Nitric oxide causes inactivation of the low molecular weight phosphotyrosine protein phosphatase.
The low M(r) phosphotyrosine protein phosphatase (PTPase) and Yersinia enterocolitica PTPase are inactivated by nitric oxide-generating compounds. Inorganic phosphate, a competitive inhibitor, protects the enzymes from inactivation, suggesting that the action of NO is directed to the active sites. Low M(r) PTPase from bovine liver lost two out of eight thiol groups present in the molecule during the inactivation with sodium nitroprusside and with other NO-producing compounds. The mass spectrometric analyses of tryptic fragments of the enzyme, performed after chemical modification of the NO-unreacted thiol groups, demonstrated that NO caused the oxidation of Cys-12 and Cys-17 to form an S-S bond. A similar reaction was described previously for the reaction of NO with N-methyl-D-aspartate receptor. The NO-inactivated low M(r) PTPase was reactivated by treating the inactive enzyme with thiol-containing reagents. Since all members of the PTPase family have the same reaction mechanism and possess a conserved active site motif that contains an essential cysteine residue, the findings on low M(r) and Yersinia PTPases are potentially interesting for all PTPases, an enzyme class that is involved in a number of important biological processes.


The expectations of generation Z regarding the university educational act in Romania: optimizing the didactic process by providing feedback.
In the university education system, the evaluation and provision of two-way teacher-student feedback are tools within the control function of educational management. Feedback can be defined as the process that takes place in the context of teacher-student interaction during university courses, both individually and in groups, for the development and achievement of the performance of the two actors involved through evaluation, appreciation, support, perception, and teaching. The research aims to develop an innovative feedback tool for the higher education engineering sector to support the improvement of learning outcome-oriented curricula and teaching activities to better meet the learning needs of Gen Z students while being relevant to the labor market and to society in general. The research had a number of subjects: 246 students (67.5% women, 32.5% men) and 7 teachers to whom two feedback instruments were applied (the SKS instrument and the standard instrument of Lucian Blaga University of Sibiu). After testing the four hypotheses, it was observed that the feedback provided by Gen Z is focused on four areas of competence: psychological, pedagogical, education management, and general impression. Each field includes a set of professional and transversal competencies. The SKS (STOP, KEEP, and START) evaluation form is more reliable in evaluating different disciplines than the standard evaluation form by providing a more homogenous type of feedback for each discipline or teacher.


Precritical abnormalities in routine blood parameters in necrotizing fasciitis.
Necrotizing fasciitis is a rare and severe infectious disease that is often fatal and is characterized by the extensive necrosis of subcutaneous tissue and fascial planes. A number of clinical parameters have been intensively investigated to diagnose and assess the severity and prognosis of necrotizing fasciitis. Since it currently remains unclear whether these parameters are also abnormal before disease onset, the present study investigated this issue. We retrospectively recruited 38 patients, including 12 and 26 patients with necrotizing fasciitis and cellulitis, respectively. The results of routine blood examinations were collected at disease onset and also at baseline, which was defined as the time point before disease onset. No significant differences were observed in age or sex between the necrotizing fasciitis and cellulitis groups. However, significant differences were noted in the levels of hemoglobin, lymphocyte count, platelet count, neutrophil-to-lymphocyte ratio, sodium, creatinine, albumin, D-dimer, and Laboratory Risk Indicator for Necrotizing Fasciitis (LRINEC) score at disease onset. Significant differences were also observed in the levels of hemoglobin, lymphocyte count, monocyte count, platelet count, creatinine, D-dimer, and LRINEC score at baseline. Hemoglobin, platelet count, C-reactive protein, creatinine, albumin, and D-dimer levels were already abnormal at baseline in the necrotizing fasciitis group. In conclusion, the present results revealed precritical abnormalities in routine blood parameters in patients with necrotizing fasciitis. Therefore, individuals predisposed to necrotizing soft tissue infection may be identified prior to disease onset.


Long-term tear volume changes after blepharoptosis surgery and blepharoplasty.
To evaluate long-term changes in tear volume by using video meniscometry following blepharoptosis surgery and upper blepharoplasty. Forty-three eyes of 27 patients with blepharoptosis and 29 eyes of 18 patients with dermatochalasis without lacrimal duct obstruction or other eyelid diseases underwent anterior approach levator advancement or blepharoplasty. Tear volume was evaluated by measurement of tear meniscus radius (R), using video meniscometry preoperatively and at 1.5, 3, and 6 months postoperatively. Margin reflex distance-1 (MRD-1) was measured before and after surgery by using photographs. After blepharoptosis surgery, the mean MRD-1 was significantly increased: 0.45 mm preoperatively, 3.64 mm at 1.5 months, 3.56 mm at 3 months, and 3.57 at 6 months postoperatively (P < 0.001), and the average R value was significantly decreased: 0.29 mm preoperatively, 0.22 mm at 1.5 months, 0.23 mm at 3months, and 0.24 mm at 6 months postoperatively (P < 0.05). Preoperative R was significantly correlated to the reduction rate of R (ΔR). A higher preoperative R was more likely to be decreased (P < 0.01). Postoperative MRD-1 and change in MRD-1 were not correlated to ΔR. After blepharoplasty, the preoperative mean MRD-1 (3.11 mm) was significantly decreased at 1.5 months (2.47 mm; P < 0.01) and 3 months (2.71 mm; P < 0.05) but recovered at 6 months (3.14 mm). However, the average R was not changed: 0.31 mm preoperatively, 0.34 mm at 1.5 months, 0.31 mm at 3 months, and 0.33 mm at 6 months postoperatively. Long-term tear volume was not changed after blepharoplasty but was decreased after blepharoptosis surgery, and even more so in cases with an initial high tear volume.


