Adipocyte-induced transdifferentiation of osteoblasts and its potential role in age-related bone loss.
Our preliminary findings have lead us to propose bone marrow adipocyte secretions as new contributors to bone loss. Indeed, using a coculture model based on human bone marrow stromal cells, we previously showed that soluble factors secreted by adipocytes induced the conversion of osteoblasts towards an adipocyte-like phenotype. In this study, microarray gene expression profiling showed profound transcriptomic changes in osteoblasts following coculture and confirmed the enrichment of the adipocyte gene signature. Double immunofluorescence microscopic analyses demonstrated the coexpression of adipogenic and osteoblastic specific markers in individual cells, providing evidence for a transdifferentiation event. At the molecular level, this conversion was associated with upregulated expression levels of reprogramming genes and a decrease in the DNA methylation level. In line with these in vitro results, preliminary immunohistochemical analysis of bone sections revealed adipogenic marker expression in osteoblasts from elderly subjects. Altogether, these data suggest that osteoblast transdifferentiation could contribute to decreased bone mass upon ageing.


Synergistic tumor suppression by a Perilla frutescens-derived methoxyflavanone and anti-cancer tyrosine kinase inhibitors in A549 human lung adenocarcinoma.
Anti-cancer tyrosine kinase inhibitors (TKIs) are effective in many types of cancers including non-small cell lung cancer, while appearance of TKI-resistant tumors suggests a need for the development of their potentiation strategies. We have previously shown that a methoxyflavanone derivative from the Asian medicinal herb Perilla frutescens (Perilla-derived methoxyflavanone; PDMF) shows a prominent anti-tumor activity against A549 human lung adenocarcinoma. Here we show that PDMF and anti-cancer TKIs (nilotinib, bosutinib, dasatinib, and ponatinib) synergistically suppress proliferation of A549 cells. Flow cytometric analysis indicated that co-stimulation with nilotinib (4 μM) and PDMF induced G2/M cell cycle arrest in low PDMF doses (10-50 μM), whereas this combination triggered de novo G1 arrest in higher PDMF dosages (50-125 μM). We also found that co-administration with nilotinib and PDMF significantly suppressed in vivo tumorigenicity of A549 cells in athymic nude mice.


Use of intravascular ultrasound as a "Quality Control" technique during carotid stent-angioplasty: are there risks to its use?
Intravascular ultrasound (IVUS) provides high-resolution vessel imaging and has been shown to improve clinical outcomes when used to assess the technical result of peripheral angioplasty procedures. Our vascular group compared anatomic and clinical outcomes of carotid artery stent-angioplasty (CAS) performed with angiogram monitoring alone, or in combination with IVUS imaging to select stent/balloon diameter and interrogate stent deployment region for residual stenosis. A retrospective review of our carotid stent registry (N=306) identified 220 CAS procedures performed with either a digital C-arm fluoroscopy alone (N=110) or in conjunction with IVUS (N=110) with at least 6-month of clinical follow-up. Outcome measures of procedure time, angioplasty balloon diameter, contrast dye volume, Duplex surveillance testing for recurrent stenosis, and procedure event (death, cardiac, neurologic) rates were compared to assess the risks and benefits of IVUS. All procedures utilized a cerebral protection device deployed prior to IVUS imaging. Procedure times were similar, but IVUS usage resulted in lower (P<0.05) contrast agent volumes due to fewer angiogram runs for stent sizing and verification of adequate stent deployment. IVUS imaging resulted in the use of larger diameter balloons (typically 6 mm) for final stent angioplasty based on distal internal carotid artery (ICA) dia measurements, and identified (P<0.01) more residual stent abnormalities (N=12, 11%) versus CAS with angiogram assessment alone (N=2, 1.8%). No procedural or 30-day cardiac events or deaths occurred. The overall stroke rate was 0.9%; two events (stroke-1; reperfusion injury-1) in the angio+IVUS group (1.8%) and none in the angio alone group. Duplex ultrasound surveillance following CAS demonstrated a higher (P<0.01) incidence of >50% diameter-reducing in-stent stenosis in the angio alone group (11% vs 7% at 1 month ; 24% vs 6% at last surveillance; mean 36 moontha; range: 6-66 months). The quality control of the CAS procedure was enhanced by IVUS imaging which directed stent /balloon sizing and was more accurate than angiography in confirming adequate stent expansion. No IVUS related adverse events occurred. Based on the anatomic information provided by IVUS, larger diameter angioplasty balloons were used which correlated with less residual stenosis after CAS based on duplex ultrasound testing.


Family Caregiver Training Needs and Medicare Home Health Visit Utilization.
Medicare home health providers are now required to deliver family caregiver training, but potential consequences for service intensity are unknown. The objective of this study was to assess how family caregiver training needs affect the number and type of home health visits received. Observational study using linked National Health and Aging Trends Study (NHATS), Outcomes and Assessment Information Set (OASIS), and Medicare claims data. Propensity score adjusted, multivariable logistic, and negative binomial regressions model the relationship between caregivers' training needs and number/type of home health visits. A total of 1217 (weighted n=5,870,905) National Health and Aging Trends Study participants receiving Medicare-funded home health between 2011 and 2016. Number and type of home health visits, from Medicare claims. Family caregivers' training needs, from home health clinician reports. Receipt of nursing visits was more likely when family caregivers had medication management [adjusted odds ratio (aOR): 3.03; 95% confidence interval (CI): 1.06, 8.68] or household chore training needs (aOR: 3.38; 95% CI: 1.33, 8.59). Receipt of therapy visits was more likely when caregivers had self-care training needs (aOR: 1.70; 95% CI: 1.01, 2.86). Receipt of aide visits was more likely when caregivers had household chore (aOR: 3.54; 95% CI: 1.82, 6.92) or self-care training needs (aOR: 2.12; 95% CI: 1.11, 4.05). Medication management training needs were associated with receiving an additional 1.06 (95% CI: 0.11, 2.01) nursing visits, and household chores training needs were associated with an additional 3.24 total (95% CI: 0.21, 6.28) and 1.32 aide visits (95% CI: 0.36, 2.27). Family caregivers' activity-specific training needs may affect home health visit utilization.


Optogenetic micro-electrocorticography for modulating and localizing cerebral cortex activity.
Spatial localization of neural activity from within the brain with electrocorticography (ECoG) and electroencephalography remains a challenge in clinical and research settings, and while microfabricated ECoG (micro-ECoG) array technology continues to improve, complementary methods to simultaneously modulate cortical activity while recording are needed. We developed a neural interface utilizing optogenetics, cranial windowing, and micro-ECoG arrays fabricated on a transparent polymer. This approach enabled us to directly modulate neural activity at known locations around micro-ECoG arrays in mice expressing Channelrhodopsin-2. We applied photostimuli varying in time, space and frequency to the cortical surface, and we targeted multiple depths within the cortex using an optical fiber while recording micro-ECoG signals. Negative potentials of up to 1.5 mV were evoked by photostimuli applied to the entire cortical window, while focally applied photostimuli evoked spatially localized micro-ECoG potentials. Two simultaneously applied focal stimuli could be separated, depending on the distance between them. Photostimuli applied within the cortex with an optical fiber evoked more complex micro-ECoG potentials with multiple positive and negative peaks whose relative amplitudes depended on the depth of the fiber. Optogenetic ECoG has potential applications in the study of epilepsy, cortical dynamics, and neuroprostheses.


A novel task for examining the neural basis of Theory of Mind deficits in bipolar disorder.
Deficits in theory of mind (ToM) processing have been observed in people with bipolar disorder (BD), but the neural basis of these deficits remains unclear. Here, we studied the relations between neural activation, dysfunctional beliefs and behavioral responses in people with BD during a second-order ToM task. Twenty-five patients and 25 healthy-control participants (HC) underwent functional magnetic resonance imaging (fMRI) while performing a novel ToM task. The Dysfunctional Attitudes scale (DAS) and the Brief Hypomanic Attitudes and Positive Predictions Inventory (BHAPPI) were used to assess dysfunctional beliefs. Significant differences in neural activation were observed between HC and BD patients in regions associated with ToM processing: medial frontal, cingulate, anterior cingulate and superior temporal gyri. Correlations between DAS scores and neural activity in medial frontal and cingulate gyri were observed for HC only. Increased activation in brain regions associated with ToM processing in patients compared to HC provides further evidence of disruption in networks controlling social-cognitive processes. Whether this results from compensatory responses to maintain appropriate behavior is unknown.


An ethnomycological review of psychoactive agarics in Australia and New Zealand.
A comprehensive review is presented of the recreational and accidental ingestion of psychoactive mushrooms in Australia and New Zealand; 15 recognized species are considered from Australia and eight from New Zealand. Common epithets, potency levels, and methods of ingestion are discussed. Legal aspects involving the use of these psychoactive fungi are noted. In addition, medical and psychoactive effects of these mushrooms and treatment for psilocybian mushroom poisoning are described. Numerous case reports, with commentary, are also presented.


Incorporating geriatrics into baccalaureate nursing curricula: laying the groundwork with faculty development.
In June 2001, the John A. Hartford Foundation of New York awarded the American Association of Colleges of Nursing (AACN) a 3.99 million dollar grant to enhance gerontology curriculum development and new clinical experiences in 20 baccalaureate and 10 graduate schools of nursing. Over the 4-year grant implementation period, AACN learned a valuable lesson from the grant's site directors: Faculty development is the single most necessary precursor to the successful implementation and maintenance of geriatric curricular enhancements. Unless faculty members foster positive attitudes toward aging, expand their geriatric nursing knowledge base, and are able to integrate geriatric content into the curricula, progress cannot be made. Enhancing Geriatric Nursing Education project directors recommend that the following steps be taken toward the creation of successful faculty development activities: (1) anoint a champion to mentor and persuade faculty members to embrace gerontology; (2) garner faculty buy-in by engaging the faculty early so that they become active participants in the curricular change process; (3) assess faculty knowledge and comfort level by administering tools developed by the John A. Hartford Foundation Institute for Geriatric Nursing and by conducting surveys based on AACN geriatric core competencies; (4) conduct faculty development workshops that include cutting-edge knowledge and research and provide the faculty with opportunities to discuss feelings and stereotypes about aging; (5) elicit the dean's support to encourage and allow time and opportunities for training; and (6) use the many excellent resources that help the faculty integrate geriatric content into their courses. This article will further elucidate such strategies and will highlight the range of faculty development activities in which grant-funded schools engaged.


