Deproteinizing effects on resin-tooth bond structures

This study investigated the effects of NaOCl on resin-tooth bonds to simulate the situations of long-term durability and caries invasion. Resin-tooth bonded specimens were produced with the use of two resin adhesives (Excite and One-Bond). Resin-tooth bonded beams (adhesive area; 0.9 mm2) were serially sectioned and the specimens were immersed in 10% NaOCl medium for 0 (control), 2, 4, and 6 h after being stored in water for 24 h. After immersion, microtensile bond tests were performed. SEM fractography was conducted to calculate each failure mode by image analysis. In addition, the adhesive interface was examined with the use of TEM. In the control specimens, enamel bond strengths had no difference between Excite (45.6 +/- 15.0) and One-Bond (56.9 +/- 12.9). On the other hand, dentin bond strengths had significant difference between Excite (80.6 +/- 21.2) and One-Bond (50.7 +/- 11.2). The bond strengths decreased with increased storage time for both systems with enamel and dentin bonds. The deteriorated mineralized dentin of beams resulted in bond-strength reduction for resin-enamel bonds. For dentin bonding, the adhesive interface was gradually dissolved from the outer to the center portion of the beam. The depletion of collagen fibrils within the demineralized dentin or hybrid layer deformation was found under SEM and TEM examinations. These morphological changes are responsible for bond strength reduction of resin-dentin bonds.     

Changes in element concentrations induced by agonist in pig pancreatic acinar cells

 Changes in electrolytes of pig pancreatic acinar cells following application of gastrin-cholecystokinin (CCK) were investigated using the technique of X-ray microanalysis of hydrated and dehydrated sections of freshly frozen pancreas. After stimulation by CCK (10^–9 M), Na and Cl increased significantly in the cytoplasm [Na, from 10 mmol/kg wet wt. (48 mmol/kg dry wt.) to 19 mmol/kg (95 mmol/kg); Cl, from 22 mmol/kg (105 mmol/kg) to 49 mmol/kg (245 mmol/kg)] as well as in the luminal interspace [Na, from 53 mmol/kg (189 mmol/kg) to 65 mmol/kg (283 mmol/kg); Cl, from 65 mmol/kg (232 mmol/kg) to 102 mmol/kg (443 mmol/kg)]. In the secretory granules Cl increased significantly from 30 mmol/kg (86 mmol/kg) to 67 mmol/kg (203 mmol/kg). K decreased significantly from 120 mmol/kg (571 mmol/kg) to 81 mmol/kg (405 mmol/kg) in the cytoplasm, while both increased from 38 mmol/kg (109 mmol/kg) to 58 mmol/kg (176 mmol/kg) in the granules and from 46 mmol/kg (164 mmol/kg) to 48 mmol/kg (209 mmol/kg) in the luminal interspace. Ca increased significantly in the cytoplasm as well as in the luminal interspace, and decreased significantly in the secretory granules. CCK evoked Ca release from secretory granules in the secretory pole of acinar cells. The values were measured from dehydrated sections, and agreed well with those from hydrated sections. The effect of furosemide, an inhibitor of the Na⁺-K⁺-2Cl^–co-transporter, on the ion transport of acinar cell was studied. When furosemide (10^–5 M) was added to the external solution, the cytoplasmic Cl and Ca concentrations decreased significantly, while there was a little decrease in Na and K concentrations under the secretory condition. These results indicate that Na⁺-K⁺-2Cl^–co-transport, and Na⁺, Cl^–and K⁺ exits into the lumen are involved in the mechanism of ion secretion in pig pancreatic acinar cells. Received: 17 January 1996 / Accepted: 12 March 1996     

Characterization of DNA sequences constituting the terminal heterochromatin of the chicken Z chromosome

