Tongue-palatal contact changes in patients with skeletal mandibular prognathism after sagittal split ramus osteotomy: an electropalatography study.

The purpose of this study was to investigate the changes in tongue-palatal contact patterns using electropalatography (EPG) before and after sagittal split ramus osteotomy (SSRO) in patients with mandibular prognathism. Nine clients who underwent SSRO for mandibular setback and seven control subjects were participated in this study. Tongue-palatal contact patterns for /t/, /s/ and /k/ production were investigated using EPG before surgery and 3 months after surgery. The mean value of whole total of palate contact (WT) in the maximum contact frame was examined before and after SSRO. The correlation quantity between the change of center of gravity (COG) value and the amount of mandibular setback was also evaluated. The mean value of WT for /t/ and /s/ significantly increased after SSRO, and the EPG pattern became normal. However, a remarkable change in WT for /k/ was not observed, and the mean value was significantly larger in the SSRO group before and after surgery than in the control group. A negative correlation between COG variation and the amount of mandibular setback for /t/ and positive correlation for /s/ was observed. This study demonstrated that tongue-palatal contact patterns for /t/ and /s/ articulation improved clearly after SSRO. There was a significant correlation between COG variation and the amount of mandibular setback. However, no significant change was detected through perceptual assessment before and after SSRO. Further investigation is needed to determine whether these results will change over time.

A new method for characterizing hand dysfunction in cervical spondylotic myelopathy: a preliminary study.

STUDY DESIGN: A case-control investigation. OBJECTIVES: The objective of this study was to quantitatively study impaired ability to appropriately adjust pinch strength while holding a small object in patients with cervical spondylotic myelopathy (CSM). SETTING: Kochi Medical School Hospital, Japan. METHODS: The subjects consisted of 19 CSM patients who had frequent episodes of failing to grasp and hold small objects in their daily life (Group A), 13 CSM patients who did not experience such episodes (Group B) and 16 healthy subjects (Control Group). We continuously measured the dynamic internal pressure of a pneumatic rubber object called a blower pinched by the subject, following two different sets of instructions: (1) pinching with eyes open and with the minimal strength required to prevent dropping; and (2) maintaining a constant pinch strength at given levels with eyes closed. RESULTS: Compared with the other two groups, Group A subjects used a significantly (P<0.01) greater pinch strength to avoid dropping the blower held with eyes open and showed a significantly (P<0.01) greater deviation in pinch strength from the baseline values with eyes closed. These tendencies in Group A showed a significant correlation with the tactile perception threshold of the digits (P<0.01) but not with impairment of rapid repetitive movements of the digits that reflects spasticity. CONCLUSION: Our technique applied to CSM patients helps assess functional integrity primarily, if not exclusively, of the fasciculus cuneatus mediating the feedback signals from proprioceptive and cutaneous receptors in the digits, which are otherwise difficult to evaluate quantitatively.

Altitudinal flower size variation correlates with local pollinator size in a bumblebee-pollinated herb, Prunella vulgaris L. (Lamiaceae).

The influence of locally different species interactions on trait evolution is a focus of recent evolutionary studies. However, few studies have demonstrated that geographically different pollinator-mediated selection influences geographic variation in floral traits, especially across a narrow geographic range. Here, we hypothesized that floral size variation in the Japanese herb Prunella vulgaris L. (Lamiaceae) is affected by geographically different pollinator sizes reflecting different pollinator assemblages. To evaluate this hypothesis, we posed two questions. (1) Is there a positive correlation between floral length and the proboscis length of pollinators (bumblebees) across altitude in a mountain range? (2) Does the flower-pollinator size match influence female and male plant fitness? We found geographic variation in the assemblage of pollinators of P. vulgaris along an altitudinal gradient, and, as a consequence, the mean pollinator proboscis length also changed altitudinally. The floral corolla length of P. vulgaris also varied along an altitudinal gradient, and this variation strongly correlated with the local pollinator size but did not correlate with altitude itself. Furthermore, we found that the size match between the floral corolla length and bee proboscis length affected female and male plant fitness and the optimal size match (associated with peak fitness) was similar for the female and male fitness. Collectively, these results suggest that pollinator-mediated selection influences spatial variation in the size of P. vulgaris flowers at a fine spatial scale.

