Study of interaction of methylene blue with DNA and albumin.

The interaction of thiazine dye methylene blue (MB) with Calf thymus DNA and human blood serum albumin (HSA) has been studied. MB was revealed to stabilize the native structure of DNA and HSA, since the melting temperature of the complexes is shifted to higher values in relation to that of both macromolecules in pure state. It was also revealed that the absorption and fluorescence spectra of the MB-DNA complexes change significantly, while those of MB-albumin complexes do not change noticeably. Analysis of the obtained data allows to conclude that MB binds to DNA by two modes, including intercalation and electrostatic mechanisms. In the case of HSA, the main binding mode of MB, conditioning the stabilization of the protein native structure, is the electrostatic mechanism.Communicated by Ramaswamy H. Sarma.

The effect of stainable drinks followed by simulated brushing on the roughness and stainability of acrylic resins polymerized with different cycles.

STATEMENT OF PROBLEM: No information is available on roughness and stainability of acrylic resins polymerized by experimental microwave cycles after immersion in stainable liquids and simulated brushing. PURPOSE: The purpose of this in vitro study was to evaluate the effect of stainable drinks and brushing on roughness and stainability of acrylic resins (Vipi Cril [CA] and Vipi Wave [MA]) polymerized with different cycles. MATERIAL AND METHODS: CA and MA specimens (n=5; diameter, 20 mm; thickness, 3 mm) were made using 4 methods recommended by the manufacturer (water bath polymerization and microwave polymerization cycles) and experimental at 550 W or 650 W for 3 or 5 minutes (M550/3 and M650/5), respectively. After storage in distilled water at 37°C for 48 hours (T0), the specimens were stored in water, coffee, or red wine (37°C) for 36 days with simulated brushing (54 000 cycles, T1). Roughness (Ra) and stainability (ΔE/National Bureau of Standards) were measured at T0 and T1. Roughness and stainability data were analyzed by 3-way repeated measures and 2-way ANOVA, respectively, followed by the Bonferroni test (α=.05). RESULTS: After storing in coffee and brushing, CA showed the highest (M550/3=2.33 ±0.72 µm) and the lowest roughness (water bath polymerization=1.22 ±0.58 µm), whereas roughness of MA specimens processed by M650/5 increased (1.57 ±0.59 µm). Storing in wine and brushing increased roughness (1.75 ±0.32 µm) in the M550/3 group. No staining was observed on MA after brushing regardless of the polymerization cycle. All values were acceptable (ΔE≤3.3), except for MA (microwave polymerization), which showed National Bureau of Standards=4.49 (appreciable change) after storing in wine and brushing. CONCLUSIONS: A slight increase in material roughness was observed after staining and brushing. Only MA polymerized following manufacturer cycles showed relevant stainability after immersion in wine and brushing.

The pattern of GPI-80 expression is a useful marker for unusual myeloid maturation in peripheral blood.

Myeloid-derived suppressor cells (MDSCs) have a wide spectrum of immunosuppressive activity; control of these cells is a new target for improving clinical outcomes in cancer patients. MDSCs originate from unusual differentiation of neutrophils or monocytes induced by inflammatory cytokines, including granulocyte-colony stimulating factor (G-CSF) and granulocyte-macrophage (GM)-CSF. However, MDSCs are difficult to detect in neutrophil or monocyte populations because they are not uniform cells, resembling both neutrophils and monocytes; thus, they exist in a heterogeneous population. In this study, we investigated GPI-80, a known regulator of Mac-1 (CD11b/CD18) and associated closely with neutrophil maturation, to clarify this unusual differentiation. First, we demonstrated that the mean fluorescence intensity (MFI) of GPI-80 and coefficient of variation (CV) of GPI-80 were increased by treatment with G-CSF and GM-CSF, respectively, using a human promyelocytic leukaemia (HL60) cell differentiation model. To confirm the value of GPI-80 as a marker of unusual differentiation, we measured GPI-80 expression and MDSC functions using peripheral blood cells from metastatic renal cell carcinoma patients. The GPI-80 CV was augmented significantly in the CD16hi neutrophil cell population, and GPI-80 MFI was increased significantly in the CD33hi monocyte cell population. Furthermore, the GPI-80 CV in the CD16hi population was correlated inversely with the proliferative ability of T cells and the GPI-80 MFI of the CD33hi population was correlated with reactive oxygen species production. These results led us to propose that the pattern of GPI-80 expression in these populations is a simple and useful marker for unusual differentiation, which is related to MDSC functions.