Stability of extemporaneous erlotinib, lapatinib, and imatinib oral suspensions.
The stability of extemporaneously prepared erlotinib, lapatinib, and imatinib oral liquid dosage forms using two commercially available vehicles when stored at 4 and 25 °C was evaluated. Three batches of extemporaneous oral suspensions were prepared for each drug. Erlotinib and lapatinib tablets were crushed and mixed in a 1:1 mixture of Ora-Plus:Ora-Sweet solution to yield 10- and 50-mg/mL suspensions, respectively. Imatinib tablets were crushed and mixed in Ora-Sweet solution to yield a 40-mg/mL suspension. Suspensions were stored in amber plastic bottles, and samples from each bottle were obtained on days 0, 1, 3, 7, 14, and 28. Erlotinib 10-mg/mL and lapatinib 50-mg/mL oral suspensions in a 1:1 mixture of Ora-Plus and Ora-Sweet retained at least 90% of their initial concentration throughout the 28-day study when stored at 25 °C. Visual inspection revealed notable viscosity changes in the erlotinib and lapatinib suspensions stored at 4 °C for 7 days and beyond. The viscosity of these preparations increased with time and was particularly evident with the erlotinib suspension, which exhibited a puddinglike texture. Imatinib 40-mg/mL oral suspension in Ora-Sweet appeared stable for up to 14 days when stored at both 25 and 4 °C. Erlotinib 10-mg/mL and lapatinib 50-mg/mL oral suspensions prepared from commercially available tablets were stable for at least 28 days when prepared in a 1:1 mixture of Ora-Plus:Ora-Sweet at 25 °C. Imatinib 40-mg/mL oral suspension prepared from commercially available tablets was stable for up to 14 days when prepared in Ora-Sweet and stored at 25 and 4 °C.


Monoclonal antibodies against Vibrio vulnificus RtxA1 elicit protective immunity through distinct mechanisms.
Vibrio vulnificus causes rapidly progressing septicemia with an extremely high mortality rate (≥50%), even with aggressive antibiotic treatment. The bacteria secrete multifunctional autoprocessing repeats-in-toxin (MARTX) toxins, which are involved in the pathogenesis of Gram-negative Vibrio species. Recently, we reported that immunization with the C-terminal region of V. vulnificus RtxA1/MARTXVv, RtxA1-C, elicits a protective immune response against V. vulnificus through a poorly defined mechanism. In this study, we generated a panel of new monoclonal antibodies (MAbs) against V. vulnificus RtxA1-C and investigated their protective efficacies and mechanisms in a mouse model of infection. Prophylactic administration of seven MAbs strongly protected mice against lethal V. vulnificus infection (more than 90% survival). Moreover, three of these MAbs (21RA, 24RA, and 47RA) demonstrated marked efficacy as postexposure therapy. Notably, 21RA was therapeutically effective against lethal V. vulnificus infection by a variety of routes. Using Fab fragments and a neutropenic mouse model, we showed that 21RA and 24RA mediate protection from V. vulnificus infection through an Fc-independent and/or neutrophil-independent pathway. In contrast, 47RA-mediated protection was dependent on its Fc region and was reduced to 50% in neutropenic mice compared with 21RA-mediated and 24RA-mediated protection. Bacteriological study indicated that 21RA appears to enhance the clearance of V. vulnificus from the blood. Overall, these studies suggest that humoral immunity controls V. vulnificus infection through at least two different mechanisms. Furthermore, our panel of MAbs could provide attractive candidates for the further development of immunoprophylaxis/therapeutics and other therapies against V. vulnificus that target the MARTX toxin.


Trends in family planning and contraception carried out in primary health care in Croatia: 1995-2012: study based on routinely collected data.
The lack of objective findings provided the reason to undertake this study with the main aim of exploring the trends in family planning and contraception carried out in primary health care in Croatia. Based on the Croatian health statistical yearbooks from 1995 to 2012, data were collected on the number and structure of visits and the number and type of prescribed contraceptives by public and private gynecological practices, and visits to family practices and school medicine services. The results showed that only a small number of fertile women received primary care for family planning issues, trends that are evident through the entire follow-up period. A trend of "migration" from the public to the private sector is also observed. Family doctors and school medicine are almost entirely excluded from the provision of family planning despite the magnitude of the family planning problem resulting in a waste of human and material resources.


Hypertension: an unstudied potential risk factor for adverse outcomes during continuous flow ventricular assist device support.
In end-stage heart failure, left ventricular assist devices (LVADs) represent an exciting new frontier in which post-device implantation survival approaches that of heart transplant. However, expansion of this technology is still limited by complications that impact morbidity and mortality. Thus, it is essential to identify and optimize modifiable predictors of poor outcomes. One such predictor may be hypertension (HTN). Not only may chronic HTN as a traditional cardiovascular risk factor be present during long-term LVAD support, but HTN may also contribute to device malfunction or device-associated complications. Although current guidelines identify blood pressure (BP) control as important to outpatient continuous flow (CF) LVAD management, there is no evidence base to support these guidelines. Indeed, our comprehensive literature search did not identify any studies that evaluated post-device implantation HTN as a potential predictor of adverse CF-LVAD outcomes. HTN among CF-LVAD patients is likely a relatively unstudied factor because of difficulties using standard noninvasive techniques to measure BP in the setting of reduced pulsatile flow. Fortunately, recent research has elucidated the meaning of Doppler BP measurements and validated a slow-cuff deflation system for BP measurements in the setting of CF-LVAD support. Therefore, CF-LVAD researchers and clinicians may (1) consider potential mechanisms relating HTN to poor outcomes, (2) realize that HTN management is a stated goal despite scarce evidence, and (3) utilize the new reliable and valid methods for outpatient BP measurement that make research and management possible. It is critical and now feasible that research on HTN in the CF-LVAD patient population move forward.


Replication-fork stalling and processing at a single psoralen interstrand crosslink in Xenopus egg extracts.
Interstrand crosslink (ICL)-inducing agents block the separation of the two DNA strands. They prevent transcription and replication and are used in clinics for the treatment of cancer and skin diseases. Here, we have introduced a single psoralen ICL at a specific site in plasmid DNA using a triplex-forming-oligonucleotide (TFO)-psoralen conjugate and studied its repair in Xenopus egg extracts that support nuclear assembly and replication of plasmid DNA. Replication forks arriving from either side stalled at the psoralen ICL. In contrast to previous observations with other ICL-inducing agents, the leading strands advanced up to the lesion without any prior pausing. Subsequently, incisions were introduced on one parental strand on both sides of the ICL. These incisions could be detected whether one or both forks reached the ICL. Using small molecule inhibitors, we found that the ATR-Chk1 pathway, but not the ATM-Chk2 pathway, stimulated both the incision step and the subsequent processing of the broken replication intermediates. Our results highlight both similarities and differences in fork stalling and repair induced by psoralen and by other ICL-forming agents.


Neuronal evidence for individual eye control in the primate cMRF.
Previous single unit recordings and electrical stimulation have suggested that separate regions of the MRF participate in the control of vergence and conjugate eye movements. Neurons in the supraoculomotor area (SOA) have been found to encode symmetric vergence [Zhang, Y. et al. (1992). J. Neurophysiol., 67: 944-960] while neurons in the central MRF, the cMRF, located ventral to the SOA and lateral to the oculomotor nucleus are associated with conjugate eye movements [Waitzman, D.M. et al. (1996). J. Neurophysiol., 75(4): 1546-1572]. However, it remains unknown if cMRF neurons are strictly associated with conjugate movements since eye movements were recorded with a single eye coil in monkeys viewing visual stimuli at a distance of at least 50 cm. In the current study we addressed whether neurons in the cMRF might also encode vergence-related information. Interestingly, electrical stimulation elicited disconjugate saccades (contralateral eye moved more than the ipsilateral eye) from locations previously thought to elicit only conjugate saccades. Single unit recordings in this same area made in two rhesus monkeys trained to follow visual stimuli moved rapidly in depth along the axis of sight of an individual eye demonstrate that cMRF neurons do not simply encode conjugate information during disconjugate saccades; in fact our findings provide evidence that cMRF neurons are most closely associated with the movement of an individual eye. These results support the hypothesis that the midbrain shapes the activity of the pre-motor saccadic neurons by encoding integrated conjugate and vergence commands.


Strategies for managing Type IV laryngotracheoesophageal clefts at Great Ormond Street Hospital for Children.
To review our strategy for the treatment of patients with Type IV laryngotracheoesophageal cleft-a very rare congenital malformation. Retrospective review of nine cases of Type IV laryngotracheoesophageal cleft managed between October 1994 and January 2004 at Great Ormond Street Hospital for Children, London. Three children were not operated upon (Group A) because of serious co-morbidity and parental decision not to opt for repair; they died at the ages of 2, 7 and 14 days, respectively. Six cases were repaired (Group B) at ages ranging from 2 to 53 days, all using an anterior cervico-thoracic approach. Two cases were operated upon using conventional ventilation, three using cardiopulmonary bypass and one using extracorporeal membrane oxygenation. There was no intra-operative mortality. The number of operative and diagnostic procedures varied from 4 to 37. Two operated cases expired at the ages of 11 days and 25 months; both were operated upon using cardiopulmonary bypass and had significant cardiac co-morbidity. Post-operative microlaryngoscopy and bronchoscopy showed dehiscence in the cleft repair ranging from 1mm to 2cm in five cases. This was observed at the distal end in three patients and just below the vocal cords in two cases. Three cases underwent further repair which was successful. The most recent case repaired using extracorporeal membrane oxygenation required less heparin when compared with those done on cardiopulmonary bypass and had the best post-operative result. Two patients were finally decannulated. The total duration of diagnostic and operative procedures ranged from 9 to 26h and 30min. The hospital stay in the operated cases varied from 9 days to 2 years 2 months 3 days, and their ICU stay ranged from 9 days to 10 months 7 days. All four of our patients with clefts which ended above the carina are alive after multiple repairs whereas all five cases with clefts extending all the way to the carina died (repair was attempted in two). Early diagnosis and repair are essential for successful treatment of Type IV laryngotracheoesophageal clefts. We believe it is best to repair this defect via an anterior cervico-thoracic approach, with or without a median sternotomy, and extracorporeal membrane oxygenation is now our preferred method of gas exchange during such repair. The longest Type IV clefts extending all the way to the carina have the worst prognosis. The decision to operate or not should be based upon the associated co-morbidity and fully informed parental choice, since treatment entails significant morbidity and mortality.