Characterization of water pollution in drainage networks using continuous monitoring data in the Citadel area of Hue City, Vietnam.

In the Citadel area of Hue City, drainage systems that include canals and ponds are considerable sources of fecal contaminants to inundated water during the rainy season because canals and ponds receive untreated wastewater. It is important to investigate the characteristics of hydraulics and water pollution in canals and ponds. At the canals and ponds, water sampling was conducted during dry and wet weather periods in order to evaluate fecal contamination and to investigate changes in water pollution caused by runoff inflow. Inundated water was also collected from streets during heavy rainfall. At the canals and ponds, concentrations of Escherichia coli and total coliform exceeded the Vietnamese regulation values for surface water in 23 and 24 out of 27 samples (85 and 89%), respectively. The water samples were categorized based on the characteristics of water pollution using cluster analysis. In the rainy season, continuous monitoring was conducted at the canals and ponds using water depth and electrical conductivity (EC) sensors to investigate the dynamic relationship between water level and water pollution. It is suggested that in the canals, high EC meant water stagnation and low EC signified river water inflow. Therefore, EC might be a good indicator of water flow change in canals.

Runoff load estimation of particulate and dissolved nitrogen in Lake Inba watershed using continuous monitoring data on turbidity and electric conductivity.

Easy-to-measure surrogate parameters for water quality indicators are needed for real time monitoring as well as for generating data for model calibration and validation. In this study, a novel linear regression model for estimating total nitrogen (TN) based on two surrogate parameters is proposed based on evaluation of pollutant loads flowing into a eutrophic lake. Based on their runoff characteristics during wet weather, electric conductivity (EC) and turbidity were selected as surrogates for particulate nitrogen (PN) and dissolved nitrogen (DN), respectively. Strong linear relationships were established between PN and turbidity and DN and EC, and both models subsequently combined for estimation of TN. This model was evaluated by comparison of estimated and observed TN runoff loads during rainfall events. This analysis showed that turbidity and EC are viable surrogates for PN and DN, respectively, and that the linear regression model for TN concentration was successful in estimating TN runoff loads during rainfall events and also under dry weather conditions.

Emdogain stimulates matrix degradation by osteoblasts.

Emdogain has been used clinically for periodontal regeneration, although the underlying molecular mechanisms are not clear at present. In this study, we hypothesized that Emdogain stimulated degradation of type I collagen via osteoblasts. We showed that Emdogain enhanced cell-mediated degradation of type I collagen in an MMP-dependent manner. Although MG-63 cells spontaneously produced a zymogen form of MMP-1, treatment with Emdogain significantly induced the generation of the active form of this enzyme. We demonstrated that MMP-3 was produced from MG63 cells in response to Emdogain in a MEK1/2-dependent manner. Concomitantly, blocking of MEK1/2 activation by U0126 significantly inhibited the generation of the active form of MMP-1 without affecting the total production of this collagenase. These results suggest that Emdogain facilitates tissue regeneration through the activation of the collagenase, MMP-1, that degrades matrix proteins in bone tissue microenvironments.

Statistical modelling of knee valgus during a continuous jump test.

Landing with the knee in a valgus position is recognized as a risk factor for anterior cruciate ligament (ACL) injury. Using linear and non-linear regression analyses, the purpose of this study was to examine the correlation between two-dimensional (2D) knee valgus and three-dimensional (3D) knee kinematics measured during a jump landing task. Twenty-eight female collegiate athletes participated. All participants were required to perform a continuous jump test. The average maximum angles of abduction and internal tibial rotation during landing were measured using the Point Cluster Technique. Average peak knee valgus angle was measured using a 2D approach. Linear and non-linear regression analyses between 2D valgus and 3D knee abduction, and between 2D valgus and 3D internal tibial rotation, were performed. The R2 value between 2D valgus and 3D knee abduction was significantly different from zero and had a moderate correlation for all models, whereas the R2 value between 2D valgus and 3D internal tibial rotation was not significantly different from zero. The 2D approach could be used to screen a specific group of individuals for risk of ACL injury; however, using frontal plane 2D analysis of valgus motion to evaluate internal tibial rotation is not advised.

