Long waiting time before tooth extraction may increase delayed wound healing in elderly Japanese.

In osteoporosis patients receiving antiresorptive medications, stopping the drug and delaying tooth extraction has been suggested to reduce the risk of osteonecrosis of the jaw (ONJ). However, postponing tooth extraction for ≥ 2 months was associated with an increased risk of delayed wound healing beyond 8 weeks after extraction, a risk factor for developing ONJ. INTRODUCTION: A long waiting time before tooth extraction could result from concern about a potential increased risk of osteonecrosis of the jaw (ONJ) in osteoporosis patients. We clarified whether a long waiting time before tooth extraction during the past year may be associated with an increased risk of delayed wound healing beyond 8 weeks after tooth extraction, which may be a risk factor of ONJ. METHODS: Of 5639 patients aged ≥ 60 years who visited our 20 clinics or hospitals and answered a structured questionnaire, 426 patients (151 men, 275 women) aged 60-96 years comprised the final participants in this study. Self-reported kyphosis was used as a surrogate marker of vertebral fractures. Stepwise logistic regression analysis, adjusted for covariates, was used to calculate the odds ratio (OR) and the 95% confidence interval (CI) for the presence of delayed wound healing longer than 8 weeks after tooth extraction during the past year based on the duration before extraction. RESULTS: Subjects who had waited > 2 months for tooth extraction had a significantly higher risk of delayed wound healing compared with those whose tooth was extracted within 1 month (OR = 7.23; 95% CI = 2.19-23.85, p = 0.001) regardless if antiresorptive medications for osteoporosis were used. The presence of self-reported kyphosis was significantly associated with an increased risk of delayed wound healing (OR = 5.08; 95% CI = 1.11-23.32, p = 0.036). CONCLUSIONS: A long waiting time before tooth extraction may be a risk factor for delayed wound healing beyond 8 weeks after extraction in patients aged ≥ 60 years.

A translocation interrupts the COL5A1 gene in a patient with Ehlers-Danlos syndrome and hypomelanosis of Ito.

Ehlers-Danlos syndrome (EDS) is a genetically and pathogenetically heterogeneous group of disorders of which at least 11 types have been described. All are connective tissue disorders characterized by defects of the skin, ligaments and blood vessels with the clinical spectrum ranging from innocuous findings to lethality. Mutations in the genes encoding the major fibrillar collagen types I and III have been demonstrated in EDS types VII and IV, respectively, while mutations in the lysyl hydroxylase and ATP7A genes, with roles in collagen cross-linking, are responsible for EDS types VI and IX. The biochemical and molecular bases for the most common forms of EDS (types I, II and III) are unknown. Here, we describe a balanced translocation between chromosome 9 and an X chromosome that disrupts the minor fibrillar collagen type V gene COL5A1 in a patient with both EDS type I and hypomelanosis of Ito. The breakpoint occurs at 9q34 within COL5A1 intron 24 and interestingly, within a LINE-1 (L1) element at Xp21.1. A fusion mRNA between COL5A1 and an Alu sequence is produced, but no aberrant protein is detectable. Rather, the amount of type V collagen is reduced in the patient's fibroblasts, suggesting haploinsufficiency as a cuase of the phenotype. This demonstrates that a mutation in a type V collagen gene, COL5A1, results in EDS type I, and shows the involvement of L1 sequences in a constitutional chromosomal translocation. Because collagen type V is a heteromorphic protein in which molecules may be composed of polypeptides encoded by three COL5A genes, this suggests all three genes as candidates for mutations in EDS.

Bone morphogenetic protein-1: the type I procollagen C-proteinase.

Bone morphogenetic proteins (BMPs) are bone-derived factors capable of inducing ectopic bone formation. Unlike other BMPs, BMP-1 is not like transforming growth factor-beta (TGF-beta), but it is the prototype of a family of putative proteases implicated in pattern formation during development in diverse organisms. Although some members of this group, such as Drosophila tolloid (TLD), are postulated to activate TGF-beta-like proteins, actual substrates are unknown. Procollagen C-proteinase (PCP) cleaves the COOH-propeptides of procollagens I, II, and III to yield the major fibrous components of vertebrate extracellular matrix. Here it is shown that BMP-1 and PCP are identical. This demonstration of enzymatic activity for a BMP-1/TLD-like protein links an enzyme involved in matrix deposition to genes involved in pattern formation.

Dual Gas Treatment With Hydrogen and Carbon Monoxide Attenuates Oxidative Stress and Protects From Renal Ischemia-Reperfusion Injury.

