Investigation and response to an outbreak of Neisseria meningitidis serogroup Y ST-1466 urogenital infections, Australia.

Abstract: In 2023, an increased number of urogenital and anorectal infections with Neisseria meningitis serogroup Y (MenY) were reported in New South Wales (NSW). Whole genome sequencing (WGS) found a common sequence type (ST-1466), with limited sequence diversity. Confirmed outbreak cases were NSW residents with a N. meningitidis isolate matching the cluster sequence type; probable cases were NSW residents with MenY isolated from a urogenital or anorectal site from 1 July 2023 without WGS testing. Of the 41 cases, most were men (n = 27), of whom six reported recent contact with a female sex worker. Five cases were men who have sex with men and two were female sex workers. Laboratory alerts regarding the outbreak were sent to all Australian jurisdictions through the laboratories in the National Neisseria Network. Two additional states identified urogenital MenY ST-1466 infections detected in late 2023. Genomic analysis showed all MenY ST-1466 sequences were interspersed, suggestive of an Australia-wide outbreak. The incidence of these infections remains unknown, due to varied testing and reporting practices both within and across jurisdictions. Isolates causing invasive meningococcal disease (IMD) in Australia are typed, and there has been no MenY ST-1466 IMD recorded in Australia to end of March 2024. Concerns remain regarding the risk of IMD, given the similarity of these sequences with a MenY ST-1466 IMD strain causing a concurrent outbreak in the United States of America.

Nonspecific effect of Mycograb on amphotericin B MIC.

Mycograb C28Y is a recombinant human antibody fragment thought to target HSP-90 and potentiate amphotericin B (AMB). Absence of in vivo efficacy led us to reevaluate its in vitro activity. Interactions between AMB and Mycograb were investigated using a checkerboard design. Addition of Mycograb or various unrelated proteins, including human serum, resulted in similar decreases in the MIC of AMB. Potentiation of AMB by Mycograb appears to be a nonspecific protein effect.

Adherence to prophylaxis in adolescents and young adults with severe haemophilia: a qualitative study with healthcare professionals.

Aim: to examine healthcare professionals' (HP) perceptions and experiences in relation to adherence to prophylactic treatment among young people living with haemophilia (YPH). Methods: All HPs in four haemophilia centres across England and Wales were invited to participate, and all HPs who agreed to take part (n = 6) were interviewed. Interviews were audio-recorded, transcribed and then analysed using Interpretative Phenomenological Analysis (IPA). Results: HPs estimate that generally young people with haemophilia keep to their treatment regimen well, although they also recognise that adherence may fluctuate with many patients going through shorter periods of non-adherence. The increasingly personalised or flexible approach to prophylaxis makes it harder to assess adherence. The main themes identified through IPA included (1) HPs' suggest that adherence fluctuates (2) Non-adherence is mainly driven by lifestyle and developmental, social and family factors, and (3) Education, HPs' sensitivity to individual needs, and psychological and peer support are key facilitators of good adherence. Conclusion: The increasingly flexible approach to prophylaxis requires a new way of thinking about, and assessment of, adherence. More personalised treatment regimen can be more complicated and may, therefore, lead to accidental non-adherence. The results of this study with HPs complement those of a previous qualitative study with patients but place greater emphasis on a broader perspective on understanding drivers of non-adherence as well as understanding strategies to improve adherence in the minority of patients who appear to struggle.

Increasing role of herpes simplex virus type 1 in first-episode anogenital herpes in heterosexual women and younger men who have sex with men, 1992-2006.

