Risk of foetal harm with letrozole use in fertility treatment: a systematic review and meta-analysis.
The aromatase inhibitor letrozole is increasingly recommended for ovulation induction, as it is more effective with fewer side-effects than other agents. But many clinicians are reluctant to use the drug for fertility treatment due to a strong-label warning against its use, which warns about congenital malformation risk to the foetus in women seeking pregnancy. The aim of this study was to determine the risks of congenital malformations and pregnancy loss with letrozole compared with clomiphene primarily, and with other fertility drugs and natural conception. A systematic review and meta-analysis using PRISMA harms guidelines. We searched MEDLINE, EMBASE and other sources from inception until January 2020, with the MeSH words for 'letrozole' and pregnancy OR foetal/neonatal outcome. We included studies reported on congenital malformations in foetuses born to mothers conceived after fertility treatment, with letrozole versus clomiphene, placebo, gonadotrophins, metformin, natural conception or other agents, from randomised trials, comparative cohort studies and non-comparative observational cohorts. Quality of the studies was assessed using Cochrane risk of bias tool and Newcastle Ottawa Scale. The McMaster tool was used to assess the quality of reported harm for foetal congenital malformations in the studies. We compared the absolute risk of events using risk difference measures and pooled the findings using a fixed-effect model. We evaluated the statistical heterogeneity using forest plots and the I2 statistic and funnel plot to assess publication bias. We assessed the strength of evidence for congenital malformation and pregnancy loss as per the GRADE recommendations and with the Fragility index. We included 46 studies (18 randomised trials; 21 comparative cohorts; 7 non-comparative cohorts). Overall 2.15% (101/4697; 95% CI 1.7 to 2.5) of babies conceived on letrozole for fertility treatment had congenital foetal malformations. We did not observe a significant increase in congenital malformations with letrozole versus clomiphene in the randomised trials (risk difference (RD) 0.01, 95% CI -0.02, 0.03; I2 = 0%; 14 studies) and found a significant reduction in the cohort studies (RD -0.02, 95% CI -0.04, -0.01; I2 = 0%, 11 studies). The fragility index was 44% (7/16) (either an increase in the intervention arm or a decrease in control arm was needed to alter the results). The risks of pregnancy loss were not increased with letrozole versus clomiphene in the 14 randomised trials (RD -0.01, 95% CI -0.06, 0.04; I2 = 0%), and the risks were reduced in the six cohort studies (RD -0.09, 95% CI -0.17, -0.00; I2 = 68%). The GRADE quality of evidence was low to moderate for congenital malformations and pregnancy loss. We did not find any increased congenital malformation risk with letrozole versus gonadotrophins, natural conception or natural cycle ART, but the number of studies was small. There is no evidence that letrozole increases the risk of congenital foetal malformation or pregnancy loss compared with clomiphene, natural conception or other fertility agents, to warrant warning against its use. Given its therapeutic benefits and lack of evidence of harm to the foetus, clinicians should consider letrozole as first-line agent for ovulation induction.


Prevention of recurrent cryptogenic stroke with percutaneous closure of patent foramen ovale; one year follow-up study with magnetic resonance imaging and Holter monitoring.
In this study, we aimed to evaluate the effect of percutaneous closure of patent foramen ovale (PFO) on the recurrence of stroke and new cardiac arrhythmia using magnetic resonance imaging (MRI) and Holter monitoring. Patients with PFO had >1 previous stroke or transient ischemic attack documented with MRI in the first event. PFO with right to left shunt was detected by transesophageal echocardiography (TEE) and transcranial Doppler ultrasound. MRI examinations were performed on patients before and one year after PFO closure was applied. A twenty-four hour Holter monitoring was performed in all patients within 1 month before and 6 months after the procedure. Percutaneous PFO closure was performed on 47 patients (25 female, mean age: 38.7 years) who had cerebral ischemic events detected by MRI. A year after the procedure, TEE showed that there was no residual interatrial right-to-left shunting. After a 14 month follow-up, no new cerebrovascular event and no new lesion on MRI were recorded. The incidence of arrhythmia did not increase significantly after the procedure on Holter monitoring (p=0.917). One-year clinical and MRI follow-up study of patients with cerebral ischemic events and percutaneous closure of PFO showed no recurrent event and no significant complication associated with the procedure. In addition, Holter monitorization demonstrated that the procedure did not increase the incidence of arrhythmias compared with pre-procedural monitoring.


An enhancer directs differential expression of the linked Mrf4 and Myf5 myogenic regulatory genes in the mouse.
The myogenic regulatory factors, Mrf4 and Myf5, play a key role in skeletal muscle formation. An enhancer trap approach, devised to isolate positive-acting elements from a 200-kb YAC covering the mouse Mrf4-Myf5 locus in a C2 myoblast assay, yielded an enhancer, A17, which mapped at -8 kb 5' of Mrf4 and -17 kb 5' of Myf5. An E-box bound by complexes containing the USF transcription factor is critical for enhancer activity. In transgenic mice, A17 gave two distinct and mutually exclusive expression profiles before birth, which correspond to two phases of Mrf4 transcription. Linked to the Tk or Mrf4 minimal promoters, the nlacZ reporter was expressed either in embryonic myotomes, or later in fetal muscle, with the majority of Mrf4 lines showing embryonic expression. When linked to the Myf5 minimal promoter, only fetal muscle expression was detected. These observations identify A17 as a sequence that targets sites of myogenesis in vivo and raise questions about the mutually exclusive modes of expression and possible promoter/enhancer interactions at the Mrf4-Myf5 locus.


Outcomes of total versus subtotal abdominal hysterectomy.
There is still controversy about the best technique for hysterectomy to reduce postoperative adverse effects. This randomized clinical study in Babol, Islamic Republic of Iran, compared some clinical complications and sexual functioning following subtotal (SAH) and total abdominal hysterectomy (TAH). A total of 150 women (50 allocated to SAH and 100 to TAH) were followed up at 6 months postoperatively. Length of hospitalization was 4.40 (SD 1.90) days after SAH and 4.48 (SD 1.67) days after TAH. Haemoglobin level, postoperative fever, symptoms of dyspareunia and frequency of sexual intercourse were not significantly different between the 2 groups of women. SAH did not show any significant benefits over TAH.


Integrator networks: illuminating the black box linking genotype and phenotype.
Emerging concepts in developmental biology, such as facilitated variation and dynamical patterning modules, address a major shortcoming of the Modern Synthesis in Biology: how genotypic variation is transduced into functional yet diverse phenotypic variation. Still, we lack a theory to explain how variation at the cellular and tissue level is coordinated into variation at the whole-organism level, especially as priority of cellular and tissue functions change over an individual's lifetime and are influenced by environmental variation. Here, we propose that interactions among a limited subset of physiological factors that we call, integrators, regulate most phenotypic variation at the organismal level. Integrators are unique among physiological factors in that they have the propensity to coordinate the expression of conserved gene modules of most types of tissues because they participate as nodes in a hierarchical network. In other words, integrator networks impose physiological epistasis, meaning that whole-organism phenotypic responses will be influenced by previous experiences, current environmental conditions, and fitness priorities as encoded by individual integrators. Below, we provide examples of how integrator networks are responsible for both profound and irreversible phenotypic changes (i.e., metamorphosis, sexual differentiation) as well as subtler, transient (e.g., pelage color, seasonal fluctuations in lymphoid and reproductive tissues) variation. The goal of this article is not to describe completely how integrator networks function, but to stimulate discussion about the role of physiology in linking genetic to phenotypic variation. To generate useful data sets for understanding integrator networks and to inform whole-organism physiology generally, we describe several useful tools including vector-field editing, response-surface regression, and experiments of life-table responses. We then close by highlighting some implications of integrator networks for conservation and biomedicine.


Association of p53 mutations with metastatic prostate cancer.
In prostate cancer, mutation of the p53 tumor suppressor gene has been associated with locally advanced disease and hormone-resistant disease that is predominantly localized to bone. However, little is known regarding the status of the p53 gene in metastatic prostate cancer that has not been treated with hormonal manipulation. We evaluated formalin-fixed, paraffin-embedded malignant tissues from 86 patients with various stages of prostate cancer, including pathologically confined, locally advanced, and metastatic disease, to detect abnormal p53 nuclear protein accumulation using immunohistochemistry. No abnormal p53 immunostaining was detected in 18 patients with prostate cancer confined to the gland. Two tumors from 21 patients with locally advanced disease (extracapsular extension and/or seminal vesicle invasion) had abnormal nuclear p53 accumulation, and a mutation in exon 7 of the p53 gene was detected in tumor DNA from one patient using single-strand conformation polymorphism-direct sequencing analysis. Of the remaining 47 patients studied in whom tissues from the prostate gland and a metastatic site (44 lymph node, 2 bone, and 1 lung) were available, only 3 had received hormonal therapy prior to obtaining metastatic tissue. In four patients both primary and metastatic tumors demonstrated accumulation of p53 protein, whereas seven additional patients exhibited p53 accumulation only at the metastatic site. In three patients the metastatic tumors harbored missense single-base substitutions in exon 5, as detected using single-strand conformation polymorphism-direct sequencing. These results indicate that p53 abnormalities are associated with lymph node metastases derived from prostate cancer patients that had not undergone hormonal therapy.


Nd:Li6Y(BO3)3 crystal waveguide properties at wavelengths of 633 and 1539 nm produced by oxygen or silicon ion implantation.
We report on the waveguide formation in the Nd:Li6Y(BO3)3 laser crystal by oxygen or silicon ion implantation, respectively, and how the waveguide structure effectively supports the fundamental mode in the visible and near-infrared telecommunication band. Compared with Si-ion, the waveguide produced by O-ion has a larger effective refractive index of fundamental mode and lower propagation loss, which shows that peak position of implanted ion nuclear energy loss has a significant impact on waveguide features. The investigation of the photoluminescence and Raman spectra demonstrates that the Nd3+ luminescence feature and crystal structure of the waveguide active region do not change significantly and gain good preservation after ion implantation.


Percutaneous lung biopsy: an ordering clinician's guide to current practice.
The discovery of a pulmonary nodule or mass usually leads to a clinical assessment of patient risk factors for malignancy and an imaging workup. The latter may include appropriately timed follow-up, computed tomography with the possible addition of a computed tomographic nodule enhancement study, positron emission tomography, and/or percutaneous biopsy of the lung lesion or associated lymph nodes or metastatic lesions. This article reviews the rationale for the imaging workup of lung lesions and the details of percutaneous needle biopsy, including risks, technical feasibility, techniques, needle systems, cytological/pathological evaluation, accuracy, complications, and complication management.