A Survey of the Management of Patients with Temporomandibular Disorders by General Dental Practitioners in Southern Brazil.

PURPOSE: Temporomandibular disorders (TMD) are recognized as one of the most controversial topics in dentistry, despite the fact that both basic science and clinical researchers have currently reached some degree of consensus. This study aimed to conduct a questionnaire-based survey about the management of TMD patients by general dental practitioners (GDPs). MATERIALS AND METHODS: One hundred fifty-one GDPs with a private practice in a city of southern Brazil were included, independent of school of origin, gender, graduation year, and curriculum content. All participants were administered a questionnaire about the management of patients with TMD, and the responses were analyzed by binomial and chi-square tests (α = 0.05). RESULTS: Of the GDPs, 88.7% received TMD patients, who were primarily diagnosed on the basis of medical history (36.6%) or physical examination (30.4%). Of these, 65.4% referred the patients elsewhere, primarily to specialists in occlusion (36.1%) or orthodontics (29.7%). Occlusal splinting was the most commonly used management modality (20.8%), followed by occlusal adjustment (18.1%) and pharmacotherapy (16.6%). Splints were fabricated in maximum habitual intercuspation or centric relation depending on individual patient (54.8%). The hard stabilization form was the most common type of appliance used (35.0%). Moreover, 73.8% of the GDPs did not employ semi-adjustable articulators, and 69.5% adjusted the appliances at the time of fixing. The duration of splint use and the frequency of follow-up were considered patient dependent by 62.1% and 72.8%, respectively. GDPs considered the two major TMD etiologic categories as multifactorial (20.8%) and occlusion (19.9%). Multidisciplinary medical and dental treatment was considered necessary by 97.9%. CONCLUSIONS: The evaluated general dental practitioners manage TMD patients according to international guidelines.

Failures in the rehabilitation treatment with removable partial dentures.

BACKGROUND: Despite being functional and having aesthetic benefits, the acceptance of patients regarding the use of removable partial dentures (RPDs) has been low. In part, this is due to the deleterious effects that causes discomfort to the patient. Success depends not only on the care expended by the patient, including daily care and oral hygiene, but also on common goals set by their professional and clinical staff, aiming beyond aesthetics, to incorporate issues of functionality and the well-being of patients. METHODS AND RESULTS: For rehabilitation treatment with RPDs to reach the desired level of success without damaging the support structure, all the steps (diagnose, cavity preparation, adaptation of the metal structures, functional of distal extension and posterior follow-up) in the rehabilitative treatment should be carefully developed. A literature review was carried out, searching through MEDLINE (PubMed) articles published between 1965 and December 2012 including clinical trials and reviews about the use of RPDs. CONCLUSIONS: This study describes factors that lead to failures and complications in oral rehabilitation through the use of RPDs and suggests possible solutions.

Vascular calcification and secondary hyperparathyroidism of severe chronic kidney disease and its relation to serum phosphate and calcium levels.

BACKGROUND AND PURPOSE: Various complications consequent on disordered calcium and phosphate homeostasis occur frequently in chronic kidney disease (CKD) patients. Particularly, vascular calcification has high morbidity and mortality rates. There is a clear need for a better CKD model to examine various aspects of this disordered homeostasis. EXPERIMENTAL APPROACH: Oral dosing with adenine induced CKD in rats in only 10 days. Serum calcium, phosphate and parathyroid hormone were measured and calcification in aorta was assessed histologically. The effects of varying phosphorus content of diet or treatment with phosphate binders or active vitamin D(3) on these parameters were examined. KEY RESULTS: After adenine dosing, significant hyperphosphatemia, hypocalcemia and secondary hyperparathyroidism (2HPT) were observed during the experimental period of 15 weeks. Aortic calcification was detected in only some of the animals even at 15 weeks (approximately 40%). Treatment with vitamin D(3) for 18 days, even at a low dose (100 ng x kg(-1), 3-4 times week(-1), p.o), caused aortic calcification in all animals and increases in serum calcium levels up to the normal range. The vitamin D(3)-induced calcification was significantly inhibited by phosphate binders which lowered serum phosphate levels and the calcium x phosphate product, although serum calcium levels were elevated. CONCLUSIONS: These data suggest that rats dosed orally with adenine provide a more useful model for analysing calcium/phosphate homeostasis in severe CKD. Controlling serum calcium/phosphate levels with phosphate binders may be better than vitamin D(3) treatment in hyperphosphatemia and 2HPT, to avoid vascular calcification.