OBJECTIVES: Herpes simplex virus (HSV) type 1 is causing an increasing proportion of anogenital herpes; however, it is unclear which populations are affected. We describe the contribution of HSV-1 to first-episode anogenital herpes and its associations. METHODS: For all cases of first-episode anogenital herpes diagnosed at the Sydney Sexual Health Centre from 1992 to 2006, medical record review was used to confirm the type and anatomical site. Age, sex, HIV status and sexual behaviour data were extracted from the clinic database. RESULTS: Overall, among 1845 confirmed cases of first-episode anogenital herpes the proportion attributable to HSV-1 increased from 29% to 42% (odds ratio (OR) per 3-year band 1.19; 95% CI 1.11 to 1.27). When stratified by gender of sexual partners the proportion of first-episode anogenital herpes due to HSV-1 increased over time, but only achieved significance in heterosexual women (p<0.01). Among men who have sex with men (MSM), HSV-1 only increased for those less than 28 years of age, 17% in 1992-4 to 76% in 2004-6 (OR per 3-year band 1.58; 95% CI 1.14 to 2.19). The proportion attributable to HSV-1 was higher for anal than genital herpes and MSM were much more likely to have anal disease. CONCLUSIONS: The proportion of first-episode anogenital herpes due to HSV-1 significantly increased among younger MSM and heterosexual women over the 15-year period. In some clinical populations, such as young MSM and women or patients with anal disease, HSV-1 may now account for the majority of first-episode anogenital herpes.

Allylamine derivatives: new class of synthetic antifungal agents inhibiting fungal squalene epoxidase.

A new class of synthetic antifungal agents, the allylamines , has been developed by modification of naftifine , a topical antimycotic. SF 86-327, the most effective of these compounds so far, is highly active in vitro against a wide range of fungi and exceeds clinical standards in the oral and topical treatment of guinea pig dermatophytoses. SF 86-327 is a powerful specific inhibitor of fungal squalene epoxidase, a key enzyme in sterol biosynthesis.

Use of oral contraception in the United States, 1965. In only 5 years oral contraception has become a major means of regulating fertility.

This is the first report from the Na-tional Fertility Study, 1965, a survey of the reproductive behavior of a national sample of married women, under the age of 55, living with their husbands. The report presents basic data on the use of oral contraception by women under the age of 45, in relation to age, parity, education, race, and religion. The study leads to certain conclusions, as follows. Present, past, and prospective use vary inversely with the age of the woman and directly with the number of years of schooling; the majority of young women with college training have already used the oral contraceptive. Use by Negroes is somewhat less extensive than use by whites, particularly for ages below 25; some of this difference is explainable by concomitant racial differences in educational level. Negroes seem less likely than whites to use oral contraception for timing early births, and more likely, when they do use it, to be attempting to terminate their fertility. The same observation holds for white Catholics in relation to white non-Catholics. Although the extent of use may be lower among Catholics than non-Catholics, the proportion of Catholics who report use is substantial indeed in view of the persisting theological controversy. The prospects for increased use of oral contraception seem very good at present, but they may be limited by further developments in the technology of fertility regulation. Meanwhile the birth rate has declined substantially. Although much sophisticated analysis of other data from the survey will be required to determine the extent of the contribution of oral contraception to this decline, the findings presented here suggest that the contribution is substantial for young married couples. The major effect on the couple's eventual number of children may be less than the effect on the time pattern of childbearing; in any event, both lower eventual parity and delayed fertility contribute to a decline in the numbers of births from year to year. Whatever the intent may be, it is apparent that young American couples have adopted a new means for achieving their reproductive goals.

Adherence to prophylaxis in adolescents and young adults with severe haemophilia A, a qualitative study with patients.

Introduction: Reported levels of adherence to prophylaxis among young people with haemophilia (YPH) vary widely and are predominately based on estimations made by healthcare professionals and parents. Reasons for (non)adherence among YPH in particular have not been evidenced. Aim: to examine experiences in relation to prophylaxis with YPH themselves, and barriers and facilitators to their adherence. Methods: 11 Participants were recruited in five haemophilia centres across England and Wales. All patients who met the inclusion criteria (aged 12-25, diagnosed with haemophilia, on prophylaxis) were approached during a routine check-up appointment, and all participants who agreed to take part were interviewed. Interviews were audio recorded, transcribed and analysed using Interpretative Phenomenological Analysis. Results: Self-reported adherence to prophylaxis was good. Few participants admitted to intentionally skipping injections although they reported sometimes forgetting. However, due to the increasingly personalised and flexible approach to prophylaxis, adherence is not straightforward to define. Barriers to adherence included a busy lifestyle, dislike of the intravenous injection, venous access issues, anxiety or stress and being out of one's normal routine. Support was an important facilitator to adherence, including support from health professionals at the haemophilia centre as well as friends. Parents appear to be very involved with their child's haemophilia management, even after they leave home. Conclusion: What this study adds is that the increasingly flexible and personalised approach to managing prophylaxis in haemophilia may sometimes lead to confusion around treatment frequency and dosing. This may lead to accidental non-adherence, which is distinct from both skipping and forgetting. Advice from haemophilia teams may not always be consistent and is likely to be interpreted differently by different individuals. Some additional training and education of patients and their families to increase their knowledge and skills around prophylaxis may reduce this confusion and therefore is likely to improve adherence further.

