Identification and characterization of a chymotrypsin-like serine protease from periodontal pathogen, Tannerella forsythia.

Tannerella forsythia is a bacteria associated with severe periodontal disease. This study reports identification and characterization of a membrane-associated serine protease from T. forsythia. The protease was isolated from T. forsythia membrane fractions and shown to cleave both gelatin and type I collagen. The protease was able to cleave both substrates over a wide range of pH values, however optimal cleavage occurred at pH 7.5 for gelatin and 8.0 for type I collagen. The protease was also shown to cleave both gelatin and type I collagen at the average reported temperature for the gingival sulcus however it showed a lack of thermal stability with a complete loss of activity by 60 °C. When treated with protease inhibitors the enzyme's activity could only be completely inhibited by serine protease inhibitors antipain and phenylmethanesulfonyl fluoride (PMSF). Further characterization of the protease utilized serine protease synthetic peptides. The protease cleaved N-succinyl-Ala-Ala-Pro-Phe p-nitroanilide but not Nα-benzoyl-dl-arginine p-nitroanilide (BAPNA) or N-methoxysuccinyl-Ala-Ala-Pro-Val p-nitroanilide indicating that the protease is a chymotrypsin-like serine protease. Since type I collagen is a major component in the gingival tissues and periodontal ligament, identification and characterization of this enzyme provides important information regarding the role of T. forsythia in periodontal disease.

Native Language Spoken as a Risk Marker for Tooth Decay.

OBJECTIVE: The purpose of this study was to assess dmft, the number of decayed, missing (due to caries), and/ or filled primary teeth, of English-speaking and non-English speaking patients of a hospital based pediatric dental clinic under the age of 72 months to determine if native language is a risk marker for tooth decay. STUDY DESIGN: Records from an outpatient dental clinic which met the inclusion criteria were reviewed. Patient demographics and dmft score were recorded, and the patients were separated into three groups by the native language spoken by their parents: English, Spanish and all other languages. RESULTS: A total of 419 charts were assessed: 253 English-speaking, 126 Spanish-speaking, and 40 other native languages. After accounting for patient characteristics, dmft was significantly higher for the other language group than for the English-speaking (p<0.001) and Spanish-speaking groups (p<0.05), however the English-speaking and Spanish-speaking groups were not different from each other (p>0.05). CONCLUSIONS: Those patients under 72 months of age whose parents' native language is not English or Spanish, have the highest risk for increased dmft when compared to English and Spanish speaking patients. Providers should consider taking additional time to educate patients and their parents, in their native language, on the importance of routine dental care and oral hygiene.

Effect of Human Milk and its Components on Streptococcus Mutans Biofilm Formation.

OBJECTIVE: This study investigated the effects of human breast milk and its components on the nutritional aspect of the caries process due to Streptococcus mutans UA159 biofilm formation. STUDY DESIGN: Human breast milk was collected from 11 mothers during 3-9 months postpartum. To test for the effect on biofilm formation, a 16-hour culture of S. mutans was treated with dilutions of human breast milk and several major components of human breast milk, lactose, lactoferrin, IgA, and bovine casein in sterile 96-well flat bottom microtiter plates for 24 hours. The biofilms were fixed, washed, stained with crystal violet, and extracted. Absorbance was measured to evaluate biofilm growth mass. RESULTS: Dilutions 1:10-1:2,560 of the human breast milk samples increased biofilm formation by 1.5-3.8 fold compared to the control. Lactoferrin decreased biofilm formation significantly in all dilutions (average milk concentration of 3 mg/ml). Lactose had no effect at average breast milk concentrations (60 mg/ml) except at its lowest concentration (15 mg/ml) where it was increased. IgA significantly decreased biofilm formation at its highest concentration of 2,400 µg/ml (average milk concentration 600 µg/ml). Casein caused significantly increased biofilm formation at all concentrations tested above the average milk content (2.3 mg/ml). CONCLUSIONS: The results of this study demonstrate an increase in S. mutans biofilm formation by human breast milk 3-9 months post partum. Among its major components, only casein significantly increased biofilm formation among the concentrations analyzed. Lactose had no effect except at 15 mg/ml. Lactoferrin and IgA significantly decreased S. mutans biofilm formation at their highest concentrations. This information expands the current knowledge regarding the nutritional influence of breastfeeding and validates the necessity to begin an oral hygiene regimen once the first tooth erupts.