Development of a genus-specific PCR assay for the molecular detection, confirmation and identification of Fusobacterium spp.

A genus-specific polymerase chain reaction (PCR)-based assay is developed for the detection and identification of clinically relevant Fusobacterium species, including F. nucleatum and F. necrophorum. Two 16S ribosomal DNA (rDNA) primers, FUSO1 (forward primer: 5'-GAG AGA GCT TTG CGT CC-3' [17-mer]) and FUSO 2 (reverse primer: 5'-TGG GCG CTG AGG TTC GAC -3' [18-mer]) are designed to target conserved regions of the 16S rDNA gene for Fusobacterium spp. Subsequent proof-of-principle studies employing this assay detected Fusobacterium spp. in the faeces of eight (10%) out of 80 patients with suspected gastrointestinal infection. This assay may be used for the genus-specific detection of Fusobacterium spp. from clinical specimens and for subsequent species identification.

A novel class of plant-specific zinc-dependent DNA-binding protein that binds to A/T-rich DNA sequences.

Complementary DNA encoding a DNA-binding protein, designated PLATZ1 (plant AT-rich sequence- and zinc-binding protein 1), was isolated from peas. The amino acid sequence of the protein is similar to those of other uncharacterized proteins predicted from the genome sequences of higher plants. However, no paralogous sequences have been found outside the plant kingdom. Multiple alignments among these paralogous proteins show that several cysteine and histidine residues are invariant, suggesting that these proteins are a novel class of zinc-dependent DNA-binding proteins with two distantly located regions, C-x(2)-H-x(11)-C-x(2)-C-x((4-5))-C-x(2)-C-x((3-7))-H-x(2)-H and C-x(2)-C-x((10-11))-C-x(3)-C. In an electrophoretic mobility shift assay, the zinc chelator 1,10-o-phenanthroline inhibited DNA binding, and two distant zinc-binding regions were required for DNA binding. A protein blot with (65)ZnCl(2) showed that both regions are required for zinc-binding activity. The PLATZ1 protein non-specifically binds to A/T-rich sequences, including the upstream region of the pea GTPase pra2 and plastocyanin petE genes. Expression of the PLATZ1 repressed those of the reporter constructs containing the coding sequence of luciferase gene driven by the cauliflower mosaic virus (CaMV) 35S90 promoter fused to the tandem repeat of the A/T-rich sequences. These results indicate that PLATZ1 is a novel class of plant-specific zinc-dependent DNA-binding protein responsible for A/T-rich sequence-mediated transcriptional repression.

Nucleotide sequencing and analysis of 16S rDNA and 16S-23S rDNA internal spacer region (ISR) of Taylorella equigenitalis, as an important pathogen for contagious equine metritis (CEM).

The primer set for 16S rDNA amplified an amplicon of about 1500 bp in length for three strains of Taylorella equigenitalis (NCTC11184(T), Kentucky188 and EQ59). Sequence differences of the 16S rDNA among the six sequences, including three reference sequences, occurred at only a few nucleotide positions and thus, an extremely high sequence similarity of the 16S rDNA was first demonstrated among the six sequences. In addition, the primer set for 16S-23S rDNA internal spacer region (ISR) amplified two amplicons about 1300 bp and 1200 bp in length for the three strains. The ISRs were estimated to be about 920 bp in length for large ISR-A and about 830 bp for small ISR-B. Sequence alignment of the ISR-A and ISR-B demonstrated about 10 base differences between NCTC11184(T) and EQ59 and between Kentucky188 and EQ59. However, only minor sequence differences were demonstrated between the ISR-A and ISR-B from NCTC11184(T) and Kentucky188, respectively. A typical order of the intercistronic tRNAs with the 29 nucleotide spacer of 5'-16S rDNA-tRNA(Ile)-tRNA(Ala)-23S rDNA-3' was demonstrated in the all ISRs. The ISRs may be useful for the discrimination amongst isolates of T. equigenitalis if sequencing is employed.