Pseudoepitheliomatous hyperplasia causing a painful plaque in a HIV-infected female.

Dermatological conditions are more common and can present atypically, in human immunodeficiency virus-infected individuals. This case report describes a 22-year-old human immunodeficiency virus-positive Caucasian female who presented with a vulval lesion eight weeks after starting antiretroviral treatment. Clinical examination revealed a 2 cm well-demarcated plaque on the outer aspect of the left labium minus. The lesion was tender, no contact bleeding or ulceration present. She was presumptively treated for chancroid and herpes simplex with 500 mg ceftriaxone IM stat, 1 g azithromycin PO stat, and valacyclovir 500 mg BD for five days. The lesion persisted despite treatment, and during follow-up, a punch biopsy was carried out. She was diagnosed with pseudoepitheliomatous hyperplasia of the epidermis. In addition to highlighting this condition that has been previously reported in human immunodeficiency virus/herpes simplex virus co-infection, this case demonstrates that unusual skin presentations must be considered in human immunodeficiency virus-infected individuals and illustrates the importance of biopsy for any non-healing lesions.

A collaborative hit-to-lead investigation leveraging medicinal chemistry expertise with high throughput library design, synthesis and purification capabilities.

High throughput screening (HTS) campaigns, where laboratory automation is used to expose biological targets to large numbers of materials from corporate compound collections, have become commonplace within the lead generation phase of pharmaceutical discovery. Advances in genomics and related fields have afforded a wealth of targets such that screening facilities at larger organizations routinely execute over 100 hit-finding campaigns per year. Often, 10(5) or 10(6) molecules will be tested within a campaign/cycle to locate a large number of actives requiring follow-up investigation. Due to resource constraints at every organization, traditional chemistry methods for validating hits and developing structure activity relationships (SAR) become untenable when challenged with hundreds of hits in multiple chemical families per target. To compound the issue, comparison and prioritization of hits versus multiple screens, or physical chemical property criteria, is made more complex by the informatics issues associated with handling large data sets. This article describes a collaborative research project designed to simultaneously leverage the medicinal chemistry and drug development expertise of the Novartis Institutes for Biomedical Research Inc. (NIBRI) and ArQule Inc.'s high throughput library design, synthesis and purification capabilities. The work processes developed by the team to efficiently design, prepare, purify, assess and prioritize multiple chemical classes that were identified during high throughput screening, cheminformatics and molecular modeling activities will be detailed.

Properties of a particulate squalene epoxidase from Candida albicans.

The properties and requirements of squalene epoxidase and effects of some inhibitors were investigated in the pathogenic yeast Candida albicans. A washed 'microsomal' fraction converted radiolabelled squalene to 2,3-oxidosqualene and lanosterol. Minimum requirements for activity were molecular oxygen, NADH or NADPH, and FAD. Epoxidase activity was stimulated by up to 100% by addition of the soluble cytoplasmic fraction, which itself contained negligible epoxidase activity. This stimulation was most powerful at low concentrations of enzyme, or high concentrations of squalene. Divalent cations did not stimulate activity and EDTA was not inhibitory. An apparent Km for squalene of 50 microM was determined in the presence of soluble cytoplasm. Epoxidase activity was destroyed by Triton X-100, deoxycholate or Cu2+, and partially inhibited by thiol reagents, rotenone and antimycin A. The enzyme was not inhibited by cyanide or by several inhibitors of cytochrome P-450.

The effect of the 3-hydroxy-3-methylglutaryl CoA reductase inhibitor ML-236b on phytosterol synthesis in Acer pseudoplatanus tissue culture.

