HIV controllers: a genetically determined or inducible phenotype?
Less than 0.5% of human immunodeficiency virus-1 (HIV-1)-infected patients maintain viral load below the threshold of detection by conventional assays for many years without antiretroviral therapy. These patients are known as HIV controllers (HICs) and offer a unique opportunity to explore and unravel efficient mechanisms of defense against HIV. An intense research effort has led to identifying viral and host factors that contribute to the control of viral replication. Although the host factors appear to be the primary determinants for HIV control, different genomic, transcriptomic, and immunological profiles have been described in HICs. This supports the existence of different mechanisms of control. We review here the most significant data in the field, including our own work, in the light of a crucial question: whether the ability to achieve and maintain the control of HIV replication is linked to genetic or immunological features specific to HIC or can be induced by vaccine or therapeutic approaches in the general population.


Clostridium difficile-associated diarrhoea.
Clostridium difficile is an important nosocomial pathogen and the most frequently diagnosed cause of infectious hospital-acquired diarrhoea. Toxigenic strains usually produce toxin A and toxin B, which are the primary virulence factors of C. difficile. Some recently described strains produce an additional toxin, an adenosine-diphosphate ribosyltransferase known as binary toxin, the role of which in pathogenicity is unknown. There has been concern about the emergence of a hypervirulent fluoroquinolone-resistant strain of C. difficile in North America and Europe. The use of fluoroquinolone antimicrobials appears to be acting as a selective pressure in the emergence of this strain. In this review, we describe the current state of knowledge about C. difficile as a cause of diarrhoeal illness.


Real-time prediction of size-resolved ultrafine particulate matter on freeways.
Ultrafine particulate matter (UFP; diameter <0.1 μm) concentrations are relatively high on the freeway, and time spent on freeways can contribute a significant fraction of total daily UFP exposure. We model real-time size-resolved UFP concentrations in summer time on-freeway air. Particle concentrations (32 bins, 5.5 to 600 nm) were measured on Minnesota freeways during summer 2006 and 2007 ( Johnson, J. P.; Kittelson, D. B.; Watts, W. F. Environ. Sci. Technol. 2009 , 43 , 5358 - 5364 ). Here, we develop and apply two-way stratified multilinear regressions, using an approach analogous to mobile-monitoring land-use regression but using real-time meteorological and traffic data. Our models offer the strongest predictions in the 10-100 nm size range (adj-R(2): 0.79-0.89, average adj-R(2): 0.85) and acceptable but weaker predictions in the 130-200 nm range (adj-R(2): 0.41-0.62, average adj-R(2): 0.52). The aggregate model for total particle counts performs well (adj-R(2) = 0.77). Bootstrap resampling (n = 1000) indicates that the proposed models are robust to minor perturbations in input data. The proposed models are based on readily available real-time information (traffic and meteorological parameters) and can thus be exploited to offer spatiotemporally resolved prediction of UFPs on freeways within similar geographic and meteorological environments. The approach developed here provides an important step toward modeling population exposure to UFP.


Monitoring the development of xenograft triple-negative breast cancer models using diffusion-weighted magnetic resonance imaging.
Evaluations of tumor growth rates and molecular biomarkers are traditionally used to assess new mouse models of human breast cancers. This study investigated the utility of diffusion weighted (DW)-magnetic resonance imaging (MRI) for evaluating cellular proliferation of new tumor models of triple-negative breast cancer, which may augment traditional analysis methods. Eleven human breast cancer cell lines were used to develop xenograft tumors in severe combined immunodeficient mice, with two of these cell lines exhibiting sufficient growth to be serially passaged. DW-MRI was performed to measure the distributions of the apparent diffusion coefficient (ADC) in these two tumor xenograft models, which showed a correlation with tumor growth rates and doubling times during each passage. The distributions of the ADC values were also correlated with expression of Ki67, a biomarker of cell proliferation, and hypoxia inducible factor (HIF)-1α and vascular endothelial growth factor receptor-2 (VEGFR2), which are essential proteins involved in regulating aerobic glycolysis and angiogenesis that support tumor cell proliferation. Although phosphatase and tensin homolog (PTEN) levels were different between the two xenograft models, AKT levels did not differ nor did they correlate with tumor growth. This last result demonstrates the complexity of signaling protein pathways and the difficulty in interpreting the effects of protein expression on tumor cell proliferation. In contrast, DW-MRI may be a more direct assessment of tumor growth and cancer cell proliferation.


Medium-term follow up of pulmonary autograft aortic valve replacement: technical advances and echocardiographic follow up.
The Ross procedure (aortic valve replacement (AVR) with pulmonary autograft and pulmonary homograft replacement of pulmonary valve) was developed as a durable aortic valve substitute that avoids the need for anticoagulation and provides young patients with a long-lasting aortic valve substitute. Our seven-year follow up echocardiography data are reviewed. Between May 1993 and March 2000, 40 adult patients (28 males, 12 females; mean age 33.3 years) underwent the Ross procedure at the Brigham and Women's Hospital for congenital aortic stenosis (n = 6), aortic insufficiency (n = 17) and mixed disease (n = 17). All patients had aortic root replacement with the pulmonary autograft and had no regurgitation after operation. Postoperative evaluation was conducted by transthoracic echocardiography, office visit and/or telephone interview. NYHA functional class, aortic and pulmonary valve function and aortic root dimensions were evaluated. One patient died postoperatively as a result of a low output state related to global left ventricular dysfunction. Four patients (10%) developed pulmonary homograft stenosis with a peak gradient >40mmHg; and six developed mild pulmonary stenosis. One patient had aortic insufficiency seven years postoperatively that required valve replacement. Eight patients developed mild dilatation (>37 mm) of the neoaortic root, and five of these had aortic insufficiency. One patient required transplantation at 40 months for restrictive cardiomyopathy. The Ross procedure is an effective means of AVR that can be accomplished with low perioperative morbidity and mortality if certain technical modifications are carried out. In this series of 40 patients with mid-term follow up, a significant number developed moderate pulmonary trunk stenosis, though echo characterization demonstrated good valve function.


Development of a genomics-based PCR assay for detection of Mycoplasma pneumoniae in a large outbreak in New York State.
A genomics-based PCR method was developed and used to test specimens from patients involved in a large outbreak of Mycoplasma pneumoniae in a closed religious community in New York State. New P1 adhesin gene primers were designed to bind to 9 of 10 target sequences in the repetitive-element sequences obtained from the whole genome sequence of M. pneumoniae. This PCR method had a sensitivity of 0.006 CFU and a specificity of 100% for M. pneumoniae. The PCR was validated by testing a subset of patient samples by culture and comparing the results to those obtained by PCR. Of the initial 280 samples tested, 73 were positive by PCR and 22 were positive by culture. All samples positive by culture were also positive by PCR. Follow-up testing of selected patients 3 to 6 weeks after antibiotic treatment revealed that eight samples remained positive by PCR and that three samples remained positive by culture. Additionally, no nonspecific PCR inhibition was detected as a result of the specimen type, transport medium, or sample preparation methodology. The study demonstrates that the PCR described here is a rapid, sensitive, and specific method for the identification of M. pneumoniae and was helpful for the detection and monitoring of the outbreak.


Recommendations from the Council of Emergency Medicine Residency Directors: Osteopathic Applicants.
The Council of Emergency Medicine Residency Directors (CORD) Advising Students Committee (ASC-EM) has previously published student advising recommendations for general emergency medicine (EM) applicants in an effort to disseminate standardized information to students and potential advisors. As the shift to a single graduate medical education system occurs by 2020, osteopathic students will continue to represent a larger portion of matched EM applicants, but data shows that their match rate lags that of their allopathic peers, with many citing a lack of access to knowledge EM advisors as a major barrier. Based on available data and experiential information, a sub-group of ASC-EM committee sought to provide quality, evidence-based advising resources for students, their advisors, and medical leadership. The recommendations advise osteopathic students to seek early mentorship and get involved in EM-specific organizations. Students should take Step 1 of the United States Medical Licensing Exam and complete two EM rotations at academic institutions to secure two Standardized Letters of Evaluation and consider regional and program-specific data on percentage of active osteopathic residents.


Removal of cationic dye pollutants from wastewater with HS loaded semi-IPN composites: kinetic and thermodynamic studies.
In this study, methylene blue (MB) pollutant in water was removed using produced hazelnut shell loaded semi-interpenetrating polymer networks (HS loaded semi-IPN) adsorbent. The physical and chemical characterizations of the adsorbents were investigated using TGA, DSC, FT-IR, BET, FE-SEM, and EDX. Experimental parameters such as temperature, swelling, dye concentration, contact time, pH solution, and adsorbent dosage for MB adsorption were thoroughly investigated. It was determined that the HS loaded semi-IPN adsorbent removed 92.1% of MB dye. Subsequently, the adsorption properties between the adsorbent and dye were investigated in detail using several different kinetic, isotherm, and thermodynamic models. As a result of the obtained data, the interaction between adsorbent and dye molecules is discussed. Moreover, studies on the industrial usability of the adsorbent have been carried out, and it has been observed that the adsorbent can be employed even after four cycles.


Increased concentrations of certain persistent organic pollutants in subjects with self-reported electromagnetic hypersensitivity--a pilot study.
Electromagnetic hypersensitivity (EHS) is used for a variety of subjective symptoms related to exposure to electromagnetic fields (EMF). The aim of this pilot study was to analyze the concentrations of certain persistent organic pollutants (POPs) in subjects with self-reported EHS. In total, 13 EHS subjects and 21 controls were included, all female. The concentration of several POPs was higher in EHS subjects than in controls. Lower concentrations were found for hexachlorobenzene and two types of chlordanes. The only significantly increased odds ratios (ORs) were found for polybrominated diphenyl ether (PBDE) #47 yielding OR=11.7, 95% confidence interval (CI)=1.45-94.7 and the chlordane metabolite MC6 with OR=11.2, 95% CI=1.18-106. The results were based on low numbers and must be interpreted with caution. This hypothesis generating study indicates the necessity of a larger investigation on this issue.


An unusual cause of recurrent pancreatitis: A gastric duplication cyst with an accessory pancreatic lobe.
Congenital anomalies of pancreas and its ductal drainage are uncommon but in general surgically correctable causes of recurrent pancreatitis. A gastric duplication cyst communicated with an accessory pancreatic lobe is an extremely rare cause of recurrent pancreatitis, but an early and accurate diagnosis of this anomaly is important because suitable surgical treatment may lead to a satisfactory outcome. Herein, we presented multidetector computed tomography and magnetic resonance imaging findings of a gastric duplication cyst communicating with an accessory pancreatic lobe via an aberrant duct in a 29-year-old woman with recurrent acute pancreatitis and also reviewed other similar cases reported in the literature.


