In adult tobacco smokers, adding e-cigarettes to standard smoking cessation counseling increased abstinence at 6 mo.

SOURCE CITATION: Auer R, Schoeni A, Humair JP, et al. Electronic nicotine-delivery systems for smoking cessation. N Engl J Med. 2024;390:601-610. 38354139.

Phosphorylation of Schizosaccharomyces pombe Dss1 mediates direct binding to the ubiquitin-ligase Dma1 in vitro.

Intrinsically disordered proteins (IDPs) are often multifunctional and frequently posttranslationally modified. Deleted in split hand/split foot 1 (Dss1-Sem1 in budding yeast) is a highly multifunctional IDP associated with a range of protein complexes. However, it remains unknown if the different functions relate to different modified states. In this work, we show that Schizosaccharomyces pombe Dss1 is a substrate for casein kinase 2 in vitro, and we identify three phosphorylated threonines in its linker region separating two known disordered ubiquitin-binding motifs. Phosphorylations of the threonines had no effect on ubiquitin-binding but caused a slight destabilization of the C-terminal α-helix and mediated a direct interaction with the forkhead-associated (FHA) domain of the RING-FHA E3-ubiquitin ligase defective in mitosis 1 (Dma1). The phosphorylation sites are not conserved and are absent in human Dss1. Sequence analyses revealed that the Txx(E/D) motif, which is important for phosphorylation and Dma1 binding, is not linked to certain branches of the evolutionary tree. Instead, we find that the motif appears randomly, supporting the mechanism of ex nihilo evolution of novel motifs. In support of this, other threonine-based motifs, although frequent, are nonconserved in the linker, pointing to additional functions connected to this region. We suggest that Dss1 acts as an adaptor protein that docks to Dma1 via the phosphorylated FHA-binding motifs, while the C-terminal α-helix is free to bind mitotic septins, thereby stabilizing the complex. The presence of Txx(D/E) motifs in the disordered regions of certain septin subunits may be of further relevance to the formation and stabilization of these complexes.

Evolution of A bHLH Interaction Motif.

Intrinsically disordered proteins and regions with their associated short linear motifs play key roles in transcriptional regulation. The disordered MYC-interaction motif (MIM) mediates interactions between MYC and MYB transcription factors in Arabidopsis thaliana that are critical for constitutive and induced glucosinolate (GLS) biosynthesis. GLSs comprise a class of plant defense compounds that evolved in the ancestor of the Brassicales order. We used a diverse set of search strategies to discover additional occurrences of the MIM in other proteins and in other organisms and evaluate the findings by means of structural predictions, interaction assays, and biophysical experiments. Our search revealed numerous MIM instances spread throughout the angiosperm lineage. Experiments verify that several of the newly discovered MIM-containing proteins interact with MYC TFs. Only hits found within the same transcription factor family and having similar characteristics could be validated, indicating that structural predictions and sequence similarity are good indicators of whether the presence of a MIM mediates interaction. The experimentally validated MIMs are found in organisms outside the Brassicales order, showing that MIM function is broader than regulating GLS biosynthesis.

Minimally invasive, sutureless, adolescent male circumcision with topical anesthetic: a field trial of Unicirc, a single-use surgical instrument.

BACKGROUND: Circumcision has been shown to reduce the rate of HIV transmission in Africa. It is most cost effective if performed in younger men. Surgical assist devices can increase the efficiency and potentially reduce the cost of performing circumcisions. METHODS: We used the Unicirc disposable instrument to perform circumcisions in an outpatient primary care clinic. The trial was non-blinded. Circumcisions were performed under topical anaesthetic and the wound was sealed with cyanoacrylate tissue adhesive. The primary outcome was intraoperative duration; secondary outcomes were intraoperative and postoperative pain; adverse events (AEs); time to healing and patient satisfaction; and, cosmetic result. RESULTS: A total of 82 adolescent boys (aged 10-15 years) were circumcised. The median intraoperative time was 10 minutes and the median blood loss was 1 mL. All wounds were healed by 4 weeks and cosmetic results were excellent. There were no AEs. CONCLUSIONS: Adolescent circumcision with Unicirc under topical anesthetic and wound sealing with cyanoacrylate tissue adhesive is safe, rapid, and heals by primary intention with excellent cosmetic results. It is cost effective and can be used for large scale programs.

IDDomainSpotter: Compositional bias reveals domains in long disordered protein regions-Insights from transcription factors.

