We tried a series of morphological and molecular approaches to identify a new species of Stellantchasmus (Digenea: Heterophyidae) originating from the wrestling half-beaked fish, Dermogenys pusillus of Thailand. Adult worm samples of the new species were recovered from hamsters experimentally infected with the metacercariae from D. pusillus in Thailand. Two isolates (Thai and Korean) of Stellantchasmus falcatus were used as comparative control groups. Worm samples of 3 Stellantchasmus groups were morphologically observed and molecularly analyzed with the mitochondrial cytochrome c oxidase 1 gene. The morphological characteristics of S. dermogenysi n. sp. are similar to S. falcatus originating from brackish water fish, but minor difference was noted including the absence of the prepharynx, position of the ovary near the ceca end, smaller body size, and shorter esophageal length. A phylogenetic tree derived from neighbor-joining and maximum-likelihood methods suggests that S. dermogenysi n. sp. is separated from S. falcatus supported by high bootstrap values. The relative divergences persist between these host-specific trematodes, which we suggest should be recognized as 2 distinct species. Comparisons of S. dermogenysi n. sp. with S. falcatus isolated from mullets in Thailand and Korea indicate a genetic divergence of mitochondrial DNA of 19.4% and 21.7%, respectively. By the present study, a new species, Stellantchasmus dermogenysi n. sp. (Digenea: Heterophyidae), is proposed in Thailand based on molecular evidences, in addition to minor morphological differences between S. falcatus and the new species.
Adult ; Animals ; Body Size ; Cricetinae ; DNA, Mitochondrial ; Electron Transport Complex IV ; Female ; Humans ; Korea ; Metacercariae ; Ovary ; Phylogeny ; Saline Waters ; Smegmamorpha ; Thailand ; Trees ; Wrestling

Currently, compared to jaw-closing (JC) α-motoneurons, the information on the distribution and morphology of glutamatergic synapses on the jaw-closing (JC) γ-motoneurons, which may help elucidate the mechanism of isometric contraction of the JC muscle, is very limited. This study investigated the distribution and ultrastructural features of vesicular glutamate transporter 1 (VGLUT1)- and VGLUT2-immunopositive (+) axon terminals (boutons) on JC γ-motoneurons by retrograde tracing with horseradish peroxidase, electron microscopic immunocytochemistry, and quantitative analysis. About 35% of the boutons on identified JC γ-motoneurons were VGLUT+, and of those, 99% were VGLUT2+. The fraction of VGLUT1+ boutons of all boutons and the percentage of membrane of JC γ-motoneurons covered by these boutons were significantly lower than those for the JC α-motoneurons, revealed in our previous work. The bouton volume, mitochondrial volume, and active zone area of the VGLUT2+ boutons on the JC γ-motoneurons were uniformly small. These findings suggest that the JC γ-motoneurons, in contrast to the JC α-motoneurons, receive generally weak glutamatergic synaptic input almost exclusively from VGLUT2+ premotoneurons that form direct synapse with motoneurons.
Animals ; Horseradish Peroxidase ; Immunohistochemistry ; Isometric Contraction ; Membranes ; Microscopy, Electron ; Mitochondrial Size ; Motor Neurons ; Presynaptic Terminals ; Rats ; Synapses ; Vesicular Glutamate Transport Protein 1

Athletic performance is a complex multifactorial trait involving genetic and environmental factors. The heritability of an athlete status was reported to be about 70% in a twin study, and at least 155 genetic markers are known to be related with athlete status. Mitochondrial DNA (mtDNA) encodes essential proteins for oxidative phosphorylation, which is related to aerobic capacity. Thus, mtDNA is a candidate marker for determining physical performance. Recent studies have suggested that polymorphisms of mtDNA are associated with athlete status and/or physical performance in various populations. Therefore, we analyzed mtDNA haplogroups to assess their association with the physical performance of Korean population. The 20 mtDNA haplogroups were determined using the SNaPshot assay. Our result showed a significant association of the haplogroup F with athlete status (odds ratio, 3.04; 95% confidence interval, 1.094 to 8.464; p = 0.012). Athletes with haplogroup F (60.64 ± 3.04) also demonstrated a higher Sargent jump than athletes with other haplogroups (54.28 ± 1.23) (p = 0.041). Thus, our data imply that haplogroup F may play a crucial role in the physical performance of Korean athletes. Functional studies with larger sample sizes are necessary to further substantiate these findings.
Athletes ; Athletic Performance ; DNA, Mitochondrial ; Genetic Markers ; Humans ; Oxidative Phosphorylation ; Sample Size