Massive intractable pericardial effusion in a patient with systemic lupus erythematosus treated successfully with pericardial fenestration alone.

Systemic lupus erythematosus (SLE) is often complicated by pericarditis with effusion, which generally responds well to glucocorticoid. We report herein a Japanese patient with SLE who showed a sign of cardiac tamponade and severe chest and back pain because of massive intractable pericardial effusion. Pulse glucocorticoid and pulse cyclophosphamide gained marginal effects. Pericardial effusion accumulated again soon after ultrasound-guided pericardiocentesis and drainage. Pericardial fenestration performed surgically as a last resort, for draining pericardial fluid into the pleural space, was very effective, and only a much smaller amount of fluid was observed in the space thereafter in comparison with the volume before the surgery. Pathological examination of the retrieved pericardium unfolded intense hyperplasia of small vessels and capillaries. Levels of IL-6 and TNF-alpha in pericardial effusion were extremely higher than those in serum. Pericardial effusion with extensive capillary hyperplasia in SLE would be resistant to medical treatment and require surgical fenestration.

Interleukin-11 induces osteoblast differentiation and acts synergistically with bone morphogenetic protein-2 in C3H10T1/2 cells.

Interleukin-11 (IL-11) is a pleiotropic cytokine that supports various types of hematopoietic cell growth and is involved in bone resorption. We report here the involvement of recombinant human IL-11 (rHuIL-11) in osteoblast differentiation in mouse mesenchymal progenitor cells, C3H10T1/2. rHuIL-11 alone increased alkaline phosphatase (ALP) activity and upregulated expression levels of osteocalcin (OC), bone sialo protein (BSP), and parathyroid hormone receptor (PTHR) mRNA. rHuIL-11 had no effect on expression of type II collagen, peroxisome proliferator-activated receptor-gamma2 (PPAR-gamma2), adipocyte fatty acid-binding protein P2 (aP2), and myogenic MyoD protein (MyoD). Recombinant human bone morphogenetic protein (rHuBMP)-2 increased ALP activity and mRNA expression of these genes except for MyoD. The expression patterns of ALP activity and osteoblast-specific or chondrocyte-specific genes suggest that rHuIL-11 may be involved in early differentiation of osteoblasts at a step earlier than that which is affected by rHuBMP-2. In support of this hypothesis, combined treatment with rHuIL-11 and rHuBMP-2 synergistically increased ALP activity and mRNA expression of OC and type II collagen, rHuIL-11 also abrogated the increased levels of PPAR-gamma2, aP2 mRNA caused by rHuBMP-2. Our results suggest that rHuIL-11 alone and in combination with rHuBMP-2 can induce osteoblastic differentiation of progenitor cells and plays an important role in osteogenesis.

Distribution and diversity of symbiotic thermophiles, Symbiobacterium thermophilum and related bacteria, in natural environments.

Symbiobacterium thermophilum is a tryptophanase-positive thermophile which shows normal growth only in coculture with its supporting bacteria. Analysis of the 16S rRNA gene (rDNA) indicated that the bacterium belongs to a novel phylogenetic branch at the outermost position of the gram-positive bacterial group without clustering to any other known genus. Here we describe the distribution and diversity of S. thermophilum and related bacteria in the environment. Thermostable tryptophanase activity and amplification of the specific 16S rDNA fragment were effectively employed to detect the presence of Symbiobacterium. Enrichment with kanamycin raised detection sensitivity. Mixed cultures of thermophiles containing Symbiobacterium species were frequently obtained from compost, soil, animal feces, and contents in the intestinal tracts, as well as feeds. Phylogenetic analysis and denaturing gradient gel electrophoresis of the specific 16S rDNA amplicons revealed a diversity of this group of bacteria in the environment.

Winter seasonality and rotavirus diarrhoea in adults.