[Treatment of acute rhinitis in neonates and infants].
The authors review current views on the treatment of acute rhinitis (AR) in infants and present their experience in AR treatment of 30 infants aged from 3 days to 1 year. Combined treatment with Physiomer baby quickly eradicated inflammation. The drug eliminates crusts and nasal discharge allowing application of other medication on a clear surface of nasal mucosa. As a result, medication becomes more affective and accelerates recovery. A convenient tip is well adapted for the nostrils. Administration of medicine by spray provides optimal lavage of the nasal cavity.


The role of infection models and PK/PD modelling for optimising care of critically ill patients with severe infections.
Critically ill patients with severe infections are at high risk of suboptimal antimicrobial dosing. The pharmacokinetics (PK) and pharmacodynamics (PD) of antimicrobials in these patients differ significantly from the patient groups from whose data the conventional dosing regimens were developed. Use of such regimens often results in inadequate antimicrobial concentrations at the site of infection and is associated with poor patient outcomes. In this article, we describe the potential of in vitro and in vivo infection models, clinical pharmacokinetic data and pharmacokinetic/pharmacodynamic models to guide the design of more effective antimicrobial dosing regimens. Individualised dosing, based on population PK models and patient factors (e.g. renal function and weight) known to influence antimicrobial PK, increases the probability of achieving therapeutic drug exposures while at the same time avoiding toxic concentrations. When therapeutic drug monitoring (TDM) is applied, early dose adaptation to the needs of the individual patient is possible. TDM is likely to be of particular importance for infected critically ill patients, where profound PK changes are present and prompt appropriate antibiotic therapy is crucial. In the light of the continued high mortality rates in critically ill patients with severe infections, a paradigm shift to refined dosing strategies for antimicrobials is warranted to enhance the probability of achieving drug concentrations that increase the likelihood of clinical success.


Suicide prevention efforts in the United States and their effectiveness.
Suicide is a serious public health problem in the United States, and suicide rates have been increasing for more than a decade. Rural areas are more impacted than urban areas, reinforcing that social, cultural, and economic factors contribute to risk. This article reviews recent work about these contributors to suicide and how they may inform prevention efforts. Current research has shown that suicide is more than a mental health problem with a psychiatric or medical solution. Universal screening and referral by gatekeepers target a large group with a low baseline risk, and there are few treatments proven to reduce death by suicide, as well as a severe shortage of mental health providers in the United States to provide them. Instead, suicide prevention polices can target various other factors that contribute to elevated suicide risk at the population level, including reducing socioeconomic deprivation and access to firearms, both of which are often higher in rural areas. Internet-based interventions also hold promise as they are highly scalable, accessible almost anywhere, and often anonymous. Understanding factors that increase suicide risk guide development of evidence-based policies targeted at high-risk groups. Population-level interventions should be developed in collaboration with the target audience for cultural appropriateness.


Role of E and CArG boxes in developmental regulation of muscle glycogen phosphorylase promoter during myogenesis.
Muscle glycogen phosphorylase (MGP) transcript and protein levels increase during skeletal muscle development in tandem with the products of other muscle genes responsible for glucose and glycogen metabolism. Previous studies demonstrated that a 269 bp region 5' to exon 1 of MGP is sufficient for developmental regulation in the C2C12 myogenic cell line (Froman et al., 1994). This genomic region (-209 to +60) contains four consensus E box motifs, a CArG-like sequence, and a GC-rich domain. Native MGP transcripts were not detected in pluripotent CH310T1/2 fibroblasts, but low levels of MGP mRNA were measured in CH310T1/2 cells that were stably transfected with MyoD. Three of the E box motifs in the MGP proximal promoter interacted with C2C12 nuclear proteins. However, cotransfection of the MGP promoter with myogenic regulatory factors, including MyoD and myogenin, produced less than 2-fold activation compared with 20-fold activation of the desmin promoter. Mutational analyses of the MGP promoter demonstrated that increased expression in C2C12 myotubes did not require any of the E box motifs or the CArG-like element. A small region (-76 to -68) upstream of GC-rich domain (-64 to -51) significantly reduced promoter activities in both myoblasts and myotubes. The functional studies suggest that MGP is developmentally regulated during myogenesis by alternative pathways that utilize unidentified regulatory elements or ancillary factors.


Fate of the antibiotic sulfamethoxazole and its two major human metabolites in a water sediment test.
Sulfonamide antibiotics are widely used in human and veterinary medicine. After their application, they are excreted in unchanged as well as in metabolized form. Due to incomplete elimination in wastewater treatment plants, they can be emitted into surface water. The environmental fate of both parent compounds and metabolites is currently unknown. The aim of this study was to determine the biodegradation potential of river sediment for the sulfonamide sulfamethoxazole (SMX) and its two major human metabolites N4-acetyl-SMX and SMX-N1-glucuronide using a water sediment test system. Each compound was tested in a separate series together with sterile and "water only" controls. All three compounds were efficiently removed from the test system by biodegradation in the sediment. Only for SMX-N1-glucuronide, a substantial removal in the absence of sediment was determined. Dissipation times from the aqueous phase (DT50) between 8.5 and 17.2 days were measured. Sorption to sediment was of minor importance due to the slightly basic pH of the test system. By the application of a mathematical model, biodegradation half-lives in sediment between 3.3 and 25.6 h were calculated for SMX and its metabolites. The results of this study highlight the capability of native river sediment for degrading sulfonamide antibiotics, but also the potential of human metabolites to retransform into parent SMX under environmental conditions. Based on this study, it is unlikely that SMX or its metabolites will persist or accumulate in river sediments under pH conditions where sorption is of minor importance.


The Role of Xanthine Oxidase Inhibitors in Patients with History of Stroke: A Systematic Review.
Xanthine oxidase inhibitors are commonly used to lower uric acid levels in patients with gout. Due to their effects on endothelial function, they have also been investigated for possible benefits for patients with cardiovascular disease. To assess the efficacy and safety of xanthine oxidase inhibitors in the treatment of patients with history of stroke. MEDLINE (1946-June 2017) and EMBASE (1947-June 2017) were queried using the search terms: "allopurinol" OR "febuxostat" OR "xanthine oxidase inhibitor" OR "xanthine oxidase/ antagonists and inhibitors" AND "stroke" OR "cerebral infarction" OR "cerebrovascular accident". Studies appropriate to the objective were evaluated, including five randomized, placebo-controlled, double-blind trials investigating the effect of allopurinol in patients with history of stroke. No articles evaluating the use of febuxostat in this setting were identified. In patients with history of stroke, treatment with allopurinol resulted in improvements in several markers of endothelial function, inflammatory markers, and scores on the Modified Rankin Scale. Study durations ranged from 6 weeks to 1 year, and studies used varying doses of allopurinol. Allopurinol was well tolerated in most studies, with some reports of gastrointestinal adverse effects, headache and rash. Based on the reviewed literature, allopurinol appears to be a promising therapy to improve vascular function and reduce disability in patients who have had a stroke. The benefits seen are in combination with current standard of care treatments with aspirin and lipid-lowering therapy. Larger trials are necessary to better understand the role of allopurinol in patients with history of stroke.


Altered gene expression of c-myc, epidermal growth factor receptor, transforming growth factor-alpha, and c-erb-B2 in an immortalized human breast epithelial cell line, HMT-3522, is associated with decreased growth factor requirements.
Activation of protooncogenes and constitutive secretion of autocrine growth factors are thought to be involved in the uncontrolled growth of cancer cells. We have attempted to elucidate the role of oncogenes and growth factors in the premalignant progression of human breast epithelial cells by using an immortalized, nontumorigenic, near-diploid human mammary epithelial cell line, HMT-3522, derived from a fibrocystic lesion and established in our laboratory. During propagation in tissue culture, the growth factor requirements of the HMT-3522 cells decreased simultaneously with an amplification and overexpression of the c-myc protooncogene. Other protooncogenes related to human breast cancer were unaltered with regard to gene copy number and expression. In passage 118, in which the most important growth factor still was epidermal growth factor (EGF), we were able to isolate an EGF-independent subline (S2). The EGF independence of S2 was accompanied by an overexpression of the mRNAs for epidermal growth factor receptor (EGF-R), transforming growth factor-alpha, and c-erb-B2 as compared to the EGF-dependent subline (S1). Moreover, by application of a blocking anti-EGF-R antibody, growth of S2 cells in EGF-free medium was inhibited significantly, indicating that EGF-R was involved in an autocrine loop probably with transforming growth factor-alpha as ligand. Neither the late passages of S1 cells nor S2 cells were tumorigenic after subcutaneous transplantation to athymic mice. Our results indicate that c-myc amplification and overexpression are correlated with a decreased requirement for growth factors. Even when these alterations are combined with immortalization and EGF independence, they are insufficient for malignant transformation of these human breast epithelial cells.


Adaptation from restricted geometries: the shell inclination of terrestrial gastropods.
The adaptations that occur for support and protection can be studied with regard to the optimal structure that balances these objectives with any imposed constraints. The shell inclination of terrestrial gastropods is an appropriate model to address this problem. In this study, we examined how gastropods improve shell angles to well-balanced ones from geometrically constrained shapes. Our geometric analysis and physical analysis showed that constantly coiled shells are constrained from adopting a well-balanced angle; the shell angle of such basic shells tends to increase as the spire index (shell height/width) increases, although the optimum angle for stability is 90° for flat shells and 0° for tall shells. Furthermore, we estimated the influences of the geometric rule and the functional demands on actual shells by measuring the shell angles of both resting and active snails. We found that terrestrial gastropods have shell angles that are suited for balance. The growth lines of the shells indicated that this adaptation depends on the deflection of the last whorl: the apertures of flat shells are deflected downward, whereas those of tall shells are deflected upward. Our observations of active snails demonstrated that the animals hold their shells at better balanced angles than inactive snails.