Uptake of acetylated LDL by peritoneal macrophages obtained from normal and Watanabe heritable hyperlipidemic rabbits, an animal model for familial hypercholesterolemia.

Acetylated low-density lipoprotein (acetyl-LDL) stimulated the incorporation of [14C]oleate into cholesteryl [14C]oleate in peritoneal macrophages from both normal and Watanabe heritable hyperlipidemic (WHHL) rabbits. A degradation study showed that macrophages from WHHL rabbits degraded the same amount of 125I-labeled acetyl-LDL as macrophages from normal rabbits. These findings indicate that macrophages of WHHL rabbits have functional acetyl-LDL receptors.

Oxygen uptake efficiency slope: a new index of cardiorespiratory functional reserve derived from the relation between oxygen uptake and minute ventilation during incremental exercise.

OBJECTIVES: We investigated the usefulness of a new variable, oxygen uptake efficiency slope (OUES), as a submaximal measure of cardiorespiratory functional reserve. The OUES is derived from the relation between oxygen uptake (Vo2 [ml/min]) and minute ventilation (VE [liters/min]) during incremental exercise and is determined by VO2 = a log VE + b, where a = OUES, which shows the effectiveness of Vo2. BACKGROUND: Maximal oxygen uptake (VO2max) is effort dependent. There is no standard submaximal measurement of cardiorespiratory reserve that provides generally acceptable results. METHODS: Exercise tests, following a standard Bruce protocol, were performed on a treadmill by 108 patients with heart disease and 36 normal volunteers. Expired gas was continuously analyzed. The OUES was calculated from data of the first 75%, 90% and 100% of exercise duration. We also determined the following submaximal variables: the ventilatory anaerobic threshold (VAT), the slope of the regression line of the minute ventilation-carbon dioxide production relation (VE-VCO2 slope) and the extrapolated maximal oxygen consumption (EMOC). We analyzed the relation of OUES and other submaximal variables against VO2max and examined the effects of submaximal exercise on OUES. RESULTS: The correlation coefficient of the logarithmic curve-fitting model was 0.978 +/- 0.016 (mean +/- SD). The OUES and VO2max had a significant correlation (r = 0.941, p < 0.0001). The correlation between VO2max and OUES was stronger than that between VO2max and VAT, the VE-VCO2 slope or EMOC. The OUES values for 100% and 90% of exercise were not different from each other (at an alpha value of 0.05 and treatment effect of 170, the power of the test [1-beta] was 0.90); OUES for 75% of exercise was slightly lower (3.5%). CONCLUSIONS: Our results suggest that OUES may provide an objective, effort-independent estimation of cardiorespiratory functional reserve that is related both to pulmonary dead space and to metabolic acidosis.

Tumor necrosis factor-alpha promotes proliferation of endometriotic stromal cells by inducing interleukin-8 gene and protein expression.

Endometriosis, a common disease among women of reproductive age, is characterized by the presence of endometrial-like tissue outside the uterus. We and others showed that several cytokine levels, including interleukin-8 (IL-8) and tumor necrosis factor-alpha (TNFalpha), are elevated in the peritoneal fluid of women with endometriosis compared with those in women without endometriosis. We also demonstrated that the addition of IL-8 to the culture medium stimulated the proliferation of cultured endometriotic stromal cells. TNFalpha is a multipotent cytokine that induces IL-8 production in various cell types. Therefore, we hypothesized that TNFalpha may also contribute to the pathogenesis of endometriosis by inducing the production of IL-8. To test this hypothesis, we analyzed the peritoneal fluid concentrations of IL-8 and TNFalpha using enzyme-linked immunosorbent assay (ELISA). We observed a significant correlation between the levels of TNFalpha and IL-8 in the peritoneal fluid of endometriosis patients. We also obtained the endometriotic stromal cells from chocolate cyst linings of the ovary. The expression of the receptors for TNFalpha (TNFR) was examined by RT-PCR. We observed the expression of both TNFR-I and TNFR-II genes in endometriotic stromal cells. The expression of IL-8 gene and protein was analyzed by Northern blot hybridization and enzyme-linked immunosorbent assay, respectively. TNFalpha induced the gene and protein expression of IL-8 in endometriotic stromal cells in a dose-dependent fashion. The addition of TNFalpha promoted the proliferation of the endometriotic stromal cells, and the stimulatory effects of TNFalpha were abolished by adding anti-IL-8 antibody. We demonstrated for the first time that TNFalpha stimulated proliferation of endometriotic stromal cells through induction of IL-8 gene and protein expression. We concluded that the TNFalpha may be one of the essential factors for the pathogenesis of endometriosis.