α-TEA cooperates with MEK or mTOR inhibitors to induce apoptosis via targeting IRS/PI3K pathways.

BACKGROUND: α-Tocopherol ether-linked acetic acid (α-TEA) is a promising agent for cancer prevention/therapy based on its antitumour actions in a variety of cancers. METHODS: Human breast cancer cells, MCF-7 and HCC-1954, were used to study the effect of α-TEA using Annexin V/PI staining, western blot analyses, and siRNA knockdown techniques. RESULTS: α-Tocopherol ether-linked acetic acid suppressed constitutively active basal levels of pAKT, pERK, pmTOR, and their downstream targets, as well as induced both cell types to undergo apoptosis. Phosphoinositide 3-kinase (PI3K) inhibitor wortmannin suppressed pAKT, pERK, pmTOR, and their downstream targets, indicating PI3K to be a common upstream mediator. In addition, α-TEA induced increased levels of pIRS-1 (Ser-307), a phosphorylation site correlated with insulin receptor substrate-1 (IRS-1) inactivation, and decreased levels of total IRS-1. Small interfering RNA (siRNA) knockdown of JNK blocked the impact of α-TEA on pIRS-1 and total IRS-1 and impeded its ability to downregulate the phosphorylated status of AKT, ERK, and mTOR. Combinations of α-TEA+MEK or mTOR inhibitor acted cooperatively to induce apoptosis and reduce basal levels of pERK and pmTOR. Importantly, inhibition of MEK and mTOR resulted in increased levels of pAKT and IRS-1, and α-TEA blocked them. CONCLUSIONS: Downregulation of IRS-1/PI3K pathways via JNK are critical for α-TEA and α-TEA+MEK or mTOR inhibitor-induced apoptosis in human MCF-7 and HCC-1954 breast cancer cells.

Geometric phase distributions for open quantum systems.

In an open system, the geometric phase should be described by a distribution. We show that a geometric phase distribution for open system dynamics is in general ambiguous, but the imposition of reasonable physical constraints on the environment and its coupling with the system yields a unique geometric phase distribution that applies even for mixed states, nonunitary dynamics, and noncyclic evolutions.

Antibacterial and physical properties of resin modified glass-ionomers combined with chlorhexidine.

The purpose of this study was to determine the effect on mechanical properties and antimicrobial activity of the addition of chlorhexidine (CHX) to a resin modified glass-ionomer (Photac-fil, ESPE, Norristown, PA, USA). Chlorhexidine diacetate was combined with a resin modified glass-ionomer material at a concentration of 5%. The samples were tested for hardness, tensile strength and erosion at 24 h and 6-week intervals and for elution of CHX and antimicrobial activity weekly for 6 weeks. At 24 h there was no significant difference in hardness between the two groups, but at 6 weeks the resin modified glass-ionomer group was significantly harder than the CHX groups (P < 0.05). The diametral tensile strength test indicated no difference between the control and CHX groups at 24 h or at 6 weeks. The jet erosion test demonstrated significantly less erosion with the CHX group at 24 h but at 6 weeks the CHX group showed significantly more erosion than the control group. The chemical assay data demonstrated a peak elution of CHX at week 1 with residual amounts at weeks 2 and 3. The microbial data demonstrated that the CHX group had a significant reduction in Streptococcus mutans numbers for weeks 1-3, but after week 4 there was no difference between the glass-ionomer with and without CHX. The addition of CHX to resin modified glass-ionomer altered hardness and erosion of the resin-modified glass-ionomer, but because there are no material specifications, it is difficult to determine clinical implications. Chlorhexidine did significantly improve the antimicrobial effect of the glass-ionomer which was consistent with the chemical assay data. The results indicated that the addition of CHX to resin modified glass-ionomer material (Photac-fil) did not seriously degrade the physical properties during the time period tested and that the addition of CHX resulted in a greater reduction in S. mutans when compared with glass-ionomer alone.