Evidence for biosurfactant-induced flow in corners and bacterial spreading in unsaturated porous media.
The spread of pathogenic bacteria in unsaturated porous media, where air and liquid coexist in pore spaces, is the major cause of soil contamination by pathogens, soft rot in plants, food spoilage, and many pulmonary diseases. However, visualization and fundamental understanding of bacterial transport in unsaturated porous media are currently lacking, limiting the ability to address the above contamination- and disease-related issues. Here, we demonstrate a previously unreported mechanism by which bacterial cells are transported in unsaturated porous media. We discover that surfactant-producing bacteria can generate flows along corners through surfactant production that changes the wettability of the solid surface. The corner flow velocity is on the order of several millimeters per hour, which is the same order of magnitude as bacterial swarming, one of the fastest known modes of bacterial surface translocation. We successfully predict the critical corner angle for bacterial corner flow to occur based on the biosurfactant-induced change in the contact angle of the bacterial solution on the solid surface. Furthermore, we demonstrate that bacteria can indeed spread by producing biosurfactants in a model soil, which consists of packed angular grains. In addition, we demonstrate that bacterial corner flow is controlled by quorum sensing, the cell-cell communication process that regulates biosurfactant production. Understanding this previously unappreciated bacterial transport mechanism will enable more accurate predictions of bacterial spreading in soil and other unsaturated porous media.


Antibiotic prescribing on admission to patients with pneumonia and prior outpatient antibiotic treatment: a cohort study on clinical outcome.
Most pneumonia treatment guidelines recommend that prior outpatient antibiotic treatment should be considered when planning inpatient antibiotic regimen. Our purpose was to study in patients admitted for community-acquired pneumonia the mode of continuing antibiotic treatment at the outpatient to inpatient transition and the subsequent clinical course. Retrospective cohort study. Dutch PHARMO Record Linkage System. 7323 patients aged >18 years and hospitalised with pneumonia in the Netherlands between 2004 and 2010. We identified all prescribed antibiotics prior to, during and after hospitalisation. In case of prior outpatient treatment, the continuation of antibiotic treatment on admission was categorised as: no atypical coverage > no atypical coverage; atypical coverage > atypical coverage; no atypical coverage > atypical coverage; and atypical coverage > no atypical coverage. Length of hospital stay, in-hospital mortality and readmission within 30 days. Twenty-two per cent of the patients had received prior outpatient treatment, of which 408 (25%) patients were switched on admission to antibiotics with atypical coverage. There were no differences in length of hospital stay, in-hospital mortality or readmission rate between the four categories of patients with prior outpatient treatment. The adjusted HR for adding atypical coverage versus no atypical coverage was 0.91 (95% CI 0.55 to 1.51) for time to discharge. For in-hospital mortality and readmission within 30 days, the adjusted ORs were 1.09 (95% CI 0.85 to 1.34) and 0.59 (95% CI 0.30 to 1.18), respectively. This study found no association between mode of continuing antibiotic treatment at the outpatient to inpatient transition and relevant clinical outcomes. In particular, adding atypical coverage in patients without prior atypical coverage did not influence the outcome.


Prostaglandin E1 opens the ductus venosus in the newborn lamb.
There is in vitro evidence that the ductus venosus of the fetal and newborn lamb relaxes to administration of E-series prostaglandins. We performed this study to determine whether the ductus venosus of the newborn lamb could be reopened by an infusion of prostaglandin E1. Fifteen lambs between 4 and 28 h of life and six lambs between 42 and 96 h of life had portal angiograms to determine whether the ductus venosus was patent. Ten of the lambs between 4 and 28 h of age had a closed ductus venosus as did all of the lambs between 42 and 96 h of age. The lambs with a closed ductus venosus received an infusion of either prostaglandin E1 or normal saline followed by repeat portal angiography. In the 4- to 28-h-old lambs, prostaglandin E1 opened the ductus venosus in each of the five lambs who received it, while saline did not in any of the five who received it. In the 42- to 96-h-old lambs, neither prostaglandin E1 in four lambs nor saline in two lambs opened the ductus venosus. Prostaglandin E1, then, reopens the ductus venosus in the newborn lamb during the first day of life. We speculate that prostaglandin E1 may be useful in reopening the ductus venosus in certain newborn infants such as those requiring catheterization of the right atrium and those with total anomalous pulmonary venous return below the diaphragm.


Targeting ROR1 inhibits epithelial-mesenchymal transition and metastasis.
Metastasis is responsible for 90% of cancer-related deaths. Strategies are needed that can inhibit the capacity of cancer cells to migrate across the anatomic barriers and colonize distant organs. Here, we show an association between metastasis and expression of a type I receptor tyrosine kinase-like orphan receptor, ROR1, which is expressed during embryogenesis and by various cancers, but not by normal postpartum tissues. We found that expression of ROR1 associates with the epithelial-mesenchymal transition (EMT), which occurs during embryogenesis and cancer metastasis. Breast adenocarcinomas expressing high levels of ROR1 were more likely to have gene expression signatures associated with EMT and had higher rates of relapse and metastasis than breast adenocarcinomas expressing low levels of ROR1. Suppressing expression of ROR1 in metastasis-prone breast cancer cell lines, MDA-MB-231, HS-578T, or BT549, attenuated expression of proteins associated with EMT (e.g., vimentin, SNAIL-1/2, and ZEB1), enhanced expression of E-cadherin, epithelial cytokeratins (e.g., CK-19), and tight junction proteins (e.g., ZO-1), and impaired their migration/invasion capacity in vitro and the metastatic potential of MDA-MB-231 cells in immunodeficient mice. Conversely, transfection of MCF-7 cells to express ROR1 reduced expression of E-cadherin and CK-19, but enhanced the expression of SNAIL-1/2 and vimentin. Treatment of MDA-MB-231 with a monoclonal antibody specific for ROR1 induced downmodulation of vimentin and inhibited cancer cell migration and invasion in vitro and tumor metastasis in vivo. Collectively, this study indicates that ROR1 may regulate EMT and metastasis and that antibodies targeting ROR1 can inhibit cancer progression and metastasis.


[Diagnosis of human lifetime height using foot length in forensic medical examination of corpse fragments].
This review of domestic and foreign literature concerns methods for the rough estimation of human height based on the results of foot measurement. The efficiency of all these methods appears to be very low since they yield differing and mostly unreliable information. Their main drawback is attributable to the poor dependence between the foot length and the height. This situation necessitated a new study in the search for more accurate estimates of the anatomic relationship between the two variables. The comprehensive anthropometric examination included a total of 3457 subjects (2073 men and 1384 women). It was shown that the two genders differ in terms of the height-to-foot length ratio; moreover, the mean value of this ratio varies depending on the foot length of individual subjects in the range from 6.2 to 7.1 (for women) and from 6.54 to 7.49 (for men). The length of the foot is negatively correlated with the coefficient describing its relationship to the height; the coefficient decreases with increasing foot length. Also, it may differ at a constant length of the foot within 0.6 and 0.62 in men and women respectively.


A detailed comparison of the 5'-end of the ovalbumin gene cloned from chicken oviduct and erythrocyte DNA.
We have examined homologous fragments of DNA cloned from two different tissues for changes in the dNA sequence which might be related to tissue specific gene expression. The 5' end of the chicken ovalbumin gene was cloned from oviduct or erythrocyte DNA DNA using cosmids as vectors. We have compared the two clones obtained by restriction enzyme digestions, analysis of heteroduplexes by electron microscopy or S1 nuclease digestion and by DNA sequencing. Our results show that whereas no alteration occured in the region of the gene assumed to be of importance for the control of transcription, a 4 nucleotide deletion/insertion was detected in the first intron of the ovalbumin gene.


Allostatic load and health disparities: a theoretical orientation.
Eliminating racial and ethnic health disparities requires restructuring the biomedical models that have focused on the individual as the level of analysis and emphasized the parts rather than the whole. A recently developed understanding of human physiology and adaptive regulation, constructs of allostasis and allostatic load, provides a theoretical orientation that needs to be explored. Thus, the purpose of this article is to present an orientation of allostasis and allostatic load as a theoretical framework for exploring health disparities. This article will (a) present a general background on the evolution of relevant physiologic theories, (b) offer the general theoretical definitions and explanations of allostasis, allostatic load, and mediation processes, (c) examine empirical evidence for the constructs, and (d) discuss the implications of this orientation for health disparities research.


Case Report: Malignant melanoma in a patient with Crohn's disease treated with ustekinumab.
The cornerstone of inflammatory bowel disease (IBD) treatment is immunomodulators. IBD patients are at increased risk of intestinal and extraintestinal malignancy. Ustekinumab is a fully humanized monoclonal anti-IL12/23 antibody with a good safety profile. Malignancies of breast, colon, head and neck, kidney, prostate, thyroid, and non-melanoma skin cancer have been reported among patients who received ustekinumab. We report the case of a 42-year-old Crohn's patient on long-term treatment with ustekinumab, who developed achromatic malignant melanoma. Crohn's was diagnosed at the age of 15, with upper and lower gastrointestinal involvement and was initially treated with azathioprine (2mg/kg for 4 years) and infliximab (5mg/kg for 6 weeks). Due to ileal obstruction, the patient underwent stricturoplasty and received adalimumab (40mg every other week) for two years. He then discontinued therapy and a year later underwent right hemicolectomy. Adalimumab was reinstituted (40mg every other week) and the patient remained in clinical remission for two years. His overall exposure to adalimumab was four years. Ustekinumab was initiated due to a relapse and after 3 years, an incident of scalp itching led to the diagnosis metastatic achromatic malignant melanoma bearing BRAF V600E mutation. He received targeted therapy with an initial good response. We aim to point out the risk of dermatologic malignancy in IBD patients on long-term immunosuppression and the lifelong and meticulous evaluation that is required.


Prosthetic arthroplasties of the shoulder.
Twenty-two Neer shoulder arthroplasties were performed in rheumatoid and osteoarthritic patients and followed for a minimum of two years, with an average of four years, seven months. In addition to the Neer humeral prosthesis, eight had Neer glenoid components and seven had a subacromial spacer inserted. Two years postoperation, 11 patients had no pain and eight had only mild discomfort. Shoulder function improved more than 100% compared with the preoperative ratings while the range of motion increased relatively little. There was one failure. Although the hemiarthroplasty group scored similarly to the total shoulder group, total shoulder arthroplasty is preferred because of possible further glenoid deterioration. A subacromial spacer is used if the rotator cuff cannot be repaired. The Neer shoulder prosthesis is recommended for the relief of rheumatoid or osteoarthritic shoulder pain.