Protein domains constitute regions of distinct structural properties and molecular functions that are retained when removed from the rest of the protein. However, due to the lack of tertiary structure, the identification of domains has been largely neglected for long (>50 residues) intrinsically disordered regions. Here we present a sequence-based approach to assess and visualize domain organization in long intrinsically disordered regions based on compositional sequence biases. An online tool to find putative intrinsically disordered domains (IDDomainSpotter) in any protein sequence or sequence alignment using any particular sequence trait is available at http://www.bio.ku.dk/sbinlab/IDDomainSpotter. Using this tool, we have identified a putative domain enriched in hydrophilic and disorder-promoting residues (Pro, Ser, and Thr) and depleted in positive charges (Arg and Lys) bordering the folded DNA-binding domains of several transcription factors (p53, GCR, NAC46, MYB28, and MYB29). This domain, from two different MYB transcription factors, was characterized biophysically to determine its properties. Our analyses show the domain to be extended, dynamic and highly disordered. It connects the DNA-binding domain to other disordered domains and is present and conserved in several transcription factors from different families and domains of life. This example illustrates the potential of IDDomainSpotter to predict, from sequence alone, putative domains of functional interest in otherwise uncharacterized disordered proteins.

Specificity of MYB interactions relies on motifs in ordered and disordered contexts.

Physical interactions between members of the MYB and bHLH transcription factor (TF) families regulate many important biological processes in plants. Not all reported MYB-bHLH interactions can be explained by the known binding sites in the R3 repeat of the MYB DNA-binding domain. Noteworthy, most of the sequence diversity of MYB TFs lies in their non-MYB regions, which contain orphan small subgroup-defining motifs not yet linked to molecular functions. Here, we identified the motif mediating interaction between MYB TFs from subgroup 12 and their bHLH partners. Unlike other known MYB-bHLH interactions, the motif locates to the centre of the predicted disordered non-MYB region. We characterised the core motif, which enabled accurate prediction of previously unknown bHLH-interacting MYB TFs in Arabidopsis thaliana, and we confirmed its functional importance in planta. Our results indicate a correlation between the MYB-bHLH interaction affinity and the phenotypic output controlled by the TF complex. The identification of an interaction motif outside R3 indicates that MYB-bHLH interactions must have arisen multiple times, independently and suggests many more motifs of functional relevance to be harvested from subgroup-specific studies.

R2R3 MYB Transcription Factors - Functions outside the DNA-Binding Domain.

Several transcription factor (TF) families, including the MYB family, regulate a wide array of biological processes. TFs contain DNA-binding domains (DBDs) and regulatory regions; although information on protein structure is scarce for plant MYB TFs, various in silico methods suggest that the non-MYB regions contain extensive intrinsically disordered regions (IDRs). Although IDRs do not fold into stable globular structures, they comprise functional regions including interaction motifs, and recent research has shown that IDRs perform crucial biological roles. We map here domain organization, disorder predictions, and functional regions across the entire Arabidopsis thaliana R2R3 MYB TF family, and highlight where an increased research focus will be necessary to shape a new understanding of structure-function relationships in plant TFs.

Sutureless Adult Voluntary Male Circumcision with Topical Anesthetic: A Randomized Field Trial of Unicirc, a Single-Use Surgical Instrument.

INTRODUCTION: The World Health Organization has solicited rapid and minimally invasive techniques to facilitate scale-up of voluntary medical male circumcision (VMMC). STUDY DESIGN: Non-blinded randomized controlled field trial with 2:1 allocation ratio. PARTICIPANTS: 75 adult male volunteers. SETTING: Outpatient primary care clinic. INTERVENTION: Open surgical circumcision under local anesthetic with suturing vs. Unicirc disposable instrument under topical anesthetic and wound sealing with cyanoacrylate tissue adhesive. PRIMARY OUTCOME: Intraoperative duration. SECONDARY OUTCOMES: Intraoperative and postoperative pain; adverse events; time to healing; patient satisfaction; cosmetic result. RESULTS: The intraoperative time was less with the Unicirc technique (median 12 vs. 25 min, p < 0.001). Wound healing and cosmetic results were superior in the Unicirc group. Adverse events were similar in both groups. CONCLUSIONS: VMMC with Unicirc under topical anesthetic and wound sealing with cyanoacrylate tissue adhesive is rapid, heals by primary intention with superior cosmetic results, and is potentially safer and more cost-effective than open surgical VMMC. TRIAL REGISTRATION: Clinicaltrials.gov NCT02443792.

