Targeted TNF-α Overexpression Drives Salivary Gland Inflammation.

Chronic inflammation of the salivary glands from pathologic conditions such as Sjögren's syndrome can result in glandular destruction and hyposalivation. To understand which molecular factors may play a role in clinical cases of salivary gland hypofunction, we developed an aquaporin 5 (AQP5) Cre mouse line to produce genetic recombination predominantly within the acinar cells of the glands. We then bred these mice with the TNF-αglo transgenic line to develop a mouse model with salivary gland-specific overexpression of TNF-α; which replicates conditions seen in sialadenitis, an inflammation of the salivary glands resulting from infection or autoimmune disorders such as Sjögren's syndrome. The resulting AQP5-Cre/TNF-αglo mice display severe inflammation in the salivary glands with acinar cell atrophy, fibrosis, and dilation of the ducts. AQP5 expression was reduced in the salivary glands, while tight junction integrity appeared to be disrupted. The immune dysregulation in the salivary gland of these mice led to hyposalivation and masticatory dysfunction.

Conditional TNF-α Overexpression in the Tooth and Alveolar Bone Results in Painful Pulpitis and Osteitis.

Tumor necrosis factor-α (TNF-α) is a proalgesic cytokine that is commonly expressed following tissue injury. TNF-α expression not only promotes inflammation but can also lead to pain hypersensitivity in nociceptors. With the established link between TNF-α and inflammatory pain, we identified its increased expression in the teeth of patients affected with caries and pulpitis. We generated a transgenic mouse model (TNF-α(glo)) that could be used to conditionally overexpress TNF-α. These mice were bred with a dentin matrix protein 1 (DMP1)-Cre line for overexpression of TNF-α in both the tooth pulp and bone to study oral pain that would result from subsequent development of pulpitis and bone loss. The resulting DMP1/TNF-α(glo) mice show inflammation in the tooth pulp that resembles pulpitis while also displaying periodontal bone loss. Inflammatory infiltrates and enlarged blood vessels were observed in the tooth pulp. Pulpitis and osteitis affected the nociceptive neurons innervating the orofacial region by causing increased expression of inflammatory cytokines within the trigeminal ganglia. With this new mouse model morphologically mimicking pulpitis and osteitis, we tested it for signs of oral pain with an oral function assay (dolognawmeter). This assay/device records the time required by a mouse to complete a discrete gnawing task. The duration of gnawing required by the DMP1/TNF-α(glo) mice to complete the task was greater than that for the controls; extended gnaw time in a dolognawmeter indicates reduced orofacial function. With the DMP1/TNF-α(glo) mice, we have shown that TNF-α expression alone can produce inflammation similar to pulpitis and osteitis and that this mouse model can be used to study dental inflammatory pain.

Plasma observations during the Mars atmospheric "plume" event of March-April 2012.

We present initial analysis and conclusions from plasma observations made during the reported "Mars plume event" of March - April 2012. During this period, multiple independent amateur observers detected a localized, high-altitude "plume" over the Martian dawn terminator [Sanchez-Lavega et al., Nature, 2015, doi:10.1038/nature14162], the cause of which remains to be explained. The estimated brightness of the plume exceeds that expected for auroral emissions, and its projected altitude greatly exceeds that at which clouds are expected to form. We report on in-situ measurements of ionospheric plasma density and solar wind parameters throughout this interval made by Mars Express, obtained over the same surface region, but at the opposing terminator. Measurements in the ionosphere at the corresponding location frequently show a disturbed structure, though this is not atypical for such regions with intense crustal magnetic fields. We tentatively conclude that the formation and/or transport of this plume to the altitudes where it was observed could be due in part to the result of a large interplanetary coronal mass ejection (ICME) encountering the Martian system. Interestingly, we note that the only similar plume detection in May 1997 may also have been associated with a large ICME impact at Mars.

Structure-based mutagenesis reveals distinct functions for Ras switch 1 and switch 2 in Sos-catalyzed guanine nucleotide exchange.