BACKGROUND: Hydrogen (H2) and carbon monoxide (CO) gas are both reported to reduce reactive oxygen species and alleviate tissue ischemia-reperfusion (I-R) injury. The present study was conducted to evaluate the effects of a mixture of H2 gas and CO gas (dual gas) in comparison with hydrogen gas (H2: 2%) alone on I-R renal injury (composition of dual gas; N2: 77.8%; O2: 20.9%; H2: 1.30%; CO: 250 parts per million). METHODS: Adult male Sprague-Dawley rats (body weight 250-280 g) were divided into 5 groups: (1) sham operation control, (2) dual gas inhalation (dual treatment) without I-R treatment, (3) I-R renal injury, (4) H2 gas alone inhalation (H2 treatment) with I-R renal injury, and (5) dual treatment with I-R renal injury. I-R renal injury was induced by clamping the left renal artery and vein for 45 minutes followed by reperfusion, and then contralateral nephrectomy was performed 2 weeks later. Renal function was markedly decreased at 24 hours after reperfusion, and thereafter the effects of dual gas were assessed by histologic examination and determination of the superoxide radical, together with functional and molecular analyses. RESULTS: Pathologic examination of the kidney of I-R rats revealed severe renal damage. Importantly, cytoprotective effects of the dual treatment in comparison with H2 treatment and I-R renal injury were observed in terms of superoxide radical scavenging activity and histochemical features. Rats given dual treatment and I-R renal injury showed significant decreases in blood urea nitrogen. Increased expression of several inflammatory cytokines (tumor necrosis factor-α, interleukin-6, intracellular adhesion molecule-1, nuclear factor-κB, hypoxia inducible factor-1α, and heme oxygenase-1) was attenuated by the dual treatment. CONCLUSIONS: Dual gas inhalation decreases oxidative stress and markedly improves I-R-induced renal injury.

Primary structure of the heparin-binding site of type V collagen.

The abilities of collagens, type I, II, III, IV, and V, to bind heparin were examined by heparin-affinity chromatography and binding studies with [35S]heparin. At a physiological pH and ionic strength, only type V collagen bound to heparin. Collagens type I and II showed higher affinities than types III and IV for heparin, but did not bind to a heparin column at a physiological ionic strength. The heparin binding site of type V collagen was located in a 30 kDa CNBr fragment of the alpha 1(V) chain, and the amino acid sequence of this fragment was determined. The 30 kDa fragment contained a cluster of basic amino acid residues, and enzymatic cleavage within this basic domain greatly reduced the heparin-binding activities of the resulting peptides. Thus this basic region is probably the heparin-binding site of type V collagen.

New echocardiographic observations in a patient with dissimilar atrial rhythms.

A patient with an impure flutter was found to have dissimilar atrial rhythms that were confirmed by direct ECG recordings. Echocardiographic studies disclosed that the a waves on the mitral echocardiogram occurred synchronously with the a waves on the esophageal ECG that, in turn, showed atrial flutter at a rate of 300 beats per minute. However, the a waves on the tricuspid echocardiogram coincided closely with the P waves on a surface ECG (lead V) that disclosed a chaotic atrial tachyarrhythmia. These results suggest that echocardiographic study, in conjunction with an esophageal ECG, may be a useful, noninvasive technique for the diagnosis of dissimilar atrial rhythms.

A yeast system for stable expression of hepatitis B surface antigen.

We have constructed a yeast strain that has integrated into its chromosomal ribosomal RNA gene site two copies of Hepatitis B virus surface (HBS) antigen gene under the control of the yeast (Saccharomyces cerevisiae) glyceraldehyde-3-phosphate dehydrogenase (GAP) promoter and terminator. The level of expression of HBS gene was low in the strain, but upon chemical and physical mutageneses, in combination with an immunological screening procedure, a mutant clone which expressed HBS protein at a high level was obtained. This mutant strain produces HBS antigen stably under non-selective conditions.

Tie-2 and angiopoietin-1 expression in human thyroid tumors.

Tie-2 is an endothelial cell-specific receptor tyrosine kinase involved in vascular maturation and remodeling. Although its expression is considered to be restricted to vascular endothelial cells and hematopoietic progenitors, our immunohistochemical and in situ hybridization studies showed that Tie-2 and its ligand, angiopoietin (Ang)-l were expressed not only in benign and malignant human thyroid tumor cells but also in hyperplastic regions of adenomatous goiter. To confirm the expression in these tissues further, we used a laser capture microdissection system to isolate epithelial tumor cells from tissue specimens selectively, and demonstrated the expression of Tie-2 and Ang-1 mRNAs in tumor cells by RT-PCR analysis. Furthermore, Tie-2 and Ang-1 mRNAs and proteins were also detected in rat thyroid cell lines, FRTL-5 and PCCL-3. Our results suggest that Ang-1/Tie-2 signaling may be involved in the proliferation of thyroid epithelial cells.