Existence of homoclinic connections in continuous piecewise linear systems.
Numerical methods are often used to put in evidence the existence of global connections in differential systems. The principal reason is that the corresponding analytical proofs are usually very complicated. In this work we give an analytical proof of the existence of a pair of homoclinic connections in a continuous piecewise linear system, which can be considered to be a version of the widely studied Michelson system. Although the computations developed in this proof are specific to the system, the techniques can be extended to other piecewise linear systems.


Demonstration of a neutral atom controlled-NOT quantum gate.
We present the first demonstration of a CNOT gate between two individually addressed neutral atoms. Our implementation of the CNOT uses Rydberg blockade interactions between neutral atoms held in optical traps separated by >8 microm. Using two different gate protocols we measure CNOT fidelities of F=0.73 and 0.72 based on truth table probabilities. The gate was used to generate Bell states with fidelity F=0.48+/-0.06. After correcting for atom loss we obtain an a posteriori entanglement fidelity of F=0.58.


Atomically Precise Clusters of Noble Metals: Emerging Link between Atoms and Nanoparticles.
Atomically precise pieces of matter of nanometer dimensions composed of noble metals are new categories of materials with many unusual properties. Over 100 molecules of this kind with formulas such as Au25(SR)18, Au38(SR)24, and Au102(SR)44 as well as Ag25(SR)18, Ag29(S2R)12, and Ag44(SR)30 (often with a few counterions to compensate charges) are known now. They can be made reproducibly with robust synthetic protocols, resulting in colored solutions, yielding powders or diffractable crystals. They are distinctly different from nanoparticles in their spectroscopic properties such as optical absorption and emission, showing well-defined features, just like molecules. They show isotopically resolved molecular ion peaks in mass spectra and provide diverse information when examined through multiple instrumental methods. Most important of these properties is luminescence, often in the visible-near-infrared window, useful in biological applications. Luminescence in the visible region, especially by clusters protected with proteins, with a large Stokes shift, has been used for various sensing applications, down to a few tens of molecules/ions, in air and water. Catalytic properties of clusters, especially oxidation of organic substrates, have been examined. Materials science of these systems presents numerous possibilities and is fast evolving. Computational insights have given reasons for their stability and unusual properties. The molecular nature of these materials is unequivocally manifested in a few recent studies such as intercluster reactions forming precise clusters. These systems manifest properties of the core, of the ligand shell, as well as that of the integrated system. They are better described as protected molecules or aspicules, where aspis means shield and cules refers to molecules, implying that they are "shielded molecules". In order to understand their diverse properties, a nomenclature has been introduced with which it is possible to draw their structures with positional labels on paper, with some training. Research in this area is captured here, based on the publications available up to December 2016.


Tritium enrichment by gas-solid chromatography: technic for low-level analysis.
A palladium chromatograph was developed that can detect less than 10 atoms of tritium per 10(18) atoms of hydrogen. Columns operated at ambient temperature have 70-liter hydrogen capacity (50-milliliter water sample) and give 60 percent recovery in the first 500 cubic centimeters of gas evolved.


A Qualitative Analysis of the Spontaneous Volunteer Response to the 2013 Sudan Floods: Changing the Paradigm.
Introduction While the concept of community resilience is gaining traction, the role of spontaneous volunteers during the initial response to disasters remains controversial. In an attempt to resolve some of the debate, investigators examined the activities of a spontaneous volunteer group called Nafeer after the Sudan floods around the city of Khartoum in August of 2013. Hypothesis Can spontaneous volunteers successfully initiate, coordinate, and deliver sustained assistance immediately after a disaster? This retrospective, descriptive case study involved: (1) interviews with Nafeer members that participated in the disaster response to the Khartoum floods; (2) examination of documents generated during the event; and (3) subsequent benchmarking of their efforts with the Sphere Handbook. Members who agreed to participate were requested to provide all documents in their possession relating to Nafeer. The response by Nafeer was then benchmarked to the Sphere Handbook's six core standards, as well as the 11 minimum standards in essential health services. A total of 11 individuals were interviewed (six from leadership and five from active members). Nafeer's activities included: food provision; delivery of basic health care; environmental sanitation campaigns; efforts to raise awareness; and construction and strengthening of flood barricades. Its use of electronic platforms and social media to collect data and coordinate the organization's response was effective. Nafeer adopted a flat-management structure, dividing itself into 14 committees. A Coordination Committee was in charge of liaising between all committees. The Health and Sanitation Committee supervised two health days which included mobile medical and dentistry clinics supported by a mobile laboratory and pharmacy. The Engineering Committee managed to construct and maintain flood barricades. Nafeer used crowd-sourcing to fund its activities, receiving donations locally and internationally using supporters outside Sudan. Nafeer completely fulfilled three of Sphere's core standards and partially fulfilled the other three, but none of the essential health services standards were fulfilled. Even though the Sphere Handbook was chosen as the best available "gold standard" to benchmark Nafeer's efforts, it showed significant limitations in effectively measuring this group. It appears that independent spontaneous volunteer initiatives, like Nafeer, potentially can improve community resilience and play a significant role in the humanitarian response. Such organizations should be the subject of increased research activity. Relevant bodies should consider issuing separate guidelines supporting spontaneous volunteer organizations. Albahari A , Schultz CH . A qualitative analysis of the spontaneous volunteer response to the 2013 Sudan floods: changing the paradigm. Prehosp Disaster Med. 2017;32(3):240-248.