Questions About Vitamin D for Primary Care Practice: Input From an NIH Conference.

There is considerable consumer and physician interest in vitamin D as a possible therapeutic agent for a range of clinical conditions and, despite mixed evidence, the interest does not appear to lessen. Some clinicians believe that consumption of vitamin D is inadequate and, in turn, advocate vitamin D supplementation to increase serum levels of the nutrient. However, evidence concerning the role of vitamin D in health and disease is conflicting, and primary care physicians have little time to sort through the data and may find it difficult to advise their patients. To better understand the challenges that primary care physicians face regarding vitamin D, and to help inform those who provide guidance for clinical decision-making, the Office of Dietary Supplements at the National Institutes of Health, with co-sponsorship from other federal health agencies, held a conference titled Vitamin D: Moving Toward Evidence-based Decision Making in Primary Care in December 2014. More than 20 invited presenters and panelists considered laboratory methods for measuring vitamin D status, discussed how clinical studies of vitamin D should be evaluated and used in developing recommendations, noted the role of values and preferences in clinical decision-making, debated the current science related to at-risk groups, and described emerging data about health risks of excessive intakes of vitamin D. Eight questions about vitamin D stem from the Conference presentations as well as other expert sources.

No-needle, single-visit adult male circumcision with Unicirc: a multi-centre field trial.

BACKGROUND: Voluntary medical male circumcision (VMMC) is a priority HIV preventive intervention. Current adult circumcision methods need improvement. METHODS: Field trial in 3 primary care centres. Minimally invasive VMMC using the Unicirc instrument following topical lidocaine/prilocaine anesthetic. Men were followed up at 1 and 4 weeks. RESULTS: We circumcised 110 healthy volunteers. Two men complained of transient burning pain during circumcision, but none required injectable anaesthesia. Median blood loss was 1ml and median procedure time was 9.0 min. There were 7 (6.3%) moderate complications (5 (4.5%) post-operative bleeds requiring suture and 2 (1.8%) post-operative infections) affecting 7 men. No men experienced significant wound dehiscence. 90.4% of men were fully healed at 4 weeks of follow-up and all were highly satisfied. CONCLUSIONS: Use of topical anaesthesia obviates the need for injectable anesthetic and makes the Unicirc procedure nearly painless. Unicirc is rapid, easy to learn, heals by primary intention with excellent cosmetic results, obviates the need for a return visit for device removal, and is potentially cheaper and safer than other methods. Use of this method will greatly facilitate scale-up of mass circumcision programs. TRIAL REGISTRATION: ClinicalTrials.gov NCT02091726.

Emergence of methicillin resistance and Panton-Valentine leukocidin positivity in hospital- and community-acquired Staphylococcus aureus infections in Beira, Mozambique.

OBJECTIVES: The objective of this study was to investigate the antibiotic resistance patterns, including methicillin resistance, inducible macrolide-lincosamide-streptogramin B (MLSB ) resistance and Panton-Valentine leukocidin (PVL) toxin gene carriage among hospital-acquired Staphylococcus aureus (HA-SA) and community-acquired S. aureus (CA-SA), in Beira, Mozambique. METHODS: In 2010-2011, two prospective surveillance studies were conducted on post-operative and burn wound infections at the Central Hospital of Beira and on skin and soft tissue abscesses at the São Lucas Health Centre. We cultured pus samples, identified suspected S. aureus isolates and performed antimicrobial susceptibility testing, including detection of MLSB resistance. Real-time polymerase chain reaction was used to detect mecA, Martineau and PVL genes. RESULTS: The prevalence of hospital-acquired methicillin-resistant S. aureus (HA-MRSA) infection among 53 inpatients was 15.1%; the prevalence of community-acquired methicillin-resistant S. aureus (CA-MRSA) infection among 100 outpatients was 1.0%. Inducible MLSB resistance was present in 41.7% and 10.7% of HA-SA and CA-SA isolates, respectively. PVL toxin gene was detected in 81.1% of methicillin-susceptible S. aureus (MSSA) compared with 11.1% of methicillin-resistant S. aureus. CONCLUSIONS: Our study shows, for the first time in Mozambique, the emergence of HA-MRSA. The prevalence of CA-MRSA was low, whereas the rate of PVL toxin gene carriage in MSSA was high. The high rate of inducible MLSB resistance indicates the importance of performing routine D-tests. Overall, our results show the need of strengthening laboratory facilities to provide microbiological data for both directed therapy and surveillance.