Comparison of electrosurgical and formocresol pulpotomy procedures in children.

OBJECTIVES: Although previous studies have examined the electrosurgical pulpotomy technique for primary teeth, no well-controlled, clinical human trials have been published. The purpose of this study was to prospectively compare electrosurgical pulpotomies vs. formocresol pulpotomies in human vital primary molar teeth. DESIGN: Fifty children were randomly divided into two groups, 25 receiving an electrosurgical pulpotomy and 25 receiving a formocresol pulpotomy. RESULTS: After at least 5 months postoperative observation time, the clinical and radiographic success rates for the electrosurgical groups were 96 and 84%, respectively; and for the formocresol group, 100 and 92%, respectively. CONCLUSION: There were no statistically significant differences between the success rates for the two groups at the P < 0.05 level as tested by Fisher's exact test. This study failed to demonstrate a difference in the success rate between the electrosurgical and formocresol pulpotomy techniques.

RRR-alpha-tocopheryl succinate induces MDA-MB-435 and MCF-7 human breast cancer cells to undergo differentiation.

RRR-alpha-Tocopheryl succinate (vitamin E succinate, VES) is a potent antitumor agent, inducing DNA synthesis arrest, differentiation, and apoptosis. Because little is known about VES-induced differentiation, studies reported here characterize VES effects on the differentiation status of human breast cancer cell lines and investigate possible molecular mechanisms involved. VES-induced differentiation of human MCF-7 and MDA-MB-435 breast cancer cells was characterized by morphological changes, induction of lipid droplets, induction of beta-casein mRNA expression, and down-regulation of Her2/neu protein. In contrast, VES treatment of normal human mammary epithelial cells, MCF-10A cells, and T-47D cells did not induce differentiation. Studies addressing mechanisms showed that neither antibody neutralization of the transforming growth factor-beta signaling pathway nor expression of a dominant-negative mutant of c-Jun N-terminal kinase blocked the ability of VES to induce differentiation; however, treatment of cells with PD 98059, a chemical inhibitor of mitogen-activated protein kinase kinase (MEK1/2), blocked the ability of VES to induce differentiation.

Requirement of optical coherence for continuous-variable quantum teleportation.

We show that the sender and the receiver each require coherent devices in order to achieve unconditional continuous variable quantum teleportation (CVQT), and this requirement cannot be achieved with conventional laser sources, linear optics, ideal photon detectors, and perfect Fock state sources. The appearance of successful CVQT in recent experiments is due to interpreting the measurement record fallaciously in terms of one preferred ensemble (or decomposition) of the correct density matrix describing the state. Our analysis is unrelated to technical problems such as laser phase drift or finite squeezing bandwidth.

Activation of extracellular signal-regulated kinase and c-Jun-NH(2)-terminal kinase but not p38 mitogen-activated protein kinases is required for RRR-alpha-tocopheryl succinate-induced apoptosis of human breast cancer cells.

