Utility of Ultrasound Elastography to Differentiate Benign from Malignant Cervical Lymph Nodes.

BACKGROUND: The purpose of this study was to evaluate the usefulness of strain elastography and acoustic radiation force impulse (ARFI) imaging in the differentiation of benign and malignant cervical lymph nodes (LNs). MATERIALS AND METHODS: In this prospective study, 50 enlarged cervical LNs (33 benign and 17 malignant) were examined by B-mode ultrasound (US), color Doppler, and strain elastography. Elastographic patterns (1-5) were categorized based on distribution of hard area within LN. The shear wave velocity (SWV) of LNs was evaluated by ARFI imaging. Diagnostic performance of sonoelastographic parameters was compared taking histopathology of LN as a reference standard. Optimal cutoff value of the mean SWV values for predicting malignancy was determined using receiver operating characteristic curve analysis. RESULTS: Among US parameters, borders of LN had the highest diagnostic accuracy (80%), while echogenicity had the least (48%). Majority of benign LNs (n = 31) had elastography patterns 1 and 2, while majority of malignant LNs (n = 16) had patterns 3-5 (P = 0.000). The sensitivity, specificity, and accuracy of elastography were 94.1%, 93.9%, and 94%, respectively. The mean SWV of benign LNs (1.670 ± 0.367 m/s) differed significantly from malignant LNs (2.965 ± 0.826 m/s; P = 0.000). A cutoff value of 2.05 m/s predicted malignancy with 88.2% sensitivity and 84.8% specificity and gave an area under the curve of 0.949 (95% confidence interval: 0.70-1.20). CONCLUSION: Elastography has high diagnostic accuracy in differentiating benign and malignant cervical LNs and can be potentially useful in selecting the LN with high probability of malignancy, on which fine-needle aspiration cytology/biopsy can be performed.

Preferences for partner notification method: variation in responses between respondents as index patients and contacts.

There have been very few studies focusing on what form of communication patients would find acceptable from a clinic. This study looks at the differences in preferences for various partner notification methods when the respondents were index patients compared with when they had to be contacted because a partner had a sexually transmitted infection (STI). There were 2544 respondents. When the clinic had to notify partners, respondents were more likely to report the method as good when a partner had an STI and they were being contacted compared with when the respondents had an infection and the partner was being contacted. The opposite was true for patient referral partner notification. Therefore, there are variations in the preferences of respondents for partner notification method, which depend on whether they see themselves as index patients or contacts.

Parathyroid hormone induction of rat interstitial collagenase mRNA in osteosarcoma cells is mediated through an AP-1-binding site.

Interstitial collagenase is secreted by the osteoblast in response to bone-resorbing agents. Previously, we cloned the rat interstitial collagenase cDNA from UMR 106-01 rat osteoblastic osteosarcoma cells. We demonstrated that induction of collagenase by PTH, a powerful resorbing agent, in UMR 106-01 cells is in part transcriptional. In the present study we isolate and characterize the rat interstitial collagenase gene. The gene consists of 10 exons and spans approximately 12 kbp. The major transcriptional start site, determined by primer extension analysis and confirmed by RNase protection assay, is 25 nucleotides upstream of the translational start site. The previously isolated cDNA was missing the 5'-untranslated sequence in addition to 17 nucleotides of the signal sequence of the preproenzyme; therefore, we also present these data. Chloramphenicol acetyl transferase (CAT) analyses were performed on the 5'-upstream region of the gene. These data indicate that PTH appears to mediate its effect through an AP-1 consensus-binding sequence (-51). Footprint analysis demonstrates protein binding to this site. Site-specific mutagenesis markedly decreased protein binding, which correlated directly with a decrease in CAT activation by PTH. Supershift data indicate that cAMP response element binding protein (CREB) is binding to this AP-1 consensus sequence. In addition we demonstrate that PTH induces phosphorylation of CREB.

