Lessons on protein structure from interleukin-4: All disulfides are not created equal.

Interleukin-4 (IL-4) is a hematopoietic cytokine composed by a four-helix bundle stabilized by an antiparallel beta-sheet and three disulfide bonds: Cys3-Cys127, Cys24-Cys65, and Cys46-Cys99. IL-4 is involved in several immune responses associated to infection, allergy, autoimmunity, and cancer. Besides its physiological relevance, IL-4 is often used as a "model" for protein design and engineering. Hence, to understand the role of each disulfide in the structure and dynamics of IL-4, we carried out several spectroscopic analyses (circular dichroism [CD], fluorescence, nuclear magnetic resonance [NMR]), and molecular dynamics (MD) simulations on wild-type IL-4 and four IL-4 disulfide mutants. All disulfide mutants showed loss of structure, altered interhelical angles, and looser core packings, showing that all disulfides are relevant for maintaining the overall fold and stability of the four-helix bundle motif, even at very low pH. In the absence of the disulfide connecting both protein termini Cys3-Cys127, C3T-IL4 showed a less packed protein core, loss of secondary structure (~9%) and fast motions on the sub-nanosecond time scale (lower S2 order parameters and larger τc correlation time), especially at the two protein termini, loops, beginning of helix A and end of helix D. In the absence of Cys24-Cys65, C24T-IL4 presented shorter alpha-helices (14% loss in helical content), altered interhelical angles, less propensity to form the small anti-parallel beta-sheet and increased dynamics. Simultaneously deprived of two disulfides (Cys3-Cys127 and Cys24-Cys65), IL-4 formed a partially folded "molten globule" with high 8-anilino-1-naphtalenesulphonic acid-binding affinity and considerable loss of secondary structure (~50%decrease), as shown by the far UV-CD, NMR, and MD data.

Molecular Insights into Female Hybrid Sterility in Interspecific Crosses between Drosophila melanogaster and Drosophila simulans.

Species of the genus Drosophila have served as favorite models in speciation studies; however, genetic factors of interspecific reproductive incompatibility are under-investigated. Here, we performed an analysis of hybrid female sterility by crossing Drosophila melanogaster females and Drosophila simulans males. Using transcriptomic data analysis and molecular, cellular, and genetic approaches, we analyzed differential gene expression, transposable element (TE) activity, piRNA biogenesis, and functional defects of oogenesis in hybrids. Premature germline stem cell loss was the most prominent defect of oogenesis in hybrid ovaries. Because of the differential expression of genes encoding piRNA pathway components, rhino and deadlock, the functional RDCmel complex in hybrid ovaries was not assembled. However, the activity of the RDCsim complex was maintained in hybrids independent of the genomic origin of piRNA clusters. Despite the identification of a cohort of overexpressed TEs in hybrid ovaries, we found no evidence that their activity can be considered the main cause of hybrid sterility. We revealed a complicated pattern of Vasa protein expression in the hybrid germline, including partial AT-chX piRNA targeting of the vasasim allele and a significant zygotic delay in vasamel expression. We arrived at the conclusion that the hybrid sterility phenotype was caused by intricate multi-locus differences between the species.

The role of pain-related disability on treatment outcome and psychosocial symptoms in patients with temporomandibular disorders - a pilot study.

OBJECTIVE: The aim of the pilot study was to investigate the association of pain-related disability with the outcome of conservative treatment of temporomandibular disorders (TMD) and with depressive and non-specific physical symptoms among TMD pain patients utilizing Graded Chronic Pain Scale 1.0 (GCPS1.0) as a screening instrument. MATERIAL AND METHODS: The study included 80 adult patients who were referred to the Oral and Maxillofacial Department, Oulu University Hospital, Finland, due to TMD pain. At baseline, pain-related disability was assessed by using the GCPS1.0 according to the Research Diagnostic Criteria for TMD (RDC/TMD), and the patients were categorized into three TMD subtypes, 1-3. Patients were given conservative TMD treatment. At follow-up visits (1, 3, 6, and 12 months), patients evaluated the pain intensity on an NRS and described the treatment outcome and the severity of the TMD symptoms on a numerical scale. RESULTS AND CONCLUSIONS: Patients with TMD subtype 3 (moderate/severe disability) had the highest NRS scores and described their symptoms as most severe at each time point, statistically significantly so at 1-month and 6-month follow-up (p < .05). The highest proportion of depressive symptoms was found in TMD subtype 3 (p < .05). The current pilot study showed that moderate/severe TMD-related disability, based on the GCPS 1.0 as a screening tool, may be linked with poor treatment outcome and depressive symptoms. Studies with larger samples are needed to confirm the results.