RRR-alpha-tocopherol succinate (vitamin E succinate, VES) is a potent, selective apoptotic agent for cancer cells but not normal cells. VES has been shown to inhibit the growth of a wide variety of tumor cells in cell culture and animal models. Studies addressing mechanisms of action of VES-induced apoptosis have identified transforming growth factor-beta, Fas/CD95-APO-1, and mitogen-activated protein kinase (MAPK) signaling pathway involvement. Here we show that MAPKs, the extracellular signal-regulated kinases (ERK), and the stress-activated protein kinases, c-Jun NH2-terminal kinases (JNK), but not p38, are critical mediators in VES-induced apoptosis of human breast cancer MDA-MB-435 cells. VES activates ERK1/2 and JNK both in level and duration of kinase activity. Expression of dominant negative mutants of ERK1, MAPK/ERK activator-1, or JNK1 but not p38 blocked phosphorylation of the substrate glutathione S-transferase-c-Jun and inhibited VES-induced apoptosis. Increased phosphorylation and transactivation activity of nuclear transcription factors c-Jun, ATF-2, and Elk-1 are observed after VES treatments; however, only c-Jun and ATF-2 appear to be involved in VES-induced apoptosis based on antisense blockage experiments. Collectively, these results imply a critical role for ERK1 and JNK1 but not p38 in VES-induced apoptosis of human MDA-MB-435 breast cancer cells.

Generation of enterococci bacteria in a coastal saltwater marsh and its impact on surf zone water quality.

Elevated levels of enterococci bacteria, an indicator of fecal pollution, are routinely detected in the surf zone at Huntington State and City Beaches in southern California. A multidisciplinary study was carried out to identify sources of enterococci bacteria landward of the coastline. We find that enterococci bacteria are present at high concentrations in urban runoff, bird feces, marsh sediments, and on marine vegetation. Surprisingly, urban runoff appears to have relatively little impact on surf zone water quality because of the long time required for this water to travel from its source to the ocean. On the other hand, enterococci bacteria generated in a tidal saltwater marsh located near the beach significantly impact surf zone water quality. This study identifies a potential tradeoff between restoring coastal wetlands and protecting beach water quality and calls into question the use of ocean bathing water standards based on enterococci at locations near coastal wetlands.

Sodium replacement and fluid shifts during prolonged exercise in humans.

In the study presented here, we examined the affects of a close to complete replacement of sweat water and Na+ losses on fluid shifts during exercise. Six cyclists performed three 4-h rides at 55% of their peak oxygen uptake in a 20 degrees C environment while consuming 3.85 l of an 8% carbohydrate solution containing 5, 50 or 100 mEq.l-1 of Na+. Increases in Na+ intake reduced renal free water clearance from around 40 ml.h-1 to -8 and -121 ml.h-1 and led to a decrease in urine volume from approximately equal to 1.0 to 0.5 l (P < 0.05). In contrast, the 3.5-3.9 l fluid and 150-190 mEq Na+ losses in sweat were similar in each trial, as were the approximately equal to 80 mEq K+ losses in sweat and urine and the 282-288 mosmol.kg-1 plasma osmolalities. During the low-Na+ trial, plasma osmolality was maintained by a approximately equal to 1.3 l contraction of extracellular fluid (ECF) with the loss of approximately equal to 200 mEq Na+. However, in the other trials, approximately equal to 1.3 l of water was lost from the intracellular fluid. During the medium-Na+ trial, a loss of only approximately equal to 40 mEq Na+ maintained ECF volume, and during the high-Na+ trial, a gain of approximately equal to 160 mEq Na+ expanded the ECF by approximately equal to 0.8 l. However, corresponding changes in plasma volumes from -0.20 to 0.15 l had no effect on cardiovascular drift or thermoregulation. These data suggest that during prolonged exercise of moderate intensity under mild environmental conditions when sweat rates are approximately equal to 0.9 l.h-1, complete Na+ replacement maintains plasma volume and reduces dehydration, but when fluid intake matches sweat rate, has little effect on plasma osmolality.

Hypertonic saline-dextran suppresses burn-related cytokine secretion by cardiomyocytes.

