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Behavioral and histopathological studies of cervical spinal cord contusion injury in rats caused by an adapted weight-drop device

Abstract

Background

Models of spinal cord injury (SCI) caused by weight-drop devices to cause contusion have been used extensively, and transient behavioral deficits after thoracic injury have been demonstrated. The severity of the injury caused by the device should be mild enough to allow recovery.

Objective

To determine whether our adapted weight-drop device with a small tip can effectively induce mild hemicontusion at the level of the fifth cervical vertebra.

Methods

We divided 15 adult male Sprague Dawley rats into groups of 5 for the following treatments: sham (SH, laminectomy only), mild (MSCI) or severe SCI (SSCI). Behavioral tests and histopathology were used before (day 1) and after the treatment on days 3, 7, 14, 21, 28, and 35 to assess the injury.

Results

Rats with SSCI showed a significant somatosensory deficit on days 3 and 7 compared with rats in the SH group, recovering by day 14. In a horizontal-ladder test of skilled locomotion, rats with SSCI showed a significant increase in error scores and percentage of total rungs used, and a decrease in the percentage of correct paw placement compared with rats in the SH group. There was greater recovery to normal paw placement by rats with MSCI than by rats with SSCI. These behavioral deficits were consistent with histopathology using hematoxylin and eosin counterstained Luxol fast blue, indicating the degree of injury and lesion area.

Conclusions

Mild hemicontusion caused by the adapted device can be used to evaluate SCI and provides a model with which to test the efficacy of translational therapies for SCI.

Open access
Changes in sperm quality and testicular structure in a rat model of type 1 diabetes

Abstract

Background

Chronic hyperglycemia is a characteristic of diabetes mellitus (DM). Long-lasting hyperglycemia can generate oxidative stress and reactive oxygen species. The effect of this condition on sperm quality and spermatogenesis leads to male infertility and reproductive dysfunction.

Objectives

To investigate changes in sperm quality, morphology of testicular structure, and stage of development of seminiferous tubules in a streptozotocin (STZ)-induced rat model of type 1 DM.

Methods

We divided 15 male Sprague Dawley rats into 2 groups. DM was induced in 7 rats using STZ (60 mg/kg intraperitoneally), while the other 8 were treated with citrate buffer as a vehicle control group. Rat semen was collected for quality measurements including motility, normal morphology, and concentration. Morphological changes in testicular structure and stage of development of seminiferous tubules were investigated by histology with hematoxylin and eosin (HE) staining.

Results

Significant decreases in all parameters of sperm quality and testicular weight were found in rats with induced DM. Moreover, abnormal morphology of seminiferous tubules including separation of the germinal epithelium, vacuolization, luminal sloughing of germ cells, and tubular atrophy was increased significantly in these rats, while the proportion of their seminiferous tubules at an early stage of development was significantly higher, but was dramatically decreased in the late stage of development when compared with that in vehicle-treated control rats.

Conclusions

DM has adverse effects on sperm quality, testicular structure, and development of seminiferous tubules. These findings may reflect the male infertility and reproductive dysfunction seen in patients with type 1 DM.

Open access
Cytotoxic responses of human chondrocytes to bupivacaine, levobupivacaine, and ropivacaine

Abstract

Background

Intra-articular injections of local anesthetics are used commonly in articular surgery. However, chondrocyte viability and metabolism may be adversely affected by various anesthetics.

Objectives

To assess the chondrotoxic effects of bupivacaine, levobupivacaine, and ropivacaine on human chondrocytes and elucidate possible mechanisms of chondrocyte death.

Methods

Cultured human chondrocytes (CHON-001) were exposed to 0.25% or 0.5% of bupivacaine, levobupivacaine, and ropivacaine in vitro. Cell viability was determined by flow cytometry after 15, 30, 60, and 120 min of exposure. Chondrocyte reactive oxygen species (ROS) production was measured every 10 min for up to 1 h using 2ʹ,7ʹ-dichlorodihydrofluorescein staining. Chondrocyte production of glycosaminoglycan was measured by capillary electrophoresis. NO production was measured using a colorimetric assay kit.

Results

We found a significant increase in chondrotoxicity dependent on exposure time and concentration of the anesthetic. At 60 min, chondrocyte viability was significantly (P < 0.05) decreased when exposed to 0.5% levobupivacaine (32.5%), or 0.25% or 0.5% bupivacaine (34.3% or 46.5%, respectively) compared with exposure to phosphate-buffered saline (PBS) vehicle as a control. Cell death at 120 min was mainly necrosis. There was no difference in viability after treatment with either concentration (0.25% or 0.5%) of ropivacaine at any time compared with exposure to PBS. We found increased production of NO, while ROS decreased after exposure to any of the anesthetics tested.

Conclusions

Ropivacaine may be safer than bupivacaine or levobupivacaine as an intra-articular analgesic. Chondrotoxicity of anesthetics in vitro may be mediated via a reactive nitrogen species-dependent pathway.