[Clinical use of sequential ultrafiltration with chronic hemodialysis patients].
Twenty one sequential ultrafiltration dialysis were performed to 14 patients with chronic hemodialysis, with a duration of the dialysis treatment from 2 months to 5 years. Dry ultrafiltration--2.1-5.2 litres filtrate was obtained within 90 minutes. The sequential dialysis has numerous advantages over the usual hemodialysis. Apart from the good tolerance of the patients with overweight, some other indications for its performance was stressed upon in the paper, namely: cardiac asthma, arterial hypertension difficult to correct by hemodialysis and drugs, osteodystrophia, itching during dialysis and after it, vomiting and headache during dialysis and in-between dialysis periods, pains in the legs. The patients tolerated the procedure without any complaints.


Tricks of the mind: Visual hallucinations as disorders of attention.
Visual hallucinations are common across a number of disorders but to date, a unifying pathophysiology underlying these phenomena has not been described. In this manuscript, we combine insights from neuropathological, neuropsychological and neuroimaging studies to propose a testable common neural mechanism for visual hallucinations. We propose that 'simple' visual hallucinations arise from disturbances within regions responsible for the primary processing of visual information, however with no further modulation of perceptual content by attention. In contrast, 'complex' visual hallucinations reflect dysfunction within and between the Attentional Control Networks, leading to the inappropriate interpretation of ambiguous percepts. The incorrect information perceived by hallucinators is often differentially interpreted depending on the time-course and the neuroarchitecture underlying the interpretation. Disorders with 'complex' hallucinations without retained insight are proposed to be associated with a reduction in the activity within the Dorsal Attention Network. The review concludes by showing that a variety of pathological processes can ultimately manifest in any of these three categories, depending on the precise location of the impairment.


[Biological characteristics of Wt1 gene in relation to Ph(+) leukemia cell line K562].
Wt1 is a dual-function gene involved in hematopoiesis, leukemogenesis and prognosis for leukemia. This gene is highly expressed in acute myeloid leukemia (AML) and the progression of chronic myelogenous leukemia (CML). It was reported elsewhere that high level of wt1 expression indicated worse prognosis for leukemia. Wt1 gene functions are different due to its subcellular localization. This study was aimed to investigate the expression and localization of wt1 mRNA and WT1 protein, and explore the effects of wt1 inhibitor, curcumin, on K562 cell proliferation, cell cycle and its possible mechanisms. MTT method was used to detect cell proliferation; flow cytometry was used to analyze the alteration of cell cycle, and the immunofluorescence and Western blot technology were performed to observe the subcellular localization of WT1 protein. The transcripts of wt1 and bcr/abl p210 was analyzed by real-time PCR. The results showed that wt1 mRNA and its protein were both highly expressed in K562 cells. The curcumin and imatinib (Glevec) both inhibit the cell proliferation resulting in the G(2)/M and G(0)/G(1) phase arrest respectively. Meanwhile, the transcripts of wt1 and bcr/ablp210 genes decreased greatly after being treated with the two inhibitors above. It is concluded that the alteration of wt1 gene affects the biological characteristics of Ph(+) K562 cells, such as cell proliferation, cell cycle and so on. Gene wt1 is expected to be further studied as a new therapy target in Ph(+) leukemias.


[ANCA(antineutrophil cytoplasmic antibodies)-associated vasculitis in a man with extreme fatigue, fever and progressive renal dysfunction].
A 55-year-old man, with no previous history, presented with extreme fatigue and fever and was admitted to hospital. He had progressive renal dysfunction and his serum anti-neutrophil cytoplasmic antibodies (ANCA) were markedly elevated. Renal histology was consistent with ANCA-associated vasculitis. The patient was successfully treated with cyclophosphamide and prednisolone. The classification and management of the ANCA-associated vasculitides are described. The classification was guided by the clinical presentation, serology and results of tissue biopsies. The ANCA inflammation had affected the middle sized and small vessels of especially the upper and lower airways, and the kidneys. The antibodies were directed at proteinase-3 (PR3) or myeloperoxidase (MPO). PR3-ANCA is predominantly found in Wegener's granulomatosis, while MPO-ANCA is related to microscopic polyangiitis. Tissue studies showed granulomatous inflammation of the airways which is typical of Wegener's disease. This type of inflammation is absent in microscopic polyangiitis. The initial treatment schedule consists of prednisone 1 mg/kg daily and oral cyclophosphamide 2 mg/kg daily. In the remission phase, the cyclophosphamide is replaced by azathioprine. It is not yet known how long maintenance treatment should be continued and which parameters have prognostic value.


Biological markers in psychiatry.
Biological markers in psychiatry are defined as measurable and quantifiable indices of psychiatric disorders. Criteria for biological markers are discussed. The criteria for the assignment of a biological marker to the pathophysiology, pathogenesis, or etiology of a disorder are proposed. None of the biological markers described in the literature for depression, schizophrenia, alcoholism or anxiety as separate disorder fulfills the specified criteria. Most of them can at best be considered as biological markers of the aspecific processes of strain and stress. Free-floating anxiety is the clinical hallmark of strain.


Whole genome sequence association and ancestry-informed polygenic profile of EEG alpha in a Native American population.
EEG alpha activity is the dominant oscillation in most adult humans, is highly heritable, and has been associated with a number of cognitive functions. Two EEG phenotypes, low- and high-voltage alpha (LVA & HVA), have been demonstrated to have high heritabilities. They have different prevalence depending on a population's ancestral origins. In the present study we assessed the influence of ancestry admixture on EEG alpha power, and conducted a whole genome sequencing association analysis and an ancestry-informed polygenic study on those phenotypes in a Native American (NA) population that has a high prevalence of LVA. Seven common variants, in LD with each other upstream from gene ASIC2, reached genome-wide significance (p = 2 × 10-8 ) having a positive association with alpha voltage. They had lower minor allele frequencies in the NAs than in a global population sample. Overall correlations between lower degrees of NA (higher degree European) ancestry and HVA, and higher degrees of NA and LVA were also found. Additionally a rare-variant gene-based study identified gene TIA1 being negatively associated with LVA. Approximately 3% of SNPs exhibited a 15-fold enrichment that explained nearly half of the total SNP-heritability for EEG alpha. These regions showed the most significant anti-correlations between NA ancestry and alpha voltage, and were enriched for genes and pathways mediating cognitive functions. Our findings suggested that these regions likely harbor causal variants for HVA, and lacking of such variants could explain the high prevalence of LVA in this NA population, possibly illuminating the ancestral origin and genetic basis for EEG alpha.


Adenosine A1 receptors (A1Rs) play a critical role in osteoclast formation and function.
Adenosine regulates a wide variety of physiological processes via interaction with one or more G-protein-coupled receptors (A(1)R, A(2A)R, A(2B)R, and A(3)R). Because A(1)R occupancy promotes fusion of human monocytes to form giant cells in vitro, we determined whether A(1)R occupancy similarly promotes osteoclast function and formation. Bone marrow cells (BMCs) were harvested from C57Bl/6 female mice or A(1)R-knockout mice and their wild-type (WT) littermates and differentiated into osteoclasts in the presence of colony stimulating factor-1 and receptor activator of NF-kappaB ligand in the presence or absence of the A(1)R antagonist 1,3-dipropyl-8-cyclopentyl xanthine (DPCPX). Osteoclast morphology was analyzed in tartrate-resistant acid phosphatase or F-actin-stained samples, and bone resorption was evaluated by toluidine blue staining of dentin. BMCs from A(1)R-knockout mice form fewer osteoclasts than BMCs from WT mice, and the A(1)R antagonist DPCPX inhibits osteoclast formation (IC(50)=1 nM), with altered morphology and reduced ability to resorb bone. A(1)R blockade increased ubiquitination and degradation of TRAF6 in RAW264.7 cells induced to differentiate into osteoclasts. These studies suggest a critical role for adenosine in bone homeostasis via interaction with adenosine A(1)R and further suggest that A(1)R may be a novel pharmacologic target to prevent the bone loss associated with inflammatory diseases and menopause.


Management of esophageal and pharyngeal perforation in the newborn infant.
Perforation of the esophagus or pharynx may occur during placement of endotracheal or nasogastric tubes in the newborn infant. Controversy exists, however, whether medical or surgical therapy is better in the management of these perforations. Nine patients who had esophageal or pharyngeal perforation in the neonatal period and were treated medically with antibiotics, nutritional support, and closed chest-tube drainage of pneumothoraces are described. All perforations healed without surgical repair. No mortality or morbidity occurred secondary to these perforations. This study, together with a review of the 73 patients described in the literature, indicate that perforations of the pharynx and esophagus can be satisfactorily managed medically. There is no apparent advantage to routine early surgical exploration. Only complications such as mediastinitis and mediastinal mass formation seem to require surgical treatment. Medical therapy with close observation for signs of sepsis and/or mediastinal changes will enable most newborn infants to avoid an operation and will identify those infants for whom surgery is definitely indicated.


Fractures of the femoral head.
Fractures of the femoral head infrequently accompany hip dislocations, but create complex therapeutic dilemmas for the orthopaedist when they do occur. The difficulties are compounded by existing incomplete classification schemes, which do not satisfactorily predict the final clinical results of a given injury or treatment. This chapter encompasses an extensive literature review of 238 published cases and analyzes the areas of confusion concerning this group of injuries. Our experience with 27 cases of femoral head fracture is presented. These examples emphasize the need for prompt, concentric reduction of the hip joint followed by individualized treatment of each type of femoral head fracture. An improved classification system to eliminate the ambiguities of previous systems and to provide the clinician with treatment guidelines for each type of femoral head fracture is proposed. These guidelines should improve the final results in these difficult and rare injuries.


Alzheimer disease as a vascular disorder: nosological evidence.
The main stumbling block in the clinical management and in the search for a cure of Alzheimer disease (AD) is that the cause of this disorder has remained uncertain until now. Evidence that sporadic (nongenetic) AD is primarily a vascular rather than a neurodegenerative disorder is reviewed. This conclusion is based on the following evidence: (1) epidemiological studies showing that practically all risk factors for AD reported thus far have a vascular component that reduces cerebral perfusion; (2) risk factor association between AD and vascular dementia (VaD); (3) improvement of cerebral perfusion obtained from most pharmacotherapy used to reduce the symptoms or progression of AD; (4) detection of regional cerebral hypoperfusion with the use of neuroimaging techniques to preclinically identify AD candidates; (5) presence of regional brain microvascular abnormalities before cognitive and neurodegenerative changes; (6) common overlap of clinical AD and VaD cognitive symptoms; (7) similarity of cerebrovascular lesions present in most AD and VaD patients; (8) presence of cerebral hypoperfusion preceding hypometabolism, cognitive decline, and neurodegeneration in AD; and (9) confirmation of the heterogeneous and multifactorial nature of AD, likely resulting from the diverse presence of vascular risk factors or indicators of vascular disease. Since the value of scientific evidence generally revolves around probability and chance, it is concluded that the data presented here pose a powerful argument in support of the proposal that AD should be classified as a vascular disorder. According to elementary statistics, the probability or chance that all these findings are due to an indirect pathological effect or to coincidental circumstances related to the disease process of AD seems highly unlikely. The collective data presented in this review strongly support the concept that sporadic AD is a vascular disorder. It is recommended that current clinical management of patients, treatment targets, research designs, and disease prevention efforts need to be critically reassessed and placed in perspective in light of these important findings.