A proposal for a database of sounds generated by temporomandibular joints and a tool for automateddiagnosisbasedonanRDC/TMDquestionnaire.

The prevalence of TMD indicates a need to develop new tools that are useful in the case of screening examinations. These methods can support diagnosis at the early stage of the disorder. The purpose of this research was to develop a comprehensive tool that would function as both a database of sounds generated by TMJ and as software which facilitates automated diagnosis. The software would also use the data from the RDC/TMD questionnaire. Such a tool may significantly reduce the time spent by dentists on making manual RDC/TMD diagnoses. Moreover, this solution would enable dentists who do not specialise in TMD to make effective diagnoses. 95 patient took part in the clinical examination: 30 man and 65 females. The mean age of the participants was 33 years. Patients participating in the clinical process were examined according to the Polish version of the RDC/TMD questionnaire (Axis I and Axis II). Subsequently, all subjects were auscultated with an electronic stethoscope. An application has been implemented based on the RDC/TMD diagnostic flow chart. This tool was used for the automated generation of RDC/TMD diagnoses for all patients. As a result of the kind permission and participation of the patients under examination, it was possible to store records of ninety-five people. Each record contains RDC/TMD questionnaire data, auscultation signals and RDC/TMD diagnoses. For the first time, a database was created that has the potential to facilitate further examination. However, the developed system is universal and can therefore be adapted to new DC/TMD criteria.

Pushing the Limits of Characterising a Weak Halogen Bond in Solution.

Detection and characterisation of very weak, non-covalent interactions in solution is inherently challenging. Low affinity, short complex lifetime and a constant battle against entropy brings even the most sensitive spectroscopic methods to their knees. Herein we introduce a strategy for the accurate experimental description of weak chemical forces in solution. Its scope is demonstrated by the detailed geometric and thermodynamic characterisation of the weak halogen bond of a non-fluorinated aryl iodide and an ether oxygen (0.6 kJ mol-1 ). Our approach makes use of the entropic advantage of studying a weak force intramolecularly, embedded into a cooperatively folding system, and of the combined use of NOE- and RDC-based ensemble analyses to accurately describe the orientation of the donor and acceptor sites. Thermodynamic constants (ΔG, ΔH and ΔS), describing the specific interaction, were derived from variable temperature chemical shift analysis. We present a methodology for the experimental investigation of remarkably weak halogen bonds and other related weak forces in solution, paving the way for their improved understanding and strategic use in chemistry and biology.

Connection of Isolated Stereoclusters by Combining 13C-RCSA, RDC, and J-Based Configurational Analyses and Structural Revision of a Tetraprenyltoluquinol Chromane Meroterpenoid from Sargassum muticum.

The seaweed Sargassum muticum, collected on the southern coast of Galicia, yielded a tetraprenyltoluquinol chromane meroditerpene compound known as 1b, whose structure is revised. The relative configuration of 1b was determined by J-based configurational methodology combined with an iJ/DP4 statistical analysis and further confirmed by measuring two anisotropic properties: carbon residual chemical shift anisotropies (13C-RCSAs) and one-bond 1H-13C residual dipolar couplings (1DCH-RDCs). The absolute configuration of 1b was deduced by ECD/OR/TD-DFT methods and established as 3R,7S,11R.

Distribution of anxiety and depression among different subtypes of temporomandibular disorder: A systematic review and meta-analysis.