N4-Methoxydeoxycytidine triphosphate is in the imino tautomeric form and substitutes for deoxythymidine triphosphate in primed poly d[A-T] synthesis with E. coli DNA polymerase I.
N4- Methoxydeoxycytidine triphosphate ( mo4dCTP ) substitutes for dTTP in poly d[A-T] synthesis with E. coli DNA polymerase I (Pol I). In parallel experiments using as template-primer, poly d[G-C], no incorporation of [14C] mo4dC was detected. This indicates that this deoxy derivative acts as the imino tautomer, as previously found for the riboderivative . Nearest neighbor analysis of transcripts of poly d[A-T] containing mo4dC shows that the derivative substitutes for only one base. In replication, singlestranded mo4dC -containing polymers gave little misincorporation, including that of dATP which can hydrogen-bond to mo4dC in the imino form, if the methoxy group is anti to the N-3. It is therefore assumed that the methoxy group is constrained anti in a polymer such as d[A-T], but can be in the syn form in singlestranded polymers and not recognized by DNA polymerase. mo4dC destabilizes the poly d[A-T] helix, as indicated by a lowered and less cooperative melting. Steric factors such as adjacent base displacement were invoked for similar findings with the doublestranded r( U61 , mo4C39 ) X r(A).


Deletions in the tL structure upstream to the rRNA genes in the E. coli rrnB operon cause transcription polarity.
A number of deletions have been constructed within the leader region of the rrnB operon from E. coli. The deletions remove a potential transcription terminator structure downstream from an antitermination recognition sequence (Box A), which precedes the structural gene for the 16S RNA. Cells harbouring plasmids, where the terminator structure was deleted, partially or totally, showed a reduction in growth rate under minimal growth conditions. Measurement of the ribosomal RNA synthesis rates of such cells determined by pulselabeling and hybridisation to appropriate DNA probes, showed that the amount of the more distally located 23S RNA was reduced compared to the promoter-proximal 16S RNA. This polarity in transcription, resulting in a non-stoichiometric synthesis of the ribosomal RNAs, is most likely the result of a defective antitermination. The reduction in the amount of 23S RNA in such cells is compensated for by an increase in the overall ribosomal RNA synthesis, in concordance with the ribosomal RNA feedback regulation model. The accumulation of transcripts of the tRNAGlu2 gene, coded in the spacer region between the 16S and 23S RNA genes, in cells with an altered rRNA stoichiometry supports this interpretation.


Moral dilemmas in (not) treating patients who feel they are a burden.
Working as clinical ethicists in an academic hospital, we find that practitioners tend to take a principle-based approach to moral dilemmas when it comes to (not) treating patients who feel like a burden, in which respect for autonomy tends to trump other principles. We argue that this approach insufficiently deals with the moral doubts of professionals with regard to feeling that you are a burden as a motive to decline or withdraw from treatment. Neither does it take into adequately account the specific needs of the patient that might underlie their feeling of being a burden to others. We propose a care ethics approach as an alternative. It focuses on being attentive and responsive to the caring needs of those involved in the care process-which can be much more specific than either receiving or withdrawing from treatment. This approach considers these needs in the context of the patient's identity, biography and relationships, and regards autonomy as relational rather than as individual. We illustrate the difference between these two approaches by means of the case of Mrs K. Furthermore, we show that a care ethics approach is in line with interventions that are found to alleviate feeling a burden and maintain that facilitating moral case deliberation among practitioners can supports them in taking a care ethics approach to moral dilemmas in (not) treating patients who feel like a burden.


Technical standards and guidelines: venous thromboembolism (Factor V Leiden and prothrombin 20210G >A testing): a disease-specific supplement to the standards and guidelines for clinical genetics laboratories.
These standards and guidelines are designed primarily as an educational resource for clinical laboratory geneticists to help them provide quality clinical laboratory genetic services. Adherence to this statement does not necessarily ensure a successful medical outcome. These standards and guidelines should not be considered inclusive of all proper procedures and tests or exclusive of other procedures and tests that are reasonably directed to obtaining the same results. In determining the propriety of any specific procedure or test, the clinical molecular geneticist should apply his or her own professional judgment to the specific clinical circumstances presented by the individual patient or specimen. It may be prudent, however, to document in the laboratory record the rationale for any significant deviation from these standards and guidelines.


Probing subdiffraction limit separations with plasmon coupling microscopy: concepts and applications.
Due to their advantageous material properties, noble metal nanoparticles are versatile tools in biosensing and imaging. A characteristic feature of gold and silver nanoparticles is their ability to sustain localized surface plasmons that provide both large optical cross-sections and extraordinary photophysical stability. Plasmon coupling microscopy takes advantage of the beneficial optical properties and utilizes electromagnetic near-field coupling between individual noble metal nanoparticle labels to resolve subdiffraction limit separations in an all-optical fashion. This Tutorial provides an introduction into the physical concepts underlying distance dependent plasmon coupling, discusses potential experimental implementation of plasmon coupling microscopy, and reviews applications in the area of biosensing and imaging.


Risk of metachronous colon cancer following surgery for rectal cancer in mismatch repair gene mutation carriers.
Despite regular surveillance colonoscopy, the metachronous colorectal cancer risk for mismatch repair (MMR) gene mutation carriers after segmental resection for colon cancer is high and total or subtotal colectomy is the preferred option. However, if the index cancer is in the rectum, management decisions are complicated by considerations of impaired bowel function. We aimed to estimate the risk of metachronous colon cancer for MMR gene mutation carriers who underwent a proctectomy for index rectal cancer. This retrospective cohort study comprised 79 carriers of germline mutation in a MMR gene (18 MLH1, 55 MSH2, 4 MSH6, and 2 PMS2) from the Colon Cancer Family Registry who had had a proctectomy for index rectal cancer. Cumulative risks of metachronous colon cancer were calculated using the Kaplan-Meier method. During median 9 years (range 1-32 years) of observation since the first diagnosis of rectal cancer, 21 carriers (27 %) were diagnosed with metachronous colon cancer (incidence 24.25, 95 % confidence interval [CI] 15.81-37.19 per 1,000 person-years). Cumulative risk of metachronous colon cancer was 19 % (95 % CI 9-31 %) at 10 years, 47 (95 % CI 31-68 %) at 20 years, and 69 % (95 % CI 45-89 %) at 30 years after surgical resection. The frequency of surveillance colonoscopy was 1 colonoscopy per 1.16 years (95 % CI 1.01-1.31 years). The AJCC stages of the metachronous cancers, where available, were 72 % stage I, 22 % stage II, and 6 % stage III. Given the high metachronous colon cancer risk for MMR gene mutation carriers diagnosed with an index rectal cancer, proctocolectomy may need to be considered.


Biomechanical properties of 3D-printed bone scaffolds are improved by treatment with CRFP.
One of the major challenges in orthopedics is to develop implants that overcome current postoperative problems such as osteointegration, proper load bearing, and stress shielding. Current implant techniques such as allografts or endoprostheses never reach full bone integration, and the risk of fracture due to stress shielding is a major concern. To overcome this, a novel technique of reverse engineering to create artificial scaffolds was designed and tested. The purpose of the study is to create a new generation of implants that are both biocompatible and biomimetic. 3D-printed scaffolds based on physiological trabecular bone patterning were printed. MC3T3 cells were cultured on these scaffolds in osteogenic media, with and without the addition of Calcitonin Receptor Fragment Peptide (CRFP) in order to assess bone formation on the surfaces of the scaffolds. Integrity of these cell-seeded bone-coated scaffolds was tested for their mechanical strength. The results show that cellular proliferation and bone matrix formation are both supported by our 3D-printed scaffolds. The mechanical strength of the scaffolds was enhanced by trabecular patterning in the order of 20% for compression strength and 60% for compressive modulus. Furthermore, cell-seeded trabecular scaffolds modulus increased fourfold when treated with CRFP. Upon mineralization, the cell-seeded trabecular implants treated with osteo-inductive agents and pretreated with CRFP showed a significant increase in the compressive modulus. This work will lead to creating 3D structures that can be used in the replacement of not only bone segments, but entire bones.


The association between substance use, condom use and sexual risk among low-income women.
Substance use is frequently assumed to be associated with higher levels of sexual risk-taking and lower levels of condom use. An analysis of 668 black, Hispanic and white low-income women at public health and public assistance facilities in Miami show that 19% engaged in risky sexual behavior over the preceding six months, 24% in substance use and 31% in condom use. Overall, substance users are nearly four and one-half times more likely to take sexual risks than nonusers, but are about half as likely to have relied on condoms. When the probability of condom use is considered in the context of both substance use and sexual risk, substance users who take sexual risks appear just as likely to rely on condoms as are nonusers who take sexual risks and those who do not (odds of 0.43-0.49). However, substance users who do not take sexual risks are much less likely to use condoms (odds of 0.15). This pattern holds among black, Hispanic and white women, and suggests that perceptions of risk and the risks that partners bring to sexual encounters may be more important determinants of condom use than substance use per se.


The relationship between the anticoccidial effects of clindamycin and the development of immunity in the Eimeria pragensis/mouse model of large intestinal coccidiosis.
The therapeutic effect of clindamycin on Eimeria pragensis (E. pragensis) infection in C57BL/6 mice was demonstrated by suppression of oocyst production and the appearance of degenerated endogenous stages of parasite in the intestine. Short-term clindamycin treatment, from 1 to 4 days or 4 to 8 days post infection (pi) at a dose of 800 mg/kg/day was effective to reduce clinical symptoms, oocyst production and schizogonic development. Interestingly, the short-term treatment schedules allowed the development of a measurable degree of protective immunity to challenge infection in the treated mice. In contrast, clindamycin treatment for the full 12 days period, which almost completely inhibited clinical symptoms and oocyst output, prevented the full development of protective immunity in the treated mice. All these data indicate that clindamycin is efficacious as an anti-eimerian agent and that both early and late endogenous developmental stages of E. pragensis exert a deep influence on the development of effective immunity to challenge infection.


