and control and between posttreatment and control. Compared with pretherapy, increased Tregs infiltration was noted in posttherapy (P<0.05). Although no difference was between pretreatment and control, compared with these two groups, decreased CD19+and CD5+CD19+B cell percentage in posttreatment was observed (P<0.05). Among these groups, no significant difference was displayed in intracellular IL-10 production and extracellular IL-10 secretion.Conclusions.131I Radioablation increased Tregs and decreased CD19+and CD5+CD19+B cells percentage after treatment. However, it has no effect on IL-10 and lymphocytes in peripheral blood. Therefore, longer follow-up of Tregs and Bregs should be further investigated.

scholarly journals Hyperactivity of cathepsin B and other lysosomal enzymes in fibroblasts exposed to azithromycin, a dicationic macrolide antibiotic with exceptional tissue accumulation

FEBS Letters ◽  
1996 ◽  
Vol 394 (3) ◽  
pp. 307-310 ◽  
Author(s):  
Cécile Gerbaux ◽  
Françoise Van Bambeke ◽  
Jean-Pierre Montenez ◽  
Jocelyne Piret ◽  
Grégory Morlighem ◽  
...  
Keyword(s):  
Cathepsin B ◽  
Lysosomal Enzymes ◽  
Macrolide Antibiotic ◽  
Tissue Accumulation

scholarly journals IL-17-Expressing CD4+and CD8+T Lymphocytes in Human Toxoplasmosis

2014 ◽  
Vol 2014 ◽  
pp. 1-7 ◽  
Author(s):  
Jéssica Líver Alves Silva ◽  
Karine Rezende-Oliveira ◽  
Marcos Vinicius da Silva ◽  
César Gómez-Hernández ◽  
Bethânea Crema Peghini ◽  
...  
Keyword(s):  
T Cells ◽  
T Lymphocytes ◽  
Mononuclear Cells ◽  
Parasite Invasion ◽  
Cytokine Pattern ◽  
Cytokine Levels ◽  
Th1 And Th2 Cytokines ◽  
Tc17 Cells ◽  
And Control ◽  
Bead Arrays

This study aimed to measure the synthesis of Th1 and Th2 cytokines by mononuclear cells after culture with liveT. gondiiand identified Th17 (CD4+) and Tc17 (CD8+) cells in toxoplasma-seronegative and toxoplasma-seropositive parturient and nonpregnant women. Cytometric bead arrays were used to measure cytokine levels (IL-2, TNF-α, IFN-γ, IL-4, IL-5, and IL-10); immunophenotyping was used to characterize Th17 and Tc17 cells, and the cells were stained with antibodies against CD4+and CD8+T cells expressing IL-17. The addition of tachyzoites to cell cultures induced the synthesis of IL-5, IL-10, and TNF-αby cells from seronegative parturient women and of IL-5 and IL-10 by cells from seropositive, nonpregnant women. We observed a lower level of IL-17-expressing CD4+and CD8+T lymphocytes in cultures of cells from seronegative and seropositive parturient and nonpregnant women that were stimulated with tachyzoites, whereas analysis of the CD4+and CD8+T cell populations showed a higher level of CD4+T cells compared with CD8+T cells. These results suggest that the cytokine pattern and IL-17-expressing CD4+and CD8+T lymphocytes may have important roles in the inflammatory response toT. gondii, thus contributing to the maintenance of pregnancy and control of parasite invasion and replication.

Histological examination of the cellular reactions around schistosomula of Schistosoma mansoni in the lungs of sublethally irradiated and unirradiated, immune and control rats

Parasitology ◽  
1989 ◽  
Vol 98 (1) ◽  
pp. 57-65 ◽  
Author(s):  
Dario A. A. Vignali ◽  
S. N. Klaus ◽  
Q. D. Bickle ◽  
M. G. Taylor
Keyword(s):  
Schistosoma Mansoni ◽  
Mononuclear Cells ◽  
Significant Proportion ◽  
Immune Serum ◽  
Immune Recognition ◽  
Cellular Infiltration ◽  
Cellular Composition ◽  
Immune Mediated ◽  
Immune Attack ◽  
And Control

SummaryHistopathological data on the cellular reactions (foci) around Schistosoma mansoni schistosomula in the lungs of both irradiated (750 rad) and unirradiated, passively immunized and normal rats were consistent with the idea that a significant proportion of immune-mediated attrition in passively immunized rats occurs in the lungs. In unirradiated rats, immune serum elicited an enhanced (i.e. larger) and accelerated (i.e. more rapidly developing) inflammatory cellular infiltration around lung-stage parasites when administered 5 days post-infection, when the parasites were already in the lungs. This demonstrated the antigenicity of lung-stage schistosomula and their potential as targets for immune attack. In irradiated rats, innate immunity was decreased as judged by an increase in the number of worms recovered by portal perfusion, and was accompanied by an overall decreased percentage of trapped parasites compared with unirradiated controls, suggesting that trapping in the lungs is involved in innate, as well as acquired immunity. In contrast to the results in unirradiated rats, passive transfer of immune serum into irradiated recipients did not result in larger lung foci than in the NRS-recipients. However, there was evidence of an accelerated response resulting in an essentially similar ratio of trapped parasites (VRS- compared with NRS-recipients) in irradiated rats, as compared with unirradiated rats, reflecting the similar levels of resistance manifested in both groups of rats. This also lent credence to the notion that it was the speed of immune recognition of the migrating schistosomula and the establishment of trapping foci that were of greater importance rather than the size of the enveloping granulomata. Investigations into the cellular composition of the foci surrounding trapped parasites in unirradiated rats revealed a predominance of mononuclear cells, with equal proportions of lymphocytes and macrophages. Eosinophils represented less than 3% of the cellular composition of the foci and were typically distant from the parasites themselves, arguing against their role in specific immunity in this model. Irradiation of recipient rats resulted in a corresponding increase in the percentage of macrophages in lung foci.

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