BACKGROUND: The evidence on the relationship between anxiety and depression and patients with distinct subtypes of temporomandibular disorder (TMD) is uncertain, so a thorough review study on the topic is still missing. OBJECTIVES: This systematic review investigated the distribution and severity of anxiety and depression in patients diagnosed with different subtypes of TMD. METHODS: The study is registered in PROSPERO (CRD42020150562) and it followed the PRISMA 2020 Statement. We searched in PubMed, Web of Science, Scopus and SciELO databases (last search: 12 March 2021) and the reference list from the included studies. Study eligibility criteria consisted of: (i) patients diagnosed with TMD using the Research Diagnostic Criteria (RDC/TMD) or Diagnostic Criteria (DC/TMD) instruments; (ii) assessment of anxiety and/or depression with validated psychological instruments and (iii) allocation of patients into a minimum of two distinct TMD subtypes with at least one group having myofascial pain (comparison group). Analyses were carried out using RevMan 5.3.5 statistical package and random- or fixed-effects models (α = 0.05). The quality of evidence was assessed based on review authors' judgment derived from a 10-item appraisal tool for prevalence studies and with the Newcastle-Ottawa scale. RESULTS: Of the 4086 records identified in total, 24 were eligible for inclusion; meta-analyses were conducted with 20 studies. In total, 3678 subjects were included in the review. Most of the studies found that patients with myofascial pain showed similar occurrence and severity of anxiety/depression as compared to other subtypes of TMD, although the average prevalence seemed to be higher among the diagnoses consisting of myofascial pain (muscular TMD). Despite the moderate-to-high heterogeneity, anxiety and depression were more frequently distributed within patients with myofascial pain (p = .001). TMD patients without myofascial pain presented less severe levels of anxiety and depression than patients with only myofascial pain (p ≤ .01). The type of psychological instrument seems to affect the assessment of both anxiety and depression emotional states. CONCLUSION: The findings of this review suggest that patients with myofascial pain are more anxious and more depressed than patients with other subtypes of TMD. IMPLICATIONS: Considering that anxiety and depression are differently distributed within the TMD population, a proper assessment of the psychological state of patients seems essential to offer an adequate treatment and management of each specific subtype of TMD.

Temporomandibular joint involvement in children with juvenile idiopathic arthritis-Symptoms, clinical signs and radiographic findings.

BACKGROUND: Although many children with juvenile idiopathic arthritis (JIA) develop arthritis and deformity of the temporomandibular joint (TMJ), many go undetected. OBJECTIVE: This study investigates whether findings from patient history and clinical examination using the Research Diagnostic Criteria for Temporomandibular Disorders (RDC/TMD) can be used to diagnose TMJ involvement. METHODS: As a part of the screening program, 59 consecutive JIA patients age 7-14 years underwent a clinical examination according to RDC/TMD including self-reported orofacial pain and pain related to jaw function, and cone beam computer tomography (CBCT). Data were obtained from the patient's medical charts. Patients were divided into two groups based on the presence or absence of TMJ deformities on CBCT. RESULTS: Self-reported TMJ symptoms before inclusion were reported by 52% of children with and 18% of children without TMJ deformities on CBCT (p = .020). On a group level, the maximum unassisted (mouth) opening (MUO) with and without pain was within the normal range, but children with TMJ deformities showed a significantly smaller MUO with pain (p = .035). A diagnosis of osteoarthritis and osteoarthrosis was more prevalent in children with TMJ deformities. CONCLUSION: Although there were few differences between children with and without radiographic TMJ deformities, self-reported previous TMJ symptoms and reduced MUO with pain could indicate the presence of TMJ involvement. However, radiographic examinations are needed to confirm TMJ involvement. Thus, this study indicates that the RDC/TMD protocol is a blunt tool when targeting TMJ involvement in JIA.

Sensing Magnetic Field and Intermolecular Interactions in Diamagnetic Solution Using Residual Dipolar Couplings of Zephycandidine.