Parathyroid hormone induces rat interstitial collagenase mRNA through Ets-1 facilitated by cyclic AMP response element-binding protein and Ca(2+)/calmodulin-dependent protein kinase II in osteoblastic cells.

Parathyroid hormone (PTH), a powerful bone-resorbing agent, is capable of stimulating interstitial collagenase (MMP-13) mRNA production in osteoblastic cells. In this study, a PEA3 consensus binding sequence (-80; AGGAAGT) in addition to a 'TRE-like' sequence (-89; CGACTCA) in the 5' upstream regulatory region of the rat MMP-13 gene were examined. In response to PTH, there was a time-dependent increase in binding of nuclear factors to an oligonucleotide containing the PEA3 region (-95 to -71). This increase in binding was first observed at 0.5 h, peaked at 4 h (7. 6-fold) then returned to basal levels by 24 h. Mutagenesis of the PEA3 site in a chloramphenicol acetyl transferase (CAT) construct containing 5' upstream regulatory sequence of the rat MMP-13 gene significantly decreased activation by PTH. PTH-mediated binding of nuclear factors to an oligonucleotide containing the mutant PEA3 sequence was decreased as compared with the wild type. Mutation or deletion of the TRE-like sequence affected basal as well as PTH-mediated induction of corresponding CAT constructs. Treatment with KN93, a Ca(2+)/calmodulin-dependent protein kinase II specific inhibitor, greatly reduced the amount of protein binding to the PEA3 region in response to PTH which correlated to a notable decrease in the amount of MMP-13 mRNA produced in response to PTH. Antibodies against Ets-1, cyclic AMP response element (CREB)-binding protein (CBP) and CREB were capable of supershifting proteins binding to the oligonucleotide containing the PEA3 region. These data suggest a possible co-operative interaction of factors binding to the PEA3 and TRE-like sequences and provide the first indication of a role for a calcium-mediated pathway in the PTH induction of MMP-13 mRNA in osteoblastic cells.

New rapid latex agglutination test for diagnosing Trichomonas vaginalis infection.

A newly developed latex agglutination test for Trichomonas vaginalis infection was compared for sensitivity, specificity, efficiency, and positive and negative predictive values with microscopy, culture, and an enzyme linked immunosorbent assay (ELISA) in the diagnosis of 395 women attending a genitourinary medicine clinic. T vaginalis infection was diagnosed in 42 (11%) women. The sensitivities of both the latex agglutination test and the ELISA were 95% compared with 74% for microscopy and 76% for culture. The latex test was specific and showed no cross reaction with a wide range of other genital tract infections. The latex agglutination test can detect antigen in both soluble and insoluble forms, and as it is simple to perform, can be undertaken during routine examination without recourse to special equipment or training. Further evaluation is required.

Treatment of poultry slaughterhouse wastewater in hybrid upflow anaerobic sludge blanket reactor packed with pleated poly vinyl chloride rings.

In this study, the performance of 5.4 L hybrid upflow anaerobic sludge blanket (HUASB) reactor for treating poultry slaughterhouse wastewater under mesophilic conditions (29-35 °C), was investigated. After starting-up, the reactor was loaded up to an OLR of 19 kg COD/m3 d and achieved varied TCOD and SCOD removal efficiencies of 70-86% and 80-92%, respectively. The biogas was varied between 1.1 and 5.2 m3/m3 d with the maximum methane content of 72%. The maximum methane yield was 0.32 m3/kg CODremoved at an OLR of 9.27 kg COD/m3 d. Black matured granules of size between 2.5 and 5 mm were observed at the end of 225 d operation. RTD study showed the flow behavior was in mixed regime at the end of performance study. Step wise polynomial regression analysis was fitted well. Methanobacterium and Methanosaeta bacteria were dominant at the end of start-up whereas Methanosarcina, Cocci and rods were predominant at the end of performance studies.

Patient preferences for partner notification.