Sex disparities in colorectal cancer incidence by anatomic subsite, race and age.
Although incidence of colorectal cancer (CRC) in the United States has declined in recent years, rates remain higher in men than in women and the male-to-female incidence rate ratio (MF IRR) increases progressively across the colon from the cecum to the rectum. Rates among races/ethnicities other than Whites or Blacks have not been frequently reported. To examine CRC rates by sex across anatomic subsite, age and racial/ethnic groups, we used the National Cancer Institute's Surveillance, Epidemiology and End Results (SEER) program for cases diagnosed among residents of 13 registries during 1992-2006. Incidence rates were expressed per 100,000 person-years and age-adjusted to the 2000 US Standard Population; MF IRR and 95% confidence intervals were also calculated. Among each racial/ethnic group, the MF IRR increased fairly monotonically from close to unity for cecal cancers to 1.81 (Hispanics) for rectal cancers. MF IRRs increased with age most rapidly for distal colon cancers from <1.0 at ages <50 years to 1.4-1.9 at older ages. The MF IRR for rectal cancers also rose with age from about 1.0 to 2.0. For proximal cancer, the MF IRR was consistently <1.5; among American Indian/Alaska Natives, it was <1.0 across all ages. The MF IRRs for CRC vary markedly according to subsite and age but less by racial/ethnic group. These findings may partially reflect differences in screening experiences and access to medical care but also suggest that etiologic factors may be playing a role.


Roles of heat shock proteins and gamma delta T cells in inflammation.
Elimination of activated inflammatory cells that infiltrate and damage host organs can reduce morbidity and mortality. A better understanding of the mechanisms by which these processes occur may lead to new approaches to prevent tissue damage. The lungs, gastrointestinal tract, and skin are particularly prone to infection and collateral damage by inflammatory cells. Specialized lymphocytes protect these organs from collateral tissue damage by eliminating neutrophils and macrophages from inflamed tissues. These lymphocytes recognize signals produced by inflammatory cells. One such signal is heat shock protein (Hsp) expressed on the cell surface of inflamed phagocytes. Mammalian Hsp molecules closely resemble their microbial equivalents, and therefore phagocytes decorated with these molecules are recognized as target cells. T lymphocytes bearing the gammadelta T cell receptor (TCR) elicit cytotoxic activity toward macrophages and neutrophils that express Hsp60 and Hsp70, respectively, protecting host organs from collateral tissue damage by phagocytes.


The relationship between proton pump inhibitor adherence and fracture risk in the elderly.
Studies suggest that long-term use of proton pump inhibitors (PPIs) may be associated with an increased risk of fracture. However, the role of medication adherence in this association is not fully understood. A retrospective cohort study was conducted to examine the relationship between PPI use/adherence and fracture risk among elderly subjects by combining administrative pharmacy claims data, survey data, and Medicare data. The study cohort included 1,604 PPI users and 23,672 nonusers who were enrolled in Pennsylvania's Pharmaceutical Assistance Contract for the Elderly program. PPI adherence was measured by the proportion of days covered (PDC). Time-dependent Cox proportional hazards models were used to estimate adjusted hazard ratios (HRs) of PPI use/adherence for fracture risk while controlling for demographics, comorbidity, body mass index, smoking, and non-PPI medication use. The overall incidence of any fracture per 100 person-years was 8.7 for PPI users and 5.0 for nonusers. A gradient in fracture risk according to PPI adherence was observed. Relative to nonusers, fracture HRs associated with the highest (PDC ≥ 0.80), intermediate (PDC 0.40-0.79), and lowest (PDC <0.40) adherence levels were 1.46 (p < 0.0001), 1.30 (p = 0.02), and 0.95 (p = 0.75), respectively. In addition, the fracture risk of PPI use was significant for hip (HR = 1.32, p = 0.04) and vertebral (HR = 1.69, p = 0.0005) fractures, and risk was similar between major osteoporotic and other fractures. These results provide further evidence that PPI use may increase fracture risk in the elderly and highlight the need for clinicians to periodically reassess elderly patients' individualized needs for ongoing PPI therapy, while weighing potential risks and benefits.


miRNA signatures in sera of patients with active pulmonary tuberculosis.
Several studies showed that assessing levels of specific circulating microRNAs (miRNAs) is a non-invasive, rapid, and accurate method for diagnosing diseases or detecting alterations in physiological conditions. We aimed to identify a serum miRNA signature to be used for the diagnosis of tuberculosis (TB). To account for variations due to the genetic makeup, we enrolled adults from two study settings in Europe and Africa. The following categories of subjects were considered: healthy (H), active pulmonary TB (PTB), active pulmonary TB, HIV co-infected (PTB/HIV), latent TB infection (LTBI), other pulmonary infections (OPI), and active extra-pulmonary TB (EPTB). Sera from 10 subjects of the same category were pooled and, after total RNA extraction, screened for miRNA levels by TaqMan low-density arrays. After identification of "relevant miRNAs", we refined the serum miRNA signature discriminating between H and PTB on individual subjects. Signatures were analyzed for their diagnostic performances using a multivariate logistic model and a Relevance Vector Machine (RVM) model. A leave-one-out-cross-validation (LOOCV) approach was adopted for assessing how both models could perform in practice. The analysis on pooled specimens identified selected miRNAs as discriminatory for the categories analyzed. On individual serum samples, we showed that 15 miRNAs serve as signature for H and PTB categories with a diagnostic accuracy of 82% (CI 70.2-90.0), and 77% (CI 64.2-85.9) in a RVM and a logistic classification model, respectively. Considering the different ethnicity, by selecting the specific signature for the European group (10 miRNAs) the diagnostic accuracy increased up to 83% (CI 68.1-92.1), and 81% (65.0-90.3), respectively. The African-specific signature (12 miRNAs) increased the diagnostic accuracy up to 95% (CI 76.4-99.1), and 100% (83.9-100.0), respectively. Serum miRNA signatures represent an interesting source of biomarkers for TB disease with the potential to discriminate between PTB and LTBI, but also among the other categories.


Exciton lifetime tuning by changing the plasmon field orientation with respect to the exciton transition moment direction: CdTe-Au core-shell nanorods.
We studied the anisotropy of the influence of plasmonic fields, arising from the optical excitation of a gold nanoshell plasmon absorption at 770 nm, on the lifetime of the bandgap state of the CdTe core in vertically aligned CdTe-Au core-shell nanorods. The previously observed decrease in the lifetime was studied as a function of the tilt angle between the long axis of the nanorod and the electric field polarization direction of the plasmon inducing exciting light. It is observed that the strongest enhancement to the exciton relaxation rate occurs when the two axes are parallel to one another. These results are discussed in terms of the coupling between the exciton transition moment of the CdTe rod and the electric field polarization direction of the gold nanoshell plasmon at 770 nm, which was determined from theoretical modeling based on the discrete dipole approximation.


Novel ginsenoside-based multifunctional liposomal delivery system for combination therapy of gastric cancer.
The clinical treatment of gastric cancer (GC) is hampered by the development of anticancer drug resistance and the unfavorable pharmacokinetics, off-target toxicity, and inadequate intratumoral accumulation of the current chemotherapy treatments. Ginsenosides combined with paclitaxel (PTX) have been shown to exert synergistic inhibition of human GC cell proliferation. In the present study, we developed a novel multifunctional liposome system, in which ginsenosides functioned as the chemotherapy adjuvant and membrane stabilizer. These had long blood circulation times and active targeting abilities, thus creating multifunctionality of the liposomes and facilitating drug administration to the GC cells. Methods: Three ginsenosides with different structures were used to formulate the unique nanocarrier, which was prepared using the thin-film hydration method. The stability of the ginsenoside liposomes was determined by particle size analysis using dynamic light scattering. The long circulation time of ginsenoside liposomes was compared with that of conventional liposome and polyethylene glycosylated liposomes in vivo. The active targeting effect of ginsenoside liposomes was examined with a GC xenograft model using an in vivo imaging system. To examine the antitumor activity of ginsenoside liposomes against GC, MTT, cell cycle, and apoptosis assays were performed on BGC-823 cells in vitro and PTX-loaded ginsenoside liposomes were prepared to evaluate the therapeutic efficacy on GC in vivo. Results: The ginsenosides stabilized the liposomes in a manner similar to cholesterol. We confirmed the successful delivery of the bioactive combination drugs and internalization into GC cells via analysis of the glucose-related transporter recognition and longer blood circulation time. PTX was encapsulated in different liposomal formulations for use as a combination therapy, in which ginsenosides were found to exert their inherent anticancer activity, as well as act synergistically with PTX. The combination therapy using these targeted liposomes significantly suppressed GC tumor growth and outperformed most reported PTX formulations, including Lipusu® and Abraxane®. Conclusion: We established novel ginsenoside-based liposomes as a tumor-targeting therapy, in which ginsenoside functioned not only as a chemotherapy adjuvant, but also as a functional membrane material. Ginsenoside-based liposomes offer a novel platform for anticancer drug delivery and may lead to a new era of nanocarrier treatments for cancer.


Effects of arterial vasodilators on cardiac hypertrophy and sympathetic activity in rats.
In spontaneously hypertensive rats (SHR), the progression (or absence of regression) of cardiac hypertrophy despite adequate blood pressure (BP) control by arterial vasodilators has been attributed to increased cardiac sympathetic activity. We evaluated changes in indices of general and cardiac sympathetic tone in relation to changes in cardiac anatomy during treatment of normotensive rats and SHR with hydralazine, 120 mg/L, or minoxidil, 120 mg/L of drinking water. In SHR, both vasodilators reduced BP rapidly and consistently. Significant increases in heart rate and plasma norepinephrine were observed only in the initial 2 days of arterial vasodilator treatment. After 5 weeks of treatment, marked increases in left and right ventricular sympathetic activity (as assessed by norepinephrine turnover rates) were present, but no increase was seen in heart rate and plasma norepinephrine. Intravascular volume expansion was observed on Day 14 of minoxidil and Day 35 of hydralazine treatment. Prolonged treatment with minoxidil induced significant increases in left ventricular internal diameter, as well as in left and right ventricular weights, but not in the wall thickness of the left ventricle. Treatment with hydralazine did not affect left ventricular weight and caused a small increase in the weight of the right ventricle. In normotensive rats, both vasodilators initially decreased BP, but tolerance developed within 1 to 2 weeks of treatment. Plasma norepinephrine and heart rate showed increases only at Day 1 of either treatment, whereas cardiac sympathetic hyperactivity persisted at 2 and 5 weeks of treatment. Changes in cardiac anatomy were qualitatively similar to those observed in SHR. We conclude that, during treatment of normotensive rats and SHR with arterial vasodilators, cardiac sympathetic hyperactivity persists and may be involved in the cardiac effects of arterial vasodilators. However, other mechanisms, such as chronic cardiac volume overload, may also play an important role, particularly with minoxidil.