Scene-Motion Thresholds Correlate with Angular Head Motions for Immersive Virtual Environments.
To better understand motion perception in immersive virtual environments, we conducted a user study to quantify perception of scene motion as subjects yawed their heads. We measured psychometric functions of scene-velocity thresholds for different head motions and then extracted 75% thresholds, creating scene-velocity thresholds as functions of three measures of head motion: 1) Angular Range, 2) Peak Angular Velocity, 3) and Peak Angular Acceleration. We also measured scene-velocity thresholds for four phases of head motion: 1) the Start of the head turn, 2) the Center of the head turn, 3) the End of the head turn, 4) and All of the head turn. Scene-velocity thresholds increased as head motion increased for all tested conditions.


Trinucleotide repeat expansion and human disease.
Trinucleotide repeat expansions have been identified as the underlying mutation in an increasing number of human genetic diseases, such as fragile site syndromes, myotonic dystrophy and several neurodegenerative disorders including Huntington's disease. By an unknown mechanism, polymorphic GC-rich triplet repeats expand in all these diseases. The expansions of a CCG repeat in fragile-site-associated disorders and the CTG repeat (in the 3'-untranslated region of the myotonin kinase gene) causing myotonic dystrophy are very large, whereas small expansions of CAG repeats have been identified in the open reading frame of genes in a number of neurological genetic disorders.


A haemorrhagic disorder in pregnancy due to an "anticoagulant" preventing the conversion of fibrinogen to fibrin.
A bleeding disorder occurring during labour and affecting mother and foetus is described. All stages of coagulation were normal until the reaction fibrinogen-->fibrin. Either there was some abnormality present in the fibrinogen molecule, or there was an "anticoagulant" acting at this stage. The abnormality was reversible by protamine sulphate and toluidine blue, but in other respects did not resemble hyperheparinaemia. Reversibility of a clotting defect by protamine or toluidine blue is not sufficient evidence on which to make a diagnosis of hyperheparinaemia. Demonstration of a prolonged clotting time occurring in late pregnancy does not necessarily justify the diagnosis of "afibrinogenaemia."


An approach for the stable immobilization of proteins.
An approach is presented for the stable covalent immobilization of proteins with a high retention of biological activity. First, chemical modification studies were used to establish enzyme structural and functional properties relevant to the covalent immobilization of an enzyme to agarose based supports. Heparinase was used as a model enzyme in this set of studies. Amine modifications result in 75-100% activity loss, but the effect is moderated by a reduction in the degree of derivatization. N-hydroxysuccinimide, 1,1,1-trifluoroethanesulfonic acid, and epoxide activated agarose were utilized to determine the effect of amine reactive supports on immobilized enzyme activity retention. Cysteine modifications resulted in 25-50% loss in activity, but free cysteines were inaccessible to either immobilized bromoacetyl or p-chloromercuribenzoyl groups. Amine reactive coupling chemistries were therefore utilized for the covalent immobilization of heparinase. Second, to ensure maximal stability of the immobile protein-support linkage, the identification and subsequent elimination of the principal sources of protein detachment were systematically investigated. By using high-performance liquid chromatography (HPLC), electrophoresis, and radiolabeling techniques, the relative contributions of four potential detachment mechanisms-support degradation, proteolytic degradation, desorption of noncovalently bound protein, and bond solvolysis-were quantified. The mechanisms of lysozyme, bovine serum albumin, and heparinase leakage from N-hydroxysuccinimide or 1,1,1-trifluoroethanesulfonic acid activated agarose were elucidated. By use of stringent postimmobilization support wash procedures, noncovalently bound protein loss. An effective postimmobilization washing procedure is presented for the removal of adsorbed protein and the complete elimination of immobilized protein loss.


Uncommon Terpenoids from Salvia Species: Chemistry, Biosynthesis and Biological Activities.
The search for new bioactive compounds from plant sources has been and continues to be one of the most important fields of research in drug discovery. However, Natural Products research has continuously evolved, and more and more has gained a multidisciplinary character. Despite new developments of methodologies and concepts, one intriguing aspect still persists, i.e., different species belonging to the same genus can produce different secondary metabolites, whereas taxonomically different genera can produce the same compounds. The genus Salvia L. (Family Lamiaceae) comprises myriad distinct medicinal herbs used in traditional medicine worldwide that show different pharmacological activities due to the presence of a variety of interesting specialized metabolites, including mono-, sesqui-, di-, sester-, tri-, tetra-, and higher terpenoids as well as phenylpropanoids, phenolic acid derivatives, lignans, flavonoids, and alkaloids. We herein summarize the research progress on some uncommon terpenoids, isolated from members of the genus Salvia, which are well recognized for their potential pharmacological activities. This review also provides a current knowledge on the biosynthesis and occurrence of some interesting phytochemicals from Salvia species, viz. C23-terpenoids, sesterterpenoids (C25), dammarane triterpenoids (C30), and uncommon triterpenoids (C20+C10). The study was carried out by searching various scientific databases, including Elsevier, ACS publications, Taylor and Francis, Wiley Online Library, MDPI, Springer, Thieme, and ProQuest. Therefore, 106 uncommon terpenoids were identified and summarized. Some of these compounds possessed a variety of pharmacological properties, such as antibacterial, antiviral, antiparasitic, cytotoxic and tubulin tyrosine ligase inhibitory activities. Due to the lack of pharmacological information for the presented compounds gathered from previous studies, biological investigation of these compounds should be reinvestigated.


Accuracy of the magnetic resonance imaging pathway in the detection of prostate cancer: a systematic review and meta-analysis.
Although magnetic resonance imaging and subsequent targeted biopsy ('MRI pathway') have been widely adopted in routine clinical practice, it is still a common practice to perform systematic biopsy concurrently, because the accuracy of the MRI pathway is yet to be fully defined. This systematic review of the literature assessed the sensitivity of the MRI pathway for detecting clinically significant prostate cancer. Multiple databases were searched up to May 2017 according to the Preferred Reporting Items for Systematic Review and Meta-analysis (PRISMA) statement for studies assessing the accuracy of MR-guided biopsy (MRGB) compared to a reference standard which consisted of both MRGB and systematic biopsy with at least 20-cores. The primary outcome was the sensitivity of detecting clinically significant prostate cancer defined as Gleason ≥7 disease. A total of 15 studies met the predefined inclusion criteria. Overall, studies were assessed to be of low quality with inadequate blinding of personnel, which could introduce performance and detection bias. The calculated summary sensitivity of the MRI pathway was 78.3% [95%CI 75.0-81.4%]. There was moderate heterogeneity between the included studies (I2 = 36%). Subgroup analysis was performed based on clinical setting, the strength of MRI magnet and mode of image fusion as factors but no interaction was identified between any of the subgroups. No publication bias was identified. The MRI pathway cannot yet be solely relied upon to diagnose clinically significant disease and hence additional systematic sampling should still be performed during the biopsy procedure.


Helix-rod host-guest complexes with shuttling rates much faster than disassembly.
Dynamic assembly is a powerful fabrication method of complex, functionally diverse molecular architectures, but its use in synthetic nanomachines has been hampered by the difficulty of avoiding reversible attachments that result in the premature breaking apart of loosely held moving parts. We show that molecular motion can be controlled in dynamically assembled systems through segregation of the disassembly process and internal translation to time scales that differ by four orders of magnitude. Helical molecular tapes were designed to slowly wind around rod-like guests and then to rapidly slide along them. The winding process requires helix unfolding and refolding, as well as a strict match between helix length and anchor points on the rods. This modular design and dynamic assembly open up promising capabilities in molecular machinery.


Physiological characterization of Streptococcus bovis mutants that can resist 2-deoxyglucose-induced lysis.
Streptococcus bovis JB1 does not normally lyse, but stationary phase lysis can be induced by including 2-deoxyglucose (2DG) in the growth medium. Isolates deficient in glucose/2DG phosphotransferase activity (PTS-) also lysed when 2DG was present (Lys+) and this result indicated that 2DG phosphorylation via the PTS was not an obligate requirement for 2DG-induced lysis. Cells and cell walls from 2DG-grown cultures lysed faster when proteinase K was added, but glucose-grown cultures and cell walls were not affected. A lipoteichoic acid (LTA) extract (aqueous phase from hot phenol treatment) from glucose-grown cells inhibited the lysis of 2DG-grown cultures, but a similar extract prepared from 2DG-grown cells was without effect. Thin-layer chromatography and differential staining indicated that wild-type and Lys+ PTS- cells incorporated 2DG into LTA, but lysis-resistant cultures (Lys- PTS+ and Lys- PTS-) did not. LTA from lysis-resistant (Lys- PTS+ and Lys- PTS-) cells grown with glucose and 2DG also prevented 2DG-dependent lysis of the wild-type. LTA could not inhibit degradation of cell walls isolated from 2DG-grown cultures, but LTA inhibited the lysis of Micrococcus lysodeikticus (Micrococcus luteus) cells that were exposed to supernatants from 2DG-grown S. bovis cultures. Group D streptococci (including S. bovis) normally have an alpha-1,2 linked glucose disaccharide (kojibiose) in their LTA, but kojibiose cannot be synthesized from 2DG. This observation suggested that the kojibiose moiety of LTA was involved in autolysin inactivation. Wild-type S. bovis had ATP- as well as PEP-dependent mechanisms of 2DG phosphorylation and one lysis-resistant phenotype (Lys- PTS-) had reduced levels of both activities. However, the Lys- PTS+ phenotype was still able to phosphorylate 2DG via ATP and PEP and this result indicated that some other step of 2DG incorporation into LTA was being inhibited. Based on these results, growth in the presence of 2DG appears to prevent synthesis of normal LTA, which is involved in the regulation of autolytic enzymes.


Resolution and brightness characteristics of short-lag spatial coherence (SLSC) images.
We previously described a novel beamforming method that images the spatial correlation of an echo wave field with demonstrated applications to clutter reduction in high-noise environments. In this paper, several characteristics of the resolution and brightness of short-lag spatial coherence (SLSC) images formed by this method are compared with B-mode images formed by conventional delay-and-sum beamforming methods. Point target widths were measured to estimate resolution, the autocorrelation of image texture was measured to estimate texture size, and the contrast (i.e., brightness ratio) of clinically relevant targets was assessed. SLSC images demonstrate improved resolution and contrast with increasing values of channel noise and clutter, whereas B-mode resolution was degraded in the presence of high noise (i.e., > -12 dB channel noise-to-signal ratios) and high clutter magnitudes (i.e., > -21 dB relative to point target magnitude). Lateral resolution in SLSC images was improved with increasing lag value, whereas axial resolution was degraded with increasing correlation kernel length. The texture size of SLSC images was smaller than that of matched B-mode images. Results demonstrate that the resolution and contrast of coherence-based images depend on a range of parameters, but are generally superior to those of matched B-mode images under challenging imaging conditions.