Ras GTPases function as binary switches in signaling pathways controlling cell growth and differentiation. The guanine nucleotide exchange factor Sos mediates the activation of Ras in response to extracellular signals. We have previously solved the crystal structure of nucleotide-free Ras in complex with the catalytic domain of Sos (Boriack-Sjodin, P. A., Margarit, S. M., Bar-Sagi, D., and Kuriyan, J. (1998) Nature 394, 337-343). The structure demonstrates that Sos induces conformational changes in two loop regions of Ras known as switch 1 and switch 2. In this study, we have employed site-directed mutagenesis to investigate the functional significance of the conformational changes for the catalytic function of Sos. Switch 2 of Ras is held in a very tight embrace by Sos, with almost every external side chain coordinated by Sos. Mutagenesis of contact residues at the switch 2-Sos interface shows that only a small set of side chains affect binding, with the most important contact being mediated by tyrosine 64, which is buried in a hydrophobic pocket of Sos in the Ras.Sos complex. Substitutions of Ras and Sos side chains that are inserted into the Mg(2+)- and nucleotide phosphate-binding site of switch 2 (Ras Ala(59) and Sos Leu(938) and Glu(942)) have no effect on the catalytic function of Sos. These results indicate that the interaction of Sos with switch 2 is necessary for tight binding, but is not the critical driving force for GDP displacement. The structural distortion of switch 1 induced by Sos is mediated by a small number of specific contacts between highly conserved residues on both Ras and Sos. Mutations of a subset of these residues (Ras Tyr(32) and Tyr(40)) result in an increase in the intrinsic rate of nucleotide dissociation from Ras and impair the binding of Ras to Sos. Based on this analysis, we propose that the interactions of Sos with the switch 1 and switch 2 regions of Ras have distinct functional consequences: the interaction with switch 2 mediates the anchoring of Ras to Sos, whereas the interaction with switch 1 leads to disruption of the nucleotide-binding site and GDP dissociation.

Acquired maxillary sinus hypoplasia: a consequence of endoscopic sinus surgery?

The effects of functional endoscopic sinus surgery (FESS) on sinus and midfacial development remain unclear. The authors report five children who, at a median age of 30 months, underwent FESS for refractory sinusitis. Three of the children had cystic fibrosis, and two had asthma. Preoperative computed tomographic (CT) scanning showed symmetric maxillary sinus development with varying degrees of mucosal disease. At a mean of 42 months after surgery, CT scans were obtained to evaluate recurrent symptoms in the five children. The scans showed unilateral maxillary hypoplasia in four children and bilateral maxillary sinus hypoplasia in one child. No child had clinically apparent facial asymmetry or midfacial hypoplasia. The authors also discuss the factors involved in maxillary sinus pneumatization, the possible effects of sinus surgery on sinus development, and the clinical implications of "acquired" maxillary sinus hypoplasia.

An open multicentre study of tropisetron for cisplatin-induced nausea and vomiting.

OBJECTIVES: (i) To assess the efficacy and tolerability of tropisetron when used for acute and delayed cisplatin-induced emesis. (ii) To investigate whether dexamethasone added to tropisetron improves the control of emesis for patients who do not achieve a complete response to tropisetron alone. (iii) To assess sex of the patient and alcohol intake as prognostic factors for nausea and vomiting. DESIGN: A prospective open label phase II trial over one or two cycles of chemotherapy. Data collection was based on observed response and patients' self-reporting. SETTING: Twenty Australian tertiary care hospitals in 1994. PATIENTS: 102 male and female patients from 18 to 75 years with histologically confirmed malignancy receiving their first chemotherapy containing > or = 50 mg/m2 cisplatin. INTERVENTION: In Cycle 1 tropisetron 5 mg was given intravenously before chemotherapy on Day 1, then 5 mg orally before breakfast on Days 2 to 6. In Cycle 2, dexamethasone 20 mg intravenously on Day 1, then 8 mg orally on Days 2 to 6 could be added to tropisetron if a complete antiemetic response had not been achieved in Cycle 1. MAIN OUTCOME MEASURES: Number of vomiting episodes and severity of nausea for 6 days after chemotherapy; severity of side effects; patient satisfaction with chemotherapy treatment; oestradiol levels in women; and past alcohol consumption in men and women. RESULTS: (i) The complete response rate (CR) for acute emesis in Cycle 1 was 64% (95% confidence interval [CI], 54%-72%), with 84% (95% CI, 76%-90%) having < or = 2 vomits. The CR for delayed emesis was 24% (95% CI, 17%-32%). The CR for acute nausea was 56% (95% CI, 47%-66%), with 97% (95% CI, 91%-99%) having < or = 2 nausea episodes. The CR for delayed nausea was 21% (95% CI, 14%-30%). Seventy-one patients received Cycle 2. The main side effects were headache (20 patients) and constipation (16 patients). The control of acute emesis was rated as "good" or "very good" by 68% of investigators; 85% rated the tolerability of treatment as "good" or "very good". Treatment was rated as "very satisfactory" or "satisfactory" by 52% of patients. (ii) The CR for acute emesis with dexamethasone added was 78% (95% CI, 64%-88%). (iii) Women with lower oestradiol levels had better control of emesis, although this difference was not statistically significant. Chronic alcohol intake and binge drinking were strongly associated with a complete acute antiemetic response. CONCLUSIONS: Tropisetron was effective for acute cisplatin-induced emesis; adding dexamethasone enhanced this response. Both single and combined therapy had less effect on delayed emesis. The impact of alcohol on control of emesis is a chronic rather than acute phenomenon which requires prospective testing.