Wall correction factors, Pwall, for thimble ionization chambers.
The EGSnrc Monte Carlo user-code CSnrc is used to calculate wall correction factors, Pwall, for thimble ionization chambers in photon and electron beams. CSnrc calculated values of Pwall give closer agreement with previous experimental results than do the values from the standard formalism used in current dosimetry protocols. A set of Pwall values, computed at the reference depth in water, is presented for several commonly used thimble chambers. These values differ from the commonly used values by up to 0.8% for megavoltage photon beams, particularly for nominal beam energies below 6 MV. The sleeve effect, which is not currently taken into account by the TG-51 dosimetry protocol, is computed to be up to 0.3% and is in some cases larger than the Pwal1 correction itself. In electron beams, where dosimetry protocols assume a wall correction of unity, CSnrc calculations show Pwall values of up to 0.6% at the reference depth, depending on the wall material. Pwall is shown to be sensitive to the depth of measurement, varying by 2.5% for a graphite-walled cylindrical Farmer-like chamber between a depth of 0.5 cm and R50 in a 6 MeV electron beam.


Restricted Spatial Windows of Visibility in Myalgic Encephalomyelitis (ME).
Myalgic encephalomyelitis (ME) is a devastating disorder marked by debilitating fatigue. It not well understood and its diagnosis is controversial. It is very important therefore that significant clinical features are investigated. Visual symptoms in ME represent a group of distinct, quantifiable, clinical features that could significantly improve diagnosis and provide insights into underlying pathology. The purpose of the present study was therefore to explore the effect of ME on spatial windows of visibility using the spatial contrast sensitivity function. Contrast sensitivity was determined for stationary luminance-defined sinusoidal gratings spanning a five-octave range of spatial frequencies (0.5 to 16 c/deg) in a group of 19 individuals with ME and a group of 19 matched (age, gender) controls. Compared to controls, the ME group exhibited a restricted spatial window of visibility for encoding stimulus contrast. This was characterised principally by a contrast sensitivity deficit at lower spatial frequencies and a narrower bandwidth. Our findings suggest that contrast sensitivity deficits may represent a visual marker of ME, and be indicative of abnormal visual processing at the level of the retina and in the cortical and subcortical visual pathways.


The effectiveness of functional electrical stimulation for the treatment of shoulder subluxation and shoulder pain in hemiplegic patients: A randomized controlled trial.
To investigate the effect of functional electrical stimulation (FES) for the treatment of shoulder subluxation and shoulder pain in hemiplegic patients. A total of 50 hemiplegic patients with shoulder subluxation and shoulder pain were included in the study. The patients were randomly divided into the study and control groups. All patients were put on a rehabilitation program using conventional methods while the study group patients were additionally applied FES to supraspinatus and posterior deltoid muscles. The shoulder pain of all patients during resting, passive range of motion (PROM) and active range of motion (AROM) was measured with the visual analog scale (VAS) while the shoulder subluxation levels were evaluated with the classification developed by Van Langenberghe and by using the millimetric measurements on anteroposterior shoulder X-ray before and after the physical treatment and rehabilitation program and compared. Comparison of the resting AROM vs. PROM VAS value changes showed no significant difference between the groups. There was a significant difference between the two groups for the amount of change in shoulder subluxation in favor of the study group. The results of our study have shown that applying FES treatment to the supraspinatus and posterior deltoid muscles in addition to conventional treatment when treating the subluxation in hemiplegic patients is more beneficial than conventional treatment by itself.


Associations of HbA1c and educational level with risk of cardiovascular events in 32,871 drug-treated patients with Type 2 diabetes: a cohort study in primary care.
To explore the association of HbA1c and educational level with risk of cardiovascular events and mortality in patients with Type 2 diabetes. A cohort of 32 871 patients with Type 2 diabetes aged 35 years and older identified by extracting data from electronic patient records for all patients who had a diagnosis of Type 2 diabetes and had glucose-lowering agents prescribed between 1999 and 2009 at 84 primary care centres in Sweden. Associations of mean HbA1c levels and educational level with risks of cardiovascular events and all-cause mortality were analysed. The associations of HbA1c with risk of all-cause and cardiovascular mortality were J-shaped, with the lowest risk observed for cardiovascular mortality at an HbA1c level of 51 mmol/mol (6.8%) for subjects on oral agents and 56 mmol/mol (7.3%) in insulin-treated patients. The lowest risk observed for all-cause mortality was at an HbA1c level of 51 mmol/mol (6.8%) for subjects on oral agents and 56 mmol/mol (7.3%) in insulin-treated patients. There was an increased risk for cardiovascular death [hazard ratio 1.6 (1.2-2.1), P = 0.0008] at the lowest HbA1c decile for subjects in the low education category. For subjects with higher education there was no evident J curve for cardiovascular death [hazard ratio 1.2 (0.8-1.6), P = 0.3873]. Our results lend support to the recent American Diabetes Association/ European Association for the Study of Diabetes position statement that emphasizes the importance of additional factors, including the propensity for hypoglycaemia, which should influence HbA1c targets and treatment choices for individual patients. (Clinical Trials Registry No; NCT 01121315).


An AF9/ENL-targted peptide with therapeutic potential in mixed lineage leukemias.
Misregulation of transcription elongation is proposed to underlie the pathobiology of MLL leukemia. AF4, AF9, and ENL, common MLL fusion partners, are found in complex with positive transcription elongation factor b (P-TEFb). AF9 and its homolog ENL directly interact with AF4 within these complexes. Previously, we designed a peptide that mimics the AF9 binding domain of AF4 and reported that MLL leukemia cell lines are inhibited by it. Extending these studies, we have modified the peptide design in order to avoid recognition by proteases. The peptide is as effective as its predecessor in vitro and enhances survival in mice bearing MLL leukemia cell lines.


Effects of far infrared acupoint stimulation on autonomic activity and quality of life in hemodialysis patients.
Patients receiving regular hemodialysis sessions have been known to suffer from fatigue and depression. This experiment was designed to determine the effects of far infrared ray (FIR) stimulation on acupoints of patients suffering from renal failure who are receiving regular hemodialysis. Patients receiving long-term and regular hemodialysis who volunteered for this procedure were randomly selected to undergo either FIR or heat pad (HP) therapy to determine the impact of FIR treatment on these patients. Both the activities of the autonomic nervous system and changes in quality of life were measured before and after treatment to determine the effectiveness of the FIR treatment. Results from this study show that FIR therapy decreases both stress and fatigue levels of these patients. It also stimulates autonomic nervous system (ANS) activity in patients who are diagnosed with end-stage renal disease (ESRD) and are receiving regular hemodialysis (HD). Therefore, benefits of FIR stimulation on these patients are clearly demonstrated in this preliminary study.


Association of the polycystic ovary syndrome with genomic variants related to insulin resistance, type 2 diabetes mellitus, and obesity.
We have evaluated the possible association of polycystic ovary syndrome (PCOS) with 15 genomic variants previously described to influence insulin resistance, obesity, and/or type 2 diabetes mellitus. Seventy-two PCOS patients and 42 healthy controls were genotyped for 15 variants in the genes encoding for paraoxonase (three variants), plasma cell differentiation antigen glycoprotein, human sorbin and SH3 domain containing 1, plasminogen activator inhibitor-1, peroxisome proliferator-activated receptor-gamma2, protein tyrosine phosphatase 1B (two variants), adiponectin (two variants), IGF1, IGF2, IGF1 receptor, and IGF2 receptor. Compared with controls, PCOS patients were more frequently homozygous for the -108T variant in paraoxonase (36.6% vs. 9.5%; P = 0.002) and homozygous for G alleles of the ApaI variant in IGF2 (62.9% vs. 38.1%; P = 0.018). Paraoxonase is a serum antioxidant enzyme and, because -108T alleles result in decreased paraoxonase expression, this increase in oxidative stress might result in insulin resistance. G alleles of the ApaI variant in IGF2 may increase IGF2 expression, and IGF2 stimulates adrenal and ovarian androgen secretion. In conclusion, the paraoxonase -108 C-->T variant and the ApaI polymorphism in the IGF2 gene are associated with PCOS and might contribute to increased oxidative stress, insulin resistance, and hyperandrogenism in this prevalent disorder.


Glial GLT-1 blockade in infralimbic cortex as a new strategy to evoke rapid antidepressant-like effects in rats.
Ketamine and deep brain stimulation produce rapid antidepressant effects in humans and rodents. An increased AMPA receptor (AMPA-R) signaling in medial prefrontal cortex (mPFC) has been suggested to mediate these responses. However, little research has addressed the direct effects of enhancing glutamate tone or AMPA-R stimulation in mPFC subdivisions. The current study investigates the behavioral and neurochemical consequences of glutamate transporter-1 (GLT-1) blockade or s-AMPA microinfusion in the infralimbic (IL) and prelimbic (PrL) cortex. Owing to the connectivity between the mPFC and raphe nuclei, the role of serotonin is also explored. The bilateral microinfusion of the depolarizing agent veratridine into IL -but not PrL- of rats evoked immediate antidepressant-like responses. The same regional selectivity was observed after microinfusion of dihydrokainic acid (DHK), a selective inhibitor of GLT-1, present in astrocytes. The DHK-evoked antidepressant-like responses appear to be mediated by an AMPA-R-driven enhancement of serotonergic activity, as (i) they were prevented by NBQX 2,3-dioxo-6-nitro-1,2,3,4-tetrahydrobenzo[f]quinoxaline-7-sulfonamide disodium salt) and mimicked by s-AMPA; (ii) DHK and s-AMPA elevated similarly extracellular glutamate in IL and PrL, although extracellular 5-HT and c-fos expression in the midbrain dorsal raphe increased only when these agents were applied in IL; and (iii) DHK antidepressant-like responses were prevented by 5-HT synthesis inhibition and mimicked by citalopram microinfusion in IL. These results indicate that an acute increase of glutamatergic neurotransmission selectively in IL triggers immediate antidepressant-like responses in rats, likely mediated by the activation of IL-raphe pathways, which then results in a fast increase of serotonergic activity.