[Metastasis of prostatic cancer (based on lymphographic data)].
A lymphographic examination of 56 patients with prostatic carcinoma established the incidence of lymphogenous metastases for different stages of primary tumor advancement: there were no metastases in patients with stage I tumors; metastases were found in 3 out of 16 cases of stage II tumors; in 4 out of 12 with stage III tumors and in 13 out of 21 patients with stage IV tumors. The prevalent site of prostatic carcinoma metastases were iliac lymph nodes. Indications for lymphographic examination depending on local advancement of tumor process are discussed.


Hyoid Bone and Thyroid Cartilage Metastases from Sigmoid Colon Adenocarcinoma: A Case Report.
Secondary tumours of the hyoid bone and thyroid cartilage are extremely rare. In this paper, we present a case of the hyoid bone and thyroid cartilage metastases in a patient treated for sigmoid colon adenocarcinoma. Four years after sigmoid colon adenocarcinoma was diagnosed and treated with surgery and chemotherapy, the patient developed bone metastases in the left sacroiliac joint and right proximal humerus. Although the patient did not complain of any related symptoms, in a bone scintigraphy the accumulation of Technetium-99m was incidentally detected in the two sites of the anterior neck. On ultrasound examination there were two hyperechoic and heterogeneous masses with calcifications placed in front of the hyoid bone and thyroid cartilage. Computerized tomography demonstrated massive hyoid bone and thyroid cartilage destruction. In patients with progressive sigmoid colon adenocarcinoma, destruction of the hyoid bone and thyroid cartilage could be suspected for metastases.


Narrative review: furosemide for hypercalcemia: an unproven yet common practice.
Although primary hyperparathyroidism is the most common cause of hypercalcemia, cancer is the most common cause requiring inpatient intervention. An estimated 10% to 20% of all patients with cancer have hypercalcemia at some point in their disease trajectory, particularly in advanced disease. Aggressive saline hydration and varying doses of furosemide continue to be the standard of care for emergency management. However, a review of the evidence for the use of furosemide in the medical management of hypercalcemia yields only case reports published before the introduction of bisphosphonates, in contrast to multiple randomized, controlled trials supporting the use of bisphosphonates. The use of furosemide in the management of hypercalcemia should no longer be recommended.


Skin microbiota: Friend or foe in pediatric skin health and skin disease.
The human integument and gastrointestinal tract host unique microbial ecosystems. Within the last decade, research has focused on understanding the contributions of the microbiota to human health and disease. The majority of skin microbiome studies involve adults. This review focuses on key studies conducted within the pediatric population and provides a framework for future skin microbiome work in this ever-expanding field. This article begins by exploring the skin microbiome at birth and reviews the impact of delivery mode on infant skin colonization. How skin microbial colonization evolves from infancy to adulthood and normal development impacts the abundance of skin commensals such as Streptococcus, Staphylococcus, and Cutibacterium is also highlighted. Finally, several skin microbiome research studies in common pediatric skin conditions are reviewed, including body odor, atopic dermatitis (AD), and acne. The bacteria involved in metabolizing sweat, the impact on body odor, and how this process evolves from childhood to adulthood is outlined. In AD, different bacteria genera that predominate in children and adults and the impact of current AD therapies on skin microbiota are explored. Finally, in acne, the understanding of how Cutibacterium acnes contributes to acne pathogenesis and how acne therapies impact the skin microbial communities is reviewed.


In silico study of amyloid beta-protein folding and oligomerization.
Experimental findings suggest that oligomeric forms of the amyloid beta protein (Abeta) play a critical role in Alzheimer's disease. Thus, elucidating their structure and the mechanisms of their formation is critical for developing therapeutic agents. We use discrete molecular dynamics simulations and a four-bead protein model to study oligomerization of two predominant alloforms, Abeta40 and Abeta42, at the atomic level. The four-bead model incorporates backbone hydrogen-bond interactions and amino acid-specific interactions mediated through hydrophobic and hydrophilic elements of the side chains. During the simulations we observe monomer folding and aggregation of monomers into oligomers of variable sizes. Abeta40 forms significantly more dimers than Abeta42, whereas pentamers are significantly more abundant in Abeta42 relative to Abeta40. Structure analysis reveals a turn centered at Gly-37-Gly-38 that is present in a folded Abeta42 monomer but not in a folded Abeta40 monomer and is associated with the first contacts that form during monomer folding. Our results suggest that this turn plays an important role in Abeta42 pentamer formation. Abeta pentamers have a globular structure comprising hydrophobic residues within the pentamer's core and hydrophilic N-terminal residues at the surface of the pentamer. The N termini of Abeta40 pentamers are more spatially restricted than Abeta42 pentamers. Abeta40 pentamers form a beta-strand structure involving Ala-2-Phe-4, which is absent in Abeta42 pentamers. These structural differences imply a different degree of hydrophobic core exposure between pentamers of the two alloforms, with the hydrophobic core of the Abeta42 pentamer being more exposed and thus more prone to form larger oligomers.


Primary Rectal Tumor With Extensive Choriocarcinoma Differentiation in a Woman With Lung, Liver and Disseminated Peritoneal Disease: A Primary Rectal Adenocarcinoma With Extensive Choriocarcinoma Differentiation or Primary Rectal Choriocarcinoma?
Primary rectal adenocarcinoma with extensive choriocarcinomatous differentiation is a rare neoplasm, with only sporadic cases reported worldwide. The prognosis is typically poor, and no standard therapy has been established for this tumor. We report a case of a 63-year-old woman who presented with lower abdominal and pelvic discomfort, as well as rectal bleeding. Endoscopy revealed a rectal tumor. She was diagnosed with primary rectal adenocarcinoma with extensive choriocarcinomatous differentiation, accompanied by liver metastasis and peritoneal carcinomatosis. The immunohistochemical profile demonstrated strong and diffuse positivity for keratin (AE1/AE3), beta-human chorionic gonadotropin (β-HCG), p53, MYC, p16, and Ki-67. Molecular analysis indicated mutations in KRAS, TP53, and PI3KCA. Despite the tumor's profile, the serum β-HCG level was not elevated. A chemotherapy regimen for metastatic colorectal adenocarcinoma was initiated, but there was a poor response, with rapid tumor progression. The patient survived for only 5 months postdiagnosis. We discuss the histopathological, immunohistochemical, and molecular findings, emphasizing their relevance to the differential diagnosis of neoplasms with choriocarcinomatous differentiation.


Amelioration of renal carcinogenesis by bee propolis: a chemo preventive approach.
The present study was designed to investigate the chemo preventive efficacy of bee propolis (BP) against diethylnitrosamine (DEN) initiated and ferric nitrilotriacetate (Fe-NTA) promoted renal carcinogenesis in Wistar rats. Chronic treatment of Fe-NTA induced oxidative stress, inflammation and cellular proliferation in Wistar rats. BP is a resinous material collected by bees from various plants which has been used from centuries in folk medicine. Renal cancer was initiated by single intraperitoneal injection of N-nitrosodiethylamine (DEN 200 mg/kg body weight) and promoted by twice weekly administration of Fe-NTA 9 mg Fe/kg body weight for 16 weeks. The chemo preventive efficacy of BP was studied in terms of lipid peroxidation (LPO), renal anti-oxidant armory such as catalase, superoxide dismustase, glutathione S-transferase, glutathione peroxidase, glutathione reductase and glutathione (GSH), serum toxicity markers, cell proliferation, tumor suppressor protein and inflammation markers. Administration of Fe-NTA enhances renal LPO, with concomitant reduction in reduced GSH content and antioxidant enzymes. It induces serum toxicity markers, viz., blood urea nitrogen, creatinine and lactate dehydrogenase. Chemo preventive effects of BP were associated with upregulation of antioxidant armory and down regulation of serum toxicity markers. BP was also able to down regulate expression of proliferative cell nuclear antigen, cyclooxygenase-2, tumor necrosis factor-alpha and upregulated p53 along with induction of apoptosis. Histopathological changes further confirmed the biochemical and immunohistochemical results. These results provide a powerful evidence for the chemo preventive efficacy of BP against renal carcinogenesis possibly by modulation of multiple molecular pathways.


A phase I study of CNI-1493, an inhibitor of cytokine release, in combination with high-dose interleukin-2 in patients with renal cancer and melanoma.
CNI-1493, an inhibitor of proinflammatory cytokines, was studied in a Phase I trial in melanoma and renal cancer patients receiving high-dose interleukin 2 (IL-2). Objectives of the study were to define the maximum tolerated dose (MTD) and toxicity of CNI-1493, to assess its pharmacological effects, and to define its pharmacokinetics. Twenty-four patients were treated in sequential cohorts with CNI-1493 doses from 2 through 32 mg/m2 daily. Patients first received only CNI-1493 daily for 5 days. After a 9-day rest, patients received two 5-day courses of IL-2 of 600,000 IU/kg every 8 h for up to 14 doses/course plus daily CNI-1493; courses were separated by a 9-day rest period. CNI-1493 administered alone was well tolerated at doses through 32 mg/m2; MTD was not reached. The only clinical toxicity attributed to CNI-1493 was occasional injection-site phlebitis. Grade 1 creatinine increases occurred in 1 of 7 patients at 4 mg/m2, in 1 of 1 patients at 25 mg/m2, and in 3 of 6 patients at 32 mg/m2 CNI-1493 alone. In combination with high-dose IL-2, CNI-1493 at > or = 25 mg/m2 seemed to exacerbate IL-2-induced nephrotoxicity: grade 3 or 4 creatinine increases developed in 3 of 6 patients at 25 or 32 mg/m2, as compared with 1 of 16 patients at doses < or = 16 mg/m2. The MTD for CNI-1493 given with high-dose IL-2 was 16 mg/m2. The dose-limiting toxicity of IL-2 was hypotension in 63% of patients; overall tolerance to IL-2 was not improved by CNI-1493. However, relative to changes seen in a reference group receiving high-dose IL-2 alone, at doses > or = 4 mg/m2 CNI-1493 did show evidence of pharmacological activity as an inhibitor of tumor necrosis factor production.