[Geographic determinants of onchocerciasis transmission in a forest-savannah transition zone: an example of 2 villages of the Mbam focus (central region, Cameroon)].
Mbam, a focus of onchocerciasis outbreaks, is located in the forest/savannah transition zone of Cameroon. Transmission of the disease in this area is similar to that sometimes observed in West Africa. Geographical factors affecting transmission must be identified to determine the areas for priority action. The locations of breeding sites for black flies (the vector of the disease) and factors favorable to their dispersion, the social behavior of these population and the way that she occupy space, must be investigated to identify areas in which the flies and man are in close contact. This geographical study shows that the closest contact between men and black flies occurs in cocoa plantations, due to the proximity of the breeding sites. The areas of closet contact for women and children are unknown, but their identification is important because women and children may be bitten by the flies during their daily activities.


Study of corneal copper deposits in Wilson's disease by in vivo confocal microscopy.
To study corneal copper deposits in Wilson's disease (WD) patients by traditional biomicroscopy and in vivo laser scanning confocal microscopy (LSCM). Twenty WD patients and 20 matched controls underwent an ophthalmic examination in one eye randomly chosen, including slit lamp biomicroscopy with Goldmann's three-mirror contact lens examination and LSCM, in order to evaluate copper deposits in the peripheral cornea. No control subjects had corneal changes at both traditional biomicroscopy and LSCM. Only 25% of WD patients had detectable slit lamp changes, compared with 75% with LSCM examination. All cases detected by slit lamp were detected by LSCM. A significant correlation (p < 0.01) was found between deposit intensity at LSCM and daily urinary copper excretion. LSCM could detect copper deposition in WD corneas in more patients than traditional examination; it may therefore provide important information in cases of suspected WD diagnosis.


Predicting blood pressure control in hypertensive patients: an approach to quality-of-care assessment.
In assessing the quality of medical care, techniques were needed which can predict which patients are likely to have good or poor health outcomes. This study describes one way of identifying a subgroup of hypertensive patients whose diastolic blood pressures (DBP) are at high risk of remaining uncontrolled while under treatment. Medical care and patient variables were measured for 123 hypertensive medical clinic patients. Stepwise discriminant analysis identified a set of six variables which best distinguished between patients with well-controlled and uncontrolled DBP. When measurements of these variables were divided into high and low risk responses for uncontrolled DBP, the number of high risk variables was directly related to the proportion of patients with uncontrolled DBP. The variables also predicted DBP control among 63 patients in an entirely different medical care setting.


[Prospective register of primary percutaneous coronary interventions in patients with acute myocardial infarction].
The present study has evaluated the immediate angiographic results of primary percutaneous interventions (PCI) in patients with acute myocardial infarction, as well as hospital and 6-month clinical outcomes. The analysis covered a total of 265 patients (females (23%) and males (77%)); their mean age was 57+/-11 years. The mean time before the first balloon dilatation during PCI was 278+/-135 minutes after the development of the pain syndrome or 109+/-94 minutes after hospital admission. PCI proved to be effective in 96% of the patients, as evidenced by angiography. TIMI 3 blood flow was achieved in 83% of cases during PCI. After primary PCI, hospital mortality was 98.9% and 95% survived 6 months. At 6-month follow-up, 22% patients had positive exercise tests, recurrent angina pectoris and/or more than 50% luminal stenosis of the infarct-related artery. Control angiography made less than 6 months later showed 11% restenosis. This prospective study has demonstrated the high immediate and long effectiveness and safety of primary interventions in acute myocardial infarction.


Evaluation of a residential AA program.
The purpose of the study was to evaluate the effectiveness of the Serenity Lodge alcohol rehabilitation program by comparing a group of 137 men who entered the program with the same number of similar men who did not enter the program after an admission to a detoxification hospital. The residential program, for which people must pay board, is based on the AA philosophy and reality therapy. Follow-up interviews were conducted with three-quarters of the men in each group. Even after allowing for spontaneous remission as measured by the improvement in the control group, it was apparent that for a wide range of outcome criteria, and not just the obtaining of abstinence, the program was having a beneficial effect for the men who stayed at Serenity Lodge.


A new mark-recapture approach for abundance estimation of social species.
Accurate estimates of population abundance are a critical component of species conservation efforts in order to monitor the potential recovery of populations. Capture-mark-recapture (CMR) is a widely used approach to estimate population abundance, yet social species moving in groups violate the assumption of CMR approaches that all individuals in the population are detected independently. We developed a closed CMR model that addresses an important characteristic of group-living species-that individual-detection probability typically is conditional on group detection. Henceforth termed the Two-Step model, this approach first estimates group-detection probability and then-conditional on group detection-estimates individual-detection probability for individuals within detected groups. Overall abundance is estimated assuming that undetected groups have the same average group size as detected groups. We compared the performance of this Two-Step CMR model to a conventional (One-Step) closed CMR model that ignored group structure. We assessed model sensitivity to variation in both group- and individual-detection probability. Both models returned overall unbiased estimates of abundance, but the One-Step model returned deceptively narrow Bayesian confidence intervals (BCI) that failed to encompass the correct population abundance an average 52% of the time. Contrary, under the Two-Step model, CI coverage was on average 96%. Both models had similar root mean squared errors (RMSE), except for scenarios with low group detection probability, where the Two-Step model had much lower RMSE. For illustration with a real data set, we applied the Two-Step and regular model to non-invasive genetic capture-recapture data of mountain gorillas (Gorilla beringei beringei). As with simulations, abundance estimates under both models were similar, but the Two-Step model estimate had a wider confidence interval. Results support using the Two-Step model for species living in constant groups, particularly when group detection probability is low, to reduce risk of bias and adequately portray uncertainty in abundance estimates. Important sources of variation in detection need to be incorporated into the Two-Step model when applying it to field data.


The genus Clathria from the Gulf of Mexico and Mexican Caribbean, with redescription and resurrection of Clathria carteri (Poecilosclerida: Microcionidae).
The present study deals with the morphologic variability of eight Clathria species from the southern Gulf of Mexico and Mexican Caribbean. Clathria (Clathria) foliacea, C. (C.) carteri, C. (Microciona) calla, C. (M.) echinata, C. (M.) spinosa, C. (Thalysias.) venosa, and C. (T.) virgultosa were collected by scuba diving and dredging from the southern Gulf of Mexico (Veracruz, Campeche Bank, Yucatan) to the Caribbean coast (Quintana Roo) from shoreline to 120 m depth. The population of Clathria seems to be neither abundant nor diverse in the studied area. However, the seven species considered here are presumed to have a slight morphologic variability, and the interspecific relationships had not been fully resolved. The present study focuses on their differentiation through comparison of external morphology, skeletal architecture, spiculation and measurements of characters as well as scanning electronic microscopy. Comparisons were made among intra- and interspecific material and with the available type material. In particular, C. carteri Topsent (1889), previously synonymized with C. foliacea, is resurrected by establishing a neotype from material from the type locality, Campeche Bank; this has not been studied since its original description and is here redescribed for the first time. Another neotype is assigned for C. foliacea from Campeche also, since the original material is no longer available, and this species is redescribed also. Significant differences in spiculation between C. foliacea and C. carteri were found by one-way ANOVA. Although C. (T.) venosa resembles C. (T.) raraechelae in spiculation, these two species differ in measurements and skeletal organization. C. (T.) virgultosa is typified by a peculiar acanthostyle that is markedly spined on distal parts of the shaft, and by two types of tiny microscleres. This study has extended the known geographic distribution of each of these Clathria spp. within the lower Gulf of Mexico.


The neurobiology, neuropharmacology, and pharmacological treatment of the paraphilias and compulsive sexual behaviour.
There has been increasing interest in the treatment of sexual disorders in recent years. Sexual disorders are classified in DSM-IV as sexual dysfunctions, paraphilias, and gender identity disorders. The sexual dysfunctions are nondeviant or nonparaphillic. The sexual dysfunction disorders should include "hyperactive sexual desire disorder" under sexual desire disorders. Further, there should be a specifier for paraphilias of "with hypersexuality" or "without hypersexuality." There is still incomplete understanding of the neurobiology of sexual disorders although functional neuroanatomy and neoropharmcological research has exposed the neurotransmitters, receptors, and hormones that are involved in sexual desire. Various pharmacological agents including serotonin reuptake inhibitors, antiandrogens, LHRH agonists, and others have been documented as reducing sexual desire. An algorithm for the use of these drugs in the treatment of the paraphilias as well nonparaphilic hypersexuality is outlined. The modes of action, dosages, aims of treatment, and usual methods of prescribing these agents is reviewed in this article. Some future directions of research in pharmacological treatment is also discussed.


Effects of insulin therapy on non-insulin-dependent diabetics with secondary oral hypoglycemic agent failure.
To search for the mechanisms of secondary oral hypoglycemic agent (OHA) failure in non-insulin-dependent diabetic (NIDD) and to identify the effects of insulin therapy on these patients, we observed their pancreatic beta-cell function and metabolic parameters. The clinical characteristics and C-peptide responses to glucagon were analyzed in 37 NIDD with secondary OHA failure. All patients received insulin therapy and we measured their fasting plasma glucose (FPG), body mass index (BMI) and glycohemoglobin (HbA1C) levels at 3 months, their serum triglyceride and cholesterol levels at 6-12 months and performed glucagon tests at 0.5-29 months. Poor beta-cell function was observed in 11 (29.7%) patients (Group A) and fair beta-cell function in 26 (70.3%) patients (Group B). The mean age at diagnosis, the number of years since diagnosis of diabetes, FPG levels and HbA1C levels were similar in both groups. However, the patients in Group A had a lower average BMI (18.9 +/- 0.7 vs 22.1 +/- 0.6 kg/m2, p = 0.006) than those of Group B. After insulin therapy, the FPG levels (215 +/- 14 vs 282 +/- 12 mg/dL, p < 0.001) and HbA1C levels 9.2 +/- 0.5 vs 11.6 +/- 0.5%, p < 0.001) of both groups decreased but the BMI (23.1 +/- 0.7 vs 21.6 +/- 0.7 kg/m2, p < 0.001) increased significantly. However, the blood lipid levels and beta-cell function did not change. The beta-cell function was still poor in 5 out of 6 patients in Group A, but it was unchanged in all patients in Group B. About 1/3 of NIDD with secondary OHA failure are permanently insulin deficient and need long-term insulin treatment.