Urban mosquito species (Diptera: Culicidae) of dengue endemic communities in the Greater Puntarenas area, Costa Rica.
Field studies were conducted to determine the mosquito species richness in the urban area of Greater Puntarenas in Costa Rica. Two cross-sectional entomological surveys were performed in seven localities of Puntarenas: one survey was performed during the wet season and the other during the dry season. The sections evaluated were determined by applying a stratified cluster sampling method using satellite imagery, and a sample of 26 cells (100 x 100m) was selected for the study. The number of cells per locality was proportional to the area of each locality. The presence of mosquito larvae and pupae in water-filled artificial and natural containers was determined in each cell. Infestation was expressed as a diversity index per type of container (Ii). Eight types of larvae were identified (Aedes aegypti, Culex quinquefasciatus, Culex interrogator, Culex nigripalpus, Culex corniger, Culex tarsalis, Limatus durhamii and Toxorhynchites theobaldi) and in two cases it was only possible to identify the genus (Culex sp. and Uranotaenia sp.). A. aegypti was the most common species followed by C. quinquefascitus. Diversity of wet environments can explain the co-occurrence of various culicid species in some localities. Although A. aegypti is the only documented disease vector in the area, C quinquefasciatus, C nigripalpus, and the other species of Culex could be considered potential vectors of other pathogens. The presence and ecology of all mosquito species should be studied to optimize surveillance and prevention of dengue and to prevent the emergence of other mosquito-transmitted diseases.


Applying three different methods of measuring CTDIfree air to the extended CTDI formalism for wide-beam scanners (IEC 60601-2-44): A comparative study.
The weighted CT dose index (CTDIw ) has been extended for a nominal total collimation width (nT) greater than 40 mm and relies on measurements of CTDIfreeair. The purpose of this work was to compare three methods of measuring CTDIfreeair and subsequent calculations of CTDIw to investigate their clinical appropriateness. The CTDIfreeair, for multiple nTs up to 160 mm, was calculated from (1) high-resolution air kerma profiles from a step-and-shoot translation of a liquid ionization chamber (LIC) (considered to be a dosimetric reference), (2) pencil ionization chamber (PIC) measurements at multiple contiguous positions, and (3) air kerma profiles obtained through the continuous translation of a solid-state detector. The resulting CTDIfreeair was used to calculate the CTDIw , per the extended formalism, and compared. The LIC indicated that a 40 mm nT should not be excluded from the extension of the CTDIw formalism. The solid-state detector differed by as much as 8% compared to the LIC. The PIC was the most straightforward method and gave equivalent results to the LIC. The CTDIw calculated with the latest CTDI formalism will differ most for 160 mm nTs (e.g., whole-organ perfusion or coronary CT angiography) compared to the previous CTDI formalism. Inaccuracies in the measurement of CTDIfreeair will subsequently manifest themselves as erroneous calculations of the CTDIw , for nTs greater than 40 mm, with the latest CTDI formalism. The PIC was found to be the most clinically feasible method and was validated against the LIC.


Comparison of Primary Midline Closure, Limberg Flap, and Karydakis Flap Techniques in Pilonidal Sinus Surgery.
BACKGROUND Pilonidal sinus (PS) is a common disease of the sacrococcygeal-natal region. There are many treatment options, but there is still no consensus on the ideal treatment. We compared the results of our PS patients who were treated with primary midline closure (PMC), Limberg flap repair (LFR), and Karydakis flap (KF). MATERIAL AND METHODS The data for 924 PS patients from 2013 to 2017 were retrospectively examined. Demographic data, surgical procedures, schedules, and recurrence rates were examined. RESULTS The mean age was 28.4 years (14-77 years), 82.5% were male (n=762), and 17.5% were female (n=162). PMC was performed on 53.7% (n=496) of the patients, 32.5% (n=300) received LFR, and 13.9% (n=128) underwent KF. PMC was the first choice among females but LFR was the first choice in recurrent patients. The recurrence rate was 10.8% in the PMC group, 8% in the LFR group, and 3.1% in the KF group. In Short Form Survey-36 (SF-36) scores, the best cosmetic outcomes were observed in cases of PMC (p<0.05). Overall, wound dehiscence (WD) was observed in 7.5%, surgical site infection (SSI) in 2.4%, and seroma in 8.5% of all patients. The KF group had the lowest complication rates (p<0.01). CONCLUSIONS According to the results of this study, the reason for preferring PMC among women is cosmetic concerns. PMC still remains important for treatment, but it should be noted that the recurrence rates due to inadequate excision are mostly observed in cases of PMC. Considering their low recurrence rates, LFR or KF should be considered first. When low recurrence rates, patient comfort, and cosmetic results are evaluated together, KF in particular emerges as a method preferred by physicians and patients.


2-5A antisense directed against telomerase RNA produces apoptosis in ovarian cancer cells.
RNase L is converted to an active form upon binding short 2',5'-oligoadenylates (2-5A). To direct RNase L to an RNA target, 2-5A is attached to an antisense oligonucleotide (2-5A antisense). This chimera can be directed against telomerase-an RNA-protein complex that elongates telomeric DNA and is involved in cellular immortalization. Our objective is to investigate the effect of 2-5A antisense by targeting telomerase RNA (hTR) in the ovarian cancer cell line, HEY-1B. Baseline RNase L levels and telomerase activities were measured in both HEY-1B and normal ovarian epithelial cells (NOE). Cells were treated daily with chimeric oligonuclotides (ODN) directed against four different hTR sites, or control ODNs including nonchimeric antisense, 2-5A fused to a mismatched sequence, or inactive 2-5A fused to antisense. At 48 h, apoptosis was evaluated using the TUNEL assay. After six daily ODN administrations, telomerase activity was redetermined, and at 7 days viability counts were obtained. Both cell lines expressed similar levels of RNase L. Hey-1B displayed telomerase activity while NOE did not. After 7 days of transfection, 2-5A antisense ODNs caused profound cell death in the HEY-1B cells, but not in the NOE cells. This effect was seen regardless of hTR target site, and ODN controls showed no significant decrease in cell viability in either cell line. HEY1B cells treated with 2-5A antisense against hTR showed a decrease in telomerase activity and a profound induction of programmed cell death. The results suggest that 2-5A antisense directed against telomerase RNA results in apoptotic cell death in ovarian cancer cells, but not normal ovarian epithelial cells. The 2-5A antisense strategy may hold a considerable advantage over the conventional antisense approach in targeting cancer-causing genes.


Root Rot of Pea and Faba Bean in Southern Sweden Caused by Phytophthora pisi sp. nov.
A root rot disease of pea and faba bean caused by a Phytophthora sp. was observed in fields and field soil samples in southern Sweden. Observations of the disease in pea root rot greenhouse assays were systematically recorded, and incidence and geographic distribution data were compared with the pea root rot caused by Aphanomyces euteiches. Following one successful isolation of the pathogen, isolation procedures and selective media were optimized to retrieve more isolates. Phylogenetic analysis showed that the isolates belong to a novel lineage, closely related to Phytophthora sojae, and proposed here as a new species, P. pisi sp. nov. In a collection of 13 isolates from separate fields, intraspecific variation was detected in both nuclear and mitochondrial loci. Pathogenicity tests on a range of crop plants and wild legumes suggest that the host range of the pathogen is restricted to a group of legumes closely related to pea which, in addition to pea, include the crop species faba bean, lentil, common vetch, and chickpea. Morphology, growth requirements, and pathogenicity traits indicate that the species may be identical to the organism previously described as P. erythroseptica var. pisi. The work characterizes a novel Phytophthora sp. causing root rot of legume crops.


Prevalence of periodontitis in the Indian population: A literature review.
Periodontitis is one of the major reasons for tooth loss in adults. India, with a population of over 1 billion, is bound to become a developed nation soon. This transition will require a population that is healthy, including in terms of periodontal health. Early studies done in India gave an indication that the population is highly susceptible to periodontitis. This paper reviews the prevalence of periodontitis in the Indian population. Review of periodontitis prevalence studies on the Indian population. After identifying articles from PubMed and hand searching, the epidemiology of periodontitis is reviewed together with the case definition, study settings, type of population, age and other factors as all these are bound to affect the prevalence rates determined in various studies. None. This review identifies very limited number of studies that provide prevalence data and faces difficulty in comparing various studies due to nonstandardization of case definition and use of nonrepresentative samples. There is a high prevalence of periodontitis among the adults and the economically weak population. There is a very urgent need for standardized population-based studies with a robust design to identify the true prevalence of periodontitis, which in turn will help in planning oral health policies and creating the necessary infrastructure.


Expression pattern of serine protease inhibitor kazal type 3 (Spink3) during mouse embryonic development.
Recent evidence shows that the serine protease inhibitor Kazal type 3 (Spink3) has more diverse functions than expected. To gain insight into its function, we analyzed the spatiotemporal expression profile of Spink3, using in situ hybridization (ISH) and a Spink3+/lacZ knock-in mouse, in which lacZ was inserted into the Spink3 locus. Spink3lacZ expression was first observed in the foregut, midgut, hindgut and the forebrain/midbrain junction region at 9.5 days post coitus (dpc). In the pancreas, Spink3 mRNA was detected at 11.5 dpc, before formation of the typical shape of the exocrine structure of the pancreas. Acinar cell expression was clearly identified by 13.5 dpc. After differentiation of the intestinal tract, Spink3lacZ expression was observed in the large intestine at 11.5 dpc, followed by expression in the small intestine at 13.5 dpc, before appearance of intestinal digestive enzymes. Spink3 mRNA and Spink3lacZ activity were also detected in other tissues, including the mesonephric tubules and the urogenital ridge at 11.5 dpc, the genital swelling at 13.5 dpc, the ductus epididymis at 17.5 dpc, and the seminal vesicle at 8 weeks. These data suggest that Spink3 may play important roles in proliferation and/or differentiation of various cell types during development.


Correlation of placental pathology with prenatal ultrasound findings.
There have been recent major advances in obstetric ultrasound, regarding both improved technologies and sonographer expertise, which have resulted in changes in antenatal obstetric management. The placenta is routinely examined to some extent at the time of the second trimester fetal anomaly sonogram, timing of delivery in pregnancies complicated by intrauterine growth restriction is primarily dependent on Doppler sonographic assessment of umbilical and uterine artery blood flow, and an increasing number of specific placental lesions have been described. Many non-specialist diagnostic histopathologists may be unfamiliar with these obstetric advances, but they are an increasingly common indication for submission of placentas for histological examination. Since the aims of pathological examination of the placenta are to determine the pathological basis for the clinical findings and advance understanding of the pathophysiology of pregnancy complications, this review therefore provides an overview of the most common prenatal sonographic techniques and their clinical relevance to the diagnostic pathologist, primarily focusing on conditions with specific placental implications. These range from abnormalities of placental site and cord insertion, to obstetric complications such as antepartum haemorrhage, through sonographic placental parenchymal lesions such as subchorionic and intervillous thrombi, or chorioangiomata. In addition, the pathophysiological basis of abnormal maternal and fetal maternal Doppler indices and intrauterine growth restriction are now described, being associated with decidual vasculopathy and villous changes associated with reduced intervillous blood flow respectively. Finally, rare but characteristic, sonographic appearances of villous cystic or hydropic change, may be associated with intrinsic developmental placental abnormalities such as hydatidiform mole and placental mesenchymal dysplasia, which require histological examination for their specific diagnosis.