Patterns of changing cancer risks with time since diagnosis of a sibling.
Family history is a well-known risk factor for many cancers. However, it is important to know if/how the familial risk of cancer changes over time. For each of four major cancers (colorectal, breast, prostate and melanoma), we identified siblings of cancer patients (case siblings) and siblings of matched cancer-free controls sampled from Swedish population-based registers. Effects of age and time since diagnosis on sibling risks were examined using Poisson regression and presented graphically as smoothed hazard ratios (HRs). Screening effects were investigated by comparing hazards before/after the introduction of mammography for breast cancer and prostate-specific antigen (PSA) testing for prostate cancer. Case siblings had higher cancer incidence than control siblings for all cancers at all ages, with overall incidence rate ratios (IRRs) of 2.41 (95% confidence interval 2.14-2.71) for colorectal cancer, 2.37 (2.24-2.52) for breast cancer, 3.69 (3.46-3.93) for prostate cancer and 3.20 (2.72-3.76) for melanoma. Risks were highest in siblings who were young when the first cancer was diagnosed in the family, with siblings aged 30-40 having IRR 9.05 (3.03-27.00) for colorectal cancer and 4.30 (2.87-6.45) for breast cancer. Smoothed HRs remained fairly constant for up to 20 years except for prostate cancer, where the HR decreased steeply during the first few years. After introduction of PSA testing, men had higher incidence of prostate cancer shortly after diagnosis in a brother, but no such screening effect was found for breast cancer. Our findings can help inform the screening and counseling of family members of cancer patients.


Real-world evidence generation from patients, their caregivers and physicians supporting clinical, regulatory and guideline decisions: an update on Disease Specific Programmes.
To update on and describe the role of Disease Specific Programmes (DSPs), a multi-perspective real-world data (RWD) source, in the context of the evolution of the value and acceptance of real-world evidence (RWE) in clinical, regulatory and guideline decision-making. DSPs are multi-national, multi-subscriber, multi-therapy cross-sectional surveys incorporating retrospective data collection from patient, caregiver and physician perspectives. Information collected covers the patient journey, including treatment/prescribing patterns and rationale, patient-reported outcomes, impact on work and everyday activities, attitudes towards and perceptions of the condition, adherence to treatment and burden of illness. Published peer-reviewed DSP papers were aligned with current key RWE themes identified in the literature, alongside their contribution to RWE. RWE themes examined were: using RWE to inform clinical practice, patient and caregiver engagement, RWE role in supporting health technology assessments and regulatory submissions, informing value-driven healthcare decisions, real-world patient subgroup differences and therapeutic inertia/unmet needs; highlighting patients' and caregivers' experience of living with a disease, disconnect from their physicians, unmet needs and educational gaps. DSPs provide a wealth of RWD in addition to evidence generated by registries, clinical trials and observational research, with wide use for the pharmaceutical industry, government, funding/regulatory bodies, clinical practice guideline insights and, most importantly, informing improvements in people's lives. The depth, breadth and heritage of information collected via DSPs since 1995 is unparalleled, extending understanding of how diseases are managed by physicians in routine clinical practice and why treatment choices are made, patients' perceptions of their disease management, and caregiver burden.


The significance of magnetic resonance imaging and computed tomography findings in sphenoid sinus agenesis.
Agenesis of the sphenoid sinuses is a very rarely encountered anatomic variation. Findings on magnetic resonance imaging can mislead the radiologist and clinician. Therefore, the gold standard for diagnosis is computed tomography (CT) of the paranasal sinuses. We present the case of a 28-year-old man who was admitted to our hospital with a 3-month history of headache. CT of the paranasal sinuses revealed isolated bilateral sphenoid sinus agenesis.


IL-1 promotes α-epithelial Sodium Channel (α-ENaC) expression in murine lung epithelial cells: involvement of NF-κB.
Intra-amniotic exposure to proinflammatory cytokines such as interleukin-1 (IL-1) correlates with a decreased incidence of respiratory distress syndrome (RDS) in infants following premature birth. At birth, inadequate absorption of fluid from the fetal lung contributes to the onset RDS. Lung fluid clearance is coupled to Na+ transport via epithelial sodium channels (ENaC). In this study, we assessed the effects of IL-1 on the expression of ENaC, particularly the α-subunit which is critical for fetal lung fluid clearance at birth. Cultured mouse lung epithelial (MLE-12) cells were treated with either IL-1α or IL-1β to determine their effects on α-ENaC expression. Changes in IL-1-induced α-ENaC levels in the presence of IL-1 receptor antagonist (IL-1ra), cycloheximide, NF-κB inhibitor, and MAP kinase inhibitors were investigated. IL-1α and IL-1β independently induced a significant increase of α-ENaC mRNA and protein after 24 h compared to untreated cells. IL-1-dependent increases in α-ENaC protein were mitigated by IL-1ra and cycloheximide. IL-1 exposure induced NF-κB binding activity. Attenuation of IL-1-induced NF-κB activation by its inhibitor SN50 decreased α-ENaC protein abundance. Inhibition of ERK 1,2 MAPK significantly decreased both IL-1α and β-induced α-ENaC protein expression whereas inhibition of p38 MAPK only blocked IL-1β-induced α-ENaC protein levels. In contrast, IL-1-induced α-ENaC protein levels were unaffected by a c-Jun N-terminal kinase (JNK) inhibitor. Our results suggest that in MLE-12 cells, IL-1-induced elevation of α-ENaC is mediated via NF-κB activation and in part involves stimulation of the ERK 1,2 and p38 MAPK signaling pathways.


Level of clinical evidence presented at the International Society for Hip Arthroscopy Annual Scientific Meeting over 5 years (2010-2014).
The International Society for Hip Arthroscopy (ISHA) Annual Scientific Meeting is at the forefront of informing today's orthopaedic surgeons and society of the rapid advances in the exponentially growing field of hip arthroscopy. The purpose of this study was to evaluate and observe any trends in the level of clinical evidence in the papers and posters presented at the ISHA Annual Scientific Meeting from 2010 to 2014. The online abstracts of the paper and poster presentations presented at the ISHA Annual Scientific Meetings were independently evaluated by two reviewers (582 total resulting presentations). Two reviewers screened these results for clinical studies and graded the quality of evidence from level I (i.e. randomized trials) to IV (i.e. case series) based on the American Academy of Orthopaedic Surgeons classification system. Four hundred and twenty-eight presentations met the inclusion criteria and were evaluated. Overall, 10.1% of the presentations were level I, 12.8% were level II, 30.1% were level III and 47.0% were level IV evidence. Over time, from 2010 to 2014, we observed an increase in the percentage of level II paper presentations, an increase in the proportion of level III poster presentations, and a decrease in the proportion of both level IV paper and poster presentations. Significant non-random improvement in the level of evidence presented was noted for the poster presentations (P = 0.012) but not for the paper presentations (P = 0.61) over the study period. Statistical trends demonstrate ISHA's increased awareness and commitment to presenting higher quality evidence as the availability of this evidence increases.


Video-assisted thoracoscopic surgery lobectomy for lung cancer is associated with a lower 30-day morbidity compared with lobectomy by thoracotomy.
Lung cancer is the most common cause of cancer-related deaths worldwide. Survival is highly dependent on surgery. Video-assisted thoracoscopic surgery (VATS) is increasingly chosen over open thoracotomy (OT) because of the possible benefits of the minimally invasive approach. Consequently, our aim was to compare the 30-day morbidity and mortality for lung cancer patients operated by VATS lobectomy or lobectomy by OT. Data were obtained from prospective national and regional databases, including patients who underwent lobectomy for lung cancer in the eastern part of Denmark from 1 January 2005 to 31 December 2011. All patients operated before 2009 were re-staged according to the latest International Association for the Study of Lung Cancer lung cancer classification. Patient characteristics, comorbidities, pathology and operative data were assessed using an independent samples t-test, Pearson's χ(2), Fisher's exact test and Mann-Whitney test. Morbidity was assessed using multinomial logistic regression adjusted for gender, age, cancer stage, forced expiratory volume in 1 s (FEV1), year of surgery and Charlson comorbidity score. In total, 1379 patients underwent lobectomy, 785 patients via VATS and 594 patients via thoracotomy. The two groups were similar in gender and FEV1. The patients operated by VATS were older (P < 0.001), and had a lower Charlson comorbidity score (P = 0.034), higher frequency of adenocarcinomas (P < 0.001) and lower cancer stage (P < 0.001). Among the VATS patients, 285 (36.3%) and among the thoracotomy patients, 288 (48.5%) had minor complications (P < 0.001); and 157 (20.0%) VATS patients and 212 (35.7%) thoracotomy patients had major complications (P < 0.001). The 30-day mortality rate was 1% in the VATS group and 1.5% in the thoracotomy group (P = 0.47). Multinomial logistic regression analysis showed that the prevalence of both minor [odds ratio (OR) = 1.51; 95% confidence interval (Cl) = 1.18-1.96] and major complications (OR = 1.91, 95% Cl = 1.44-2.53) was significantly higher for patients who underwent lobectomy via thoracotomy compared with VATS. Patients undergoing lobectomy via VATS were less likely to have at least one minor complication within the first 30 postoperative days and less likely to have at least one major complication, compared with patients operated by thoracotomy. These findings remained after adjusting for gender, age, FEV1, cancer stage, year of surgery and Charlson comorbidity score.


A disintegrin-like and metalloproteinase with thrombospondin-1 (ADAMTS-1) levels in gingival crevicular fluid correlate with vascular endothelial growth factor-A, hypoxia-inducible factor-1α, and clinical parameters in patients with advanced periodontitis.
ADAMTS (a disintegrin-like and metalloproteinase with thrombospondin) are a family of proteinases that are structurally similar to the family of matrix metalloproteinases with critical roles in damage and repair of the extracellular matrix. Their functions are closely related to inflammation, hypoxia, and vascularization. Our aim was to determine levels of ADAMTS-1 in gingival crevicular fluid (GCF) in patients with advanced periodontal diseases and identify their association with hypoxia-inducible factor-1alpha (HIF-1α), vascular endothelial growth factor (VEGF-A), and clinical parameters of periodontitis. The study consisted of three groups: healthy individuals (control; n = 20), generalized chronic periodontitis (CP; n = 21), and generalized aggressive periodontitis (GAgP; n = 20). Clinical parameters were measured. Levels of ADAMTS-1, VEGF-A, and HIF-1α in GCF and serum were quantified by enzyme-linked immunosorbent assay (ELISA) and reported as total amounts and concentration. ADAMTS-1 total amount in GCF were significantly higher in patients with CP and GAgP compared with healthy individuals (P < 0.05). HIF-1α total amount in GCF were also higher in periodontitis groups compared with the control group (P < 0.05). GCF total VEGF-A content was significantly higher in the GAgP group compared with the CP and the controls (respectively; P = 0.023, P = 0.003). There was a significant correlation between ADAMTS-1, VEGF-A, and HIF-1α levels in the GCF and clinical periodontal parameters (probing depth [PD], bleeding on probing [BOP], and clinical attachment loss (CAL); P < 0.05). ADAMTS-1 may play a role in advanced periodontal disease pathogenesis in correlation with tissue hypoxia and vascularization.