Immunochemical identification of exposed regions of the Na+,K(+)-ATPase alpha-subunit.

A polyclonal antibody against the Na+,K(+)-ATPase holoenzyme was prepared. This antibody recognized native Na+,K(+)-ATPase and inhibited its activity. The peptide fragments corresponding to various regions of the Na+,K(+)-ATPase alpha-subunit, which were synthesized from the cDNA, were immunoprecipitated with the antibody, and the M32-D75, M158-D197 and M470-V552 fragments (the latter included K508, a putative ATP binding site) were identified as the epitopes. The M267-I442 fragment, which included a phosphorylation site at D376, and the C-terminal one-third of the alpha-subunit from M615 to the C-terminus, were not detected using this antibody. These results suggest that at least three regions of the Na+,K(+)-ATPase alpha-subunit, M32-D75, M158-D197 and M470-V552, cover its exposed regions, and that some of them are essential for ATPase activity.

Detection of small GTP-binding proteins in the outer envelope membrane of pea chloroplasts.

We found small GTP-binding proteins in the outer envelope membrane of pea chloroplasts. The proteins in this membrane were separated by SDS-PAGE, transferred to a nitrocellulose filter, and incubated with [alpha-32P]GTP. Three GTP-binding proteins with the molecular weight of 24,000 were found. Binding was prevented by 10(-8)-10(-7) M GTP or by 10(-7) M guanosine 5'-[gamma-thio]triphosphate or GDP; binding was unaffected by 10(-8)-10(-6) M ATP. Thermolysin treatment of intact chloroplasts resulted in the loss of GTP-binding activity, suggesting that these proteins were in the cytosolic side of the outer envelope membrane.

Chloroplast envelope protein encoded by chloroplast genome.

The gene product of an open reading frame of chloroplast genome, ORF 231 in pea, was immunochemically detected in chloroplast and etioplast envelopes. This is the first protein of a Chloroplast Envelope Membrane encoded by a chloroplast genome. It was named CEM A and the gene, cem A. CEM A is an acidic protein having an apparent molecular mass of 34 kDa on SDS-PAGE, and a minor component detected in the fractionated inner envelope.

Cholesterol efflux effect of high density lipoprotein is impaired by whole cigarette smoke extracts through lipid peroxidation.

It has been reported that high density lipoprotein (HDL) plays an anti-atherogenic role by stimulating cholesterol efflux from the foam cells in the atheromatous lesion. In this study, we prepared a novel modified form of HDL (CS-HDL) by incubating HDL with whole cigarette smoke (CS) extracts containing both particulate matter and gas-phase smoke, and examined its effect on cholesterol efflux. CS-HDL showed a marked increase of conjugated dienes and denaturation of apoA-I, a major protein component of HDL. The cholesterol efflux effect of CS-HDL was remarkably reduced to the same level as that of oxidatively modified HDL induced by copper ion (Ox-HDL). Addition of 20 microg/ml superoxide dismutase (SOD) during the CS-modification of HDL caused retrieval of cholesterol efflux activity by 53% and a remarkable decrease in the conjugated dienes level. SOD, however, had no ameliorative effect on apoA-I denaturation. When HDL was incubated only with gas-phase smoke (gasCS-HDL), neither increase of conjugated dienes nor impairment of the cholesterol efflux effect was observed, whereas apoA-I was denaturated to the same extent as seen in CS-HDL. These results indicate that whole CS-extracts, but not gas-phase smoke, reduces cholesterol efflux effect of HDL and that lipid peroxidation associated with superoxide anion is involved in this functional impairment.