Virulence of Bordetella bronchiseptica in the porcine respiratory tract.
The virulence of Bordetella bronchiseptica in gnotobiotic piglets was studied by intranasal infection with 11 cultures derived from eight strains isolated from pigs (4), dogs (2), a human subject and a monkey. Six of the cultures contained organisms in phase I and five contained phenotypically different phase-III or -IV organisms. Of the phase-III and -IV cultures, four were derived from strains that had been isolated in phase I. Colonisation of the nasal cavity was investigated by counting bacteria in nasal swabs and washings. The toxigenicity of cell extracts from each strain and variant was determined by tests of lethality in mice or of cytopathogenicity in cell cultures. The results showed that two phase-I cultures from pigs colonised the nasal cavity and respiratory tract of gnotobiotic piglets better than did four phase-I cultures from other species. Phase-I organisms invariably produced capsules, fimbriae and mannose-resistant haemagglutination of guinea-pig erythrocytes. Four of five cultures in phases III and IV consisted of organisms that did not produce capsules, fimbriae or haemagglutination and colonised the nasal cavity poorly. Phase variation from I to III occurred in culture and in vivo, but variation from III to I occurred in vivo only and was accompanied by enhanced colonisation. Gnotobiotic piglets infected with porcine phase-I organisms exhibited atrophy of the nasal turbinate bones after 28 days; these organisms produced significantly more toxin than did bacteria in phase I from other species, or those in phases III and IV.(ABSTRACT TRUNCATED AT 250 WORDS)


MANTIS: a phylogenetic framework for multi-species genome comparisons.
Practitioners of comparative genomics face huge analytical challenges as whole genome sequences and functional/expression data accumulate. Furthermore, the field would greatly benefit from a better integration of this wealth of data with evolutionary concepts. Here, we present MANTIS, a relational database for the analysis of (i) gains and losses of genes on specific branches of the metazoan phylogeny, (ii) reconstructed genome content of ancestral species and (iii) over- or under-representation of functions/processes and tissue specificity of gained, duplicated and lost genes. MANTIS estimates the most likely positions of gene losses on the true phylogeny using a maximum-likelihood function. A user-friendly interface and an extensive query system allow to investigate questions pertaining to gene identity, phylogenetic mapping and function/expression parameters. MANTIS is freely available at http://www.mantisdb.org and constitutes the missing link between multi-species genome comparisons and functional analyses.


Macrophage-derived Hedgehog ligands promotes fibrogenic and angiogenic responses in human schistosomiasis mansoni.
Schistosomiasis mansoni is a major cause of portal fibrosis and portal hypertension. The Hedgehog pathway regulates fibrogenic repair in some types of liver injury. Determine if Hedgehog pathway activation occurs during fibrosis progression in schistosomiasis and to determine if macrophage-related mechanisms are involved. Immunohistochemistry was used to characterize the cells that generate and respond to Hedgehog ligands in 28 liver biopsies from patients with different grades of schistosomiasis fibrosis staged by ultrasound. Cultured macrophages (RAW264.7 and primary rat Kupffer cells) and primary rat liver sinusoidal endothelial cells (LSEC) were treated with schistosome egg antigen (SEA) and evaluated using qRT-PCR. Inhibition of the Hedgehog pathway was used to investigate its role in alternative activation of macrophages (M2) and vascular tube formation. Patients with schistosomiasis expressed more ligands (Shh and Ihh) and target genes (Patched and Gli2) than healthy individuals. Activated LSEC and myofibroblasts were Hedgehog responsive [Gli2(+)] and accumulated in parallel with fibrosis stage (P < 0.05). Double IHC for Ihh/CD68 showed that Ihh(+) cells were macrophages. In vitro studies demonstrated that SEA-stimulated macrophages to express Ihh and Shh mRNA (P < 0.05). Conditioned media from such macrophages induced luciferase production by Shh-LightII cells (P < 0.001) and Hedgehog inhibitors blocked this effect (P < 0.001). SEA-treated macrophages also up-regulated their own expression of M2 markers, and Hh pathway inhibitors abrogated this response (P < 0.01). Inhibition of the Hedgehog pathway in LSEC blocked SEA-induced migration and tube formation. SEA stimulates liver macrophages to produce Hh ligands, which promote alternative activation of macrophages, fibrogenesis and vascular remodelling in schistosomiasis.


The Drosophila retinoblastoma binding protein 6 family member has two isoforms and is potentially involved in embryonic patterning.
The human retinoblastoma binding protein 6 (RBBP6) is implicated in esophageal, lung, hepatocellular and colon cancers. Furthermore, RBBP6 was identified as a strong marker for colon cancer prognosis and as a predisposing factor in familial myeloproliferative neoplasms. Functionally, the mammalian protein interacts with p53 and enhances the activity of Mdm2, the prototypical negative regulator of p53. However, since RBBP6 (known as PACT in mice) exists in multiple isoforms and pact-/- mice exhibit a more severe phenotype than mdm2-/- mutants, it must possess some Mdm2-independent functions. The function of the invertebrate homologue is poorly understood. This is complicated by the absence of the Mdm2 gene in both Drosophila and Caenorhabditis elegans. We have experimentally identified the promoter region of Snama, the Drosophila homologue, analyzed potential transcription factor binding sites and confirmed the existence of an additional isoform. Using band shift and co-immunoprecipitation assays combined with mass spectrometry, we found evidence that this gene may be regulated by, amongst others, DREF, which regulates hundreds of genes related to cell proliferation. The potential transcription factors for Snama fall into distinct functional groups, including anteroposterior embryonic patterning and nucleic acid metabolism. Significantly, previous work in mice shows that pact-/- induces an anteroposterior phenotype in embryos when rescued by simultaneous deletion of p53. Taken together, these observations indicate the significance of RBBP6 proteins in carcinogenesis and in developmental defects.


Noninvasive Doppler-derived myocardial performance index in rats with myocardial infarction: validation and correlation by conductance catheter.
The rodent model of myocardial infarction (MI) is extensively used in heart failure studies. However, long-term follow-up of echocardiographic left ventricular (LV) function parameters such as the myocardial performance index (MPI) and its ratio with the fractional shortening (LVFS/MPI) has not been validated in conjunction with invasive indexes, such as those derived from the conductance catheter (CC). Sprague-Dawley rats with left anterior descending coronary artery ligation (MI group, n = 9) were compared with a sham-operated control group (n = 10) without MI. Transthoracic echocardiography (TTE) was performed every 2 wk over an 8-wk period, after which classic TTE parameters, especially MPI and LVFS/MPI, were compared with invasive indexes obtained by using a CC. Serial TTE data showed significant alterations in the majority of the noninvasive functional and structural parameters (classic and novel) studied in the presence of MI. Both MPI and LVFS/MPI significantly (P < 0.05 for all reported values) correlated with body weight (r = -0.58 and 0.76 for MPI and LVFS/MPI, respectively), preload recruitable stroke work (r = -0.61 and 0.63), LV end-diastolic pressure (LVEDP) (r = 0.82 and -0.80), end-diastolic volume (r = 0.61 and -0.58), and end-systolic volume (r = 0.46 and -0.48). Forward stepwise linear regression analysis revealed that, of all variables tested, LVEDP was the only independent determinant of MPI (r = 0.84) and LVFS/MPI (r = 0.83). We conclude that MPI and LVFS/MPI correlate strongly and better than the classic noninvasive TTE parameters with established, invasively assessed indexes of contractility, preload, and volumetry. These findings support the use of these two new noninvasive indexes for long-term analysis of the post-MI LV remodeling.


Caring to Learn and Learning to Care.
The increasing numbers of aging and chronically ill prisoners incarcerated in Western nations is well-documented, as is the growing need for prison-based palliative and end-of-life care. Less often discussed is specifically how end-of-life care can and should be provided, by whom, and with what resources. One strategy incorporates prisoner volunteers into end-of-life services within a peer-care program. This article reports on one such program based on focused ethnographic study including in-depth interviews with inmate hospice volunteers, nursing staff, and corrections officers working in the hospice program. We describe how inmate volunteers learn hospice care through formal education and training, supervised practice, guidance from more experienced inmates, and support from correctional staff. We discuss how emergent values of mentorship and stewardship are seen by volunteers and staff as integral to prison hospice sustainability and discuss implications of this volunteer-centric model for response-ability for the end-of-life care of prisoners.


Mentalizing or submentalizing in a communication task? Evidence from autism and a camera control.
In the director task (DT), participants are instructed to move objects within a grid of shelves while ignoring those objects that cannot be seen by a human figure, the "director," located beyond the shelves. It is widely assumed that, since they are explicitly instructed to do, participants use mentalizing in this communicative task; they represent what the director can see, and therefore the DT provides important information about how and when mentalizing is used in adult life. We tested this view against a "submentalizing" hypothesis suggesting that DT performance depends on object-centered spatial coding, without mentalizing. As predicted by the submentalizing account, we found that DT performance was unchanged when the director was replaced by an inanimate object, a camera, and that participants with autism spectrum disorders were unimpaired, relative to matched control participants, in both the director and camera conditions. In combination with recent critical analyses of "implicit mentalizing," these findings support the view that adults use mentalizing sparingly in psychological experiments and in everyday life.


Ventromedial prefrontal volume predicts understanding of others and social network size.
Cognitive abilities such as Theory of Mind (ToM), and more generally mentalizing competences, are central to human sociality. Neuroimaging has associated these abilities with specific brain regions including temporo-parietal junction, superior temporal sulcus, frontal pole, and ventromedial prefrontal cortex. Previous studies have shown both that mentalizing competence, indexed as the ability to correctly understand others' belief states, is associated with social network size and that social group size is correlated with frontal lobe volume across primate species (the social brain hypothesis). Given this, we predicted that both mentalizing competences and the number of social relationships a person can maintain simultaneously will be a function of gray matter volume in these regions associated with conventional Theory of Mind. We used voxel-based morphometry of Magnetic Resonance Images (MRIs) to test this hypothesis in humans. Specifically, we regressed individuals' mentalizing competences and social network sizes against gray matter volume. This revealed that gray matter volume in bilateral posterior frontal pole and left temporoparietal junction and superior temporal sucus varies parametrically with mentalizing competence. Furthermore, gray matter volume in the medial orbitofrontal cortex and the ventral portion of medial frontal gyrus, varied parametrically with both mentalizing competence and social network size, demonstrating a shared neural basis for these very different facets of sociality. These findings provide the first fine-grained anatomical support for the social brain hypothesis. As such, they have important implications for our understanding of the constraints limiting social cognition and social network size in humans, as well as for our understanding of how such abilities evolved across primates.