Large-Scale Modeling of Wordform Learning and Representation.
The forms of words as they appear in text and speech are central to theories and models of lexical processing. Nonetheless, current methods for simulating their learning and representation fail to approach the scale and heterogeneity of real wordform lexicons. A connectionist architecture termed the sequence encoder is used to learn nearly 75,000 wordform representations through exposure to strings of stress-marked phonemes or letters. First, the mechanisms and efficacy of the sequence encoder are demonstrated and shown to overcome problems with traditional slot-based codes. Then, two large-scale simulations are reported that learned to represent lexicons of either phonological or orthographic word-forms. In doing so, the models learned the statistics of their lexicons as shown by better processing of well-formed pseudowords as opposed to ill-formed (scrambled) pseudowords, and by accounting for variance in well-formedness ratings. It is discussed how the sequence encoder may be integrated into broader models of lexical processing.


Restriction enzyme analysis of mitochondrial DNA of members of the genus Aspergillus as an aid in taxonomy.
Restriction endonuclease patterns of mitochondrial DNA from seven species of the genus Aspergillus (A. nidulans, A. wentii, A. awamori, A. niger, A. oryzae, A. tamarii and A. echinulatus) have been compared in order to test the phylogenetic relationships between these species. The fraction of restriction fragments common between all pairs of species from this set ranged from 0 to 1. The use of this approach for the taxonomy of the genus Aspergillus is discussed. The postulated phylogenetic relationships differ significantly in two cases from the classification of the genus presented by Raper & Fennell (1965).


Tuberculosis: evidence review for newly arriving immigrants and refugees.
The foreign-born population bears a disproportionate health burden from tuberculosis, with a rate of active tuberculosis 20 times that of the non-Aboriginal Canadian-born population, and could therefore benefit from tuberculosis screening programs. We reviewed evidence to determine the burden of tuberculosis in immigrant populations, to assess the effectiveness of screening and treatment programs for latent tuberculosis infection, and to identify potential interventions to improve effectiveness. We performed a systematic search for evidence of the burden of tuberculosis in immigrant populations and the benefits and harms, applicability, clinical considerations, and implementation issues of screening and treatment programs for latent tuberculosis infection in the general and immigrant populations. The quality of this evidence was assessed and ranked using the GRADE approach (Grading of Recommendations Assessment, Development and Evaluation). Chemoprophylaxis with isoniazid is highly efficacious in decreasing the development of active tuberculosis in people with latent tuberculosis infection who adhere to treatment. Monitoring for hepatotoxicity is required at all ages, but close monitoring is required in those 50 years of age and older. Adherence to screening and treatment for latent tuberculosis infection is poor, but it can be increased if care is delivered in a culturally sensitive manner. Immigrant populations have high rates of active tuberculosis that could be decreased by screening for and treating latent tuberculosis infection. Several patient, provider and infrastructure barriers, poor diagnostic tests, and the long treatment course, however, limit effectiveness of current programs. Novel approaches that educate and engage patients, their communities and primary care practitioners might improve the effectiveness of these programs.


Luteinizing release hormone tests in impotent diabetic males.
Assessment of pituitary-gonadal function was made in impotent diabetic males using luteinizing hormone-release hormone tests. Serum testosterone, sex-hormone-binding globulin capacity, and basal and incremental gonadotrophin concentrations in the impotent diabetics were similar to those in control diabetics, suggesting a primary neurologic rather than an endocrine defect as the cause of the impotence.


Chronic Inflammatory Demyelinating Polyneuropathy With Concurrent Membranous Nephropathy: An Anti-paranode and Podocyte Protein Antibody Study and Literature Survey.
Background: Several case reports have described the concurrence of chronic inflammatory demyelinating polyneuropathy (CIDP) and membranous nephropathy (MN). The presence of autoantibodies against podocyte antigens phospholipase A2 receptor (PLA2R) and thrombospondin type 1 domain containing 7A (THSD7A) in MN suggests an autoimmune mechanism. Some CIDP patients also harbor autoantibodies against paranodal proteins such as neurofascin 155 (NF155) and contactin-1 (CNTN1). We investigated the relationship between CIDP and MN by assaying autoantibodies against paranodal and podocyte antigens in a CIDP patient with MN, and by a literature survey on the clinical features of CIDP with MN. Methods: Anti-CNTN1 and NF155 antibodies were measured by flow cytometry using HEK293 cell lines stably expressing human CNTN1 or NF155. Binding capacity of antibodies was validated by immunostaining mouse teased sciatic nerve fibers. Anti-PLA2R antibodies were measured by enzyme-linked sorbent assay and anti-THSD7A antibodies by indirect immunofluorescence assay. Clinical features between 14 CIDP with MN cases including two with anti-CNTN1 antibodies and 20 anti-CNTN1 antibody-positive CIDP cases were compared. Results: A patient whose ages was in the late 70 s complained of progressive weakness and superficial and deep sensory impairment in four extremities over 6 months. Nerve conduction studies showed prominent demyelination patterns. The patient presented with nephrotic syndrome. Renal biopsy disclosed basement membrane thickening with local subepithelial projections and glomerular deposits of IgG4, compatible with MN. Autoantibody assays revealed the presence of IgG4 and IgG1 anti-CNTN1 antibodies, but an absence of anti-NF155, anti-PLA2R, and anti-THSD7A antibodies. The patient's serum stained paranodes of teased sciatic nerves. CIDP with MN and anti-CNTN1 antibody-positive CIDP commonly showed male preponderance, relatively higher age of onset, acute to subacute onset in 35-50% of cases, distal dominant sensorimotor neuropathy, proprioceptive impairment leading to sensory ataxia, and very high cerebrospinal fluid protein levels. However, 11 of 13 CIDP patients with MN had a favorable response to mono- or combined immunotherapies whereas anti-CNTN1 antibody-positive CIDP was frequently refractory to corticosteroids and intravenous immunoglobulin administration. Conclusion: CIDP with MN and anti-CNTN1 antibody-positive CIDP show considerable overlap but are not identical. CIDP with MN is probably heterogeneous and some cases harbor anti-CNTN1 antibodies.


Mutational analysis of active-site residues in the Mycobacterium leprae RecA intein, a LAGLIDADG homing endonuclease: Asp(122) and Asp(193) are crucial to the double-stranded DNA cleavage activity whereas Asp(218) is not.
Mycobacterium leprae recA harbors an in-frame insertion sequence that encodes an intein homing endonuclease (PI-MleI). Most inteins (intein endonucleases) possess two conserved LAGLIDADG (DOD) motifs at their active center. A common feature of LAGLIDADG-type homing endonucleases is that they recognize and cleave the same or very similar DNA sequences. However, PI-MleI is distinctive from other members of the family of LAGLIDADG-type HEases for its modular structure with functionally separable domains for DNA-binding and cleavage, each with distinct sequence preferences. Sequence alignment analyses of PI-MleI revealed three putative LAGLIDADG motifs; however, there is conflicting bioinformatics data in regard to their identity and specific location within the intein polypeptide. To resolve this conflict and to determine the active-site residues essential for DNA target site recognition and double-stranded DNA cleavage, we performed site-directed mutagenesis of presumptive catalytic residues in the LAGLIDADG motifs. Analysis of target DNA recognition and kinetic parameters of the wild-type PI-MleI and its variants disclosed that the two amino acid residues, Asp(122) (in Block C) and Asp(193) (in functional Block E), are crucial to the double-stranded DNA endonuclease activity, whereas Asp(218) (in pseudo-Block E) is not. However, despite the reduced catalytic activity, the PI-MleI variants, like the wild-type PI-MleI, generated a footprint of the same length around the insertion site. The D122T variant showed significantly reduced catalytic activity, and D122A and D193A mutations although failed to affect their DNA-binding affinities, but abolished the double-stranded DNA cleavage activity. On the other hand, D122C variant showed approximately twofold higher double-stranded DNA cleavage activity, compared with the wild-type PI-MleI. These results provide compelling evidence that Asp(122) and Asp(193) in DOD motif I and II, respectively, are bona fide active-site residues essential for DNA cleavage activity. The implications of these results are discussed in this report.


Risk factors for parasuicide among psychiatric inpatients.
To improve suicide prevention during hospitalization, this study examined risk factors for parasuicide among psychiatric inpatients. A group of 58 psychiatric inpatients (19 with schizophrenia, 20 with depression, and 19 with other diagnoses) who displayed suicidal behavior during hospitalization were compared with two control groups of nonparasuicidal inpatients. For inpatients with schizophrenia, a history of parasuicide was a risk factor; for those with depression, it was suicidal behavior on admission; and for patients with other diagnoses, it was violence during hospitalization.


Detecting SARS-CoV-2 neutralizing immunity: highlighting the potential of split nanoluciferase technology.
The coronavirus disease 2019 (COVID-19) pandemic has progressed over 2 years since its onset causing significant health concerns all over the world and is currently curtailed by mass vaccination. Immunity acquired against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) can be following either infection or vaccination. However, one can never be sure whether the acquired immunity is adequate to protect the individual from subsequent infection because of three important factors: individual variations in humoral response dynamics, waning of protective antibodies over time, and the emergence of immune escape mutants. Therefore, a test that can accurately differentiate the protected from the vulnerable is the need of the hour. The plaque reduction neutralization assay is the conventional gold standard test for estimating the titers of neutralizing antibodies that confer protection. However, it has got several drawbacks, which hinder the practical application of this test for wide-scale usage. Hence, various tests have been developed to detect protective immunity against SARS-CoV-2 that directly or indirectly assess the presence of neutralizing antibodies to SARS-CoV-2 in a lower biosafety setting. In this review, the pros and cons of the currently available assays are elaborated in detail and special focus is put on the scope of the novel split nanoluciferase technology for detecting SARS-CoV-2 neutralizing antibodies.


Neuropathic pain in a cat post-amputation.
Phantom limb pain is a form of neuropathic pain experienced by human patients who have had amputations. To date there are no reported cases of phantom limb pain in the veterinary literature. A cat presented with signs consistent with neuropathic or phantom limb pain 42 days after iatrogenic sciatic nerve injury and 38 days after subsequent hind limb amputation. Multimodal analgesic therapy resulted in resolution of signs of pain.


Earthquakes in El Salvador: a descriptive study of health concerns in a rural community and the clinical implications, part I.
This is the first article in a series that evaluates the health concerns of people living in a Salvadoran rural community after major earthquakes. Part I reviews the background, methods, and results of post-earthquake conditions with regards to healthcare, access to healthcare, housing, food, water and sanitation. Part II reviews the implications of these results and recommendations for improvements within the community. Part III investigates the psychosocial and mental health consequences of the earthquakes and provides suggestions for improved mental health awareness, assessment, and intervention. El Salvador experienced 2 major earthquakes in January and February 2001. This study evaluates the effects of the earthquakes on the health practices in the rural town of San Sebastian. The research was conducted with use of a convenience sample survey of subjects affected by the earthquakes. The sample included 594 people within 100 households. The 32-question survey assessed post-earthquake conditions in the areas of health care and access to care, housing, food and water, and sanitation. Communicable diseases affected a number of family members. After the earthquakes, 38% of households reported new injuries, and 79% reported acute exacerbations of chronic illness. Rural inhabitants were 30% more likely to have an uninhabitable home than were urban inhabitants. Concerns included safe housing, water purification, and waste elimination. The findings indicate a need for greater public health awareness and community action to adapt living conditions after a disaster and prevent the spread of communicable disease.