Sycamore cell cultures were incubated with various labelled sterol precursors. ML-236B, a fungal metabolite, caused virtually total inhibition of acetate or leucine incorporation into sterols, while mevalonate incorporation was unaffected. Sterol synthesis from endogenous precursors, measured by incorporation of [Me-14C] methionine into the side chain, continued at a reduced rate for at least 6 h after addition of the inhibitor.

Characterization of prenyl protein transferase enzymes in a human keratinocyte cell line.

Prenylation is a post-translational modification of proteins that involves the attachment of an isoprenoid group derived from mevalonic acid, either 15-carbon farnesyl or 20-carbon geranylgeranyl, to a specific carboxy-terminal domain of acceptor proteins. Three prenyl transferase enzymes have been identified so far. In this paper we report the presence of two prenyl transferases in the HaCaT human keratinocyte cell line. Chromatography of a cytosolic extract from these cells resolved a farnesyl protein transferase (FPT) and geranylgeranyl protein transferase-I (GGPT-I) whose activities were measured using a novel peptide-based assay. Both enzymes were inhibited dose dependently by zaragozic acids A and C. Zaragozic acid C was more active towards the FPT than GGPT-I while zaragozic acid A inhibited both enzymes with similar potency. Incubation of HaCaT cell homogenates with [3H] prenyl precursors resulted in the labelling of a number of proteins which was increased when the cells were pretreated with an inhibitor of hydroxymethylglutaryl CoA reductase. Given the role of prenylated proteins in proliferative and inflammatory processes, our finding that prenyl transferases capable of prenylating endogenous substrates are also present in keratinocytes suggests that these enzymes might provide novel therapeutic targets of dermatological importance.

Phosphatidic acid and phospholipase D both stimulate phosphoinositide turnover in cultured human keratinocytes.

Phosphatidic acid (PA) induced a rapid dose-dependent increase in production of inositol phosphates in cultured adult human keratinocytes, peaking at 30 s. Natural and dioleoyl PA were equally effective, while other phospholipid classes had no effect. Lipid A was also active. Lyso-PA also induced inositol phosphate production, but contamination of the PA preparation by lyso-PA could not account for the effect of PA. The effect of PA could not be reproduced by treatment of cells with calcium ionophore. PA-induced inositol phosphate production could be inhibited (> 50%) by pre-treatment of cells with either pertussis toxin or 12-O-tetradecanoylphorbol 13-acetate, suggesting the involvement of a GTP-binding protein and a protein kinase C-mediated negative feedback mechanism. PA also stimulated release of arachidonic acid from keratinocytes. Treatment of cells with exogenous phospholipase D similarly induced inositol phosphate production in the keratinocytes. Since PA may be formed by receptor-mediated activation of phospholipase D, or by phosphorylation of diacylglycerol, the results suggest that PA may play a significant role in signalling mechanisms of human keratinocytes.

Clinical audit: adherence to sexually transmitted infection screening guidelines for men who have sex with men.

Australian guidelines recommend regular screening of men who have sex with men (MSM) for sexually transmitted infections (STIs). This audit was performed to determine STI testing rates in Sydney Sexual Health Centre before and after the development of the guidelines, and to describe characteristics of those not tested. The electronic clinic database and a manual file review were used to determine testing rates and reasons for not testing for the years 2000 and 2002. Overall testing rates were high, with 61% of MSM having had all recommended tests within the past year in 2002. There was a significant increase in testing rates for most tests after the development of the guidelines. Asymptomatic men were more likely to be tested than symptomatic men, and HIV-positive men were less likely to be tested for syphilis.

Cloning and expression of squalene epoxidase from the pathogenic yeast Candida albicans.