The ultrastructural parameters related to synaptic release of endings which are presynaptic to tooth pulp afferent terminals (p-endings) were analyzed to understand the underlying mechanism for presynaptic modulation of tooth pulp afferents. Tooth pulp afferents were labelled by applying wheat-germ agglutinin conjugated horseradish peroxidase to the rat right lower incisor, whereafter electron microscopic morphometric analysis with serial section and reconstruction of p-endings in the trigeminal oral nucleus was performed. The results obtained from 15 p-endings presynaptic to 11 labeled tooth pulp afferent terminals were as follows. P-endings contained pleomorphic vesicles and made symmetrical synaptic contacts with labeled terminals. The p-endings showed small synaptic release-related ultrastructural parameters: volume, 0.82 ± 0.45 µm³ (mean ± SD); surface area, 4.50 ± 1.76 µm²; mitochondrial volume, 0.15 ± 0.07 µm³; total apposed surface area, 0.69 ± 0.24 µm²; active zone area, 0.10 ± 0.04 µm²; total vesicle number, 1045 ± 668.86; and vesicle density, 1677 ± 684/µm². The volume of the p-endings showed strong positive correlation with the following parameters: surface area (r=0.97, P<0.01), mitochondrial volume (r=0.56, P<0.05), and total vesicle number (r=0.73, P<0.05). However, the volume of p-endings did not positively correlate or was very weakly correlated with the apposed surface area (r=-0.12, P=0.675) and active zone area (r=0.46, P=0.084). These results show that some synaptic release-related ultrastructural parameters of p-endings on the tooth pulp afferent terminals follow the "size principle" of Pierce and Mendell (1993) in the trigeminal nucleus oralis, but other parameters do not. Our findings may demonstrate a characteristic feature of synaptic release associated with p-endings.
Animals ; Horseradish Peroxidase ; Incisor ; Mitochondrial Size ; Rats ; Tooth* ; Trigeminal Nuclei

PURPOSE: Mitochondrial disease (MD) and Duchenne muscular dystrophy (DMD) are often associated with cardiomyopathy, but the myocardial variability has not been isolated to a specific characteristic. We evaluated the left ventricular (LV) mass by echocardiography to identify the general distribution and functional changes of the myocardium in patients with MD or DMD. METHODS: We retrospectively evaluated the echocardiographic data of 90 children with MD and 42 with DMD. Using two-dimensional echocardiography, including time-motion (M) mode and Doppler measurements, we estimated the LV mass, ratio of early to late mitral filling velocities (E/A), ratio of early mitral filling velocity to early diastolic mitral annular velocity (E/Ea), stroke volume, and cardiac output. A "z score" was generated using the lambda-mu-sigma method to standardize the LV mass with respect to body size. RESULTS: The LV mass-for-height z scores were significantly below normal in children with MD (-1.02+/-1.52, P<0.001) or DMD (-0.82+/-1.61, P=0.002), as were the LV mass-for-lean body-mass z scores. The body mass index (BMI)-for-age z scores were far below normal and were directly proportional to the LV mass-for-height z scores in both patients with MD (R=0.377, P<0.001) and those with DMD (R=0.330, P=0.033). The LV mass-for-height z score correlated positively with the stroke volume index (R=0.462, P<0.001) and cardiac index (R=0.358, P<0.001). CONCLUSION: LV myocardial atrophy is present in patients with MD and those with DMD and may be closely associated with low BMI. The insufficient LV mass for body size might indicate deterioration of systolic function in these patients.
Atrophy* ; Body Mass Index ; Body Size ; Cardiac Output ; Cardiomyopathies ; Child* ; Echocardiography ; Humans ; Mitochondrial Diseases* ; Muscular Dystrophy, Duchenne* ; Myocardium ; Retrospective Studies ; Stroke Volume