Epidermal growth factor alters metabolism of inositol lipids and activity of protein kinase C in mouse embryo palate mesenchyme cells.
Epidermal growth factor (EGF) stimulated mouse embryo palate mesenchyme (MEPM) cells (1) to incorporate [32P]O4(3-) into phosphatidylinositol (PI), phosphatidylcholine, and phosphatidic acid over a period of 60 min; 2) to incorporate [32P]O4(3-) into polyphosphoinositides as a function of time; and 3) to incorporate [32P]O4(-3) into PI, only, as a function of concentration when the period of stimulation was kept short. EGF stimulated the release of radiolabeled inositol phosphates from MEPM cells that had been radiolabeled with [3H]myoinositol. The release of inositol 1-phosphate was sustained over a period of at least 60 min, whereas the release of inositol 1,4-bisphosphate and inositol trisphosphate peaked during the first 10 min of stimulation. EGF also stimulated phosphorylation of an Mr 80,000 protein whose pI, phosphopeptide map, and phosphoamino acid pattern were identical to those of an Mr 80,000 protein phosphorylated in response to phorbol 12-myristate 13-acetate. Mobilization or metabolism of arachidonic acid was not stimulated under the same conditions that permitted EGF to alter inositol lipid metabolism. We interpret these data to mean that 1) in contrast to the findings with some cell lines, alterations in inositol lipid metabolism may be part of the signalling mechanism for EGF in embryonic cells; 2) EGF is capable of activating inositol-dependent signalling pathways leading to activation of protein kinase C in MEPM cells; and 3) mobilization and metabolism of arachidonic acid are not an inherent part of this signalling mechanism.


Control of Escherichia coli lysyl-tRNA synthetase expression by anaerobiosis.
Escherichia coli lysyl-tRNA synthetase was previously shown to occur as two distinct species encoded by either the lysS or the lysU gene. The expression of one of these genes, lysU, is under the control of cell growth conditions. To study the regulation of lysU, delta lysS strains were constructed. During aerobic growth at 37 degrees C or below, the amount of the lysU product in the cell is so reduced that delta lysS bacteria grow only poorly. The reduced expression of lysU is not related to the steady-state lysyl-tRNA synthetase concentration in the cell, since the expression of a lysU::lacZ fusion is insensitive to the absence of either lysS or lysU or to the addition of a multi-copy plasmid carrying either lysU or lysS. During anaerobic growth in rich medium, the lysU gene becomes strongly expressed and, in cell extracts, the amount of lysyl-tRNA synthetase activity originating from lysU may become seven times greater than the activity originating from lysS. In minimal medium, lysU expression is only slightly induced. Evidence that the sensitivity of lysU expression to anaerobiosis, as well as to low external pH conditions (E. W. Hickey and I. N. Hirshfield, Appl. Environ. Microbiol. 56:1038-1045, 1990), is governed at the level of transcription is provided.


Novel poxvirus infection in 2 patients from the United States.
Some human poxvirus infections can be acquired through zoonotic transmission. We report a previously unknown poxvirus infection in 2 patients, 1 of whom was immunocompromised; both patients had known equine contact. The patients were interviewed and clinical information was abstracted from the patients' medical files. Biopsies of the skin lesions were collected from both patients for histopathology, immunohistochemistry, and transmission electron microscopy analysis. Oral and skin swabs were collected from animals with frequent contact with the patients, and environmental sampling including rodent trapping was performed on the farm where the immunosuppressed patient was employed. "Pan-pox and high Guanine-cytosine" polymerase chain reaction assays were performed on patient, animal, and environmental isolates. Amplicon sequences of the viral DNA were used for agent identification and phylogenetic analysis. Specimens from both human cases revealed a novel poxvirus. The agent shares 88% similarity to viruses in the Parapoxvirus genus and 78% to those in the Molluscipoxvirus genus but is sufficiently divergent to resist classification as either. All animal and environmental specimens were negative for poxvirus and both patients had complete resolution of lesions. This report serves as a reminder that poxviruses should be considered in cutaneous human infections, especially in individuals with known barnyard exposures. The clinical course of the patients was similar to that of parapoxvirus infections, and the source of this virus is currently unknown but is presumed to be zoonotic. This report also demonstrates the importance of a comprehensive approach to diagnosis of human infections caused by previously unknown pathogens.


Adalimumab: Another Medication Related to Osteonecrosis of the Jaws?
Objective. The acronym MRONJ has been created in order to identify "Medication-Related Osteonecrosis of the Jaw," observed after the use of Bisphosphonates, RANK ligand inhibitor, and antiangiogenic medications. Only a case of osteonecrosis of the jaw in a Chron's disease patient following a course of Bisphosphonate and Adalimumab therapy has been recently described, so that it has been supposed that also this medication could promote manifestation of osteonecrosis. Clinical Case. On August, 2014, a 63-year-old female with a history of idiopathic arthritis treated with medical treatment with Adalimumab from 2010 to 2013 presented referring pain in the right mandible. Results. This patient presented with nonexposed osteonecrosis of the jaw after placement, on September, 2010, of four titanium fixtures in the mandible. Conclusions. The authors suggest that the biologic therapy with an anti-TNF-α antibody might promote the manifestation of osteonecrosis and compromise oral healing capacity of the bone.


Uterotonic agents for preventing postpartum haemorrhage: a network meta-analysis.
Postpartum haemorrhage (PPH) is the leading cause of maternal mortality worldwide. Prophylactic uterotonic agents can prevent PPH, and are routinely recommended. The current World Health Organization (WHO) recommendation for preventing PPH is 10 IU (international units) of intramuscular or intravenous oxytocin. There are several uterotonic agents for preventing PPH but there is still uncertainty about which agent is most effective with the least side effects. This is an update of a Cochrane Review which was first published in April 2018 and was updated to incorporate results from a recent large WHO trial. To identify the most effective uterotonic agent(s) to prevent PPH with the least side effects, and generate a ranking according to their effectiveness and side-effect profile. We searched the Cochrane Pregnancy and Childbirth's Trials Register, ClinicalTrials.gov, the World Health Organization (WHO) International Clinical Trials Registry Platform (ICTRP) (24 May 2018), and reference lists of retrieved studies. All randomised controlled trials or cluster-randomised trials comparing the effectiveness and side effects of uterotonic agents with other uterotonic agents, placebo or no treatment for preventing PPH were eligible for inclusion. Quasi-randomised trials were excluded. Randomised trials published only as abstracts were eligible if sufficient information could be retrieved. At least three review authors independently assessed trials for inclusion and risk of bias, extracted data and checked them for accuracy. We estimated the relative effects and rankings for preventing PPH ≥ 500 mL and PPH ≥ 1000 mL as primary outcomes. Secondary outcomes included blood loss and related outcomes, morbidity outcomes, maternal well-being and satisfaction and side effects. Primary outcomes were also reported for pre-specified subgroups, stratifying by mode of birth, prior risk of PPH, healthcare setting, dosage, regimen and route of administration. We performed pairwise meta-analyses and network meta-analysis to determine the relative effects and rankings of all available agents. The network meta-analysis included 196 trials (135,559 women) involving seven uterotonic agents and placebo or no treatment, conducted across 53 countries (including high-, middle- and low-income countries). Most trials were performed in a hospital setting (187/196, 95.4%) with women undergoing a vaginal birth (71.5%, 140/196).Relative effects from the network meta-analysis suggested that all agents were effective for preventing PPH ≥ 500 mL when compared with placebo or no treatment. The three highest ranked uterotonic agents for prevention of PPH ≥ 500 mL were ergometrine plus oxytocin combination, misoprostol plus oxytocin combination and carbetocin. There is evidence that ergometrine plus oxytocin (RR 0.70, 95% CI 0.59 to 0.84, moderate certainty), carbetocin (RR 0.72, 95% CI 0.56 to 0.93, moderate certainty) and misoprostol plus oxytocin (RR 0.70, 95% CI 0.58 to 0.86, low certainty) may reduce PPH ≥ 500 mL compared with oxytocin. Low-certainty evidence suggests that misoprostol, injectable prostaglandins, and ergometrine may make little or no difference to this outcome compared with oxytocin.All agents except ergometrine and injectable prostaglandins were effective for preventing PPH ≥ 1000 mL when compared with placebo or no treatment. High-certainty evidence suggests that ergometrine plus oxytocin (RR 0.83, 95% CI 0.66 to 1.03) and misoprostol plus oxytocin (RR 0.88, 95% CI 0.70 to 1.11) make little or no difference in the outcome of PPH ≥ 1000 mL compared with oxytocin. Low-certainty evidence suggests that ergometrine may make little or no difference to this outcome compared with oxytocin meanwhile the evidence on carbetocin was of very low certainty. High-certainty evidence suggests that misoprostol is less effective in preventing PPH ≥ 1000 mL when compared with oxytocin (RR 1.19, 95% CI 1.01 to 1.42). Despite the comparable relative treatment effects between all uterotonics (except misoprostol) and oxytocin, ergometrine plus oxytocin, misoprostol plus oxytocin combinations and carbetocin were the highest ranked agents for PPH ≥ 1000 mL.Misoprostol plus oxytocin reduces the use of additional uterotonics (RR 0.56, 95% CI 0.42 to 0.73, high certainty) and probably also reduces the risk of blood transfusion (RR 0.51, 95% CI 0.37 to 0.70, moderate certainty) when compared with oxytocin. Carbetocin, injectable prostaglandins and ergometrine plus oxytocin may also reduce the use of additional uterotonics but the certainty of the evidence is low. No meaningful differences could be detected between all agents for maternal deaths or severe morbidity as these outcomes were rare in the included randomised trials where they were reported.The two combination regimens were associated with important side effects. When compared with oxytocin, misoprostol plus oxytocin combination increases the likelihood of vomiting (RR 2.11, 95% CI 1.39 to 3.18, high certainty) and fever (RR 3.14, 95% CI 2.20 to 4.49, moderate certainty). Ergometrine plus oxytocin increases the likelihood of vomiting (RR 2.93, 95% CI 2.08 to 4.13, moderate certainty) and may make little or no difference to the risk of hypertension, however absolute effects varied considerably and the certainty of the evidence was low for this outcome.Subgroup analyses did not reveal important subgroup differences by mode of birth (caesarean versus vaginal birth), setting (hospital versus community), risk of PPH (high versus low risk for PPH), dose of misoprostol (≥ 600 mcg versus < 600 mcg) and regimen of oxytocin (bolus versus bolus plus infusion versus infusion only). All agents were generally effective for preventing PPH when compared with placebo or no treatment. Ergometrine plus oxytocin combination, carbetocin, and misoprostol plus oxytocin combination may have some additional desirable effects compared with the current standard oxytocin. The two combination regimens, however, are associated with significant side effects. Carbetocin may be more effective than oxytocin for some outcomes without an increase in side effects.