Two clones, pCZTH5-8 and pCZTH12-8, were isolated from a female chicken genomic library by screening with sequences obtained from genomic libraries which had been constructed from a terminal region of a single Z chromosome of chicken utilizing laser microbeam irradiation and PCR amplification. Fluorescencein situ hybridization to the mitotic Z chromosome and the lampbrush ZW bivalent of chicken demonstrated that both the cloned sequences are located in the heterochromatic region of the Z chromosome at the end opposite to the pairing region with the W chromosome. The sequences pCZTH5-8 and pCZTH12-8 are distributed widely on both the telomeric bow-like loops (TBL) and the region I (short loops region) of the Z lampbrush chromosome. These clones, pCZTH12-8 particularly notably, hybridized also to the TBLs of lampbrush bivalents 1–4 of chicken. Both sequence are transcribed in the lampbrush stage oocytes on the Z chromosome and on other macrobivalents. The subfragment of pCZTH5-8 which hybridizes to the TBLs and the insert of pCZTH12-8 contain regions that are closely similar in sequence. The pCZTH5-8 sequence has no internal repeats and may be part of the 24-kb macrosatellite repeating unit that is evident afterNhel digestion of the genomic DNA. A cloned 24-kb unit, pFN-1, does not show significant DNA curvature, but cytosines of its CpG dinucleotides may be highly methylatedin vivo. This contrasts with the repeat sequences of the W heterochromatin which not only have highly methylated CpG but are also strongly curved. The 24-kb unit is repeated about 830 times in the diploid genome of a female chicken, suggesting that nearly the entire terminal heterochromatin on the Z chromosome consists of this macrosatellite family. Sequences of the greater part of the pCZTH5-8 are restricted to the genusGallus but the sequence of one subregion which hybridizes to TBLs is present in the genomes of the order Galliformes.accepted for publication by M. Schmid     

Concanavalin A binding sites of rough endoplasmic reticulum containing intracisternal microtubules in canine neurones

Lectin histochemistry was used to identify sugar residues of IM-containing RER in elderly canine sympathetic ganglionic neurones. IM-inclusions stained with ConA-peroxidase conjugate, but not with soybean agglutinin (SBA), wheat germ agglutinin (WGA), peanut agglutinin (PNA), Dolichos biflorus agglutinin (DBA), Ricinus communis agglutinin (RCA I) and Ulex europaeus agglutinin (UEA I). ConA-binding sites were visualized within cisternae of RER containing IM; reaction product was localized in IM-containing RER cisternae and IM. Inhibition with specific sugars (0.1 M alpha-methyl-D-mannoside or 0.5 M D-glucose) blocked the binding of ConA to IM-inclusions and normal Nissl substance. When a low sugar concentration (5 x 10(-3) M alpha-methyl-D-mannoside or 0.2 M D-glucose) was employed, IM-inclusions were still strongly ConA-positive, but normal Nissl substance was not. These results demonstrate that IM-containing RER have an excessive amount of carbohydrates (mannose or glucose-rich sugars) which are essentially detected in flattened RER under normal conditions and further indicate that glycoproteins in IM differ from those in cytoplasmic microtubules.     

Effect of iron concentrations on the DNA-degradation reaction by bleomycin

DNA-degradation reaction by bleomycin was conducted with form I of SV40 DNA as the substrate under "iron-limited conditions" and "iron-sufficient conditions". For "iron-limited conditions", the iron ions in reaction mixtures were derived from contaminant iron in the bleomycin preparation, the molar ratio of bleomycin/iron being 1/0.05. For "iron-sufficient conditions", Fe(II) was added to reaction mixtures to attain the molar ratio of bleomycin/iron at 1/1 to 1/29. DNA-degradation reaction by Fe(II) without bleomycin, or "Fe(II) alone", was also run for comparison. In any case, the reaction was allowed to proceed at such low rates that other products than form II and form III, due to further fragmentation, were negligible. The mass of DNA in each form was quantitatively determined by spectrofluorometry after agarose gel electrophoresis and staining with ethidium bromide. "Iron-limited conditions" and "iron-sufficient conditions" showed similar reaction characteristics as follows. As form I decreased with incubation time, form II increased faster than form III, hit a plateau (ca. 65% of total DNA) and decreased gradually thereafter. Form III kept increasing throughout the incubation. The reaction was dependent on incubation time and on doses of both bleomycin and Fe(II) but rather independent of temperature. The reaction was inhibited by acidic pH, Tiron (a specific chelator of iron) and Cu(II) but not EDTA. Octanucleotides with specific base sequence inhibited the reaction. DNA-degradation reaction by "Fe(II) alone" showed different characteristics as follows. The reaction proceeded rapidly at first, but ceased within 2 minutes. The extent of reaction was dependent on dose of Fe(II). The reaction product was form II accompanied by little amount of form III. The reaction was highly dependent on temperature, inhibited by EDTA, and not inhibited by Cu(II) and acidic pH. Octanucleotides, irrespective of their base sequences, inhibited the reaction.     