Whereas hypertonic saline-dextran (HSD, 7.5% NaCl in 6% D70) improves cardiac contractile function after burn trauma, the mechanisms of HSD-related cardioprotection remain unclear. We recently showed that cardiomyocytes secrete tumor necrosis factor-alpha (TNF-alpha), a response that was enhanced by burn trauma. This study addressed the question: does HSD modulate cardiac contraction/relaxation by altering cardiomyocyte TNF-alpha secretion? Wistar-Furth rats (325 g) were given a burn injury over 40% of the total body surface area and were then randomized to receive a bolus of either isotonic saline or HSD (4 ml/kg, n = 14 rats/group). Sham burn rats were given either isotonic saline or HSD (n = 14 rats/group) to provide appropriate controls for the two burn groups. Hearts were isolated 24 h postburn for either Langendorff perfusion (n = 8 hearts/group) or to prepare cardiomyocytes (n = 6 hearts/group). Myocytes were stimulated with lipopolysaccharide (LPS) (0, 10, 25, or 50 microg for 18 h) to measure cytokine secretion. Burn trauma increased myocyte TNF-alpha and interleukin-1 beta and -6 secretion, exacerbated cytokine response to LPS stimulus, and impaired cardiac contraction. HSD treatment of burns decreased cardiomyocyte cytokine secretion, decreased responsiveness to LPS challenge with regard to cytokine secretion, and improved ventricular function. These data suggest that HSD mediates cardioprotection after burn trauma, in part, by downregulating cardiomyocyte secretion of inflammatory cytokines.

Geometric phase of three-level systems in interferometry.

We present the first scheme for producing and measuring an Abelian geometric phase shift in a three-level system where states are invariant under a non-Abelian group. In contrast to existing experiments and proposals for experiments, based on U(1)-invariant states, our scheme geodesically evolves U(2)-invariant states in a four-dimensional SU(3)/U(2) space and is physically realized via a three-channel optical interferometer.

Munchausen syndrome presenting in a patient who has undergone temporomandibular joint surgery.

Munchausen syndrome is a rare psychiatric disorder. Patients with Munchausen syndrome insist on and repeatedly undergo unnecessary investigations and operative treatments. No organic pathosis is demonstrated, and treatment consistently fails to alleviate the symptoms. This article presents a case report and a brief discussion to facilitate recognition and management techniques.

Inflammatory pseudotumor of the alimentary tract: clinical and surgical experience.

BACKGROUND/PURPOSE: Initially described in 1937, inflammatory pseudotumor (IPT) inflammatory myofibroblastic tumor (IMT) or plasma cell granulomas are synonymous for an inflammatory solid tumor that contains spindle cells, myofibroblasts, plasma cells, and histocytes. Common sites of presentation include lung, mesentary, liver, and spleen; intestinal presentations are rare, and the etiology remains obscure. This report details the clinical and surgical experiences in 4 children with alimentary tract IPT at a single institution. METHODS: A retrospective chart review was conducted of pediatric patients with the pathologic diagnosis of IPT. RESULTS: Between 1990 and 1999, 4 patients (4 girls, ages 5 to 15 years) were identified with gastrointestinal tract origins of IPT. Symptoms at presentation included anemia (n = 4), intermittent abdominal pain (n = 3), fever (n = 3), weight loss (n = 2), diarrhea (n = 2), dysphagia (n = 1). Two patients had comorbid conditions of juvenile rheumatoid arthritis and mature B cell lymphoma. Three of 4 patients had elevated sedimentation rates. The sites of origin were the gastroesophageal junction, the colon, the rectum, and the appendix, with the referral diagnosis achalasia, perforated appendix, inflammatory bowel disease, and recurrent lymphoma, respectively. All were treated with aggressive surgical resection, and 3 girls have had no recurrences since the initial surgery. One patient had 3 recurrences within 8 months of presentation; she remains disease free 8 years later. CONCLUSIONS: IPT, although rare in the gastrointestinal tract, mimics more common problems. Successful surgical management is possible even in cases of multiple recurrences.