We investigated the aetiological role of group A rotavirus in adults with acute diarrhoea in a 4-year prospective study. Of 683 patients with acute diarrhoea, 97 (14%) shed rotavirus as a sole agent, whereas six (5%) of 115 patients without diarrhoea shed rotavirus. Half of patients with rotavirus diarrhoea required admission to hospital. Unlike rotavirus diarrhoea in children, the occurrence of rotavirus-positive cases did not show a significant winter seasonality. Rotavirus infection should be included in the differential diagnosis of diarrhoeal diseases in adults.

Twenty-six-week carcinogenicity study of sulfamethoxazole in CB6F1-Tg-rasH2 mice.

Sulfamethoxazole (SMX), a hormone-mediated rodent-specific nongenotoxic carcinogen, was administered to CB6F1 mice carrying a human prototype c-Ha-ras gene (Tg-rasH2) at doses of 0, 25, 100 or 400 mg/kg/day and to the wild-type mice at a dose of 400 mg/kg/day in feed for 26 weeks to evaluate the carcinogenicity and to validate the Tg-rasH2 model. N-Methyl-N-nitrosourea was administered at an intraperitoneal dose of 75 mg/kg to Tg-rasH2 as a positive control and the experimental system was confirmed to be valid. Histopathological examination revealed adenomas of the lung and Harderian gland and hemangiosarcoma of the spleen at low frequencies in the Tg-rasH2 treated with SMX; however, no statistically significant differences were observed either in the onset or prevalence rates of these neoplasms compared with that in the control group. Between the wild-type mice and Tg-rasH2, the onset rate and prevalence of the neoplasms were not significantly different, but the neoplasms tended to be more frequent in Tg-rasH2 mice showing a sensitivity to tumorigenicity. Follicular epithelial cell hyperplasia was observed in the thyroid gland in the groups of Tg-rasH2 given 100 mg/kg SMX or more, but no neoplastic lesion was observed. SMX was judged to be negative for carcinogenic potential in Tg-rasH2 in the present study.

[Toxicity study of cefmatilen hydrochloride hydrate (S-1090) (8)--Nephrotoxicity study in rabbits by single oral administration].

A nephrotoxicity study of Cefmatilen hydrochloride hydrate (S-1090) was conducted in rabbits at single oral doses of 250, 500 and 1000 mg potency/kg. All treated groups showed a decreased food consumption and a tendency for the body weight to decrease. Urinary protein and glucose were detected and slight increases of plasma creatinine and urea nitrogen were observed in the 500 mg potency/kg group. Urinary protein was also detected in the 1000 mg potency/kg group. In the histopathological examination of the kidney, tubular necrosis was observed in the 500 and 1000 mg potency/kg groups. No nephrotoxic signs were observed in the 250 mg potency/kg group. The NOAEL on the nephrotoxicity of S-1090 in rabbits was estimated to be 250 mg potency/kg.

Subarachnoid hemorrhage and systemic lupus erythematosus.

The frequency, clinical profile, treatment and outcome of subarachnoid hemorrhage (SAH) in patients with systemic lupus erythematosus (SLE) were assessed retrospectively, based on the case records of SLE of the Jichi Medical School Hospital over a 20 year period. Clinically defined SAH was found in 10 (3.9%) out of 258 SLE patients, which represented a frequency higher than previously assumed. Five patients had active SLE and lacked an apparent cause of SAH, other than SLE. A high mortality rate (5/5), no visible aneurysm on angiogram (3/4), and an onset during intractable SLE or after discontinued or no steroid therapy because of medical noncompliance (4/5) were characteristic of patients with active SLE, and thus an earlier successful suppression of SLE, if possible, might have prevented their SAH. In contrast, in the 5 patients with inactive SLE, 2 out of 3 saccular aneurysms were successfully clipped and small bleeding of one patient without aneurysms remitted spontaneously without the need for additional steroid therapy. When one death, which occurred outside of medical care, was excluded, the survival ratio of the hospitalized SAH patients with inactive SLE was significantly better than that with active SLE (3/4 versus 0/5, P=0.0476). In conclusion, the relatively common occurrence of SAH in SLE patients, and a significantly different clinical impact of SAH in respect to active and inactive SLE, were suggested from the results.

Effects of a fixation target on torsional optokinetic nystagmus.