Symptomatic glenoid loosening complicating total shoulder arthroplasty.
Glenoid component loosening is one of the most common causes of failed total shoulder arthroplasty. Previous reports indicate that it is desirable to reimplant the glenoid component during revision shoulder arthroplasty. The purpose of our study was to retrospectively evaluate the satisfaction of patients undergoing glenoid revision (reimplantation or resection) following total shoulder replacement specifically for symptomatic glenoid loosening. Twenty-eight shoulders that developed symptomatic glenoid loosening following primary total shoulder arthroplasty were included in the study. Patients were retrospectively evaluated at a minimum of 2 years postoperatively. Patients either underwent resection followed by reimplantation of the glenoid component (13) or resection of the component with or without bone grafting (15). Each patient was evaluated with the UCLA Shoulder Scale and the Constant-Murley Shoulder Assessment. There were seven excellent, 13 good, five fair and three poor results on the UCLA score. Functional outcome scores trended higher in the reimplantation group but were not statistically significant. Both groups reported equal pain relief and satisfaction. Five out of 15 patients underwent arthroscopic resection of the glenoid, and these patients scored as well on the UCLA and Constant scores as the reimplantation group. When symptomatic glenoid loosening is the indication for revision total shoulder replacement, patients tend to achieve good to excellent results. Though functional scores were slightly higher in the reimplantation group, satisfaction was equally high in both groups. Resection, when indicated, should be performed arthroscopically as this improved functional outcome in our series.


Prenatal exposure to BPA alters the epigenome of the rat mammary gland and increases the propensity to neoplastic development.
Exposure to environmental estrogens (xenoestrogens) may play a causal role in the increased breast cancer incidence which has been observed in Europe and the US over the last 50 years. The xenoestrogen bisphenol A (BPA) leaches from plastic food/beverage containers and dental materials. Fetal exposure to BPA induces preneoplastic and neoplastic lesions in the adult rat mammary gland. Previous results suggest that BPA acts through the estrogen receptors which are detected exclusively in the mesenchyme during the exposure period by directly altering gene expression, leading to alterations of the reciprocal interactions between mesenchyme and epithelium. This initiates a long sequence of altered morphogenetic events leading to neoplastic transformation. Additionally, BPA induces epigenetic changes in some tissues. To explore this mechanism in the mammary gland, Wistar-Furth rats were exposed subcutaneously via osmotic pumps to vehicle or 250 µg BPA/kg BW/day, a dose that induced ductal carcinomas in situ. Females exposed from gestational day 9 to postnatal day (PND) 1 were sacrificed at PND4, PND21 and at first estrus after PND50. Genomic DNA (gDNA) was isolated from the mammary tissue and immuno-precipitated using anti-5-methylcytosine antibodies. Detection and quantification of gDNA methylation status using the Nimblegen ChIP array revealed 7412 differentially methylated gDNA segments (out of 58207 segments), with the majority of changes occurring at PND21. Transcriptomal analysis revealed that the majority of gene expression differences between BPA- and vehicle-treated animals were observed later (PND50). BPA exposure resulted in higher levels of pro-activation histone H3K4 trimethylation at the transcriptional initiation site of the alpha-lactalbumin gene at PND4, concomitantly enhancing mRNA expression of this gene. These results show that fetal BPA exposure triggers changes in the postnatal and adult mammary gland epigenome and alters gene expression patterns. These events may contribute to the development of pre-neoplastic and neoplastic lesions that manifest during adulthood.


Insomnia: conceptual issues in the development, persistence, and treatment of sleep disorder in adults.
This paper critically reviews the evidence base for previously reported conceptual models of the development and persistence of insomnia. Although a number of perspectives have some empirical support, no one approach emerges as preeminent. Importantly, the efficacy of any particular psychological intervention cannot be taken as confirmation of presumed, underlying mechanisms. An integrated psychobiological inhibition model of insomnia is developed that accounts for the research data. The model views insomnia as arising from inhibition of de-arousal processes associated with normal sleep. It is proposed that sleep homeostatic and circadian factors are compromised by impairment of the automaticity and plasticity associated with good sleep, and that cognitive/affective processes activate the clinical complaint of insomnia. Common pathways for the action of cognitive-behavioral interventions are identified, and a research agenda is set for further conceptual and clinical study.


Rational learning and information sampling: on the "naivety" assumption in sampling explanations of judgment biases.
Recent research has argued that several well-known judgment biases may be due to biases in the available information sample rather than to biased information processing. Most of these sample-based explanations assume that decision makers are "naive": They are not aware of the biases in the available information sample and do not correct for them. Here, we show that this "naivety" assumption is not necessary. Systematically biased judgments can emerge even when decision makers process available information perfectly and are also aware of how the information sample has been generated. Specifically, we develop a rational analysis of Denrell's (2005) experience sampling model, and we prove that when information search is interested rather than disinterested, even rational information sampling and processing can give rise to systematic patterns of errors in judgments. Our results illustrate that a tendency to favor alternatives for which outcome information is more accessible can be consistent with rational behavior. The model offers a rational explanation for behaviors that had previously been attributed to cognitive and motivational biases, such as the in-group bias or the tendency to prefer popular alternatives.


The Spatial Landscape of Progression and Immunoediting in Primary Melanoma at Single-Cell Resolution.
Cutaneous melanoma is a highly immunogenic malignancy that is surgically curable at early stages but life-threatening when metastatic. Here we integrate high-plex imaging, 3D high-resolution microscopy, and spatially resolved microregion transcriptomics to study immune evasion and immunoediting in primary melanoma. We find that recurrent cellular neighborhoods involving tumor, immune, and stromal cells change significantly along a progression axis involving precursor states, melanoma in situ, and invasive tumor. Hallmarks of immunosuppression are already detectable in precursor regions. When tumors become locally invasive, a consolidated and spatially restricted suppressive environment forms along the tumor-stromal boundary. This environment is established by cytokine gradients that promote expression of MHC-II and IDO1, and by PD1-PDL1-mediated cell contacts involving macrophages, dendritic cells, and T cells. A few millimeters away, cytotoxic T cells synapse with melanoma cells in fields of tumor regression. Thus, invasion and immunoediting can coexist within a few millimeters of each other in a single specimen. The reorganization of the tumor ecosystem in primary melanoma is an excellent setting in which to study immunoediting and immune evasion. Guided by classic histopathology, spatial profiling of proteins and mRNA reveals recurrent morphologic and molecular features of tumor evolution that involve localized paracrine cytokine signaling and direct cell-cell contact. This article is highlighted in the In This Issue feature, p. 1397.


Recombinant human granulocyte colony-stimulating factor infusion and/or autologous peripheral blood stem cell transplantation as a rescue for sequential high-dose combination chemotherapy: a preliminary report.
Sequential cycles of combination chemotherapy with high-dose cyclophosphamide, etoposide and cisplatin (sHDCEP) can largely increase the total dose (TD) of drug delivered. If granulocyte colony-stimulating factor (G-CSF) and/or autologous peripheral blood stem cell (PBSC) rescue can shorten the duration of cytopenia between cycles of sHDCEP, the dose intensity (DI) can be increased as well. In order to explore the feasibility of delivering maximal TD and DI by administration of sHDCEP with G-CSF and/or PBSC rescue, this trial is undertaken to investigate the hematologic and nonhematologic toxicity observed with sHDCEP by G-CSF and/or PBSC rescue. Patients with refractory malignancy and well preserved physiologic function for whom no available therapy is likely to cure or prolong the survival were eligible for the study. Each cycle of high-dose chemotherapy consisted of: cyclophosphamide 5,000 mg/m2, etoposide 1,500 mg/m2 and cisplatin 150 mg/m2. G-CSF and/or PBSC were administered alternatively after each cycle as rescue for myelosuppression. The next cycle was given to patient who showed response to the previous cycle after recovery from toxicity for a maximal of 4 cycles. Two cases of refractory malignancy with progressive disease were treated by sHDCEP for 7 cycles, including 4 cycles with G-CSF rescue, 2 cycle with PBSC rescue, and 1 cycle with G-CSF + PBSC rescue. In the 4 cycles rescued by G-CSF alone, we observed a slightly slower granulocyte and markedly prolonged platelet recovery in the subsequent cycle. By comparing the effect of G-CSF and/or PBSC rescue on hematologic recovery with the preceding cycle in the same patient, we found that G-CSF rescue provided faster granulocyte recovery than PBSC, but PBSC rescue provided faster platelet recovery than G-CSF. Rescue by larger number of PBSCs provided only faster platelet but not granulocyte recovery than rescue by adding G-CSF to a very small number of PBSCs. However, G-CSF plus the very small number of PBSCs provided shorter duration of both granulocytopenia and thrombocytopenia than rescue by G-CSF alone. The most common nonhematologic toxicity from sHDCEP included transient nausea, vomiting, diarrhea and mild impairment of liver function but we observed no significant or irreversible major organ damage. The side effect from PBSC collection was mild and toxicity from reinfusion of the thawed PBSCs was not obvious. Using G-CSF and/or PBSC rescue, sHDCEP was delivered repeatedly in no more than 4 weeks for the next-cycles except for patient 1 who had cycle 4 delayed because of prolonged platelet recovery by only G-CSF rescue in cycle 3. Our initial experience has shown that the nonhematologic toxicity from sHDCEP, G-CSF and PBSC rescue was well tolerated. Prolonged platelet recovery after sequential cycles of HDCEP by only G-CSF rescue delayed the next cycle of chemotherapy. Although the next cycle was delivered within 4 weeks by only PBSC rescue, concurrent infusion of G-CSF and larger number of PBSCs should provide the most rapid hematologic recovery. Sequential high-dose chemotherapy administered by this model is likely to provide the maximal delivery of TD and DI, and is worthy of further clinical trials.