The allylamine antimycotic terbinafine prevents the formation of sterols by specifically inhibiting squalene epoxidase (SE). The biological and biochemical action of terbinafine on fungal pathogens has been well investigated, but little is known at the molecular level. Here we report the cloning, sequencing and expression of the target of terbinafine from the major pathogen Candida albicans. A C. albicans genomic DNA library was constructed in gamma ZAP Express and screened with a DNA fragment obtained by polymerase chain reaction with two primers designed from sequences common to Saccharomyces cerevisiae and rodent SEs. Two types of clone, approximately 3.9 kbp and 4.1 kbp, were isolated. Both contained an identical open reading frame of 1488 nucleotides, while a few sequence differences were found in the flanking regions, suggesting an allelic heterogeneity. The deduced protein sequence of C. albicans SE, 496 amino acids (55324 Da), is 54% and 41% identical to those of S. cerevisiae and rat, respectively. A 1.8-kb transcript was observed on Northern blots of C. albicans mRNA. Polyclonal antibodies, raised against an internal peptide of C. albicans SE, recognized a protein associated with the particulate fraction of M(r) 55000 on Western blots of C. albicans extracts. C. albicans SE was overexpressed in S. cerevisiae with the expression vector pYES2. In homogenates from S. cerevisiae overexpressing the C. albicans protein SE activity was 10-fold higher than the endogenous activity from controls.

Inhibition of fungal and mammalian sterol biosynthesis by 2-aza-2,3-dihydrosqualene.

2-Aza-2,3-dihydrosqualene (I) and a quaternary ammonium derivative (II) inhibited ergosterol biosynthesis in cells and cell-free extracts of the pathogenic yeast Candida albicans as measured by incorporation of radiolabelled precursors. The compounds inhibited squalene epoxidase and 2,3-oxidosqualene cyclase to varying degrees in microsomes from C. albicans and from rat liver. The rat liver epoxidase was 50% inhibited by I at 2.4 microM. In C. albicans cells, but not in cell-free extracts, I also inhibited lanosterol demethylation and led to accumulation of an unidentified polar product.

Synthesis of new polyoxin derivatives and their activity against chitin synthase from Candida albicans.

Two analogues of L-alanylpolyoxin C with a modified peptide bond were synthesized and tested for inhibition of chitin synthase in Candida albicans. N-Methylation of the peptide bond (compound 13) or the replacement of it by NH2CH2 (compound 9) led to loss of activity in the enzyme assay. A novel analogue (compound 5) of nikkomycin was synthesized from uracil polyoxin C and (2S,3R)-3-hydroxyhomotyrosine, a component of echinocandin C. Despite high activity in the chitin synthase assay, 5 had no inhibitory effect on cells of C. albicans.

Primary duodenal adenocarcinoma: a 40-year experience.

HYPOTHESIS: In patients with duodenal adenocarcinoma, certain pathologic features of the tumor will have prognostic significance. DESIGN: Retrospective case series. PATIENTS: Forty-nine patients diagnosed with duodenal adenocarcinoma between 1957 and 1998. RESULTS: The tumors of 31 (63%) of the 49 patients underwent resection, 18 (37%) had surgical palliation or underwent biopsy. Mean (+/- SEM) survival for all patients was 49 +/- 9 months. The patients whose tumors were resected had longer survival than those who underwent palliation (mean +/- SEM, 66 +/- 13 months vs 18 +/- 6 months, P =.02). Multivariate analysis revealed large tumor size (P =.01), transmural invasion (P =.004), and moderate to poor tumor grade (P =.03) were negatively correlated with survival. Lymph node status did not influence survival. CONCLUSIONS: Our 40-year experience with duodenal adenocarcinoma demonstrates that large tumor size, advanced histological grade, and transmural invasion are associated with decreased survival. These results underscore the importance of early diagnosis, and suggest the presence of nodal spread is not a contraindication to resection.

Bradykinin upregulates immediate-early gene mRNA in human keratinocytes.

We investigated the effect of bradykinin on the expression of the proto-oncogenes c-fos, c-jun and c-myc and on cell proliferation in the human keratinocyte cell line, HaCaT. Analysis of mRNAs was done by Northern blotting with single-stranded DNA hybridization probes. Bradykinin caused a rapid and transient accumulation in c-fos and c-jun mRNAs. In contrast, c-myc mRNA increased more slowly. Moreover, we report that bradykinin was a weak stimulator of HaCaT cell proliferation whereas epidermal growth factor, which induced the same degree of mRNA elevation, was shown as a powerful mitogen. Thus, while in HaCaT cells bradykinin promotes the expression of the proto-oncogenes c-fos, c-jun and c-myc, other biochemical events appear to be necessary for cell division.