Prospective study of provocative angiograms to localize functional islet cell tumors of the pancreas.
Controversy exists concerning the use of preoperative imaging studies in patients with islet cell tumors. Since 1993 we have evaluated the use of provocative angiography in patients with insulinoma or Zollinger-Ellison syndrome (ZES). Twelve patients with a working diagnosis of insulinoma (n = 4) or ZES (n = 8) were studied. Of the eight patients with ZES, four were known to have multiple endocrine neoplasia type 1. All patients underwent conventional imaging studies followed by provocative angiography. Angiograms were graded based on the ability to detect tumor and regionalize it within the pancreas. Of the three patients with a working diagnosis of ZES but equivocal results of biochemical studies, none had arteriographic imaging of an islet cell tumor or a positive provocative study result (true negative result). Of the nine patients with documented islet cell tumor, seven (78%) underwent arteriographic imaging of the tumor and eight (89%) had correct regional localization by provocative angiography. Sensitivity and specificity for imaging were 78% and 100%, respectively, and for regional localization 89% and 100%, respectively. Provocative angiography is the localization study of choice for both gastrinoma and insulinoma. Having few false-negative results, it can be used to corroborate the diagnosis and, having few false-positive results, it detects tumor and biochemically confirms localization in nearly every patient.


Staging of hip osteoarthritis for clinical trials on femoroacetabular impingement.
Future clinical trials investigating the natural history and treatment of femoroacetabular impingement (FAI) will require multimodal staging systems for hip osteoarthritis because the optimal system will differ based on the size of the study population, the specific objective in question, and the time frame in which the investigator expects to see the specified end point. Plain radiographs are readily available, low in cost, and of unquestioned validity, but they are relatively insensitive to early joint damage. MRI allows assessment of both bony and soft-tissue pathology within the joint, and it is much more sensitive for early joint damage because cartilage is visualized directly. Biochemical imaging techniques such as delayed gadolinium-enhanced MRI of cartilage, T2 mapping, and T1rho offer the potential to identify biochemical damage to cartilage before the onset of irreversible tissue loss. In the future, biomarkers may allow earlier detection of osteoarthritis before the development of radiographic evidence of disease.


Using SNOMED-CT to encode summary level data - a corpus analysis.
Extracting and encoding clinical information captured in free text with standard medical terminologies is vital to enable secondary use of electronic medical records (EMRs) for clinical decision support, improved patient safety, and clinical/translational research. A critical portion of free text is comprised of 'summary level' information in the form of problem lists, diagnoses and reasons of visit. We conducted a systematic analysis of SNOMED-CT in representing the summary level information utilizing a large collection of summary level data in the form of itemized entries. Results indicate that about 80% of the entries can be encoded with SNOMED-CT normalized phrases. When tolerating one unmapped token, 96% of the itemized entries can be encoded with SNOMED-CT concepts. The study provides a solid foundation for developing an automated system to encode summary level data using SNOMED-CT.


Cytogenetic and molecular cytogenetic findings in 43 aneurysmal bone cysts: aberrations of 17p mapped to 17p13.2 by fluorescence in situ hybridization.
Aneurysmal bone cyst is a benign, cystic lesion of bone composed of blood-filled spaces separated by fibrous septa. Relatively few cases of aneurysmal bone cyst have been cytogenetically characterized, yet abnormalities of the short arm of chromosome 17 appear to be recurrent. In this study, conventional cytogenetic analysis of 43 aneurysmal bone cyst specimens from 38 patients over a 12-year period revealed clonal chromosomal abnormalities in 12 specimens. Karyotypic anomalies of 17p, including a complex translocation and inversion, were identified in eight of these 12 specimens. In an effort to further define the aberrant 17p breakpoint, fluorescence in situ hybridization (FISH) analyses were performed using a series of probe combinations spanning a 5.1 Mb region between the TP53 (17p13.1) and Miller-Dieker lissencephaly syndrome (17p13.3) gene loci. These studies revealed the critical breakpoint locus at 17p13.2, flanked proximally by an RP11-46I8, RP11-333E1, and RP11-457I18 bacterial artificial chromosome (BAC) probe cocktail and distally by an RP11-198F11 and RP11-115H24 BAC and RP5-1050D4 P1 artificial chromosome (PAC) probe cocktail. Overall, abnormalities of the 17p13.2 locus were identified by metaphase and/or interphase cell FISH analysis in 22 of 35 (63%) aneurysmal bone cyst specimens examined including 26 karyotypically normal specimens. These cytogenetic and molecular cytogenetic findings expand our knowledge of chromosomal alterations in aneurysmal bone cyst, further localize the critically involved 17p breakpoint, and provide an alternative approach (ie FISH) for detecting 17p abnormalities in nondividing cells of aneurysmal bone cysts. The latter could potentially be utilized as an adjunct in diagnostically challenging cases.


Chronic kidney disease and coronary atherosclerosis: evidences from intravascular imaging.
Introduction: In CKD subjects, atherosclerotic cardiovascular disease (ASCVD) is a major cause of their morbidity and mortality (1-3). Current therapeutic guidelines recommend lowering LDL-C level to prevent ASCVD (4, 5). However, it remains uncertain how this therapeutic approach effectively modifies atherosclerosis of CKD. These findings suggest the need to further understand pathophysiology of atherosclerosis. Given that intravascular imaging modalities have contributed to characterize the natural history of coronary atherosclerosis (13-23), the integration of plaque imaging is expected to help to elucidate targets associated with stabilizing atheroma.Areas covered: This review will outline atherogenic stimuli in patients with CKD. In addition, the characteristics of their coronary atheroma will be presented from finding in clinical studies using a variety of intravascular imaging modalities. The efficacy of current guideline recommended anti-atherosclerotic therapies in CKD patients will be also summarized. All related articles were searched through PubMed.Expert opinion: Integration of intravascular imaging has a great potential to establish effective therapies for slowing progression of atherosclerosis in subjects with CKD. More efforts toward searching therapeutic target associated with atherosclerosis of CKD are required. In particular, identifying drivers associated with plaque calcification will lead to the development of new agents which regress calcium accumulation.


High-Intensity Forward-Backward Plyometrics After the Warm-Up Entail Better Sprint and Change-of-Direction Performance Than Low-Intensity Side-to-Side Plyometrics.
This study aimed to determine the acute effects of high-intensity forward-backward and low-intensity side-to-side plyometric jumps performed following the warm-up on sprint (5, 10, and 15 m) and change-of-direction (COD) (T-half test and repeated T-half tests) performance in youth volleyball players. After a familiarization week, 30 male volleyball players (age = 12.04 ± 1.03 years) performed three randomized conditions (no-plyometrics, high-intensity plyometrics, and low-intensity plyometrics) in three sessions. In a within-subject design, three sets of six repetitions of forward-backward 30-cm hurdle jumps (high-intensity) and side-to-side 20-cm hurdle jumps (low-intensity) were completed. Sprint and COD were tested after each of the conditions with a 2-min rest. A significant effect of the plyometric condition was observed on sprint (p < .001, ηp2 range: .56-.70) and COD (p < .01, ηp2=.24), but not on repeated COD. More specifically, the high-intensity plyometric condition exhibited significantly better results compared with the low-intensity plyometric (Cohen's d range: 0.73-1.21) and control conditions (Cohen's d range: 0.91-2.21). Due to the importance of speed and COD in volleyball, these results suggest that young volleyball players may benefit from high-intensity forward-backward plyometric protocols following the warm-up to improve subsequent performance.


Optical Measurement of the Stoichiometry of Thin-Film Compounds Synthetized From Multilayers: Example of Cu(In,Ga)Se2.
The properties of centimeter-sized thin-film compound semiconductors depend upon the morphology and chemical composition of the multiple submicrometer-thick elemental and alloy precursor layers from which they are synthesized. The challenge is to characterize the individual precursor layers over these length scales during a multistep synthesis without altering or contaminating them. Conventional electron and X-ray-based morphological and compositional techniques are invasive, require preparation, and are thus incompatible with in-line synthesis processes. In a proof-of-concept study, we applied confocal laser scanning microscopy (CLSM) as a noninvasive optical imaging technique, which measures three-dimensional surface profiles with nanoscale resolution, to this challenge. Using an array of microdots containing Cu(In,Ga)Se2 semiconductor layers for solar cells as an example, we performed CLSM correlative studies to quantify morphological and layer thickness changes during four stages of a thin-film compound synthesis. Using simple assumptions, we measured the micrometer-scale spatially resolved chemical composition of stacked precursor layers to predict the final material phases formed and predict relative device performance. The high spatial resolution, coupled with the ability to measure sizeable areas without influencing the synthesis at high speed, makes CLSM an excellent prospect for research and quality control tool for thin films.


Barriers to screening and isoniazid preventive therapy for child contacts of tuberculosis patients.
India's guidelines recommend tuberculosis (TB) screening of household contacts aged <6 years and isoniazid preventive therapy (IPT) for children without active disease. We evaluated the current status and barriers to screening and IPT provision among the child contacts of TB patients. Questionnaire and health record data were collected from index cases and health care providers (HCPs) at Sassoon General Hospital, Pune, India. Of 80 adult TB cases, 24 (30%) reported that an HCP recommended TB screening of their child contacts; 49/178 (28%) child contacts were screened. Sixteen (33%) children had active TB, and 28 (85%) of those who screened negative were prescribed IPT. Nineteen (76%) HCPs reported recommending child contact screening. Only 8 (32%) reported ever prescribing IPT. Lack of TB screening and IPT provision for child contacts was associated with inadequate HCP counseling (aOR 19.5, P < 0.001), a non-parent index case (aOR 3.72, P = 0.008) and lack of postgraduate HCP qualification (aOR 19.12, P = 0.04). TB screening and IPT provision for child contacts of adults with TB were infrequent. Many screened children had active TB. Universal, timely TB screening and IPT for exposed children are urgently needed to reduce pediatric TB in India.


