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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
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
Unusual accessory peroneal muscles, peroneus quartus, peroneus digiti quinti, and their association with peroneus brevis tendon tear

Abstract

Background

Anatomic variation and supernumerary contents in the superior peroneal tunnel, and the prominence of the retrotrochlear eminence and peroneal tubercle are related to peroneal tendon disorders.

Objectives

To investigate the prevalence, origin, and insertion of accessory peroneal muscles, the prominence of the retrotrochlear eminence and peroneal tubercle, and their association with peroneal tendon tears.

Methods

We examined 109 formalin-embalmed legs of cadavers from Thai donors. Accessory peroneal muscles and peroneal tendon tears were noted. Associations with peroneal tendon tears were evaluated using a χ2 test.

Results

We found 48 accessory peroneal muscles comprising 13 peroneus quartus (PQ), 33 peroneus digiti quinti (PDQ), and 2 unusual muscles. All PDQ originated from the PB tendon and inserted on various parts of the 5th toe. The PQ originated mostly from the PB muscle belly and less from the tendinous part with various insertions on the retrotrochlear eminence, peroneal tubercle, cuboid, and dorsolateral surface of the 5th metatarsal base. Two unusual accessory muscles were identified, 1 coexisting with the PQ. A PB tendon tear was found in 13% of specimens. We found no association between the peroneal tendon tears and the accessory peroneal muscles, or prominence of the retrotrochlear eminence or peroneal tubercle.

Conclusions

The prevalence of PQ, PDQ, and unusual accessory peroneal muscles was concordant with previous findings. We noted a new type of unusual accessory peroneal muscle coexisting with the PQ. No association was found between peroneal tendon tears and the PQ, PDQ, or prominence of the retrotrochlear eminence or peroneal tubercle.

Open access
21st Century era of anatomy
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
Open access
Anatomy of the vasculature supplying hepatobiliary structures and celiac trunk branching patterns in the Thai population

Abstract

Background

Knowledge of the anatomy of the celiac trunk (CT) and arterial supply of the hepatobiliary system is essential for surgical and interventional radiological treatment of upper abdominal diseases.

Objectives

To determine the branching patterns of the CT and variation in origin and type of the right hepatic artery (RHA), left hepatic artery (LHA), and cystic artery (CA).

Methods

The anatomy of the CT in 100 cadavers from Thai adult donors was observed in 3 aspects: its branching pattern, the origin of the RHA and LHA, and the origin of the CA and its relation to the common bile duct (CBD) and common hepatic duct (CHD).

Results

The majority of the CT branching pattern was categorized as the type II classical pattern, which has 3 branches: the left gastric artery (LGA), splenic artery (SA), and common hepatic artery (CHA). The RHA branched from proper hepatic artery in 67 cadavers. The origin of the accessory RHA was either from the abdominal aorta or superior mesenteric artery (SMA), whereas the replaced RHA originated from the CHA, SMA, or CT. The accessory LHA ramified from CHA (2 cases) and LGA (1 case). The replaced LHA was found in 30 cadavers and 29 arose from the CHA. The single and double types of CA were found in 94 and 4 cadavers, respectively. In all, 57% of single CA passed posteriorly and 39% passed anteriorly to the CBD and CHD.

Conclusions

To lower posttreatment complications, variations in the anatomy and the vascular supply of hepatobiliary structures should be considered.

Open access
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