An unusual residual dipolar coupling of methylene protons was recorded in NMR spectra because aromatic zephycandidine has preferential orientation at the external magnetic field. The observed splitting contains contribution from the dipole-dipole D-coupling and the anisotropic component of J-coupling. Absolute values of the anisotropy of magnetic susceptibility |Δχax| are larger for protic solvents because of the hydrogen-bonding compared to aprotic solvents for which polar and dispersion forces are more important. The energy barrier for the reorientation due to hydrogen-bonding is 1.22 kJ/mol in methanol-d4, 0.85 kJ/mol in ethanol-d6 and 0.87 kJ/mol in acetic acid-d6. In dimethyl sulfoxide-d6, 1.08 kJ/mol corresponds to the interaction of solvent lone pair electrons with π-electrons of zephycandidine. This energy barrier decreases for acetone-d6 which has smaller electric dipole moment. In acetonitrile-d3, there is no energy barrier which suggests solvent ordering around the solute due to the solvent-solvent interactions. The largest absolute values of the magnetic anisotropy are observed for aromatic benezene-d6 and tolune-d8 which have their own preferential orientation and enhance the order in the solution. The magnetic anisotropy of "isolated" zephycandidine, not hindered by intermolecular interaction could be estimated from the correlation between Δχax and cohesion energy density.

Structures Controlled by Entropy: The Flexibility of Strychnine as Example.

To study the flexibility of strychnine, we performed molecular dynamics simulations with orientational tensorial constraints (MDOC). Tensorial constraints are derived from nuclear magnetic resonance (NMR) interaction tensors, for instance, from residual dipolar couplings (RDCs). Used as orientational constraints, they rotate the whole molecule and molecular parts with low rotational barriers. Since the NMR parameters are measured at ambient temperatures, orientational constraints generate conformers that populate the whole landscape of Gibbs free energy. In MDOC, structures are populated that are not only controlled by energy but by the entropy term TΔS of the Gibbs free energy. In the case of strychnine, it is shown that ring conformers are populated, which has not been discussed in former investigations. These conformer populations are not only in accordance with RDCs but fulfill nuclear Overhauser effect (NOE)-derived distance constraints and 3JHH couplings as well.

Effect of the solvent on the conformation of monocrotaline as determined by isotropic and anisotropic NMR parameters.

The conformation in solution of monocrotaline, a pyrrolizidine alkaloid presenting an eleven-membered macrocyclic diester ring, has been investigated using a combination of isotropic and anisotropic nuclear magnetic resonance parameters measured in four solvents of different polarity (D2 O, DMSO-d6 , CDCl3 , and C6 D6 ). Anisotropic nuclear magnetic resonance parameters were measured in different alignment media, based on their compatibility with the solvent of interest: cromoglycate liquid crystal solution was used for D2 O, whereas a poly (methyl methacrylate) polymer gel was chosen for CDCl3 and C6 D6 , and a poly (hydroxyethyl methacrylate) gel for DMSO-d6 . Whereas the pyrrolizidine ring shows an E6 exo-puckered conformation in all of the solvents, the macrocyclic eleven-membered ring adopts different populations of syn-parallel and anti-parallel relative orientation of the carbonyl groups according to the polarity of the solvent.

Comparison of Axis II psychosocial assessment methods of RDC/TMD and DC/TMD as part of DC/TMD-FIN phase II validation studies in tertiary care Finnish TMD pain patients.

BACKGROUND: The Research Diagnostic Criteria for Temporomandibular Disorders (RDC/TMD) and Diagnostic Criteria for TMD (DC/TMD) include Axis II instruments for psychosocial assessment. OBJECTIVES: The aims were to compare the Finnish versions of Axis II psychosocial assessment methods of the RDC/TMD and DC/TMD and to study their internal reliability. METHODS: The sample comprised 197 tertiary care referral TMD pain patients. The associations between RDC/TMD [Graded Chronic Pain Scale (GCPS) 1.0, Symptom Check List 90-revised (SCL-90R)] and DC/TMD (GCPS 2.0, Patient Health Questionnaire-9 (PHQ-9), PHQ-15) assessment instruments were evaluated using Spearman correlation coefficients, Wilcoxon Signed Rank s, chi-squared test and gamma statistics. The internal reliability and internal inter-item consistency of SCL-90-R, PHQ-9, PHQ-15 and Generalized Anxiety Disorder-7 (GAD-7) were evaluated using Cronbach's alpha coefficient values. RESULTS: The DC/TMD and RDC/TMD Axis II psychosocial instruments correlated strongly (p < .001). GCPS 1.0 and GCPS 2.0 grades were similarly distributed based on both criteria. The RDC/TMD psychological instruments had a higher tendency to subclassify patients with more severe symptoms of depression and non-specific physical symptoms compared to DC/TMD. The internal reliability and internal inter-item consistency were high for the psychological assessment instruments. CONCLUSION: The Finnish versions of the RDC/TMD and DC/TMD Axis II psychosocial instruments correlated strongly among tertiary care TMD pain patients. Furthermore, the Axis II psychological assessment instruments indicated high validity and internal inter-item consistency and are applicable in Finnish TMD pain patients as part of other comprehensive specialist level assessments, but further psychometric and cut-off evaluations are still needed.