Endocrinologic regulation of carbohydrate metabolism. Amyotrophic lateral sclerosis and Parkinsonism-dementia on Guam.

Studies of the endocrinologic control of carbohydrate metabolism were conducted in Guamanians with parkinsonism-dementia (PD) or amyotrophic lateral sclerosis (ALS) and in Guamanian control patients who had various other neuromuscular disorders. Intravenously infused arginine tended to produce a more prolonged elevation in serum glucose levels in PD and ALS patients than in control subjects. On the other hand, the serum insulin response to arginine was significantly less in both PD and ALS patients than in controls. Arginine stimulated the release of growth hormone to a similar degree in all three patient groups. These observations support and extend previous reports of endocrinologic abnormalities in parkinsonism and ALS and might suggest that a defect in pancreatic islet cell function attends these disorders.

Abnormalities of erythrocyte membranes in myotonic muscular dystrophy manifested in lipid vesicles.

Sodium flux was investigated in lipid vesicles derived from erythrocyte membranes of patients with myotonic dystrophy (MyD) and matched health controls. The lipid vesicles were prepared with phospholipid from erythrocyte membranes and "spectrin" protein preparations. Abnormal sodium permeability was demonstrated in vesicles prepared from phospholipid from the MyD patients.

Different expression of modified low density lipoprotein receptors in rabbit peritoneal macrophages and Kupffer cells.

We have previously reported that mouse peritoneal macrophages have three types of modified low density lipoprotein (LDL) receptors. One is specific for acetylated LDL (Ac-LDL), the second is for oxidized LDL (Ox-LDL), and the third recognizes both (Arai, H. et al. (1989) Biochem. Biophys. Res. Commun. 159, 1375-1382). In the current study, the characteristics of modified LDL receptors in rabbit peritoneal macrophages and Kupffer cells from rabbits were investigated. Cross-competition studies of the degradation assay between Ox-LDL and Ac-LDL in rabbit peritoneal macrophages showed that the degradation of 125I-labeled Ox-LDL was almost completely inhibited by an excess amount of unlabeled Ac-LDL. On the other hand, an excess amount of unlabeled Ox-LDL suppressed 125I-labeled Ac-LDL degradation only partially. In contrast, in Kupffer cells an excess amount of unlabeled Ox-LDL inhibited the degradation of 125I-labeled Ac-LDL almost completely, whereas the degradation of 125I-labeled Ox-LDL was inhibited only partially by Ac-LDL. Scatchard analysis of binding assay showed that rabbit peritoneal macrophages have a single class of receptor for Ox-LDL, which binds maximally 0.31 microgram/mg cellular protein (Bmax) with an apparent dissociation constant (Kd) of 19.3 micrograms/ml, and two classes of receptors for Ac-LDL; one with high affinity (Bmax 0.025 microgram/mg cellular protein, Kd 0.040 micrograms/ml) and the other with low affinity (Bmax 0.08 microgram/mg cellular protein, Kd 11.31 micrograms/ml). On the other hand, Kupffer cells have two classes for Ox-LDL; one is a high affinity receptor (Bmax 0.53 microgram/mg cellular protein, Kd 0.99 microgram/ml) and the other is a low affinity receptor (Bmax 3.71 micrograms/mg cellular protein, Kd 16.2 micrograms/ml) and a single class for Ac-LDL (Bmax 0.60 microgram/mg cellular protein, Kd 7.24 micrograms/ml). These results indicate that rabbit peritoneal macrophages have two kinds of modified LDL receptors; one is specific for Ac-LDL, and the other recognizes both Ox-LDL and Ac-LDL.