Open access
Neural cell adhesion molecule (NCAM) and polysialic acid–NCAM expression in developing ICR mice

Abstract

Background

Coexpression of polysialic acid (PSA)–neuronal cell adhesion molecule (NCAM) with immature neuronal markers is used to indicate the developmental state of neurons generated in the subgranular zone (SGZ) of adult hippocampus. PSA–NCAM is highly expressed throughout the embryonic and juvenile mammalian brain, but heavily downregulated in adult brain.

Objective

To visualize the expression profiles of NCAM/PSA–NCAM in the dentate SGZ of the hippocampus in developing ICR mice.

Methods

Cellular distribution, expression, and developmental changes of NCAM/PSA–NCAM were studied in ICR mice at embryonic age 17 days (E17); and similarly at postnatal ages P3, P5, and P7. The SGZ was studied using NCAM and PSA–NCAM immunoreactive staining with or without hematoxylin counterstaining. Western blotting was used to confirm protein expression levels.

Results

NCAM expression was localized to the surface of neurons and glia and was higher in postnatal mice than it was in embryonic mice. PSA–NCAM was found in cytoplasm and membrane of neural cells, more densely staining in the dentate SGZ at P7, but no staining found at E17. Western blotting of brain tissues also showed expression of both PSA–NCAM and NCAM increased significantly at P5 and P7 compared with expression at P3.

Conclusions

Progressive increase in NCAM expression occurs in the SGZ during embryogenic and postnatal development. PSA–NCAM was not expressed in embryonic ICR mice, but was increased after birth and highly localized in the SGZ at P7. This NCAM expression pattern in the developing brain indicating structural plasticity and neurogenesis may be useful for study of brain repair.

Open access
Proteomics study of the antifibrotic effects of α-mangostin in a rat model of renal fibrosis

Abstract

Background

Renal fibrosis is a consequence of a “faulty” wound-healing mechanism that results in the accumulation of extracellular matrix, which could lead to the impairment of renal functions. α-Mangostin (AM) may prevent the formation of liver fibrosis, but there has yet to be a conclusive investigation of its effect on renal fibrosis.

Objectives

To investigate the renoprotective effect of AM against thioacetamide (TAA)-induced renal fibrosis in rats at the morphological and proteomic levels.

Methods

We divided 18 male Wistar rats into 3 groups: a control group, a TAA-treated group, and a TAA + AM group. The various agents used to treat the rats were administered intraperitoneally over 8 weeks. Subsequently, the morphology of renal tissue was analyzed by histology using Sirius Red staining and the relative amount of stained collagen fibers quantified using ImageJ analysis. One-dimensional gel liquid chromatography with tandem mass spectrometry (GeLC-MS/MS) was used to track levels of protein expression. Proteomic bioinformatics tools including STITCH were used to correlate the levels of markers known to be involved in fibrosis with Sirius Red-stained collagen scoring.

Results

Histology revealed that AM could reduce the relative amount of collagen fibers significantly compared with the TAA group. Proteomic analysis revealed the levels of 4 proteins were modulated by AM, namely CASP8 and FADD-like apoptosis regulator (Cflar), Ragulator complex protein LAMTOR3 (Lamtor3), mitogen-activated protein kinase kinase kinase 14 (Map3k14), and C-Jun-amino-terminal kinase-interacting protein 3 (Mapk8ip3).

Conclusion

AM can attenuate renal fibrosis by the suppression of pathways involving Cflar, Lamtor3, Map3k14, and Mapk8ip3.

Open access
Open access
Anatomic variations of coronary arteries: origins, branching patterns, and abnormalities

Abstract

Background

Anatomic variations in orifices, courses, branching patterns, and abnormalities of coronary arteries could affect blood supply, hemodynamic characteristics, and clinical symptoms, and could be a risk of atherosclerosis.

Objectives

To investigate the location and number of both coronary orifices in the aortic cusps, branching patterns of left main trunk, dominant pattern of posterior interventricular artery (PIA), prevalence of right posterior diagonal artery (RPDA), myocardial bridge, and other abnormalities.

Methods

We dissected 95 heart specimens from cadavers of Thai donors without the history of surgery, and the dominant patterns, location and number of orifices in the aortic cusps, branching patterns, origin and number of conal arteries, and occurrence of RPDA were determined.

Results

Dual aortic origin of the coronary orifice was the most common condition. Anomalous 2 orifices in the left aortic cusp were found in one specimen in which the right coronary artery (RCA) arose from aortic cusp and had an interarterial course. Right dominance and trifurcated form of left main trunk were found more frequently. Most frequently 2 conal arteries were found. RPDA was found in 45% and mostly originated from RCA. The prevalence of myocardial bridge was 62% and located mostly on the anterior interventricular artery (AIA).

Conclusions

The prevalence of right dominance, RPDA, the atypical origin of RCA from the left sinus, and the prevalence of myocardial bridges was more frequent than reported by others, whereas the dual aortic origin from both cusps and the prevalence of bifurcated left main trunk was less frequent.