Increased usability, algorithmic improvements and incorporation of data mining for structure calculation of proteins with REDCRAFT software package.

BACKGROUND: Traditional approaches to elucidation of protein structures by Nuclear Magnetic Resonance spectroscopy (NMR) rely on distance restraints also known as Nuclear Overhauser effects (NOEs). The use of NOEs as the primary source of structure determination by NMR spectroscopy is time consuming and expensive. Residual Dipolar Couplings (RDCs) have become an alternate approach for structure calculation by NMR spectroscopy. In previous works, the software package REDCRAFT has been presented as a means of harnessing the information containing in RDCs for structure calculation of proteins. However, to meet its full potential, several improvements to REDCRAFT must be made. RESULTS: In this work, we present improvements to REDCRAFT that include increased usability, better interoperability, and a more robust core algorithm. We have demonstrated the impact of the improved core algorithm in the successful folding of the protein 1A1Z with as high as ±4 Hz of added error. The REDCRAFT computed structure from the highly corrupted data exhibited less than 1.0 Å with respect to the X-ray structure. We have also demonstrated the interoperability of REDCRAFT in a few instances including with PDBMine to reduce the amount of required data in successful folding of proteins to unprecedented levels. Here we have demonstrated the successful folding of the protein 1D3Z (to within 2.4 Å of the X-ray structure) using only N-H RDCs from one alignment medium. CONCLUSIONS: The additional GUI features of REDCRAFT combined with the NEF compliance have significantly increased the flexibility and usability of this software package. The improvements of the core algorithm have substantially improved the robustness of REDCRAFT in utilizing less experimental data both in quality and quantity.

Frequency and management of non-obstetric fistula in the Democratic Republic of Congo: experience from the Fistula Care Plus project.

OBJECTIVE: To describe the frequency, causes and post-repair outcomes of NOF in hospitals supported by the Fistula Care Plus (FC+) project in the Democratic Republic of Congo. METHODS: Retrospective cohort study from 1 January 2015 to 31 December 2017 in three FC + supported fistula repair sites. RESULTS: Of 1984 women treated for female genital fistula between 2015 and 2017 in the three FC + supported hospitals, 384 (19%) were considered to be non-obstetric fistula (NOF) cases. 49.3% were married/in a relationship at the time of treatment vs. 69% before the fistula, P < 0.001. Type III (n = 247; 64.3%) and type I (n = 121; 31.5%) fistulas according to Kees/Waaldijk classification were the most common. The main causes of NOF were medical procedure (n = 305; 79.4%); of these, caesarean section (n = 234; 76.7%) and hysterectomy (n = 54; 17.7%) were the most common. At hospital discharge, the fistula was closed and dry in 353 women (95.7%). CONCLUSION: Non-obstetric fistula, particularly due to iatrogenic causes, was relatively common in the DRC, calling for more prevention that includes improved quality of care in maternal health services.

OBJECTIF: Décrire la fréquence, les causes et les résultats post-réparation de la fistule non obstétricale (FNO) dans les hôpitaux soutenus par le projet Fistula Care Plus (FC+) en République Démocratique du Congo. MÉTHODES: Etude de cohorte rétrospective du 1er janvier 2015 au 31 décembre 2017 dans trois sites de réparation de fistules soutenus par FC+. RÉSULTATS: Sur 1984 femmes traitées pour une fistule génitale féminine entre 2015 et 2017 dans les trois hôpitaux soutenus par FC+, 384 (19%) étaient considérées comme des cas de FNO. 49,3% étaient mariées/en couple au moment du traitement contre 69% avant la fistule, p <0,001. Les fistules de type III (n = 247; 64,3%) et de type I (n = 121; 31,5%) selon la classification de Kees/Waaldijk étaient les plus courantes. Les principales causes de FNO étaient la procédure médicale (n = 305; 79,4%); parmi lesquelles les césariennes (n = 234; 76,7%) et l'hystérectomie (n = 54; 17,7%) étaient les plus courantes. A la sortie de l'hôpital, la fistule était fermée et sèche chez 353 femmes (95,7%). CONCLUSION: La FNO, en particulier due à des causes iatrogènes, était relativement courante en RDC, appelant à plus de prévention qui comprend l'amélioration de la qualité des soins dans les services de santé maternelle.