Comparative effectiveness of revascularization strategies.
Questions persist concerning the comparative effectiveness of percutaneous coronary intervention (PCI) and coronary-artery bypass grafting (CABG). The American College of Cardiology Foundation (ACCF) and the Society of Thoracic Surgeons (STS) collaborated to compare the rates of long-term survival after PCI and CABG. We linked the ACCF National Cardiovascular Data Registry and the STS Adult Cardiac Surgery Database to claims data from the Centers for Medicare and Medicaid Services for the years 2004 through 2008. Outcomes were compared with the use of propensity scores and inverse-probability-weighting adjustment to reduce treatment-selection bias. Among patients 65 years of age or older who had two-vessel or three-vessel coronary artery disease without acute myocardial infarction, 86,244 underwent CABG and 103,549 underwent PCI. The median follow-up period was 2.67 years. At 1 year, there was no significant difference in adjusted mortality between the groups (6.24% in the CABG group as compared with 6.55% in the PCI group; risk ratio, 0.95; 95% confidence interval [CI], 0.90 to 1.00). At 4 years, there was lower mortality with CABG than with PCI (16.4% vs. 20.8%; risk ratio, 0.79; 95% CI, 0.76 to 0.82). Similar results were noted in multiple subgroups and with the use of several different analytic methods. Residual confounding was assessed by means of a sensitivity analysis. In this observational study, we found that, among older patients with multivessel coronary disease that did not require emergency treatment, there was a long-term survival advantage among patients who underwent CABG as compared with patients who underwent PCI. (Funded by the National Heart, Lung, and Blood Institute.).


Stability and Clinical Outcome after Reconstruction of Complete Triangular Fibrocartilage Disruption.
Purpose Patients with symptomatic instability of the distal radioulnar joint (DRUJ) after traumatic complete disruption of the triangular fibrocartilage complex (TFC) are best treated by anatomic reconstruction of the TFC. Postoperative clinical results from the literature are known but the improvement of DRUJ instability remains still challenging to quantify. We recently published a reliable and validated method to measure the instability of the DRUJ. This sonographic method was used to quantify the pre- and postoperative instability in correlation with clinical outcome in patients with complete TFC disruption. Methods 11 patients with complete disruption of the TFC resulting in symptomatic instability of the DRUJ underwent open reconstruction of the TFC. The instability was measured with sonography preoperatively and one year postoperatively including the Patient-Rated Wrist Evaluation score (PRWE Score). Results By subjective measurement, 9 patients showed comparable stability to the contralateral side. By objective measurements, DRUJ stability was completely restored in 6 patients. Seven patients had a very good and good clinical outcome. The dorsovolar shift (preoperative mean 5.2mm, min 2.4, max 7.1; postoperative 3.6mm, min 1.2, max 6.2) was significantly decreased (p < 0.05) and was postoperatively not different to the contralateral healthy side (p > 0.1). PRWE score in the 1 year follow up was 13.8. Three patients remained with significant pain, sonographically two of them were still more lax and one tighter compared to the contralateral side. Conclusions The sonographic measurement technique allows evaluation of the stability pre- and postoperatively and does not always correlate with the qualitative clinical assessment. The described operation technique is effective for treatment of irreplaceable TFC ruptures and significantly improves the DRUJ stability and wrist function one year after surgery, which could be quantified objectively by ultrasound.


Comparison of the outcomes of non-trapdoor-type blowout fracture repair according to the time of surgery.
We classified the outcomes of non-trapdoor-type blowout fracture repair by surgical indications and then compared the outcomes according to the time of treatment. The medical records of 591 patients with orbital fractures that were treated surgically within 30 days of trauma were included in the study. The enrolled patients were classified into 2 groups by the major surgical indications: 1 group included patients with diplopia or limited extraocular motion, and the other group of patients had significant enophthalmos (>2 mm) or a large fracture on computed tomography (>½). The clinical outcomes were compared between the patients who received surgical repair within 14 days of trauma (early) and those who received treatment from 15 to 30 days after the trauma (delayed) in each group. Two hundred thirty-three patients received surgical repair because of diplopia or limited extraocular motion. Both the early repaired group (n = 195) and the delayed repaired group (n = 38) showed significant improvement after surgeries. The degree of preoperative and postoperative diplopia and limited extraocular motion was not associated with differences between the 2 groups. Four hundred one patients received surgical repair because of enophthalmos (>2 mm) or a large fracture on computed tomography (>½). Both the early repaired group (n = 328) and the delayed repaired group (n = 73) showed significant improvement of the enophthalmos after surgeries. The degree of preoperative/postoperative enophthalmos did not show differences between the 2 groups. If the blowout fracture repairs were performed within a month, the surgical outcomes did not differ according to the time of surgery in the cases of nontrapdoor blowout fracture.


Outcomes After Hemiarthroplasty for Proximal Humerus Fracture Are Significantly Affected by Hand Dominance.
Hand dominance has been reported to be an important factor affecting outcomes after upper extremity trauma but remains unstudied after hemiarthroplasty for fracture. This study determined whether dominance affected outcomes after hemiarthroplasty for proximal humerus fractures. Retrospective cohort study. Tertiary care referral center. Sixty-one patients, after hemiarthroplasty for proximal humerus fracture, returned for comprehensive assessment and were divided into 2 groups: dominant (DOM) shoulder affected (n = 25) and non-dominant (non-DOM) shoulder affected (n = 36). Fracture-specific proximal humeral hemiarthroplasty for displaced proximal humerus fractures. Patients were assessed with self-reported outcomes (visual analog scale pain, American Shoulder and Elbow Surgeons shoulder score, disability of the arm, shoulder, and hand questionnaire, simple shoulder test, and short form 12) and objective (range-of-motion and hand-held dynamometer strength) testing. At 49 months of mean follow-up, there were no significant differences between groups for gender, age, follow-up time, or visual analog scale pain (P > 0.256). The DOM-affected group had significantly worse scores for American Shoulder and Elbow Surgeons shoulder score (P = 0.043), disability of the arm, shoulder, and hand questionnaire (P = 0.039), and simple shoulder test (P = 0.021). The DOM-affected group also had consistently higher correlations between self-reported and objective outcomes than the non-DOM group. Patients who underwent hemiarthroplasty for fracture on their DOM shoulders had significantly poorer outcomes than patients with non-DOM-sided injuries. Although positive outcomes can be expected after hemiarthroplasty, patients should be instructed that they may have less satisfactory function and strength if their injury was on the DOM side. Prognostic Level II. See Instructions for Authors for a complete description of levels of evidence.


Identification of a new locus for autosomal recessive Charcot-Marie-Tooth disease with focally folded myelin on chromosome 11p15.
Autosomal recessive Charcot-Marie-Tooth disease type 4B (CMT4B) is a demyelinating hereditary motor and sensory neuropathy characterized by abnormal folding of myelin sheaths. A locus for CMT4B has previously been mapped to chromosome 11q23 in a southern Italian pedigree. We initially excluded linkage in two Tunisian families with CMT4B to chromosome 11q23, demonstrating genetic heterogeneity within the CMT4B phenotype. Subsequently, using homozygosity mapping and linkage analysis in the largest Tunisian pedigree, we mapped a new locus to chromosome 11p15. A maximum two-point lod score of 6.05 was obtained with the marker D11S1329. Recombination events refined the CMT4B locus region to a 5.6-cM interval between markers D11S1331 and D11S4194. The second Tunisian CMT4B family was excluded from linkage to the new locus, demonstrating the existence of at least a third locus for the CMT4B phenotype.


Impact of Dengue Surveillance Workers on Community Participation and Satisfaction of Dengue Virus Control Measures in Semarang Municipality, Indonesia: A Policy Breakthrough in Public Health Action.
The aim of this study was to assess community participation in Dengue virus control measures, and community satisfaction in the Dengue surveillance workers (DSWs) performance in Semarang municipality after 3 years of empowerment. A cross-sectional survey involved 1,018 selected participants from 12 groups of social roles in 141 villages in Semarang municipality, Indonesia. A direct interview was performed using a structured questionnaire to evaluate the acceptance, and satisfaction of the community towards the DSWs. The data were analyzed descriptively. The majority of the members of the community considered that the DSWs play an important role in reducing Dengue cases, and vectors of the Dengue virus, as well as increasing the community participation in Dengue control measures. The survey showed that DSWs performance, attitudes, and abilities regarding their main tasks were perceived to be good. Overall, people in Semarang municipality were satisfied with the performance of the DSWs, and considered them important enough to be maintained and strengthened in the future so that Dengue could be controlled. This new policy needs to be disseminated to other regions that may encounter the problems associated with Dengue virus.


Comparison of phosphodiesterase type V inhibitors use in eight European cities through analysis of urban wastewater.
In this work a step forward in investigating the use of prescription drugs, namely erectile dysfunction products, at European level was taken by applying the wastewater-based epidemiology approach. 24-h composite samples of untreated wastewater were collected at the entrance of eight wastewater treatment plants serving the catchment within the cities of Bristol, Brussels, Castellón, Copenhagen, Milan, Oslo, Utrecht and Zurich. A validated analytical procedure with direct injection of filtered aliquots by liquid chromatography-tandem mass spectrometry was applied. The target list included the three active pharmaceutical ingredients (sildenafil, tadalafil and vardenafil) together with (bio)transformation products and other analogues. Only sildenafil and its two human urinary metabolites desmethyl- and desethylsildenafil were detected in the samples with concentrations reaching 60 ng L-1. The concentrations were transformed into normalized measured loads and the estimated actual consumption of sildenafil was back-calculated from these loads. In addition, national prescription data from five countries was gathered in the form of the number of prescribed daily doses and transformed into predicted loads for comparison. This comparison resulted in the evidence of a different spatial trend across Europe. In Utrecht and Brussels, prescription data could only partly explain the total amount found in wastewater; whereas in Bristol, the comparison was in agreement; and in Milan and Oslo a lower amount was found in wastewater than expected from the prescription data. This study illustrates the potential of wastewater-based epidemiology to investigate the use of counterfeit medication and rogue online pharmacy sales.


Resistance training, insulin sensitivity and muscle function in the elderly.
Ageing is associated with a loss in both muscle mass and in the metabolic quality of skeletal muscle. This leads to sarcopenia and reduced daily function, as well as to an increased risk for development of insulin resistance and type 2 diabetes. A major part, but not all, of these changes are associated with an age-related decrease in the physical activity level and can be counteracted by increased physical activity of a resistive nature. Strength training has been shown to improve insulin-stimulated glucose uptake in both healthy elderly individuals and patients with manifest diabetes, and likewise to improve muscle strength in both elderly healthy individuals and in elderly individuals with chronic disease. The increased strength is coupled to improved function and a decreased risk for fall injuries and fractures. Elderly individuals have preserved the capacity to improve muscle strength and mass with training, but seem to display a reduced sensitivity towards stimulating protein synthesis from nutritional intake, rather than by any reduced response in protein turnover to exercise.