The atria of the fetal lateral ventricles: a sonographic study of normal atrial size and choroid plexus volume.

OBJECTIVE: Evaluation of the cerebral ventricular system is a routine part of all fetal sonographic examinations. Ventriculomegaly and decreased choroid volume are indicators of poor fetal outcome, so it is important to know the normal variation of these parameters. The present study describes the normal size of the fetal lateral ventricular atrium and the normal amount of choroid plexus within the atrium during the second and third trimesters. SUBJECTS AND METHODS: Six hundred eight consecutive, healthy, singleton fetuses from 13 to 42 weeks' menstrual age were examined prospectively. The atrium of the lateral ventricle was measured in the axial and coronal planes. For each axial measurement, the width of the choroid plexus was determined. RESULTS: An axial atrial measurement was obtained in 88% of the fetuses, and a coronal atrial measurement was obtained in 84%. The mean measurement and one standard deviation are 6.5 +/- 1.5 mm in the axial plane and 6.6 +/- 1.5 mm in the coronal plane. Ninety-eight percent of all measured atria were at least 60% filled by choroid plexus, and in all cases the atria were at least 50% filled. Complete filling by choroid plexus occurred in 58% of cases. CONCLUSION: This large prospective study confirms previous observations of mean atrial size. However, four standard deviations above the mean is 12 mm, suggesting currently used cutoffs for normal atrial size are too low. Other parameters, such as choroid plexus filling, may be helpful markers of normalcy in fetuses with atrial measurements between 8 and 12 mm.

The impact of AIDS on recruitment in the health professions.

The occupational risk of AIDS has been identified as a possible factor in the decline of applicants to health professions programs. This study examined data obtained from 914 Philadelphia public and parochial high school students concerning their knowledge and attitudes about AIDS, and their knowledge and attitudes toward the health professions. The survey revealed that the majority of respondents demonstrated a good, basic understanding of the transmission of the virus. While fear of exposure to the virus may negatively impact some prospective students, the majority in this study did not express this concern. Rather, the results indicated that the respondents' lack of knowledge and understanding of the health professions is a major reason that they may not be selecting health careers. Notably, more than 50% of those surveyed responded that high schools do not provide enough information about the health professions. These findings reinforce the need for creative recruitment and career counseling programs that will foster an interest in the health professions.

Schistosoma japonicum: monoclonal antibodies to the Mr 26,000 schistosome glutathione S-transferase (Sj26) in an assay for circulating antigen in infected individuals.