PURPOSE: To investigate the effects of an imaginary and a visual target on torsional optokinetic nystagmus (tOKN) and directional symmetry of tOKN. METHODS: Torsional OKN was induced by a rotating random dot pattern (52 degrees in diameter, constant angular velocity: +/-30 deg/sec to +/-52 deg/sec) with an imaginary or a visual target in 11 eyes of 10 healthy humans by dual-search coil methods. RESULTS: Intorsional OKN and extorsional OKN were symmetrical in their slow-phase gain. The mean slow-phase gain (0. 037/0.041, intorsion/extorsion) of tOKN during fixation on a visual target at the center of the rotating random dot pattern was significantly (P: < 0.002) smaller than that (0.051/0.052, intorsion/extorsion) during fixation on an imaginary target at the center of the rotating random dot pattern. The mean tOKN slow-phase beat duration (840 msec/724 msec, intorsion/extorsion) during fixation on the visual target was significantly (P: < 0.002) longer than that (585 msec/543 msec, intorsion/extorsion) during fixation on the imaginary target. In seven eyes of six subjects, the mean slow-phase gain and beat duration (0.034 and 812 msec) of tOKN during fixation on a visual target 6.5 degrees left or right from the center of the rotating random dot pattern were not significantly different from those (0.037 and 825 msec) with a visual target at the center of the rotating random dot pattern (P: > 0.3). CONCLUSIONS: A visual target spot suppresses tOKN by a nonpursuit visual system. Intorsional and extorsional OKNs were symmetrical.

[Rapidly progressed secondary amyloidosis in a patient with Still's disease with gamma-allele in his SAA 1 gene].

A fifteen-year-old boy was admitted to our hospital because of lower abdominal pain, watery diarrhea and mucobloody stool. Two years before admission, he was diagnosed to have Still's disease presenting with polyarthritis, sore throat, remittent fever and typical skin rash. He had been treated with non-steroidal anti-inflammatory agents, oral prednisolone and low-dose methotrexate. Although he was almost free of symptoms during the next two years, serum C-reactive protein (CRP) levels continued to be elevated moderately. He began to complain of lower abdominal pain and loose stool in May 1997 and came down with mucous-bloody diarrhea in June. Laboratory data on admission showed an elevated level of serum CRP (13.9 mg/dl). The biopsy of the stomach, ileum, sigmoid colon and rectum revealed the deposition of amyloid protein of AA type, which confirmed the diagnosis of secondary amyloidosis. The dose of prednisolone was increased and dimethyl sulfoxide per os or rectum was instituted, which improved his gastro-intestinal symptoms to some extent. However, fever, arthritis and diarrhea recurred along with tapered prednisolone dosage. In addition to gastro-intestinal symptoms, arrhythmia and proteinuria appeared. These symptoms were considered to reflect general deposition of amyloid in his body. He is now on immunosuppressive agent and high-dose prednisolone. Several studies report the higher frequency of gamma-allele of SAA 1 gene in the cases of rheumatoid arthritis with AA-amyloidosis than in those without. In the patient presented here, molecular biological analysis revealed that his SAA 1 gene was composed of beta- and gamma-allele. The presence of gamma-allele in his SAA 1 gene might be one of the factors that predisposed him for generalized deposition of amyloid protein in such a short period of time.

[Three patients with systemic lupus erythematosus complicated by cytomegalovirus colitis].

We report three female patients with systemic lupus erythematosus complicated by massive intestinal hemorrhage during the recovery from lupus nephritis (case 1, 2) or central nervous system lupus (case 3) on high dose corticosteroid therapy. Large number of cytomegalovirus (CMV) antigen-positive leukocytes and cessation of bleeding with concurrent disappearance of the viral antigens after ganciclovir therapy indicated CMV colitis in all of the three patients. No recurrence of the symptom and a favorable response to ganciclovir without reduction in steroid regimen was common to these patients.

[Three patients with systemic sclerosis complicated by microangiopathic hemolytic anemia and thrombocytopenia].

Clinical profiles and the treatment process of three female patients with systemic sclerosis (cases 1, 2, and 3) complicated by thrombotic microangiopathic hemolytic anemia (TMHA) were described. Thrombocytopenia preceded renal damage and hypertension in cases 1 and 2, although the chronological relationship between these parameters were unknown in case 3. Plasma exchange therapy using fresh frozen plasma was beneficial in cases 1 and 2. Cases land 3 presented with delirium and fluctuating psychosis, respectively. Early detection of thrombocytopenia and insidious hemolysis might be essential for starting effective plasmapheresis treatment in a part of patients with scleroderma kidney who present with thrombotic thrombocytopenic purpura (TTP) like disorder.