Fundamental and clinical studies of ceftizoxime suppositories in pediatrics

Fundamental and clinical studies were carried out on ceftizoxime suppository (CZX-S), and the following results were obtained in pediatrics. In 4 patients of the CZX-S 125 mg-administered group (9.4-9.9 mg/kg), the serum concentration of CZX reached a peak of 5.55 micrograms/ml on the average, 30 minutes after dosing, i.e. at the time of initial blood collection, and decreased gradually to 0.20 microgram/ml 6 hours after dosing. The half-life was 1.09 hours. In 5 patients of the CZX-S 250 mg-administered group (8.4-18.1 mg/kg), the serum concentration of CZX peaked at 7.07 micrograms/ml on the average and then gradually declined to 0.16 microgram/ml 6 hours after dosing. The half-life was 1.00 hour. The urinary recovery rate varied as widely as 6.5-38.0% in all the patients of both groups. CZX-S was given to total 19 patients; 8 patients with urinary tract infection (UTI), 3 with pharyngitis or tonsillitis, 4 with bronchitis, 2 with pneumonia, 1 with otitis media and 1 with staphylococcal scalding skin syndrome. The overall effect of CZX-S in 15 patients was "effective" or better response, with an effectiveness rate of 83.3%, except one who discontinued the drug because of side effects. CZX-S was given to most of the patients weighing 15 kg or higher in a dose of 250 mg 3-4 times a day and frequently to patients weighing less than 15 kg in a dose of 125 mg 3-4 times a day. As to side effects, slight diarrhea was encountered in 1 patient. Laboratory examinations disclosed an increase in GOT in 1 patient, which returned to normal after continual insertion of the suppository.     

A comparative, well-controlled study of ceftizoxime suppository against ceftizoxime intravenous injection in infantile acute pneumonia

We have attempted to clinically define the therapeutic usefulness of ceftizoxime suppository (CZX-S) in children with bacterial pneumonia, in a randomized trial. Intravenous injection of ceftizoxime (CZX) was used as the control. The results are summarized below. Subjects were inpatients with bacterial pneumonia, ranging in age from 9 months to 7 years and 10 months. As a rule, the daily dose was either four 250 mg (in potency) suppositories given at 6-hour intervals or 60 mg/kg body weight intravenous CZX (control) given in 4 injections at 6-hour intervals over a period of 7 days. The number of children in the study was 67. These children were divided into 2 dosage groups (suppository, 35; injection, 32) with matching pretreatment background factors. The severity of the target disease in the majority of the children was "moderate". The rate of therapeutic effectiveness was 97.1% for the suppository and 93.8% for the injection, and did not differ significantly between the 2 groups. Rates of efficacy by severity, presence or absence of underlying diseases, daily dose and/or complications were high without exception, and did not differ significantly between the 2 groups. Eradication rates for causative microorganisms, as studied in 16 children of each group, were both 93.8%. The 2 most frequently isolated causative organisms were Haemophilus influenzae and Streptococcus pneumoniae. Side effects were examined for 36 children of each group. The frequency of side effects did not differ significantly between the suppository group (2 with diarrhea and 1 with abdominal pain) and the injection group (1 with urticaria), and 8.3% and 2.8%, respectively. The frequency of abnormal laboratory test findings differed significantly (P less than 0.01) with respect to eosinophilia which occurred in 7 (20.6%) of the injected subjects but was not encountered in the subjects treated with suppositories. Other abnormal laboratory findings included thrombocytosis in 3 (14.3%) of the injection group and increased GOT in 1 (3.2%) of the suppository group. The suppository formulation of CZX appears to be a highly useful substitute for the injectable form, and should find a special use in children whose treatment with injections experiences some difficulty.     