Temporomandibular Disorders and Oral Features in Systemic Lupus Erythematosus Patients: An Observational Study of Symptoms and Signs.

Aims: Systemic Lupus Erythematosus (SLE) is a connective tissue disease characterized by a wide range of pleomorphic pictures, including mucocutaneous, renal, musculoskeletal and neurological symptoms. It involves oral tissues, with hyposalivation, tooth decay, gingivitis, angular cheilitis, ulcers and glossitis. Temporomandibular disorders represent a heterogeneous group of inflammatory or degenerative diseases of the stomatognatic system, with algic and/or dysfunctional clinical features involving temporomandibular joint (TMJ) and related masticatory muscles. The aim of this study was to investigate the prevalence of oral manifestations and temporomandibular disorders (TMD) in SLE patients (Lp) compared with a control group. Methods: Fifty-five patients (9 men and 46 women) with diagnosed Lupus were recruited in the study group. A randomly selected group of 55 patients, matched by sex and age, served as control group. The examination for TMD symptoms and signs was based on the standardized Research Diagnostic Criteria for Temporomandibular Disorders (RDC/TMD) through a questionnaire and clinical examination. Results: Lupus patients complained more frequently (95.8%) of oral and TMJ symptoms (dysgeusia, stomatodynia, masticatory muscle pain during function, neck and shoulder muscles pain and presence of tinnitus) but only xerostomia (χ2=4,1548 p=0,0415), temple headache (χ2=4,4542 p=0,035) and the sensation of a stuck jaw (Mid-p-test p=0,043) were significant. About signs, cheilitis (p=0,0284) oral ulcers (χ2=4,0104 p=0,045) and fissured tongue are significantly more frequent in study group. The salivary flow was significantly decreased in the study group respect to the control one (p<0.0001). As regard to the oral kinematics, restricted movements (RM) in protrusion and left lateral movement were significantly different between study group and controls. In particular, 85,2% of Lp showed limited protrusion versus 56,4% of controls (χ2= 10,91 p<0,001); 59,3% of Lp had also a limitation during left lateral movement versus 47,3% of controls (T=2,225 p=0,0282). About bruxism, only the indentations on the lateral edges of the tongue were found in Lp group (72,7%), with a significant difference respect to controls (χ2=7,37 p=0,007). Conclusions: While masticatory muscles have an overlapping behavior in both groups, the findings collected show a more severe TMJ kinematic impairment in Lp than in controls, with protrusion and left lateral movements significantly different. In addition, a remarkable reduction of salivary flow has been detected in Lp compared to controls. In conclusion, this autoimmune disease seems to play a role in oral manifestations and TMJ disorders, causing an increase in orofacial pain and an altered chewing function.

Partial alignment, residual dipolar couplings and molecular symmetry in solution NMR.

Residual dipolar couplings (RDCs) and residual anisotropic chemical shifts (RACSs) are produced by the partial alignment of solution NMR samples. RDCs and RACSs yield high-resolution structural and dynamic information on the orientation of bonds and chemical groups in molecules. Many molecules form oligomers or have intrinsic symmetries, which may simplify the analysis of their partial alignment datasets. In this report, we explore the theory of partial alignment using an irreducible spherical representation, and we investigate the impact of molecular symmetry on the alignment of molecules. Though previous studies have reported simplified relationships on the partial alignment of molecules bearing different symmetry groups, we show that these simplified relationships may not be universal and only apply to a limited set of systems.

Varicella in Tshuapa Province, Democratic Republic of Congo, 2009-2014.