[Immorals III. Midwives].
This article is the third and last part of a complex anthropological study on 17th and 18th century southern Hungarian sexual customs and deviations as they appeared in witch trials. The first two parts came out in our Com. Art. Hist. Med. vols. 117-120 (1987) pp 87-102, and 133-140 (1991-1992) pp 171-216. The author has studied the papers of 1040 cases and compared it to the most important issues of Hungarian secondary literature of folklore, cultural and social anthropology, and occasionally to history as well. The three papers presented different subjects of these themes. Whereas the first paper dealt with aggression and especially aggression during sexual intercourses, the second one collected those cases which revealed other various forms of deviated sexual behaviour, and prostitution. In this essay the author studied another related area, the job of midwives in village society including not only their main activities in helping labouring women, but their additional work in giving sexual advice, curing various illnesses, and providing baby care. His main concern in this essay, nevertheless, is about witch charges against medwives. ...


Image-guided pleural biopsies: indications, technique, and results in 23 patients.
Twenty-six pleural biopsies were performed on 23 patients over a 3-year period. Twenty-three biopsies were performed guided with ultrasound; one, with computed tomography; and two, with fluoroscopy. Indications for an image-guided pleural biopsy were (a) pleural masses or thickening that were either not seen on chest radiographs or seen only on one view and (b) small or loculated pleural effusions of unknown cause with no mass seen. If only pleural fluid was present, reverse bevel needles were used for biopsy (n = 15). If a discrete pleural mass or thickening was seen with cross-sectional imaging, standard (16-20 gauge) biopsy needles were used (n = 11). In the 23 patients, biopsy results were true positive in ten (nine with malignancy, one with tuberculous pleurisy), true negative in ten (confirmed either at subsequent thoracotomy or clinical follow-up), and false negative in three. Complications were few, with a significant pneumothorax occurring in two patients (8.7%). Image-guided biopsy of small pleural lesions and small pleural effusions can be performed by the radiologist who understands the special needles and techniques involved.


[Epidemiology of osteoporosis].
Epidemiological studies have identified osteoporosis as a disease of significant public health impact and have delineated numerous potential risk factors. Nevertheless, it has proven difficult to establish preventive strategies for several reasons. First, there has been no final agreement on the definition of osteoporosis, which has hampered efforts to characterize the magnitude of the problem as a whole. Secondly, as osteoporosis is a multifactorial chronic disorder, effective programs for risk assessment and intervention depend on the development of complex disease models. In summarizing the contributions of epidemiological studies to the current understanding of osteoporosis this review intends to outline the scientific background for the European Vertebral Osteoporosis Study (EVOS) and its successors.


[Effect of tetracycline hydrochloride on the development of the thymus structure in the rat].
Histological, electron microscopy and morphological methods were used to study the structure of the thymus of Wistar rats in the antenatal and early postnatal ontogenesis after administering to their mothers of the therapeutic doses of tetracycline hydrochloride at varying times of pregnancy (days 1-6, 8-13, 14-19). A study was also made of the alterations in the thymus of one-month-old rats exposed to the antibiotic prenatally in response to the administration of bacterial antigen.


Synthesis and Characterization of an Environmentally Friendly Phenol-Formaldehyde Resin Modified with Waste Plant Protein.
To develop a lower-cost, excellent-performance, and environmentally friendly phenol-formaldehyde (PF) resin, soybean meal was used to modify PF resin, and soybean meal-phenol-formaldehyde (SMPF) resins were prepared. This reveals the effect of soybean meal on the structural, bonding, and curing properties of PF resin, which are very important for its applications in the wood industry. The resins' physicochemical properties and curing performance were investigated, showing that SMPF resins have higher curing temperatures than PF resin. The Fourier transform infrared spectroscopy results indicated that a cross-linking reaction occurred between the amino groups of soybean protein and the hydroxymethyl phenol. Moreover, with the addition of soybean meal, the viscosity of SMPF increased while the gel time decreased. It is worth mentioning that SMPF-2 resin has favorable viscosity, short gel time, low curing temperature (135.78 °C), and high water resistance and bonding strength (1.01 MPa). Finally, all the plywoods bonded with SMPF resins have good water resistance and bonding strength, which could meet the standard (GB/T 17657-2013, type I) for plywood. The optimized SMPF resins showed the potential for application to partially replace PF resin in the wood industry.