OBJECTIVE: To identify patient preferences for notification of sexual contacts when a sexually transmitted infection (STI) is diagnosed. METHODS: A questionnaire survey of 2544 patients attending three large genitourinary clinics at Derby, Birmingham, and Coventry in the United Kingdom. RESULTS: The median age of the respondents was 24 with 1474 (57.9%) women, 1835 (72.1%) white, 1826 (71.8%) single. The most favoured method of partner notification was patient referral, which was rated a "good" method by 65.8% when they had to be contacted because a sexual partner has an STI. Notifying contacts by letter as a method of provider partner notification is more acceptable than phoning, text messaging, or email. Respondents with access to mobile telephones, private emails, and private letters were more likely to rate a method of partner notification using that mode of communication as "good" compared to those without. With provider referral methods of partner notification respondents preferred to receive a letter, email, or text message asking them to contact the clinic rather than a letter, email or text message informing them that they may have an STI. CONCLUSION: Most respondents think that being informed directly by a partner is the best method of being notified of the risk of an STI. Some of the newer methods may not be acceptable to all but a significant minority of respondents prefer these methods of partner notification. The wording of letters, emails, or text messages when used for partner notification has an influence on the acceptability of the method and may influence success of the partner notification method. Services should be flexible enough to utilise the patients' preferred method of partner notification.

Use of slide latex agglutination test for rapid diagnosis of vaginal candidosis.

A new slide latex agglutination test was compared with microscopy and culture for diagnosing vaginal candidosis in 367 women attending a genitourinary medicine clinic. Vaginal candidosis occurred in 10% of the patients, and 7% had commensal carriage of Candida spp. The slide latex agglutination test was superior to microscopy in immediate diagnosis and was rapid and simple to perform. Of the patients with vaginal candidosis, 72% were detected by slide latex agglutination test compared with 38% by microscopy. The test discriminated well between patients yielding cultures of Candida spp who had symptoms and signs and those who showed commensal carriage.

Developmental changes in ob gene expression and circulating leptin peptide concentrations.

We examined the developmental changes in murine white and brown adipose tissue leptin and circulating immunoreactive total leptin concentrations. The approximately 4.4 kb leptin mRNA levels were higher at 2 and 7d postnatal ages, but declined to adult levels by the 14d stage and remained so until 160d. Paralleling the mRNA concentrations, leptin peptide levels also were higher at 2d, 7d, and 14d, declining to adult values by the 21d weaning stage. No difference in mRNA levels was observed between brown-enriched and white adipose tissue. No sexual dimorphism was observed in the leptin mRNA or peptide levels between 14 and 160d; however, at 2 and 7d, while no sex related differences were observed in the peptide levels, adipose mRNA concentrations were mildly higher in males than in the females. We conclude that leptin mRNA and peptide levels are higher during consumption of a high fat milk diet. High levels of leptin with increasing food intake and body weight gain signify hypothalamic leptin receptor resistance during the immediate postnatal period.

Sp1 and Sp3 regulate transcriptional activity of the facilitative glucose transporter isoform-3 gene in mammalian neuroblasts and trophoblasts.

The murine facilitative glucose transporter isoform 3 (Glut 3) is developmentally regulated and is predominantly expressed in neurons and trophoblasts. Employing the primer extension and RNase protection assays, the transcription start site (denoted as +1) of the murine Glut 3 gene was localized to 305 base pairs (bp) 5' to the ATG translation start codon. Transient transfection assays in N2A, H19-7 neuroblasts, and HRP.1 trophoblasts using sequential 5'-deletions of the murine Glut 3-luciferase fusion gene indicated that the -203 to +237 bp region with reference to the transcriptional start site contained promoter activity. Repressor function was limited to the -137 to -130 bp region within the transcriptional activation domain. The nuclear factors Sp1 and Sp3 bound this GC-rich region in N2A, H19-7, and HRP.1 cells. Dephosphorylation of Sp1 was essential for Glut 3 DNA binding. The related Sp3 protein also bound this same region of mouse Glut 3 in all three cell lines. Mutations of the Sp1-binding site employed in transient transfection and mobility shift assays confirmed the nature of the DNA-binding proteins, while supershift assays with anti-Sp1 and anti-Sp3 IgGs characterized the differences in the two DNA-binding proteins. Co-transfection of the Glut 3-luciferase fusion gene with or without mutations of the Sp1-binding site along with the Sp1 or Sp3 expression vectors in Drosophila SL2 cells confirmed a reciprocal effect, with Sp1 suppressing and Sp3 activating Glut 3 gene transcription.