OBJECTIVE: To describe varicella cases in Tshuapa Province of the Democratic Republic of the Congo identified during monkeypox surveillance. METHODS: Demographic, clinical and epidemiological data were collected from each suspected monkeypox case 2009-2014. Samples were tested by PCR for both Orthopoxviruses and varicella-zoster virus (VZV); a subset of VZV-positive samples was genotyped. We defined a varicella case as a rash illness with laboratory-confirmed VZV. RESULTS: There were 366 varicella cases were identified; 66% were ≤19 years old. Most patients had non-typical varicella rash with lesions reported as the same size and stage of evolution (86%), deep and profound (91%), on palms of hands and/or soles of feet (86%) and not itchy (49%). Many had non-typical signs and symptoms, such as lymphadenopathy (70%) and sensitivity to light (23%). A higher proportion of persons aged ≥20 years than persons aged ≤19 years had ≥50 lesions (79% vs. 65%, P = 0.007) and were bedridden (15% vs. 9%, P = 0.056). All VZV isolates genotyped from 79 varicella cases were clade 5. During the surveillance period, one possible VZV-related death occurred in a 7-year-old child. CONCLUSIONS: A large proportion of patients presented with non-typical varicella rash and clinical signs and symptoms, highlighting challenges identifying varicella in an area with endemic monkeypox. Continued surveillance and laboratory diagnosis will help in rapid identification and control of both monkeypox and varicella and improve our understanding of varicella epidemiology in Africa.

OBJECTIF: Décrire les cas de varicelle identifiés dans la province de Tshuapa en République Démocratique du Congo (RDC) au cours de la surveillance de la variole du singe (monkeypox). MÉTHODES: Des données démographiques, cliniques et épidémiologiques ont été recueillies pour chaque cas présumé de monkeypox entre 2009 et 2014. Les échantillons ont été testés par PCR pour les orthopoxvirus et le virus varicelle-zona (VZV); un sous-ensemble d'échantillons positifs au VZV a été génotypé. Nous avons défini un cas de varicelle comme une éruption cutanée avec confirmation du VZV en laboratoire. RÉSULTATS: 366 cas de varicelle ont été identifiés; 66% avaient 19 ans ou moins. La plupart des patients présentaient une éruption non typique de varicelle avec des lésions rapportées de la même taille et le même stade d'évolution (86%), profonds (91%), sur la paume des mains et/ou la plante des pieds (86%), sans démangeaisons (49%). Nombre d'entre eux présentaient des signes et des symptômes inhabituels, tels qu'une adénopathie lymphatique (70%) et une sensibilité à la lumière (23%). Une proportion plus élevée de personnes âgées de 20 ans et plus que de personnes âgées de 19 ans et moins avaient 50 lésions ou plus (79% contre 65%, p = 0,007) et étaient alitées (15% contre 9%; p = 0,056). Tous les isolats de VZV génotypés chez 79 cas de varicelle appartenaient au clade 5. Au cours de la période de surveillance, un décès possible lié au VZV est survenu chez un enfant de 7 ans. CONCLUSIONS: Une forte proportion de patients ont présenté une éruption de varicelle ainsi que des signes et symptômes cliniques non typiques, soulignant les difficultés rencontrées pour identifier la varicelle dans une zone endémique pour le monkeypox. Une surveillance continue et des diagnostics de laboratoire aideront à identifier et à contrôler rapidement le monkeypox et la varicelle et à améliorer notre compréhension sur l'épidémiologie de la varicelle en Afrique.

Complex macrocycle exploration: parallel, heuristic, and constraint-based conformer generation using ForceGen.