Mycoplasma hyopneumoniae (M. hyopneumoniae) is one of the causative bacteria that can induce chronic enzootic pneumonia, resulting in low production in the swine industry. Potentiation of porcine reproductive and respiratory syndrome virus-induced pneumonia by M. hyopneumoniae has also been recognized. Although some available vaccines have been developed for prevention of M. hyopneumoniae infection, protective immunity is still poor. In this study, in order to provide valuable information on vaccine antigen, we investigated the immunogenicity of M. hyopneumoniae on mouse spleen cells. Concanavalin A (ConA) and lipopolysaccharide (LPS) were used for generation of activated T and B lymphocytes. M. hyopneumoniae made clusters of spleen cells and also affected the cellular activity and viability of spleen cells by alone or with mitogens. Of particular interest, it induced a significant increase in production of TNF-alpha in ConA-treated spleen cells, meaning T helper 1 response. In addition, cell size and mitochondrial membrane potential of M. hyopneumoniae-treated spleen cells were measured by flow cytometric analysis. M. hyopneumoniae did not affect the cell size by alone, whereas ConA or LPS profoundly increased the cell size. Taken together, M. hyopneumoniae significantly affect the cellular activity and cytokine production of spleen cells by alone or in a combination of ConA. This study provides valuable information for production of the vaccine against M. hyopneumoniae.
Animals ; B-Lymphocytes ; Bacteria ; Cell Size ; Concanavalin A ; Membrane Potential, Mitochondrial ; Mice* ; Mitogens ; Mycoplasma hyopneumoniae* ; Mycoplasma* ; Pneumonia ; Porcine Reproductive and Respiratory Syndrome ; Spleen* ; Swine ; Tumor Necrosis Factor-alpha ; Vaccines

OBJECTIVETo investigate the antiobesity effect of Jueming Prescription (JMP), a Chinese herbal medicine formula, and its influence on mRNA expressions of beta3 adrenergic receptor (beta3-AR) and uncoupling protein-2 (UCP-2) in adipose tissue of diet-induced obese rats.

METHODSFifty male Sprague-Dawley rats were randomly divided into the normal control group (n =8) that was on a standard chow diet, and the obese model group (n =42) that was on a diet of high fat chow. Two weeks after the high fat diet, 29 obese rats in the obese model group were further randomly divided into 3 groups: the untreated obese model group (n =9), the metformin group (n =10, metformin 300 mg kg⁻¹ day)⁻¹, and the JMP group (n =10, JMP 4 g kg⁻¹ day⁻¹). After 8-week treatment, body weight, wet weight of visceral fat, and percentage of body fat (PBF) were measured. The levels of fasting blood glucose, serum lipids, and insulin were assessed, and insulin sensitivity index (ISI) was calculated. The adipose tissue section was stained with hematoxylin-Eosin, and the cellular diameter and quantity of adipocytes were evaluated by light microscopy. The mRNA expressions of beta3-AR and UCP-2 from the peri-renal fat tissue were determined by real-time reverse transcription polymerase chain reaction (RT-PCR).

RESULTSCompared with the obese model group, treatment with JMP resulted in significantly lower body weight, wet weight of visceral fat, PBF, and diameter of adipocytes, and significantly higher level of high-density lipoprotein cholesterol, ISI (all P<0.01), JMP increased the mRNA expressions of beta3-AR and UCP-2 from perirenal fat tissue (P <0.05, P<0.01).

CONCLUSIONSJMP could reduce body weight and adipocyte size; and the effect was associated with the up-regulation of beta3-AR and UCP-2 expressions in the adipose tissue and improvement of insulin sensitivity.

Adipocytes ; drug effects ; metabolism ; pathology ; Adiposity ; drug effects ; Animals ; Blood Glucose ; metabolism ; Body Weight ; drug effects ; Cell Size ; drug effects ; Diet, High-Fat ; Drugs, Chinese Herbal ; pharmacology ; Epididymis ; drug effects ; pathology ; Fasting ; blood ; Gene Expression Regulation ; drug effects ; Insulin ; blood ; Intra-Abdominal Fat ; drug effects ; metabolism ; pathology ; Ion Channels ; genetics ; metabolism ; Lipids ; blood ; Male ; Mitochondrial Proteins ; genetics ; metabolism ; Obesity ; blood ; genetics ; pathology ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Receptors, Adrenergic, beta-3 ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Uncoupling Protein 2 ; Weight Loss ; drug effects