Overcoming resistance against managed care - insights from a bargaining model.
Recent healthcare reforms have sought to increase efficiency by introducing managed care (MC) while respecting consumer preferences by admitting choice between MC and conventional care. This article proposes an institutional change designed to let German consumers choose between the two settings through directing payments from the Federal Health Fund to social health insurers (SHIs) or to specialized MC organizations (MCOs). To gauge the chance of success of this reform, a game involving a SHI, a MCO, and a representative insured (RI) is analyzed. In a "three-player/three-cake" game the coalitions {SHI, MCO}, {MCO, RI}, and {SHI, RI} can form. Players' possibility to switch between coalitions creates new outside options, causing the conventional bilateral Nash bargaining solution to be replaced by the so-called von Neumann-Morgenstern triple. These triples are compared to the status quo (where the RI has no threat potential) and related to institutional conditions characterizing Germany, the Netherlands, and Switzerland.


Epibulbar osseous choristoma: Two case reports.
Choristoma is a rare, benign, congenital proliferative tumor, with osseous choristoma being the rarest. Although the tumor is benign, effective identification is needed for its diagnosis and treatment. Here, we report the diagnosis and successful surgical treatment of two patients with osseous choristoma. Two patients, a young female and young male patient, were found to have a mass on the ocular surface. The tumor presented on the superior temporal bulbar conjunctiva in the first patient and on the upper eyelid in the second patient. Ultrasound biomicroscopy detected a strong echo with clear boundaries covering the lower echo, and computed tomography examination revealed calcification. Both patients underwent surgery, and histopathological evaluation of the mass showed osseous choristoma. They were treated by excision and subsequently cured. Osseous choristomas are usually asymptomatic. Our patients were cured immediately after surgery, suggesting that surgical treatment is an effective strategy.


Developmental changes in presynaptic Ca(2 +) clearance kinetics and synaptic plasticity in mouse Schaffer collateral terminals.
Presynaptic Ca(2+) influx pathways, cytoplasmic Ca(2+) buffering proteins and Ca(2+) extrusion processes undergo considerable change during the first postnatal month in rodent neurons. These changes may be critical in establishing short-term plasticity at maturing presynaptic terminals where neurotransmitter release is directly dependent on the dynamics of cytoplasmic residual Ca(2+) ([Ca(2+)](res)). In particular, the robust paired-pulse facilitation characteristic of adult neurons is almost entirely lacking in newborns. To examine developmental changes in processes controlling [Ca(2+)](res), we measured the timecourse of [Ca(2+)](res) decay in presynaptic terminals of Schaffer collateral to CA1 synapses in acute hippocampal slices following single and paired orthodromic stimuli in the stratum radiatum. Developmental changes were observed in both the rise time and slow exponential decay components of the response to single stimuli such that this decay was larger and faster in the adult. Furthermore, we observed a greater caffeine-sensitive basal Ca(2+) store, which was differentially affected when active uptake into the endoplasmic reticulum was blocked, in the presynaptic fields of the Schaffer collateral to CA1 terminals of P6 and younger mice when compared to adults. These transitions in [Ca(2+)](res) dynamics occurred gradually over the first weeks of postnatal life and correlated with changes in short-term plasticity.


The implications of health literacy on patient-provider communication.
Limited health literacy has been associated with a range of adverse health outcomes including decreased use of preventive health services, poorer disease-specific outcomes for certain chronic conditions and increased risk of hospitalisation and mortality. Although the majority of research has been conducted in the adult population, there is a small and growing body of research on this subject in the paediatric literature. In this article, we will review the research on health literacy, consider the range of other communication skills associated with limited health literacy and explore strategies to improve patient-provider communication for clinicians who care for families with limited health-literacy skills.


Hepatocyte nuclear factor-1alpha is required for expression but dispensable for histone acetylation of the lactase-phlorizin hydrolase gene in vivo.
Hepatocyte nuclear factor-1alpha (HNF-1alpha) is a modified homeodomain-containing transcription factor that has been implicated in the regulation of intestinal genes. To define the importance and underlying mechanism of HNF-1alpha for the regulation of intestinal gene expression in vivo, we analyzed the expression of the intestinal differentiation markers and putative HNF-1alpha targets lactase-phlorizin hydrolase (LPH) and sucrase-isomaltase (SI) in hnf1alpha null mice. We found that in adult jejunum, LPH mRNA in hnf1alpha(-/-) mice was reduced 95% compared with wild-type controls (P < 0.01, n = 4), whereas SI mRNA was virtually identical to that in wild-type mice. Furthermore, SI mRNA abundance was unchanged in the absence of HNF-1alpha along the length of the adult mouse small intestine as well as in newborn jejunum. We found that HNF-1alpha occupies the promoters of both the LPH and SI genes in vivo. However, in contrast to liver and pancreas, where HNF-1alpha regulates target genes by recruitment of histone acetyl transferase activity to the promoter, the histone acetylation state of the LPH and SI promoters was not affected by the presence or absence of HNF-1alpha. Finally, we showed that a subset of hypothesized intestinal target genes is regulated by HNF-1alpha in vivo and that this regulation occurs in a defined tissue-specific and developmental context. These data indicate that HNF-1alpha is an activator of a subset of intestinal genes and induces these genes through an alternative mechanism in which it is dispensable for chromatin remodeling.