Mutational analysis of the BMP-1 gene in patients with gastroschisis.

BACKGROUND: Gastroschisis is a rare abdominal wall defect. Although the pathogenesis of gastroschisis is unknown, there is some evidence of the genetic etiology of gastroschisis. Recently, a functionally null deletion of the mouse bone morphogenic protein-1 (BMP-1) gene resulted in a phenotype that resembled a human neonate with gastroschisis. BMP-1 thus became the first potential candidate gene for gastroschisis. METHODS: To explore this possibility the authors collected blood samples from 11 patients who had gastroschisis. Mutational analysis of exons 2 to 15 of the human BMP-1 gene was performed using genomic polymerase chain reaction, single-strand conformation polymorphism analysis and direct sequencing methods. RESULTS: No mutation of the human BMP-1 gene was observed in any of these patients. CONCLUSION: Although heterogeneous etiologies might be proposed for gastroschisis, our results provide further evidence of a nongenetic etiology for gastroschisis. J Pediatr Surg 36:885-887.

The analysis of lipids and glycosaminoglycans of low-density lipoprotein-glycosaminoglycans complexes isolated from normal, fatty streaks and fibrous plaques of human aortic intima.

The low-density lipoproteins (LDL)-glycosaminoglycans (GAG) complexes were isolated from fibrous plaques, fatty streaks and normal of human aortic intima and analysed for lipids and GAGs. The LDL-GAG complexes formed a precipitation line against antihuman apoprotein B, which is a major component of plasma LDL, and the lipids constituents of LDL-GAG complexes resembled those of plasma LDL. It might be suggested from these findings that the lipoproteins (LP) bound to GAG in arterial tissue was originated from plasma LDL penetrated into arterial tissue from circulating system. From the analysis of GAGs in LDL-GAG complexes by two-dimensional electrophoresis, the % composition of GAG in fibrous plaques showed higher in chondroitin-4-sulfate (CS-4-S) plus chondroitin-6-sulfate (CS-6-S) and marked lower in dermatan sulfate (DS) when compared with those of fatty streaks. There are no differences in the % composition of hyaluronic acid (HA) in normal and atherosclerotic lesions. It would be suggested from these observations that the formation of saline extractable LDL-GAG complexes would be the early events in aortic intima when plasma LDL penetrated from circulation system and that not only DS and/or heparan sulfate (HS) but also CS-4-S and/or CS-6-S have an important role on the progression of atherosclerosis.

Effects of nisoldipine on cytosolic calcium, platelet aggregation, and coagulation/fibrinolysis in patients with coronary artery disease.

The effect of nisoldipine, a dihydropyridine Ca2+ antagonist, on the platelet cytosolic Ca2+ concentration ([Ca2+]i), platelet aggregation, and various coagulation and fibrinolysis parameters was assessed in normotensive patients with coronary artery disease (CAD). Eleven patients with angiographically confirmed CAD (4 men, 7 women aged 67.3 +/- 5.4 years) were administered nisoldipine at 10 mg/day for two weeks. The [Ca2+]i was determined by use of fura2-loaded platelets, platelet aggregation was measured with an aggregometer, and coagulation/fibrinolysis parameters were measured by standard methods. Nisoldipine did not significantly affect blood pressure or heart rate. However, the [Ca2+]i decreased significantly (P<0.05) and platelet aggregation was also significantly inhibited. Plasma D-dimer levels decreased significantly (P<0.01). Thus, nisoldipine not only suppressed platelet activation but also affected the coagulation system, suggesting that it is not only a vasodilator and platelet inhibitor but also an antithrombotic agent.

Effect of LDL-apheresis on the pharmacokinetics of the lipophilic antilipidemic agent probucol.