Rapid, minimally invasive adult voluntary male circumcision: a randomised trial of Unicirc, a novel disposable device.

BACKGROUND: Voluntary medical male circumcision (VMMC) is a priority HIV preventive intervention. To facilitate VMMC scale-up, the World Health Organization is seeking circumcision techniques that are faster, easier, and safer than open surgical methods. OBJECTIVE: To compare open surgical circumcision with suturing v. the Unicirc disposable instrument plus tissue adhesive. METHODS: We conducted a non-blinded randomised controlled trial at an outpatient primary healthcare clinic in Cape Town, South Africa, with 2:1 allocation ratio of 150 male volunteers who were at least 18 years of age. Our primary outcome was intraoperative time and secondary outcomes were ease of performance, post-operative pain, adverse events, time to healing, patient satisfaction and cosmetic result. RESULTS: The intraoperative time was less with the Unicirc/adhesive technique (median 13 v. 22.6 min, respectively; p<0.001). The intraoperative suturing rate was 17% using the Unicirc device. Other adverse events and wound healing outcomes were similar in both groups, but the cosmetic result was superior in the Unicirc group. Doctors found the Unicirc procedure easier to perform and preferred it to the open surgical technique. CONCLUSIONS: This study has important implications for the scale-up of VMMC services. Excising the foreskin with the Unicirc instrument and sealing the wound with cyanoacrylate tissue adhesive in adults is quicker, easier to learn, and is potentially safer than open surgical VMMC. Further studies should be conducted with the optimised device. This new instrument has the potential to facilitate more rapid scale-up and save costs.

Open-source point-of-care electronic medical records for use in resource-limited settings: systematic review and questionnaire surveys.

BACKGROUND: Point-of-care electronic medical records (EMRs) are a key tool to manage chronic illness. Several EMRs have been developed for use in treating HIV and tuberculosis, but their applicability to primary care, technical requirements and clinical functionalities are largely unknown. OBJECTIVES: This study aimed to address the needs of clinicians from resource-limited settings without reliable internet access who are considering adopting an open-source EMR. STUDY ELIGIBILITY CRITERIA: Open-source point-of-care EMRs suitable for use in areas without reliable internet access. STUDY APPRAISAL AND SYNTHESIS METHODS: The authors conducted a comprehensive search of all open-source EMRs suitable for sites without reliable internet access. The authors surveyed clinician users and technical implementers from a single site and technical developers of each software product. The authors evaluated availability, cost and technical requirements. RESULTS: The hardware and software for all six systems is easily available, but they vary considerably in proprietary components, installation requirements and customisability. LIMITATIONS: This study relied solely on self-report from informants who developed and who actively use the included products. CONCLUSIONS AND IMPLICATIONS OF KEY FINDINGS: Clinical functionalities vary greatly among the systems, and none of the systems yet meet minimum requirements for effective implementation in a primary care resource-limited setting. The safe prescribing of medications is a particular concern with current tools. The dearth of fully functional EMR systems indicates a need for a greater emphasis by global funding agencies to move beyond disease-specific EMR systems and develop a universal open-source health informatics platform.

Circumcision--what's wrong with plastic rings?

A key issue facing countries that are scaling up circumcision services is the technical difficulty, resources used, complications, and time to healing using open surgical techniques, the only methods approved by the major external funding agency, PEPFAR. The WHO has developed a framework for evaluating new circumcision devices, and two promising disposable plastic devices that have been partially evaluated are the Shang Ring and the Prepex system. However, given South Africa's disastrous experience with the Tara KLamp, healthy scepticism about plastic ring devices is justified. The Gomco clamp has been used in children and adults since 1935 in the USA, but there are no published studies demonstrating its use in adults. Cyanoacrylate tissue adhesive, widely used in all areas of medicine, has been shown to be superior to sutures in circumcision in terms of safety, ease of use, operative time, and cosmetic results. Our experience in Mozambique suggests that Gomco clamp circumcision plus tissue adhesive closure meets all the WHO criteria for the ideal circumcision technique, and we strongly recommend that African researchers conduct clinical trials to compare it with open surgical circumcision.

Minimally invasive childhood and adult circumcision.

Using the Gomco circumcision clamp and sealing the wound with tissue adhesive results in a minimally invasive circumcision suitable for all age groups beyond the neonatal period. It is easy to perform and can be performed by generalists with local anaesthetic and standard instruments.