Risk Factors for an Iatrogenic Mallory-Weiss Tear Requiring Bleeding Control during a Screening Upper Endoscopy.
Background and Aim. In some cases of iatrogenic Mallory-Weiss tears (MWTs), hemostasis is needed due to severe mucosal tearing with bleeding. Therefore, we aimed to evaluate the risk factors for severe iatrogenic MWTs and the methods of endoscopic bleeding control. Materials and Methods. Between January 2008 and December 2012, 426,085 cases of screening upper endoscopy were performed at the Asan Medical Center. We retrospectively analyzed the risk factors for severe iatrogenic MWTs requiring an endoscopic procedure and the treatment modalities of bleeding control. Results. Iatrogenic MWTs occurred in 546 cases (0.13%) of screening upper endoscopy in 539 patients. Bleeding control due to severe bleeding was applied in 71 cases (13.0%), and rebleeding after initial bleeding control occurred in 1 case. Multivariate analysis showed that old age, a history of distal gastrectomy, and a less-experienced endoscopist (fewer than 2,237.5 endoscopic procedures at the time of the MWT) were associated with severe iatrogenic MWTs requiring an endoscopic procedure. Among 71 cases requiring bleeding control, a hemoclip was used in 81.7% (58 cases). Conclusions. Screening endoscopy procedures should be carefully performed when patients are in their old age and have a history of distal gastrectomy, particularly if the endoscopist is less experienced.


Carbonic anhydrase II-positive pancreatic cells are progenitors for both endocrine and exocrine pancreas after birth.
The regenerative process in the pancreas is of particular interest because diabetes results from an inadequate number of insulin-producing beta cells and pancreatic cancer may arise from the uncontrolled growth of progenitor/stem cells. Continued and substantial growth of islet tissue occurs after birth in rodents and humans, with additional compensatory growth in response to increased demand. In rodents there is clear evidence of pancreatic regeneration after some types of injury, with proliferation of preexisting differentiated cell types accounting for some replacement. Additionally, neogenesis or the budding of new islet cells from pancreatic ducts has been reported, but the existence and identity of a progenitor cell have been debated. We hypothesized that the progenitor cells are duct epithelial cells that after replication undergo a regression to a less differentiated state and then can form new endocrine and exocrine pancreas. To directly test whether ductal cells serve as pancreatic progenitors after birth and give rise to new islets, we generated transgenic mice expressing human carbonic anhydrase II (CAII) promoter: Cre recombinase (Cre) or inducible CreER(TM) to cross with ROSA26 loxP-Stop-loxP LacZ reporter mice. We show that CAII-expressing cells within the pancreas act as progenitors that give rise to both new islets and acini normally after birth and after injury (ductal ligation). This identification of a differentiated pancreatic cell type as an in vivo progenitor of all differentiated pancreatic cell types has implications for a potential expandable source for new islets for replenishment therapy for diabetes.


A new approach to studying ion uptake by actinomycetes.
A streptomycete that had the ability to avidly sequester iron via siderophores was previously isolated from environmental soil samples. The chelating agent expressed by this organism is confirmed by HPLC as desferrioxamine E. Although the traditional chromo azuerol sulphate (CAS) assay for detection of siderophores is based upon the chelation of iron we were interested to examine the relationship of these iron-capturing molecules with other ions. Consequently, a new approach was employed that enabled the assessment of the affinity of the siderophore moieties for other ions by adapting the CAS assay. The present study reveals that the isolate produced a siderophore that was capable of sequestering a range of ions including Mn, Co, Cd, Ni, Al, Li, Cu, Zn and Mg. On the basis of the assay described it would appear that the organism sequesters copper more readily than iron. This raises an age-old debate surrounding the replacement of copper as a fundamentally essential element with iron as a consequence of the evolution of the di-oxygen environment.


Risk factors for dermatitis in submariners during a submerged patrol: an observational cohort study.
The aim of this pilot study was to determine risk factors, including Staphylococcus aureus nasal carriage, for dermatitis in submariners during a submarine patrol. 36 submariners undertaking a submerged 6-week patrol participated in the study. Severity of dermatitis and its impact was assessed using visual analogue scales and questionnaires at baseline and weekly throughout the patrol. S. aureus carriage levels in submariners were determined by nasal swabbing at baseline and shortly before disembarking the submarine. Occurrence of any skin or soft tissue infections (SSTI) were reported to the medical officer and swabs of the area were taken for subsequent analysis. S. aureus carriers were significantly more likely than non-carriers to have previously received treatment for a cutaneous abscess (39% vs 5%, OR=13 (95% CI 1.3 to 130)) with a trend to being submariners longer (p=0.051). Skin scores at baseline and on patrol were not significantly associated with carriage status. Higher dermatitis scores were observed in those who had been submariners longer (p=0.045). Smoking and allergies were not found to be linked to carriage status or skin health score in this cohort. This small pilot study investigates S. aureus carriage status and skin health in submariners. Length of submarine service but not S. aureus carriage was identified as a risk factor for worsening skin health in this small cohort during a 6-week patrol. This does not support S. aureus decolonisation to improve skin health in this population. Further investigation into causes of dermatitis in submariners is required. This data supports a better understanding of the potential impact of exposure to environmental factors that could affect skin health in submariners.