Synthetic inositol 1,3,4,5-tetrakisphosphate analogues.
Inositol 1,3,4,5-tetrakisphosphate [Ins(1,3,4,5)P4] analogues were synthesized and their effects on [3H]Ins(1,3,4,5)P4 5-phosphatase, [3H]Ins(1,3,4,5)P4 3-phosphatase and [3H]inositol 1,4,5-trisphosphate [3H]Ins(1,4,5)P3] 5-phosphatase activities were examined. The Ins(1,3,4,5)P4 analogue with the aminobenzoyl group at the 2-position of Ins(1,3,4,5)P4 inhibited the hydrolysis of 5-phosphate of [3H]Ins(1,3,4,5)P4 catalysed by erythrocyte ghosts, with a lower Ki value than seen with Ins(1,3,4,5)P4, whereas the analogue with the aminocyclohexanecarbonyl group at the same position had a higher Ki value. The Ins(1,4,5)P3 analogues that we had previously synthesized were also capable of inhibiting this process, with the same tendency as Ins(1,3,4,5)P4 analogues. Such differences in the potency among Ins(1,3,4,5)P4 and Ins(1,4,5)P3 analogues were applicable to other phosphatase activities, namely [3H]Ins(1,3,4,5)P4 3-phosphatase and [3H]Ins(1,4,5)P3 5-phosphatase. These results suggest that the active sites of these enzymes may catalyse the dephosphorylation in a similar fashion.


Identifying changes in gait waveforms following a strengthening intervention for women with knee osteoarthritis using principal components analysis.
Lower limb strengthening exercise is pivotal for the management of symptoms related to knee osteoarthritis (OA). Though improvement in clinical symptoms is well documented, concurrent changes in gait biomechanics are ill-defined. This may occur because discrete analyses miss changes following an intervention, analyses limited to the knee undermine potential mechanical trade-offs at other joints, or strengthening interventions not been designed based on biomechanical principles. The purpose of this study was to characterize differences in entire gait waveforms for sagittal plane ankle, knee, and hip angles and external moments; the knee adduction moment; and frontal plane hip angle and moment following 12-weeks of a previously designed novel lower limb strengthening program. Forty women with knee OA completed two laboratory visits: one at baseline and one immediately following intervention (follow-up). Self-report measures, strength, and gait analyses were completed at each visit. Principal components analyses were completed for sagittal angles and external moments at the ankle, knee, and hip joints, as well as frontal plane angle and moment for the hip. Participants improved self-report and strength (p≤0.004). Two significant, yet subtle differences in principal components were identified between baseline and follow-up waveforms (p<0.05) pertaining to the knee and hip sagittal external moments. The subtle changes in concert with the lack of differences in other joints and planes suggest the lower limb strengthening program does not translate to changes in the gait waveform. It is likely this program is improving symptoms without worsening mechanics.


Defective DNA-mismatch repair: a potential mediator of leukemogenic susceptibility in therapy-related myelodysplasia and leukemia.
We investigated the potential role of defective DNA-mismatch repair (MMR) as a mediator of leukemogenic susceptibility in patients with therapy-related myelodysplasia (t-MDS) and leukemia (t-leuk). Thirty-seven individuals with t-MDS/t-leuk were analyzed for microsatellite instability (MSI), the hallmark of defective DNA-MMR. Using standardized international criteria, 5/37 (14%) patients displayed high MSI, whereas 3 other patients had low MSI (8%). To determine the stage at which MSI had developed, we analyzed the primary tumors of 12 patients. Three of 4 patients with high MSI t-MDS/t-leuk also had microsatellite unstable primary tumors. Conversely, MSI was not detected in any primary malignancy of patients with low MSI or microsatellite stable t-MDS/t-leuk (P = 0.0182). In the high MSI group, we further investigated genes targeted by defective DNA-MMR (BAX, TGFBRII, IGFIIR, Caspase-5, APC, PTEN, E2F4, MBD4, MSH6, and MSH3) in both primary tumor and t-MDS/t-leuk. However, no mutation was found in any gene. The significant association of MSI in t-MDS/t-leuk and corresponding primary tumors suggests that defective DNA-MMR confers leukemogenic susceptibility to this cohort of patients.


Inhibition of chemotactic factor-activated Na+/H+ exchange in human neutrophils by analogues of amiloride: structure-activity relationships in the amiloride series.
The ability of a number of analogues of the diuretic, amiloride, to inhibit chemotactic factor-stimulated Na+/H+ exchange in human neutrophils was investigated. Intracellular pH (pHi) changes were measured from the equilibrium distribution of 14C-labeled 5,5-dimethyloxazolidine-2,4-dione (DMO). Exposure of cells to 10 nm N-formyl-methionyl-leucyl-phenylalanine (FMLP) caused activation of Na+/H+ exchange: in 140 mM Na+ medium (extracellular pH 7.40), the pHi rose from a resting value of approximately 7.25 to reach a new steady state of approximately 7.75 by 10-15 min. This intracellular alkalinization was sensitive to amiloride (apparent Ki approximately 75 microM), a known inhibitor of Na+/H+ countertransport. The structure-activity relationships in the amiloride series were characterized by testing the effect of these compounds on the DMO-derived pHi changes and on the FMLP-stimulated rate of 22Na+ efflux from the cells. Substitutions of the guanidino group of amiloride resulted in relatively inactive products (Ki greater than or equal to 1 mM). Replacement of the 6-Cl group of amiloride by other halogen atoms had only modest effects on drug efficacy. However, replacement of one or both H atoms of the 5-amino group by short alkyl groups led to a 10-500-fold increase in potency for inhibition of Na+/H+ exchange. Amiloride and three of its more potent derivatives (compounds I, O, and MM, the 5-N,N-dimethyl, 5-N,N-diethyl, and 5-N,N-hexamethylene analogues, respectively) caused parallel inhibition of FMLP-activated 22Na+ efflux and the rate of intracellular alkalinization, with apparent Ki values of approximately 75, 8, 1, and 0.2 microM, respectively. In each instance, the inhibitory effects of the drugs were readily reversible on washing the cells. None of the compounds altered the binding of 3H-labeled FMLP to its cell surface receptors. The development of potent derivatives of amiloride should provide powerful tools for assessing the role of FMLP-activated Na+/H+ exchange and the resultant pHi transients on stimulated neutrophil functions.


Nanoscopic micelle delivery improves the photophysical properties and efficacy of photodynamic therapy of protoporphyrin IX.
Nanodelivery systems have shown considerable promise in increasing the solubility and delivery efficiency of hydrophobic photosensitizers for photodynamic therapy (PDT) applications. In this study, we report the preparation and characterization of polymeric micelles that incorporate protoporphyrin IX (PpIX), a potent photosensitizer, using non-covalent encapsulation and covalent conjugation methods. Depending on the incorporation method and PpIX loading percentage, PpIX existed as a monomer, dimer or aggregate in the micelle core. The PpIX state directly affected the fluorescence intensity and (1)O(2) generation efficiency of the resulting micelles in aqueous solution. Micelles with lower PpIX loading density (e.g. 0.2%) showed brighter fluorescence and higher (1)O(2) yield than those with higher PpIX loading density (e.g. 4%) in solution. However, PDT efficacy in H2009 lung cancer cells showed an opposite trend. In particular, 4% PpIX-conjugated micelles demonstrated the largest PDT therapeutic window, as indicated by the highest phototoxicity and relatively low dark toxicity. Results from this study contribute to the fundamental understanding of nanoscopic structure-property relationships of micelle-delivered PpIX and establish a viable micelle formulation (i.e. 4% PpIX-conjugated micelles) for in vivo evaluation of antitumor efficacy.


Taylor Dispersion of Polydisperse Nanoclusters and Nanoparticles: Modeling, Simulation, and Analysis.
The dimensions of ultrasmall inorganic nanoparticles (US-NPs) is in the heart of the design of diagnostic and therapeutic efficacy; yet its accurate measurement is challenging for most experimental techniques. We show here how to design and analyze Taylor dispersion experiments to characterize the two most sought-after parameters describing size distributions: the number-averaged mean size and polydispersity index. To demonstrate the power of the method, we simulated and analyzed taylograms corresponding to gold US-NPs distributed normally. By using simulation and including experimental noise, we had the advantage that the true values describing size distribution were known exactly, and thus, we were able test the absolute accuracy of our analysis and its robustness against noise. Theory and computational experiments were found in very good agreement, providing a significant step in the analysis of ultrasmall inorganic nanoparticles and Taylor dispersion experiments.


Survival analysis of temporary anchorage devices: A retrospective analysis in a Nigerian orthodontic patient population.
Temporary anchorage devices (TADs) are skeletal anchorage devices. They are minimally invasive and placed by the orthodontist to prevent unwanted tooth movement. This study evaluated the survival rate of orthodontic TADs at 6 months. This study also assessed the effect of age, gender, side, site, dental arch of placement, and length of the TADs on its survival rate. Ethical approval was obtained from the Health Research Ethics Committee of the hospital. The study sample comprised orthodontic patients who required the placement of TADs during treatment at a private dental facility in Lagos. Data for the study were obtained from the case files of the study subjects and included the subjects' age, gender, date of placement of the TADs, the site, side and arch of placement, the length of the TADs, and the survival rate of 6 months after placement. We reviewed 90 placed TADs and observed a survival rate of 88.9%. Most TAD failures occurred in the first month of placement (p = 001). There was no observable statistically significant effect of all other variables assessed (age, gender, arch, site, side, or implant length) on the survival rate of the TADs. The survival rate of TADs was high. Most TAD failures significantly occurred within one month of placement. There was no significant association between all other clinical variables and orthodontic mini-implant survival.


Respiratory markers of conversational interaction.
Respiratory movements were recorded from 10 dyads (20 subjects) during quiet breathing, reading aloud, spontaneous monologue, scripted dialog, and spontaneous conversation. Timing measures of inspiratory, expiratory, and total cycle duration were used to compare respiratory function during quiet breathing, listening, and speech. Cross-correlation analyses of the respiratory movements of conversational partners provided an index of conversational synchrony. Inspiratory duration was found to be the most consistent and sensitive measure for discriminating quiet breathing from speech breathing. In the scripted dialog and spontaneous conversation conditions, respiratory kinematics changed during listening to more closely resemble speech, and systematic changes were observed in anticipation of turn-taking speech onset. For the breathing cycles immediately surrounding turn changes and simultaneously produced vocal events, the kinematic signals of conversational partners were strongly correlated. Results are discussed in the context of similar findings concerning conversational interactions and motor preparation for speech.


Full flex-grid asynchronous multiplexing demonstrated with Nyquist pulse-shaping.
We demonstrate full flex-grid operation with Nyquist frequency division multiplexing. The technique supports high spectral efficiency, asynchronous operation of channels, variable channel loading with different modulation formats and dynamic bandwidth allocation. Data from different sources with different bit and symbol rates are encoded onto electrical Nyquist pulses with different electrical subcarrier frequencies, and then transmitted optically. We give details on the transceiver design with digital signal processing and investigate the implementation penalty as a function of several design parameters such as limited filter length and effective number of bits. Finally, experiments are performed for receivers with direct detection, intradyne and remote heterodyne reception.