Artificial Liver: Present and Future

Early in 1956, the first model of a biological artificial liver, using a live dog's liver incorporated in a cross-hemodialyzer, was placed in an experimental animal with portocaval encephalopathy. This "biological artificial liver," a hybrid artificial liver in the present terminology, was the first in the world. In October 1958, the first human patient, a young male patient in hepatic coma due to liver cirrhosis, was placed on the laboratory-made biological artificial liver composed of four parabiotic cross-hemodialyzers connected with four live dogs' livers to which the "hepatic reactors" for ammonium adsorption and acid-base balance were additionally equipped. This first case was very successful, resulting in the patient's recovery from coma. This article introduces the past history of the artificial liver, research of which has mainly been conducted in Japan since the early 1950s by the author, M. Mito, and Y. Nosé. Until recently, little progress has been made in this field through the application of blood purification principles such as hemoadsorption, plasmapheresis, and other modifications and combinations. Accumulation of clinical experiences with such conventional methods has stimulated the third generation of the artificial liver to a return to a hybrid organ applying modern science and technology. A concept of hybrid organs in comparison with organ transplants is introduced. The Japanese national project of developing a new artificial liver system, as conducted by the author as the chairman and his associates, is introduced.     

Association of feces sign with prognosis of non-emergency adhesive small bowel obstruction

Background/Objective: The feces sign has been reported as a possible predictive factor for non-operative treatment of small bowel obstruction. However, its relationship with prognosis of non-emergency adhesive small bowel obstruction remains unclear. This study aimed to clarify the relationship between the feces sign and prognosis of non-emergency adhesive small bowel obstruction.MethodsNinety-two patients with non-emergency adhesive small bowel obstruction with the transitional zone visible on computed tomography were included. Patients were categorized into two groups: feces sign positive (n = 40) and negative (n = 52). Clinical features and prognosis were compared between the two groups. Cox proportional hazards regression models incorporating the feces sign were used to analyze odds of diet resumption and discharge.ResultsPatients with feces sign were younger (p = 0.015), had a higher body mass index (p = 0.027), and a lower white blood cell count (p = 0.019) on admission. More patients with feces sign were successfully treated with fasting and/or nasogastric tube placement (p < 0.001), and no patient with feces sign suffered from recurrent obstruction after diet resumption. Kaplan–Meier analysis showed that patients with feces sign took less time for diet resumption (p = 0.007) and discharge (p = 0.004) than those without it. Using Cox proportional hazards regression model, the feces sign was reported as an independent predictor of diet resumption (odds ratio 1.685, p = 0.018) and discharge (odds ratio 1.861, p = 0.007).ConclusionsThe feces sign is associated with improved odds for diet resumption and discharge.     

High HNRNPA3 expression is associated with lymph node metastasis and poor prognosis in patients treated with radical cystectomy

ObjectiveWe sought to identify heterogeneous nuclear ribonucleoprotein A3 (HNRNPA3) expression in bladder cancer and its relationship to clinicopathological findings and prognosis.MethodsImmunohistochemical staining for HNRNPA3 was performed on 122 archived radical cystectomy specimens, with immunoreactivity being stratified on a 0 to 3 scale. The percentage of HNRNPA3 expressing tumor cells was calculated and multiplied by the staining score over an average of 5 areas to obtain a semiquantitative H-score (maximum value: 300). HNRNPA3 expression was categorized as high (≥80) or low (<80).ResultsThe patients’ median age was 70 years, and the median follow-up period was 39.4 months. High HNRNPA3 expression was significantly associated with lymph node metastasis (P= 0.014) and S100A8, S100A9 and uroplakin III expression (P= 0.028, 0.002, and 0.047, respectively). Log-rank tests indicated that high HNRNPA3 expression was significantly associated with disease progression and cancer-specific death (P= 0.013 and 0.006, respectively). In the Cox proportional hazards regression analysis, only lymph node metastasis was associated with disease progression and cancer-specific survival.ConclusionHNRNPA3 may be a new biomarker to predict biologically aggressive cancers and determine the appropriate treatment modality in patients after radical cystectomy.