Analysis of metachronous colorectal neoplasms and survival following segmental or extended resection in patients with hereditary non-polyposis colorectal cancer.
The high incidence of metachronous colorectal tumours in patients with hereditary non-polyposis colorectal cancer (HNPCC) encourages extended resection (ER); however, the optimal surgical approach remains unclear. We evaluated the incidences of metachronous colorectal neoplasms following curative colorectal cancer segmental resection (SR) vs ER in patients with HNPCC and investigated patients' oncologic outcomes according to surgical modality and mismatch repair status. We retrospectively investigated medical records of patients with HNPCC (per the Amsterdam II criteria) treated for primary colon cancer at our institution between 2001 and 2017. All patients underwent intensive endoscopic surveillance. We included 87 patients (36 who underwent SR and 51 who underwent ER). The cumulative incidence of metachronous adenoma was higher in the SR group. One patient in the SR group (2.8%) and 3 in the ER group (5.9%) developed metachronous colon cancer; the difference was not significant (P = 0.693). Four patients in the SR group (11.1%) and 1 in the ER group (2.0%) developed distant recurrences; again, the difference was not significant (P = 0.155). Moreover, no significant differences were observed in the 5-year overall survival rates of patients in the SR and ER groups (88.2% vs 95.5%, P = 0.446); the same was true for 5-year disease-free survival rates (79.5% vs 91.0%, P = 0.147). The incidence of metachronous cancer was not significantly different between the ER and SR groups; however, that of cumulative metachronous adenoma was higher in the SR group. Hence, intensive surveillance colonoscopy may be sufficient for patients with HNPCC after non-extensive colon resection.


Hsp90 inhibitor gedunin causes apoptosis in A549 lung cancer cells by disrupting Hsp90:Beclin-1:Bcl-2 interaction and downregulating autophagy.
Hsp90 is regarded as an important therapeutic target in cancer treatment. Client proteins of Hsp90 like Beclin-1, PI3K, and AKT, are associated with tumor development, poor prognosis, and resistance to cancer therapies. This study aims to analyze the role of Gedunin, an Hsp-90 inhibitor, in mediation of crosstalk between apoptosis and autophagy by targeting Beclin-1:Bcl-2 interaction, and ER stress. A549 cells were treated with different concentrations of gedunin, and inhibitory rate was evaluated by MTT assay. Effect of gedunin on generation of reactive oxygen species, mitochondrial membrane potential, and chromatin condensation was studied by staining methods like DCFH-DA, MitoTracker, and DAPI. Expression of EGFR, PIK3CA, AKT, marker genes for apoptosis and autophagy were studied using semi-quantitative RT-PCR. Interaction study of Hsp90:Beclin-1:Bcl-2 was done by immunoprecipitation analysis. Protein expression of autophagy and apoptosis markers along with Grp78, Hsp70, and Hsp90 was analyzed by immunoblotting. Gedunin exerts cytotoxic effects, causes increase in ROS generation, downregulates mitochondrial membrane potential and induces loss in DNA integrity. mRNA expression analysis revealed that gedunin sensitized A549 cells towards apoptosis by downregulating EGFR, PIK3CA, AKT, and autophagy. Gedunin also inhibited interaction between Hsp90:Beclin-1:Bcl-2, leading to downregulation of autophagy (Beclin-1, Atg5-12 complex, and LC3) and antiapoptotic protein Bcl-2, which may result in ER stress-induced apoptosis. Moreover, Hsp90 inhibition by gedunin did not cause upregulation of Hsp70 expression. Gedunin induces apoptosis in lung cancer cells by disrupting Hsp90:Beclin-1:Bcl-2 interaction and autophagy downregulation, thus making gedunin a good drug lead for targeting lung cancer.


Effectiveness of clinical practice change strategies in improving dietitian care for head and neck cancer patients according to evidence-based clinical guidelines: a stepped-wedge, randomized controlled trial.
Best practice guidelines make a number of recommendations regarding dietitian management of head and neck cancer (HNC) patients. Randomized trials assessing the effectiveness of clinical practice change strategies for improving the nutritional management of HNC patients have not previously been conducted. The purpose of this study was to evaluate the effect of practice change strategies on improving the implementation of best practice guideline recommendations for the nutritional management of HNC patients. Four Australian radiotherapy departments participated in a stepped-wedge, randomized controlled trial. Baseline data were collected across all sites simultaneously, and the intervention was then introduced to each site sequentially, in a randomly determined order. During the intervention phase, sites received a range of supportive clinical practice change strategies to facilitate dietitian adherence to clinical practice guidelines. To assess the associated practice change by dietetic staff, we evaluated the change in implementation of six guideline recommendations for dietitians from preintervention to postintervention periods. Adherence to the clinical practice guidelines during the preintervention period was generally very low. The clinical practice change strategies significantly improved the odds of provision of four of the six guideline recommendations. The study found the intervention significantly enhanced dietitian provision of recommended care for HNC patients during the postintervention period. This finding holds clinical importance for clinician and health service effective implementation of guideline recommendations as well as HNC patient treatment outcomes. Trial registration number ACTRN12613000320752, https://www.anzctr.org.au.