Accurate, strong, and stable reporting of choroid plexus epithelial cells in transgenic mice using a human transthyretin BAC.
Choroid plexus epithelial cells express high levels of transthyretin, produce cerebrospinal fluid and many of its proteins, and make up the blood-cerebrospinal fluid barrier. Choroid plexus epithelial cells are vital to brain health and may be involved in neurological diseases. Transgenic mice containing fluorescent and luminescent reporters of these cells would facilitate their study in health and disease, but prior transgenic reporters lost expression over the early postnatal period. Human bacterial artificial chromosomes in which the transthyretin coding sequence was replaced with DNA for tdTomato or luciferase 2 were used in pronuclear injections to produce transgenic mice. These mice were characterized by visualizing red fluorescence, immunostaining, real-time reverse transcription polymerase chain reaction, and luciferase enzyme assay. Reporters were faithfully expressed in cells that express transthyretin constitutively, including choroid plexus epithelial cells, retinal pigment epithelium, pancreatic islets, and liver. Expression of tdTomato in choroid plexus began at the appropriate embryonic age, being detectable by E11.5. Relative levels of tdTomato transcript in the liver and choroid plexus paralleled relative levels of transcripts for transthyretin. Expression remained robust over the first postnatal year, although choroid plexus transcripts of tdTomato declined slightly with age whereas transthyretin remained constant. TdTomato expression patterns were consistent across three founder lines, displayed no sex differences, and were stable across several generations. Two of the tdTomato lines were bred to homozygosity, and homozygous mice are healthy and fertile. The usefulness of tdTomato reporters in visualizing and analyzing live Transwell cultures was demonstrated. Luciferase activity was very high in homogenates of choroid plexus and continued to be expressed through adulthood. Luciferase also was detectable in eye and pancreas. Transgenic mice bearing fluorescent and luminescent reporters of transthyretin should prove useful for tracking transplanted choroid plexus epithelial cells, for purifying the cells, and for reporting their derivation from stem cells. They also should prove useful for studying transthyretin synthesis by other cell types, as transthyretin has been implicated in many functions and conditions, including clearance of β-amyloid peptides associated with Alzheimer's disease, heat shock in neurons, processing of neuropeptides, nerve regeneration, astrocyte metabolism, and transthyretin amyloidosis.


Gender and Black-White Race Differences in Young Adult Exercise Dependence Prevalence and Associated Disordered Eating.
Purpose: Exercise dependence, an unhealthy preoccupation with exercise that results in physiological and psychological symptoms, may be particularly prevalent among college students given sociocultural exercise and body ideals in this population. Yet few studies have examined this phenomenon in diverse college samples. The aims of the present study were to examine gender and Black-White race differences in the prevalence of exercise dependence and associated disordered eating. Method: Undergraduates (N = 839) completed online measures of exercise dependence and disordered eating. Chi square tests were used to test for differences in prevalence, and linear regression was used to examine race and gender moderating effects on the exercise dependence-disordered eating association. Results: More men reported exercise dependence symptoms than women. More White participants reported symptoms than Black participants. Of the four gender and race combinations examined, White men had the highest proportion and Black women the lowest proportion experiencing symptoms. Gender, but not race, moderated the exercise dependence-disordered eating association. Conclusions: Although more men experience exercise dependence symptoms, women experience stronger associations between exercise dependence symptoms and disordered eating. The number of students who reported exercise dependence symptoms underscores the need for further research in this population and the development of culturally sensitive interventions.


p47phox and reactive oxygen species production modulate expression of microRNA-451 in macrophages.
The production of microRNAs (miRNA) is influenced by various stimuli, including environmental stresses. We hypothesized that reactive oxygen species (ROS)-associated stress could regulate macrophage miRNA synthesis. miRNAs undergo unique steps of maturation processing through either one of two pathways of cytoplasmic processing. Unlike the canonical pathway, the regulation of alternative cytoplasmic processing of miRNA has not been fully elucidated yet. We cultured bone marrow derived macrophages (BMDM) from wild type (WT) and p47(phox-/-) mice and profiled miRNA expression using microarrays. We analyzed 375 miRNAs including four endogenous controls to normalize the data. At resting state, p47(phox-/-) BMDM has the markedly reduced expression of miR-451 compared to WT BMDM, without other significant differences. Unlike majority of miRNAs, miR-451 goes through the unique alternative processing pathway, in which Ago2 plays a key role. In spite of significant reduction of mature miR-451, however, its precursor form, pre-mir-451, was similar in both BMDMs, suggesting that the processing of pre-mir-451 is impaired in p47(phox-/-) BMDM. Moreover, p47(phox-/-) BMDM expressed significantly reduced level of Ago2. In contrast, Ago2 mRNA levels were similar in WT and p47(phox-/-) BMDM, suggesting a post-transcriptional defect of Ago2 production in p47(phox-/-) macrophages, which resulted in impaired processing of pre-miR-451. In order to examine the functional significance of miR-451 in macrophages, we cultured BMDMs from miR-451 knock-out mice. Of interest, miR-451-deficient BMDM exhibited reduced ROS generation upon zymosan stimulation, compared to WT BMDM. Our studies suggest functional crosstalk between ROS and miR-451 in the regulation of macrophage oxidant stress.


Effect of pathological high-risk features on cancer-specific mortality in non-metastatic clear cell renal cell carcinoma: a tool for optimizing patient selection for adjuvant therapy.
Adjuvant therapies for non-metastatic renal cell carcinoma (nmRCC) are being tested to improve outcomes in patients with high-risk (hR) nmRCC. The objective of the current study is to test the ability of three hR features to identify patients who are at the highest risk of cancer-specific mortality (CSM) after partial or radical nephrectomy. Within the Surveillance Epidemiology and End Results (SEER) database (1988-2013), we identified 23,632 nm "clear cell" RCC partial or radical nephrectomy patients with hR features: Fuhrman grade (FG) 3 or 4 or pathological classifications T3a or T3b or lymph node invasion (LNI), or combination of these. Kaplan-Meier analyses (KM) and multivariable Cox's regression models (CRM) evaluated the effect of hR features on CSM. Overall 11,568 (48.9%) patients harbored FG3-4, 5575 (23.6%) pT3a/b, 140 (0.6%) LNI, 5366 (22.7%) FG3-4 and pT3a/b, 183 (0.8%) LNI and pT3a/b, 203 (0.9%) LNI and FG3-4 and 597 (2.5%) LNI, FG3-4 and pT3a/b. Median CSM-free survival was 51, 58 and 22 months for LNI and pT3a/b, for LNI and FG3-4 and for LNI, FG3-4 and pT3a/b and was not reached for the other groups. These results remained unchanged in multivariable CRMs, where all hR features represented independent predictors. Individuals with combination of LNI with FG3-4 or pT3a/b and patients with all three hR features are at highest risk of CSM. In consequence, these patients may represent ideal candidates for adjuvant therapy either in clinical practice or future prospective trials.


Structural and catalytic diversity within the amidohydrolase superfamily.
The amidohydrolase superfamily comprises a remarkable set of enzymes that catalyze the hydrolysis of a wide range of substrates bearing amide or ester functional groups at carbon and phosphorus centers. The most salient structural landmark for this family of hydrolytic enzymes is a mononuclear or binuclear metal center embedded within the confines of a (beta/alpha)(8)-barrel structural fold. Seven variations in the identity of the specific amino acids that function as the direct metal ligands have been structurally characterized by X-ray crystallography. The metal center in this enzyme superfamily has a dual functionality in the expression of the overall catalytic activity. The scissile bond of the substrate must be activated for bond cleavage, and the hydrolytic water molecule must be deprotonated for nucleophilic attack. In all cases, the nucleophilic water molecule is activated through complexation with a mononuclear or binuclear metal center. In the binuclear metal centers, the carbonyl and phosphoryl groups of the substrates are polarized through Lewis acid catalysis via complexation with the beta-metal ion, while the hydrolytic water molecule is activated for nucleophilic attack by interaction with the alpha-metal ion. In the mononuclear metal centers, the substrate is activated by proton transfer from the active site, and the water is activated by metal ligation and general base catalysis. The substrate diversity is dictated by the conformational restrictions imposed by the eight loops that extend from the ends of the eight beta-strands.


A direct comparison of wet, dry and insulating bioelectric recording electrodes.
Alternatives to conventional wet electrode types are keenly sought for biomedical use and physiological research, especially when prolonged recording of biosignals is demanded. This paper describes a quantitative comparison of three types of bioelectrode (wet, dry and insulating) based on tests involving electrode impedance, static interference and motion artefact induced by various means. Data were collected simultaneously, and in the same physical environment for all electrode types. Results indicate that in many situations the performance of dry and insulating electrodes compares favourably with wet electrodes. The influence of non-stationary electric fields on shielded dry and insulating electrode types was compared to wet types. It was observed that interference experienced by dry and insulating electrode types was 40 dB and 34 dB less than that experienced by wet electrode types. Similarly, the effect of motion artefact on dry and insulating electrodes was compared to wet types. Artefact levels for dry and insulating electrodes were significantly higher than those for wet types at the beginning of trials conducted. By the end of the trial periods artefact levels for dry and insulating types were lower than wet electrodes by an average of 8.2 dB and 6.8 dB respectively. The reservations expressed in other studies regarding the viability of dry and insulating electrodes for reliable sensing of biosignals are not supported by the work described here.