Open access
Association of SLC1A2 and SLC17A7 polymorphisms with major depressive disorder in a Thai population

Abstract

Background

Major depressive disorder (MDD) is a common psychiatric disorder with high prevalence and high risk of suicide. Genetic variation of glutamate transporters may associate with MDD and suicide attempt.

Objectives

To evaluate polymorphisms of excitatory amino acid transporter 2 gene (SLC1A2; rs752949, rs1885343, rs4755404, and rs4354668) and vesicular glutamate transporter 1 gene (SLC17A7; rs1043558, rs2946848, and rs11669017) in patients with MDD with and without suicide attempt, and determine the association of these polymorphisms with age of onset and severity of MDD.

Methods

DNA was extracted from blood taken from patients with MDD (n = 100; including nonsuicidal [n = 50] and suicidal [n = 50] subgroups) and controls (n = 100). Genotyping was conducted using TaqMan single-nucleotide polymorphism (SNP) genotyping.

Results

We found a significant difference in SLC17A7 rs2946848 genotype distribution between patients in the MDD and control groups (P = 0.016). Moreover, significant differences in SLC1A2 rs752949 (P = 0.022) and SLC17A7 rs2946848 (P = 0.026) genotype distributions were observed between patients in the nonsuicidal MDD and suicidal MDD groups. SLC1A2 rs1885343 A allele carriers showed significantly lower age of onset than GG genotype (P = 0.049). Furthermore, the severity of MDD indicated by the Hamilton Depression Rating Scale (HDRS) score of G allele carriers of SLC1A2 rs4755404 was significantly greater than the CC genotype (P = 0.013).

Conclusions

Polymorphisms of SLC1A2 and SLC17A7 may contribute to the risk of MDD and/or suicide attempt. An association of an SLC1A2 polymorphism with the severity of MDD was apparent.

Open access
Evaluation of SHP1-P2 methylation as a biomarker of lymph node metastasis in patients with squamous cell carcinoma of the head and neck

Abstract

Background

Hypermethylation of Src homology region 2 domain-containing protein-tyrosine phosphatase 1 promoter 2 (SHP1-P2) has been proven as an epithelial-specific marker. This marker has been used for the detection of lymph node metastasis in patients with lung cancer or colon cancer.

Objectives

To investigate SHP1-P2 methylation in patients with squamous cell carcinoma of the head and neck (HNSCC) and determine its potential for micrometastasis detection in the lymph nodes of patients with HNSCC.

Methods

SHP1-P2 methylation levels were analyzed by combined methylation-specific primer TaqMan real-time PCR in 5 sample groups: normal tonsils (n = 10), microdissected squamous cell carcinoma epithelia (n = 9), nonmetastatic head and neck cancer lymph nodes (LN N0, n = 15), metastatic HNSCC histologically negative for tumor cells (LN–, n = 18), and matched cases histologically positive for tumor cells (LN+, n = 18).

Results

SHP1-P2 methylation of 10.27 ± 4.05% was found in normal tonsils as a lymphoid tissue baseline, whereas it was 61.31 ± 17.00% in microdissected cancer cell controls. In the 3 lymph node groups, the SHP1-P2 methylation levels were 9.99 ± 6.61% for LN N0, 14.49 ± 10.03% for LN- Nx, and 41.01 ± 24.51% for LN+ Nx. The methylation levels for LN- Nx and LN+ Nx were significantly different (P = 0.0002). Receiver operating characteristic curve analysis of SHP1-P2 methylation demonstrated an area under the curve of 0.637 in distinguishing LN N0 from LN– Nx.

Conclusions

SHP1-P2 methylation was high in HNSCC, and low in lymphoid tissues. This methylation difference is concordant with lymph node metastasis.

Open access
Sex determination in Northern Thai from crania by using computer-aided design software and conventional caliper methods

Abstract

Background

Identification of sex from skeletal remains is an essential step in forensic anthropology. The skull is the second choice, after the pelvis, to estimate sex by osteometric methods.

Objective

To evaluate the process of identification of sex in Northern Thai from crania by using computer-aided design (AutoCAD) software and conventional caliper methods.

Methods

Dry skulls of 86 men and 74 women were examined. AutoCAD software and digital calipers were used to measure dimensions. Eleven of the 15 parameters were created for this study.

Results

Men are significantly larger than women in all parameters, except in the nasospinale–prosthion measurement. There were no significant differences in the intraobserver error test and between the AutoCAD and digital caliper measurements. The logistic regression analysis yielded a sex classification accuracy rate of 92.9% in men, 93.4% in women, and 93.1% of overall accuracy for AutoCAD software. When using digital calipers, there was an accuracy rate of 89.3% in men, 94.7% in women, and 91.9% for overall accuracy.

Conclusions

AutoCAD software is a reliable method to predict the sex and provide high accuracy in sex determination from crania.

Open access