Social capital, income inequality, and self-rated health in 45 countries.
There has been growing interest in the relationship between the social environment and health. Among the concepts that have emerged over the past decade to examine this relationship are socio-economic inequality and social capital. Using data from the World Values Survey and the World Bank, we tested the hypothesis that self-rated health is affected by social capital and income inequality cross-nationally. The merit of our approach was that we used multilevel methods in a larger and more diverse sample of countries than used previously. Our results indicated that, for a large number of diverse countries, commonly used measures of social capital and income inequality had strong compositional effects on self-rated health, but inconsistent contextual effects, depending on the countries included. Cross-level interactions suggested that contextual measures can moderate the effect of compositional measures on self-rated health. Sensitivity tests indicated that effects varied in different subsets of countries. Future research should examine country-specific characteristics, such as differences in cultural values or norms, which may influence the relationships between social capital, income inequality, and health.


Distribution of hydroxyapatite crystallite orientation and ultrasonic wave velocity in ring-shaped cortical bone of bovine femur.
At the nanoscopic level, bone consists of calcium phosphate, which forms incomplete hydroxyapatite (HAp) crystals. The preferred orientation of the c-axis of HAp crystallites induces anisotropy and inhomogeneity of elastic properties in bone. In this study, the effect of the preferred orientation of HAp crystallites on the spatial distribution of ultrasonic wave velocity was experimentally investigated, considering bone mineral density (BMD) and microstructure. Three ring-shaped cortical bone samples were made from a 36-month-old bovine femur. Longitudinal wave velocity was measured by a conventional ultrasonic pulse system, using self-made polyvinylidene fluoride transducers. The integrated intensity of the (0002) peak obtained using X-ray diffraction was estimated to evaluate the amount of preferred orientation. The velocity distribution pattern was similar to the distribution of integrated intensity of (0002). The effect of the preferred orientation of HAp crystallites on velocity was clearly observed in the plexiform structure, despite the fact that the BMD value was almost independent of the preferred orientation of HAp crystallites. Velocity measurement of cortical bone can reveal information about HAp crystallite orientation.


Respiratory disease of workers harvesting grain.
The incidence of respiratory symptoms caused by grain dust during harvesting was surveyed in a group of Lincolnshire farmers. A quarter complained of respiratory distress after working on combine harvesters or near grain driers and elevators, with cough, wheezing, and breathlessness, sometimes so severe as to prevent work. The airborne dust around combine harvesters contained up to 200 million fungus spores/m3 air with Cladosporium predominant while drivers were exposed to up to 20 million spores/m3 air. Verticillium/Paecilomyces type spores, mostly from Verticillium lecanii, Aphanocladium album, and Paecilomyces bacillosporus, were abundant in the dust. Extracts of these species produced immediate weal reactions in skin tests, precipitin reactions with sera, and rapid decreases in FEV1 when inhaled by affected workers. There was no delayed reactions. Results suggest type I immediate hypersensitivity to the spores although the physical effect of a heavy dust deposit could be important. Drivers could be protected by cabs ventilated with filtered air.


Specialisation of extracellular matrix for function in tendons and ligaments.
Tendons and ligaments are similar structures in terms of their composition, organisation and mechanical properties. The distinction between them stems from their anatomical location; tendons form a link between muscle and bone while ligaments link bones to bones. A range of overlapping functions can be assigned to tendon and ligaments and each structure has specific mechanical properties which appear to be suited for particular in vivo function. The extracellular matrix in tendon and ligament varies in accordance with function, providing appropriate mechanical properties. The most useful framework in which to consider extracellular matrix differences therefore is that of function rather than anatomical location. In this review we discuss what is known about the relationship between functional requirements, structural properties from molecular to gross level, cellular gene expression and matrix turnover. The relevance of this information is considered by reviewing clinical aspects of tendon and ligament repair and reconstructive procedures.


Modulation of emission properties of phosphine-sulfonate ligand containing copper complexes: playing with solvato-, thermo-, and mechanochromism.
Modulations of various neutral phosphine containing copper(i) complexes exhibiting phosphorescence and TADF are easily achieved by solvent modification restricting intramolecular rotation or by modifying the L-X phosphine-sulfonate chelate. This concept is not limited to solvents as the addition of metallic salts also led to the formation of original well-defined photoluminescent K2CuLX3 and KCuL2X2 cuprate complexes.


Battered woman syndrome: empirical findings.
The construct of Battered Woman Syndrome (BWS) has been conceptualized as a subcategory of posttraumatic stress disorder (PTSD). It is composed of the following symptoms: (a) re-experiencing the battering as if it were reoccurring even when it is not, (b) attempts to avoid the psychological impact of battering by avoiding activities, people, and emotions, (c) hyperarousal or hypervigilance, (d) disrupted interpersonal relationships, (e) body image distortion or other somatic concerns, and (f) sexuality and intimacy issues. This article presents empirical data derived from administering the Battered Woman Syndrome Questionnaire (BWSQ) to women of four countries--United States, Spain, Greece, and Russia. The data support a theory of BWS.