The effect of LDL-apheresis on the pharmacokinetics of antilipidemic agents has not been evaluated thoroughly. In this study, we investigated the effect of LDL-apheresis on the pharmacokinetics of probucol, a lipophilic antilipidemic agent, by studying its distribution and changes in the blood concentration of probucol after LDL-apheresis. The concentrations of lipoproteins were measured before and after LDL-apheresis in eight patients with familial hypercholesterolemia taking probucol. Concentrations of probucol in the various lipoprotein fractions and plasma were measured by HPLC. The serum concentrations of probucol before and after LDL-apheresis were 39.8 +/- 3.3 and 16.5 +/- 1.6 micrograms/ml, and the correlation coefficient between the changes in the serum probucol concentration and those in the serum cholesterol concentration before and after LDL-apheresis was significant (r = 0.73, P < 0.01). Changes in the probucol and cholesterol concentrations after LDL-apheresis were mainly found in the LDL fraction. The calculated reductions in the serum contents of probucol and cholesterol were similar to the contents of probucol and cholesterol in the irrigation fluid of the dextran sulfate column. These data suggest that changes of probucol concentration in plasma by LDL-apheresis are mainly due to reductions in the LDL fraction.

Unusual findings of M-mode echocardiogram in a case with left ventricular thrombus.

The unusual echo findings of the left ventricular thrombus by M-mode echocardiogram is reported. The patient was a 41-year-old man with hypokinesis of the inferior and inferolateral wall due to acute myocardial infarction. The findings of M-mode echocardiogram revealed dense thick linear echoes just below the anterior wall of the apex, which had a higher density than the myocardium and looked "band like". These abnormal echoes obtained by M-mode echocardiogram were also detected through examinations by 2-dimensional echocardiogram, computed tomogram and left ventriculogram, and were assumed to arise from the index finger-sized thrombus in the left ventricle at the time of surgical procedure. Although this abnormal echo, revealed to have a "band like" feature by M-mode echocardiogram, is usually seen in the left ventricular myxoma, we assume that this "band like" echo by M-mode echocardiogram arose from the left ventricular thrombus under conditions such as myocardial infarction, ventricular aneurysm, dilated cardiomyopathy and long-term congestive heart failure.

[The significance of obesity in UOEH medical students--multiple regression analysis of the annual physical checkup data in 1991].

For the purpose of evaluating the significance of obesity in a young population as a risk factor toward various chronic diseases, a multiple regression analysis was performed on the data from the annual physical checkup of medical students of UOEH in 1991. The following results were obtained. (1) The average obesity index of the students showed a progressive and significant increase in the past 13 years from 1979 to 1991. (2) A close correlation between the obesity index and serum GPT was recognized by elevation of the standard partial regression coefficient of serum GPT to obesity index and that of obesity index to serum GPT when the data from all 617 students was analysed in one group. This finding was intensified in 142 obese students with an obesity index of over 10%, but it was not seen in the remaining non-obese students. (3) The correlation between the obesity index and serum GPT was found solely in the group of students with constant obesity; i.e., whose obesity index was always more than 10% during a maximum of 9 years in the past. (4) Systolic blood pressure was related to the red blood cell count rather than to the obesity index in this young population. (5) No particularly close correlation to serum cholesterol was found with any of 10 representative items examined in this physical checkup, including the obesity index, indicating that the other factors should be related to serum cholesterol levels. From the above findings, it can be concluded that constant obesity in students is related to liver dysfunction, probably due to fatty liver frequently seen in the precise examination of these individuals.

Platelet aggregation in variant angina: its relation to plasma catecholamines.

We concurrently measured plasma catecholamine levels and platelet aggregation in patients with variant angina, effort angina and old myocardial infarction. Platelet aggregability was lower in variant angina than in effort angina and old myocardial infarction. Plasma concentrations of epinephrine and norepinephrine in the variant angina group showed higher values than those in the effort angina group and the old myocardial infarction group. Moreover, there was a statistically significant negative correlation between plasma catecholamine concentration and ADP-induced platelet aggregation. These findings suggest that the lower platelet aggregation is related to the higher concentration of plasma catecholamine in variant angina.

Physical training augments plasma catecholamines and natural killer cell activity.

To elucidate the effects of a 10-week exercise training period on physical fitness and plasma catecholamine concentration at rest, and on natural killer cell activity in young healthy untrained females, subjects (20-22 years old. N = 16) carried out physical training by a protocol which consisted of treadmill jogging at a work-intensity of 50% of their VO2max, two hours a day three times a week for ten weeks. VO2max was increased significantly from 33.1 +/- 3.4 ml/kg/min to 38.3 +/- 6.6 ml/kg/min by the ten weeks of physical training (P < 0.005), and natural killer cell activity was also increased significantly from 31.9 +/- 14.3% to 46.4 +/- 18.4% (P < 0.05). The concentrations of epinephrine and norepinephrine before and after physical training were 18.3 +/- 8.7 pg/ml and 20.4 +/- 8.9 pg/ml, and 134.1 +/- 52.2 pg/ml and 248.1 +/- 106.8 pg/ml (P < 0.005), respectively. Changes in norepinephrine and epinephrine correlated with the change in VO2max (r = 0.780, P < 0.005; r = 0.556, P < 0.05). While the change in natural killer cell activity correlated with the change in epinephrine (r = 0.623, P < 0.01), the correlation of the change in natural killer cell activity with change in norepinephrine did not reach statistical significance (r = 0.497, P = 0.0503). From these results we concluded that physical training augments plasma catecholamine levels and natural killer cell activity at rest in young healthy females.