ForceGen is a template-free, non-stochastic approach for 2D to 3D structure generation and conformational elaboration for small molecules, including both non-macrocycles and macrocycles. For conformational search of non-macrocycles, ForceGen is both faster and more accurate than the best of all tested methods on a very large, independently curated benchmark of 2859 PDB ligands. In this study, the primary results are on macrocycles, including results for 431 unique examples from four separate benchmarks. These include complex peptide and peptide-like cases that can form networks of internal hydrogen bonds. By making use of new physical movements ("flips" of near-linear sub-cycles and explicit formation of hydrogen bonds), ForceGen exhibited statistically significantly better performance for overall RMS deviation from experimental coordinates than all other approaches. The algorithmic approach offers natural parallelization across multiple computing-cores. On a modest multi-core workstation, for all but the most complex macrocycles, median wall-clock times were generally under a minute in fast search mode and under 2 min using thorough search. On the most complex cases (roughly cyclic decapeptides and larger) explicit exploration of likely hydrogen bonding networks yielded marked improvements, but with calculation times increasing to several minutes and in some cases to roughly an hour for fast search. In complex cases, utilization of NMR data to constrain conformational search produces accurate conformational ensembles representative of solution state macrocycle behavior. On macrocycles of typical complexity (up to 21 rotatable macrocyclic and exocyclic bonds), design-focused macrocycle optimization can be practically supported by computational chemistry at interactive time-scales, with conformational ensemble accuracy equaling what is seen with non-macrocyclic ligands. For more complex macrocycles, inclusion of sparse biophysical data is a helpful adjunct to computation.

Temporomandibular Disorders and Oral Features in Early Rheumatoid Arthritis Patients: An Observational Study.

Aims: Temporomandibular disorders (TMD) represent a heterogeneous group of inflammatory or degenerative diseases of the stomatognatic system, with algic and/or dysfunctional clinical features involving temporomandibular joint (TMJ) and related masticatory muscles. Rheumatoid Arthritis (RA) is an autoimmune polyarthritis characterized by the chronic inflammation of synovial joints and oral implications such as hyposalivation, difficulty in swallowing and phoning, feeling of burning mouth, increased thirst, loss of taste or unpleasant taste and smell, dental sensitivity. The aim of this observational study was to investigate the prevalence of TMD symptoms and signs as well as oral implications in patients with Early Rheumatoid Arthritis (ERA), that is a RA diagnosed within 12 months, compared with a control group. Methods: The study group included 52 ERA patients (11 men, 41 women) diagnosed according to the 2010 ACR/EULAR Classification Criteria for Rheumatoid Arthritis. A randomly selected group of 52 patients not affected by this disease, matched by sex and age, served as the control group. The examination for TMD signs and symptoms was based on the standardized Research Diagnostic Criteria for Temporomandibular Disorders (RDC/TMD) by means of a questionnaire and through clinical examination. Results: Regarding the oral kinematics, the left lateral excursion of the mandible was restricted in statistically significant way in ERA patients (p=0.017). The endfeel values were significantly increased in ERA group (p=0.0017), thus showing the presence of a higher muscle contracture. On the other side, the study group complained less frequently (67.3%) of TDM symptoms (muscle pain on chewing, pain in the neck and shoulders muscles, difficulty in mouth opening, arthralgia of TMJ, tinnitus) than controls (90.4%) (χ2= 8.301 p=0.0039). The presence of TMJ noises was significantly lower in the study group (χ2= 3.869 p=0.0049), as well as presence of opening derangement (χ2= 14.014 p=0.0002). The salivary flow was significantly decreased in the study group respect to the control one (p<0.0001). Conclusions: The data collected show a weak TMJ kinematic impairment, a paucisymptomatic muscle contracture (positive endfeel) and a remarkable reduction of salivary flow in ERA patients. Myofacial pain (MP) evoked by palpation was more frequent and severe in the control group than in the study one, this result being highly significant.

Structure of a low-population binding intermediate in protein-RNA recognition.

The interaction of the HIV-1 protein transactivator of transcription (Tat) and its cognate transactivation response element (TAR) RNA transactivates viral transcription and represents a paradigm for the widespread occurrence of conformational rearrangements in protein-RNA recognition. Although the structures of free and bound forms of TAR are well characterized, the conformations of the intermediates in the binding process are still unknown. By determining the free energy landscape of the complex using NMR residual dipolar couplings in replica-averaged metadynamics simulations, we observe two low-population intermediates. We then rationally design two mutants, one in the protein and another in the RNA, that weaken specific nonnative interactions that stabilize one of the intermediates. By using surface plasmon resonance, we show that these mutations lower the release rate of Tat, as predicted. These results identify the structure of an intermediate for RNA-protein binding and illustrate a general strategy to achieve this goal with high resolution.