Prevalence of chlamydial infection in acute epididymo-orchitis.

The prevalence of sexually transmitted infection was studied in 40 men presenting with acute epididymo-orchitis in Leeds. Chlamydia trachomatis infection was identified in 13 of 29 men (45%) aged under 35 years. Neisseria gonorrhoeae was isolated from four of these 13 men with chlamydial urethritis. C trachomatis was isolated from the urethra of only one of 11 men (9%) aged over 35. Appreciable bacteriuria was found in six of these 11 men (55%). Noting details of sexual history and screening for sexually acquired pathogens is advocated in younger men with acute epidiymo-orchitis.

Developmental regulation of genes mediating murine brain glucose uptake.

We examined the molecular mechanisms that mediate the developmental increase in murine whole brain 2-deoxyglucose uptake. Northern and Western blot analyses revealed an age-dependent increase in brain GLUT-1 (endothelial cell and glial) and GLUT-3 (neuronal) membrane-spanning facilitative glucose transporter mRNA and protein concentrations. Nuclear run-on experiments revealed that these developmental changes in GLUT-1 and -3 were regulated posttranscriptionally. In contrast, the mRNA and protein levels of the mitochondrially bound glucose phosphorylating hexokinase I enzyme were unaltered. However, hexokinase I enzyme activity increased in an age-dependent manner suggestive of a posttranslational modification that is necessary for enzymatic activation. Together, the postnatal increase in GLUT-1 and -3 concentrations and hexokinase I enzymatic activity led to a parallel increase in murine brain 2-deoxyglucose uptake. Whereas the molecular mechanisms regulating the increase in the three different gene products may vary, the age-dependent increase of all three constituents appears essential for meeting the increasing demand of the maturing brain to fuel the processes of cellular growth, differentiation, and neurotransmission.

Effect of primary congenital hypothyroidism upon expression of genes mediating murine brain glucose uptake.

Using hyt/hyt mice that exhibit naturally occurring primary hypothyroidism (n = 72) and Balb/c controls (n = 66), we examined the mRNA, protein, and activity of brain glucose transporters (Glut 1 and Glut 3) and hexokinase I enzyme at various postnatal ages (d 1, 7, 14, 21, 35, and 60). The hyt/hyt mice showed an age-dependent decline in body weight (p < 0.04) and an increase in serum TSH levels (p < 0.001) at all ages. An age-dependent translational/posttranslational 40% decline in Glut 1 (p = 0.02) with no change in Glut 3 levels was observed. These changes were predominant during the immediate neonatal period (d 1). A posttranslational 70% increase in hexokinase enzyme activity was noted at d 1 alone (p < 0.05) with no concomitant change in brain 2-deoxy-glucose uptake. This was despite a decline in the hyt/hyt glucose production rate. We conclude that primary hypothyroidism causes a decline in brain Glut 1 associated with no change in Glut 3 levels and a compensatory increase in hexokinase enzyme activity. These changes are pronounced only during the immediate neonatal period and disappear in the postweaned stages of development. These hypothyroid-induced compensatory changes in gene products mediating glucose transport and phosphorylation ensure an adequate supply of glucose to the developing brain during transition from fetal to neonatal life.