Two monoclonal antibodies have been produced that bind to separate epitopes on the Mr 26,000 glutathione S-transferase (GST) of Schistosoma japonicum worms (Sj26). Both antibodies have been used in an enzyme immunoassay (EIA) with sera from infected individuals from the Philippines. Relatively high signals were obtained with sera from some, but not all, individuals who are positive for fecal eggs. Evidence was obtained that the material detected by the monoclonal antibodies was present in minute amounts and in some sera was bound in a complex with phosphorylcholine-containing molecules. It could not be absorbed by reaction with glutathione-agarose columns. There was no detectable immunoglobulin in the complex. The possibility exists that the complexes are composed of schistosome GST, or fragments, and damaged tegumental lipids shed as a result of surface immune attack. However, the presence of the native Sj26 molecule has not been proven. More detailed longitudinal studies in endemic areas are required to determine whether the assay can be used as an indicator of acquired resistance ("concomitant immunity") and whether it will be useful in the search for immunological correlates of this resistance in humans.

Comparison of flow cytometry and retrospectively applied static cytometry on lymphoid tissue.

Twenty cases of non-Hodgkin's malignant lymphoma were examined using both flow cytometry (FCM) and static cytometry (SCM) DNA analysis to detect aneuploidic cell populations. FCM was performed on fresh cell suspensions whilst SCM was performed retrospectively on formalin-fixed, paraffin-embedded samples of the same tissue. A total of 34 aneuploidic cell lines were detected by FCM compared to a total of 41 detected by SCM. Of the 20 cases 14 showed DNA indexes within 10% of each other for each method, indicating a reasonable degree of comparability between the two methods. Of the 6 cases which did not show comparable DNA indexes, 2 displayed similar cell lines for both methods, but the major abnormal cell populations were of different ploidy by SCM as compared to FCM. A qualitative comparison is thus possible between these two cases. The results suggest that retrospectively applied SCM ploidy analysis of lymphoid tissue is comparable to FCM on fresh tissue samples and can thus be used in retrospective studies of both prognostic and diagnostic significance.

Cytogenetic abnormalities in benign lymphoid hyperplasia: a dual-parameter study using chromosome analysis and flow cytometry.

This is a prospective study of lymphoid tissue showing benign reactive hyperplasia (18 lymph nodes and 2 tonsils), using cytogenetic analysis of cells stimulated with T- or B-cell mitogens. The reason for this study was the detection of an abnormal chromosomal population in cells from an enlarged lymph node excised from a 7-year-old female who on further investigation was found to be clinically well and after one year's close follow-up had not developed any further signs or symptoms of malignancy. In addition, DNA content was measured by flow cytometry (FCM) on fresh cell suspensions in 17 cases and fixed cell suspensions in 3 cases. Structural and numerical clonal chromosome abnormalities were found in 9 of the 20 samples, but no common specific defect was identified. FCM showed an abnormal DNA content in 10 of the 20 samples studied; 3 of these showed clonal chromosome abnormalities. Surface membrane immunoglobulin studies were carried out on 15 of the 20 samples using cell suspensions and frozen tissue sections. In 5 of the 15 cases, monoclonal surface immunoglobulin was detected. There was no direct correlation between the surface membrane immunoglobulin studies and the chromosome and FCM analyses. We conclude that aneuploidy is a common feature in reactive lymphoid tissue, but both cytogenetics and FCM are needed to identify it.

A prospective single-blind trial of minocycline and doxycycline in the treatment of genital Chlamydia trachomatis infection in women.

A total of 2124 women who attended the Richmond Family Planning Association Clinic in Melbourne consecutively were screened for the presence or absence of Chlamydia trachomatis. One hundred and three women were found to have Chl. trachomatis infection of the cervix and were invited to participate in a clinical trial of minocycline and doxycycline for the treatment of chlamydial infection. A 10-day course of either drug resulted in a negative result of a chlamydial culture for all patients at the follow-up assessment, which occurred between 11 days to 12 weeks after the therapy. Minocycline and doxycycline showed equal effectiveness in the eradication of mycoplasmas in over 80% of the treated patients. Minocycline appeared to have a slight advantage with respect to the resolution of the gynaecological symptoms that were associated with the chlamydial infection. The number of adverse events that were recorded during the trial was similar for both treatment regimens. Gynaecological symptoms were associated with chlamydial infection in approximately 50% of the women in the study. The lack of association between chlamydial infection and gynaecological symptoms has led to the instigation of routine testing for the presence of Chlamydia spp. in young women who have more than one sexual partner.

Microbiological profile of the cervix in 1,000 sexually active women.