This paper reviews the advances of DNA detection on three types of difficult biological specimens including degraded samples, trace evidences and mixed samples. The source of different samples, processing methods and announcements were analyzed. New methods such as mitochondrial test system, changing the original experimental conditions, low-volume PCR amplification and new technologies such as whole genome amplification techniques, laser capture micro-dissection, and mini-STR technology in recent years are introduced.
Biomarkers ; Body Fluids/chemistry* ; DNA/genetics* ; DNA Fingerprinting/methods* ; DNA, Mitochondrial/genetics* ; Forensic Medicine/methods* ; Genome, Human ; Humans ; Laser Capture Microdissection/methods* ; Microsatellite Repeats ; Nucleic Acid Amplification Techniques/methods* ; Polymerase Chain Reaction/methods* ; Reproducibility of Results ; Sample Size

Ultrastructural parameters related to synaptic release and their correlation with synaptic connectivity were analyzed in the low-threshold mechanoreceptive vibrissa afferent boutons in laminae III and IV of the trigeminal caudal nucleus (Vc). Rapidly adapting vibrissa afferents were intra-axonally labeled, and quantitative ultrastructural analyses with serial sections were performed on the labeled boutons and their presynaptic endings (p-endings). The volume of the labeled boutons was widely distributed from small to large ones (0.8~12.3 microm3), whereas the p-endings were small and uniform in size. The volume of the labeled boutons was positively correlated with the ultrastructural parameters such as mitochondrial volume (correlation coefficient, r=0.96), active zone area (r=0.82) and apposed surface area (r=0.79). Vesicle density (r=-0.18) showed little correlation to the volume of labeled boutons, suggesting that the total vesicle number of a bouton is proportional to its volume. In addition, the bouton volume was positively correlated with the number of p-endings (r=0.52) and with the number of dendrites postsynaptic to the labeled bouton (r=0.83). These findings suggest that low-threshold mechanoreception conveyed through vibrissa afferents is processed in a bouton size-dependent manner in the Vc, which may contribute to the sensory-motor function of laminae III/IV in Vc.
Animals ; Cats ; Dendrites ; Mitochondrial Size ; Synapses ; Trigeminal Caudal Nucleus

PURPOSE: Biphasic calcium phosphates have been of great interest recently. Mixing adequate ratios of hydroxyapatite(HA) and beta-tricalcium phosphate(beta-TCP) allowed to control the resorption rate without distorting its osteoconductive property. This study evaluated the bone formation effect of newly developed biphasic calcium phosphate(BCP) in calvarial defect of rabbits. MATERIALS AND METHODS: 6 male New Zealand rabbits were used. Four defects with 8mm in diameter were created on each animal. BCP with HA/beta-TCP ratio of 7:3 and particle size of 0.5~1.0 mm was used as the test group and bovine bone with 0.25~1.0 mm particle size, as the control group. Both test and control group materials were randomly implanted in the calvarial defects and were covered witha polymer membrane. The animals were sacrificed after 12, 24, and 48 weeks of implantation under general euthanasia. Resin blocks were obtained and were stained by masson's trichrome for histological observation. RESULTS: Overall results were uneventful without any defect exposure or inflammation. The amount of new bone formation and bone maturity increased with increase in healing period at both groups. New bone in test group was mostly formed along the material particle surrounded by osteoblasts, and observation of osteoblastic stream was also present. Bone maturity increased as it was closer to thedefect margins. Under the same healing period, the test group showed more bone formation than the control group with more stable bovine bone particles remaining even after 48 weeks, whereas considerable resorption took place in BCP. Almost total defect closure was observed in test group with new bone formation in the central part of the defect. However, limited new bone formation was observed in the control group. CONCLUSION: Within the limits of the study, the present study reveals the newly developed BCP to be a good osteoconductive material. However, further studies are needed to be conducted in a different study model with a larger sample size.
Animals ; Calcium ; Calcium Phosphates ; Durapatite ; Euthanasia ; Humans ; Hydroxyapatites ; Hypogonadism ; Inflammation ; Male ; Membranes ; Mitochondrial Diseases ; Ophthalmoplegia ; Osteoblasts ; Osteogenesis ; Particle Size ; Pilot Projects ; Polymers ; Rabbits ; Rivers ; Sample Size