[Autonomic nerve function expressed by the baroreflex sensitivity (BRS) in elderly adults].

We examined 24 hour heart rate variability (HRV) components and baroreflex sensitivity (BRS) in elderly adults. Forty-eight subjects, aged 65-69 years old (24 men, 24 women) were examined in this study. BRS was measured in the morning (8:00-9:00) using noninvasive cross spectral analysis. There were significant correlations between BRS and low-frequency (LF) power (r = 0.593, p < 0.001) of HRV, and between BRS and high-frequency (HF) power (r = 0.402, p < 0.005). BRS values were lower in women compared with men (p < 0.01), and LF/HF was significantly lower in women than in men (p < 0.05).

Effects of NAD kinase 2 overexpression on primary metabolite profiles in rice leaves under elevated carbon dioxide.

The concentration of carbon dioxide (CO2) in the atmosphere is projected to double by the end of the 21st century. In C3 plants, elevated CO2 concentrations promote photosynthesis but inhibit the assimilation of nitrate into organic nitrogen compounds. Several steps of nitrate assimilation depend on the availability of ATP and sources of reducing power, such as nicotinamide adenine dinucleotide phosphate (NADPH). Plastid-localised NAD kinase 2 (NADK2) plays key roles in increasing the ATP/ADP and NADP(H)/NAD(H) ratios. Here we examined the effects of NADK2 overexpression on primary metabolism in rice (Oryza sativa) leaves in response to elevated CO2. By using capillary electrophoresis mass spectrometry, we showed that the primary metabolite profile of NADK2-overexpressing plants clearly differed from that of wild-type plants under ambient and elevated CO2. In NADK2-overexpressing leaves, expression of the genes encoding glutamine synthetase and glutamate synthase was up-regulated, and the levels of Asn, Gln, Arg, and Lys increased in response to elevated CO2. The present study suggests that overexpression of NADK2 promotes the biosynthesis of nitrogen-rich amino acids under elevated CO2.

The influence of growth hormone/insulin-like growth factor deficiency on prostatic dysplasia in pbARR2-Cre, PTEN knockout mice.

BACKGROUND: Elevated insulin-like growth factor-I (IGF-I) serum levels and phosphatase and tensin homolog (PTEN) loss are prostate cancer (PCa) risk factors that enhance androgen-responsive and castration-resistant PCa xenografts growth. METHODS: The impact of suppressed growth hormone (GH)/IGF-I levels on neoplastic initiation of PTEN-deficient prostate epithelia was assessed histologically and by epithelial-to-mesenchymal marker expression in Ghrhr D60G homozygous (lit/lit) and heterozygous (lit/+) pbARR2-Cre, PTEN(fl/fl) (PTEN-/-) mice. How suppressed GH/IGF-I levels impacted growth of PTEN-/- mouse-derived prostate cells (MPPK) was examined by growth and survival signaling of cells cultured in lit/+ or lit/lit serum. RESULTS: Body weight, prostate weight and serum GH and IGF-I levels were reduced in lit/lit relative to lit/+ PTEN-/- littermates. While the anterior lobes of lit/+ PTEN-/- prostates consistently presented swollen, indicative of ductal blockage, the degree of prostatic dysplasia in 15- and 20-week-old lit/lit and lit/+ PTEN-/- mice was indistinguishable as measured by normalized prostatic weight, tissue histology, or probasin, PSP94, E-cadherin, N-cadherin and vimentin expression. However, growth and AKT activation of MPPK cells was decreased when cultured in lit/lit serum as compared with lit/+ serum and restored in lit/lit serum supplemented with IGF-I and, to a lesser extent, GH. CONCLUSIONS: These results suggest that initiation of prostate carcinogenesis by loss of PTEN is not influenced by germline variation of genes encoding signaling molecules in the GH/IGF-I axis, but suggests that these factors may affect the progression of dysplastic phenotype and supports previous studies, indicating that the GH/IGF milieu does impact the growth of PTEN-deficient dysplastic prostatic cells once transformed.