One thousand consecutive women who attended the Richmond Family Planning Association Clinic and who were to undergo a vaginal examination were asked to participate in a large prospective microbiological study. Participants were questioned about their sexual activity during the previous 12 months and any apparent signs of sexually transmitted disease. On examination the cervix was inspected for evidence of inflammation, ectopy or discharge and cervical swabs were taken microbiological assessment. Chlamydia trachomatis was isolated in 5.1% of women tested while Ureaplasma urealyticum and Mycoplasma hominis were found in 48.8% and 16.4% of women respectively. Significant associations were found between the number of sexual partners during the previous 12 months and the incidence of all 3 organisms. The carriage rate of the genital mycoplasmas was significantly affected by the type of contraception. In addition the association between the presence of the genital mycoplasmas and pelvic and cervical abnormalities was determined.

Mercapturic acid formation in the marmoset (Callithrix jacchus).

Benzylmercapturic acid is a major metabolite of [methylene-14C]benzyl chloride in the marmoset, as in the rat. The excretion of the minor metabolites benzylmercapturic acid sulphoxide and benzylcysteine accounted for a greater proportion of the dose than in the rat. Excretion of hippuric acid as a metabolite of benzyl chloride was variable in the marmoset. Acetylation of S-benzyl- and S-pentyl-L-cysteine to the corresponding mercapturic acids was extensive in the marmoset. Trace amounts of the sulphoxides of these acids were also excreted.

A comparison of some methods for identifying amelanotic and oligomelanotic melanoma metastases in paraffin sections.

Four methods were compared for identifying amelanotic and oligomelanotic melanomas in paraffin sections of formalin-fixed metastases from subjects with primary cutaneous melanomas. Of the amelanotic and oligomelanotic metastases a characteristic pattern of fluorescence was seen with an incident-light fluorescence microscope in 11 of 25 (44%); the Warthin-Starry stain at pH 3.2 was positive in 14 of 25 (56%); these two procedures together were a little more effective, 63% positive; S100 protein was revealed by immunoperoxidase staining in 26 of 28 (93%); the monoclonal antibody NKI/C-3 against a human melanoma antigen gave positive immunoperoxidase staining in 24 of 27 cases (89%). Of the pigmented metastases S100 protein was demonstrated in 18 of 21 (86%) and NKI/C-3 gave positive staining in all 20 tested. These antibodies are not specific for melanoma but a metastasis which does not react with either antibody is unlikely to be melanoma.

Some pathways of xenobiotic metabolism in the adult and neonatal marmoset (Callithrix jacchus).

1. An oral dose of [carboxyl-14C]benzoic acid was excreted as hippuric acid or benzoylglucuronide by the adult marmoset Callithrix jacchus, and the relative proportions of the metabolites were dose-dependent over the range 1-100mg/kg body weight, the proportion of glucuronideincreasing with increasing dose level. 2. [14C]Aspirin was rapidly absorbed by the marmoset, giving peak blood levels within 1 h of dosing. Unchanged aspirin (trace to 5% dose) was excreted up to 3 h after dosage. Salicylic acid was the major metabolite, relatively little asalicyluric acid was excreted; glucuronic acid conjugates accounted for about 30% of the urinary metabolites and gentisic acid about 2%. 3. p-aminobenzoic acid administered to adult marmosets was excreted as p-acetamidobenzoic acid, the major urinary metabolite, unchanged p-aminobenzoates, p-aminobenzoyl glucuronide and p-aminohippuric acid each representing about 10% of the urinary metabolities; one-fifth of the p-aminohippurate was acetylated. 4. After oral administration of [35S]sulphadiazine to marmosets, peak blood levels of 35S occurred within 5 h dosing, and immediately fell to near-zero values at 24 h. Sulphadiazine and N-acetylsulphadiazine comprised 36% and 34% respectively of the urine 35S while the N1-glucuronide (tentative identification) accounted for 22%. Minor metabolites were the N4-glucuronide and N4-sulphate of sulphadiazine. 5. The extent of acetylation of sulphadimidine by marmosets corresponded to that of fast acetylators in man. 6. The metabolism of [14C]benzoate and of p-aminobenzoic acid in the neonatal marmoset was compared with that in similarly dosed neonatal rats.