Sesamin (a compound from sesame oil) increases tocopherol levels in rats fed ad libitum.
Six groups of rats were fed diets low, but adequate, in alpha-tocopherol but high in gamma-tocopherol. The six diets differed only in their contents (0, 0.25, 0.5, 1.0, 2.0, and 4.0 g/kg, respectively) of sesamin, a lignan from sesame oil. After four weeks of ad libitum feeding, the rats were sacrificed and the concentrations of alpha- and gamma-tocopherols were measured in the plasma, livers, and lungs. Sesamin-feeding increased gamma-tocopherol and gamma-/alpha-tocopherol ratios in the plasma (P < 0.05), liver (P < 0.001), and lungs (P < 0.001). The increase was non-significant for alpha-tocopherol. Thus, sesamin appears to spare gamma-tocopherol in rat plasma and tissues, and this effect persists in the presence of alpha-tocopherol, a known competitor to gamma-tocopherol. This suggests that the bioavailability of gamma-tocopherol is enhanced in phenol-containing diets as compared with purified diets.


Geometrical optics modeling of the grating-slit test.
A novel optical testing method termed the grating-slit test is discussed. This test uses a grating and a slit, as in the Ronchi test, but the grating-slit test is different in that the grating is used as the incoherent illuminating object instead of the spatial filter. The slit is located at the plane of the image of a sinusoidal intensity grating. An insightful geometrical-optics model for the grating-slit test is presented and the fringe contrast ratio with respect to the slit width and object-grating period is obtained. The concept of spatial bucket integration is used to obtain the fringe contrast ratio.


Two biomarker-directed randomized trials in European and Chinese patients with nonsmall-cell lung cancer: the BRCA1-RAP80 Expression Customization (BREC) studies.
In a Spanish Lung Cancer Group (SLCG) phase II trial, the combination of BRCA1 and receptor-associated protein 80 (RAP80) expression was significantly associated with outcome in Caucasian patients with nonsmall-cell lung cancer (NSCLC). The SLCG therefore undertook an industry-independent collaborative randomized phase III trial comparing nonselected cisplatin-based chemotherapy with therapy customized according to BRCA1/RAP80 expression. An analogous randomized phase II trial was carried out in China under the auspices of the SLCG to evaluate the effect of BRCA1/RAP80 expression in Asian patients. Eligibility criteria included stage IIIB-IV NSCLC and sufficient tumor specimen for molecular analysis. Randomization to the control or experimental arm was 1 : 1 in the SLCG trial and 1 : 3 in the Chinese trial. In both trials, patients in the control arm received docetaxel/cisplatin; in the experimental arm, patients with low RAP80 expression received gemcitabine/cisplatin, those with intermediate/high RAP80 expression and low/intermediate BRCA1 expression received docetaxel/cisplatin, and those with intermediate/high RAP80 expression and high BRCA1 expression received docetaxel alone. The primary end point was progression-free survival (PFS). Two hundred and seventy-nine patients in the SLCG trial and 124 in the Chinese trial were assessable for PFS. PFS in the control and experimental arms in the SLCG trial was 5.49 and 4.38 months, respectively [log rank P = 0.07; hazard ratio (HR) 1.28; P = 0.03]. In the Chinese trial, PFS was 4.74 and 3.78 months, respectively (log rank P = 0.82; HR 0.95; P = 0.82). Accrual was prematurely closed on the SLCG trial due to the absence of clinical benefit in the experimental over the control arm. However, the BREC studies provide proof of concept that an international, nonindustry, biomarker-directed trial is feasible. Thanks to the groundwork laid by these studies, we expect that ongoing further research on alternative biomarkers to elucidate DNA repair mechanisms will help define novel therapeutic approaches. NCT00617656/GECP-BREC and ChiCTR-TRC-12001860/BREC-CHINA.


The natural history of biocatalytic mechanisms.
Phylogenomic analysis of the occurrence and abundance of protein domains in proteomes has recently showed that the α/β architecture is probably the oldest fold design. This holds important implications for the origins of biochemistry. Here we explore structure-function relationships addressing the use of chemical mechanisms by ancestral enzymes. We test the hypothesis that the oldest folds used the most mechanisms. We start by tracing biocatalytic mechanisms operating in metabolic enzymes along a phylogenetic timeline of the first appearance of homologous superfamilies of protein domain structures from CATH. A total of 335 enzyme reactions were retrieved from MACiE and were mapped over fold age. We define a mechanistic step type as one of the 51 mechanistic annotations given in MACiE, and each step of each of the 335 mechanisms was described using one or more of these annotations. We find that the first two folds, the P-loop containing nucleotide triphosphate hydrolase and the NAD(P)-binding Rossmann-like homologous superfamilies, were α/β architectures responsible for introducing 35% (18/51) of the known mechanistic step types. We find that these two oldest structures in the phylogenomic analysis of protein domains introduced many mechanistic step types that were later combinatorially spread in catalytic history. The most common mechanistic step types included fundamental building blocks of enzyme chemistry: "Proton transfer," "Bimolecular nucleophilic addition," "Bimolecular nucleophilic substitution," and "Unimolecular elimination by the conjugate base." They were associated with the most ancestral fold structure typical of P-loop containing nucleotide triphosphate hydrolases. Over half of the mechanistic step types were introduced in the evolutionary timeline before the appearance of structures specific to diversified organisms, during a period of architectural diversification. The other half unfolded gradually after organismal diversification and during a period that spanned ∼2 billion years of evolutionary history.


Rhinofilling with hyaluronic acid thought as a cartilage graft.
There is great demand for nonsurgical aesthetic procedures. In this case series, the authors describe their stepwise technique of injecting hyaluronic acid (HA) into the nose to yield pleasing, stable results. A total of 148 patients underwent minimally invasive rhinoplasty by injection of HA filler. Anatomic contraindications for the procedure were a large dorsal hump, extreme tip projection, or excessive nasal deviation. HA injections were made in a specific order to reshape and stabilize the nose; the surgical plan was conceptually similar to placement of cartilage grafts. Patients indicated their satisfaction with the surgical results on a visual analog scale. Immediately after the procedure, all patients had transient redness and slight swelling at the injection site; this resolved spontaneously by 24 h posttreatment. Vascular impairment developed in 1 patient and was managed with hyaluronidase. In general, patients expressed a high level of satisfaction. Thirty-two patients returned for a "touch-up" filler injection 1 year posttreatment. Nasal reshaping by HA injection can be carried out quickly, safely, and effectively with minimal patient discomfort or downtime. Care must be given to select patients who are indicated for this procedure, and meticulous pretreatment planning is necessary.


The Role of Genome Accessibility in Transcription Factor Binding in Bacteria.
ChIP-seq enables genome-scale identification of regulatory regions that govern gene expression. However, the biological insights generated from ChIP-seq analysis have been limited to predictions of binding sites and cooperative interactions. Furthermore, ChIP-seq data often poorly correlate with in vitro measurements or predicted motifs, highlighting that binding affinity alone is insufficient to explain transcription factor (TF)-binding in vivo. One possibility is that binding sites are not equally accessible across the genome. A more comprehensive biophysical representation of TF-binding is required to improve our ability to understand, predict, and alter gene expression. Here, we show that genome accessibility is a key parameter that impacts TF-binding in bacteria. We developed a thermodynamic model that parameterizes ChIP-seq coverage in terms of genome accessibility and binding affinity. The role of genome accessibility is validated using a large-scale ChIP-seq dataset of the M. tuberculosis regulatory network. We find that accounting for genome accessibility led to a model that explains 63% of the ChIP-seq profile variance, while a model based in motif score alone explains only 35% of the variance. Moreover, our framework enables de novo ChIP-seq peak prediction and is useful for inferring TF-binding peaks in new experimental conditions by reducing the need for additional experiments. We observe that the genome is more accessible in intergenic regions, and that increased accessibility is positively correlated with gene expression and anti-correlated with distance to the origin of replication. Our biophysically motivated model provides a more comprehensive description of TF-binding in vivo from first principles towards a better representation of gene regulation in silico, with promising applications in systems biology.


Does sperm origin-Ejaculated or testicular-Affect embryo morphokinetic parameters?
It is unclear whether sperm origin, either ejaculated or testicular, in couples diagnosed with male factor infertility, affects the timing of the embryo's developmental events evaluated by time-lapse monitoring and implantation rates. To examine the effect of sperm origin on embryo morphokinetics in couples diagnosed with male factor infertility. This study included a retrospective analysis of morphokinetic parameters performed by time-lapse monitoring between 2013 and 2017. The developmental processes and morphokinetic parameters of 419 embryos obtained from couples with male factor infertility attributed to oligo-astheno-teratozoospermia, 158 embryos derived from surgically extracted testicular spermatozoa from couples diagnosed with non-obstructive azoospermia, and 190 embryos from couples with normal ejaculated spermatozoa and female mechanical factor-related infertility, were evaluated. A comparison of morphokinetic parameters, implantation, and clinical pregnancy rates was performed between the groups with additional analysis in accordance with implantation status. Embryos from the normal ejaculated spermatozoa and oligo-astheno-teratozoospermia patients reached the later morphokinetic milestones-synchronous division (S3) and time to morula (tM)-faster than embryos obtained from testicular spermatozoa. Implantation rate was similar in the normal ejaculated spermatozoa and oligo-astheno-teratozoospermia groups (41.9% vs. 45.8%, NS), with higher implantation rate in the oligo-astheno-teratozoospermia group compared to the testicular spermatozoa group (45.8% vs. 33.6%, p = 0.02). Comparison of Known Implantation Data (KID) positive (KIDp) and KID negative (KIDn) embryos in each group revealed more rapid development in KIDp embryos in the normal ejaculated spermatozoa and the oligo-astheno-teratozoospermia groups, while in the testicular spermatozoa group implanted embryos reached the late morphokinetic milestones (time to 8 cell stage-t8, ECC3, S3, and tM) significantly faster than embryos that failed to implant. In a multivariate logistic regression analysis of the male factor infertility population, (oligo-astheno-teratospermia) (OR = 2.54, p = 0.003) and t8 (OR = 0.95, p = 0.027) were predictive of successful implantation. Male factor infertility embryos that reached the t8 milestone within 48-56 h had favorable implantation rates (p < 0.001). The study results may highlight another pathophysiology by means of which sperm origin affects embryo developmental kinetics. Selecting embryos demonstrating a faster developmental rate at t8 and specifically the 48- to 56 h interval following time of pronuclei fading (tPNf) may improve implantation rates in cases of male factor infertility. This study showed that ejaculated spermatozoa is associated with faster late cell divisions, more rapid compaction, and higher implantation rates compared to testicular spermatozoa. Additionally, t8 is an important predictor for implantation in the male factor infertility population.