Hereditary cancer-associated mutations in women diagnosed with two primary cancers: an opportunity to identify hereditary cancer syndromes after the first cancer diagnosis.
Patients with hereditary cancer syndromes are at high risk for a second primary cancer. Early identification of these patients after an initial cancer diagnosis is the key to implementing cancer risk-reducing strategies. A commercial laboratory database was searched for women with a history of both breast and ovarian or colorectal and endometrial cancer who underwent genetic testing for hereditary breast and ovarian cancer (HBOC) or Lynch syndrome (LS). Among women with both breast and ovarian cancer, 22.4% (2,237/9,982) had a BRCA1 or BRCA2 mutation. Among women with both colorectal and ovarian cancer, 28.1% (264/941) had a mutation associated with LS. In 66.6% of BRCA1 or BRCA2 mutation carriers and in 58.3% of LS mutation carriers, >5 years passed between the cancer diagnoses. Of patients with HBOC and LS, 56 and 65.2%, respectively, met the National Comprehensive Cancer Network guidelines for hereditary cancer testing after their initial diagnosis based on their personal cancer history alone. A substantial number of women tested for LS or HBOC after being diagnosed with two successive primary cancers were diagnosed with a hereditary cancer syndrome. In many cases, the time interval between the diagnoses was long enough to allow for the implementation of surveillance and/or prophylactic measures.


Sera from habitual aborters induce monocyte procoagulant activity: a lymphocyte-dependent event.
Placental thrombosis is a prominent feature in patients with unexplained recurrent fetal loss. To determine whether induction of monocyte procoagulant activity might be a relevant mechanism for unexplained recurrent fetal loss, peripheral blood mononuclear cells isolated from normal healthy controls were cocultured with (a) sera from normal healthy controls (n = 16), (b) sera from habitual aborters (n = 41), and (c) lipopolysaccharide as a positive control. Sera from three patients were fractionated on Sephracryl S-300 and the inducing molecule(s) characterized. Sera from normal healthy controls failed to induce procoagulant activity above basal levels of 21 +/- 4.6 mU/10(5) peripheral blood mononuclear cells. Of the sera from 41 habitual aborters examined 26 (63%) induced procoagulant to a mean value of 410 +/- 48 mU/10(5) peripheral blood mononuclear cells (P < 0.01). Sera from 15 patients failed to augment procoagulant activity. The induction of procoagulant activity was maximal after 6 hr of incubation and was lymphocyte dependent. Fractionation of serum from the three patients on Sepharcryl S300 revealed the procoagulant activity (PCA)-inducing factor(s) to have a molecular weight of between 300,000 and 800,000 Da. The serum factor was found to be heat, alkaline, and acid sensitive. Both anti-IgM and anti-IgA immunoabsorbents reduced the PCA-inducing factor. We conclude that IgM and IgA from some patients with unexplained recurrent fetal loss are capable of inducing procoagulant activity and could contribute to the development of placental microthrombi and infarction, prominent features of this syndrome.


The removable buried retention suture: a new technique.
Through-and-through retention sutures are inherently inefficient and prone to produce skin and subcutaneous damage. Most surgeons are reluctant to permanently bury non-absorbable retention sutures. This new technique uses tube conduits to bring the retention sutures from deep fascial level to above the skin-surface, where they are available for removal. Thus, the sutures do not cross the skin and subcutaneous tissue, and they exert maximal efficiency at the deep fascial layers. This method has been used in over 60 cases without deep-wound separation, and complications have been minimal.


Occupational safety, health, and hygiene in the urban informal sector of sub-Saharan Africa: an application of the Prevention and Control Exchange (PACE) program to the informal-sector workers in healthy city projects.
In October 2000, at an international meeting in Pretoria, South Africa, chaired by the World Health Organization (WHO), 45 delegates from Sub-Saharan Africa, Europe, and the United States discussed the results of the Healthy City project in the region and the possibilities for incorporating occupational safety, health, and hygiene items into the project. Working conditions play only a minor role in current Healthy City projects. This is partly the result of a low profile of occupational safety, health, and hygiene on political agendas, and partly a result of a lack of convincing demonstration projects. To tackle these points WHO will start two initiatives. First, a document for decision makers will be issued, to highlight the subject and bring it to the attention of local, national, and international bodies. Second, demonstration projects will be started, making use of the methods developed in the Work Improvement In Small Enterprises (WISE) projects and the experience of the Prevention and Control Exchange (PACE) project.


Nutritional studies of the embryo during early organogenesis with normal embryos and embryos exhibiting yolk sac dysfunction.
In 1961 we reported that heterologous kidney antiserum when injected into pregnant rats resulted in wide spectrum of congenital malformations. Further studies identified that it was the IgG component of the antiserum that was teratogenic and that complement was not necessary to produce the teratogenic effect. Labeled antibody studies demonstrated that the kidney antiserum localized in the kidney and in the visceral yolk sac (VYS) and parietal yolk sac placentas. Preparation of yolk sac (YS) antiserum proved to be more potent than the kidney antiserum. Adsorption studies with VYS and parietal yolk sac antiserum revealed that the site of the teratogenic process was located in the VYS. In vitro embryo culture experiments demonstrated that direct injection of the teratogenic antibody into the amniotic or YS cavity did not injure the embryo, thus indicating that the teratogenic antibody had to come in contact with the absorptive surface of the VYS. Collaboration with Dr. John Lloyd demonstrated that teratogenic antibody interfered with the process of pinocytosis and the delivery of amino acids (AA) to the developing embryo. Our studies into the nature of the source of AA for the embryo indicated that in some instances > 95% of the AA present in the developing embryo were derived from maternal proteins and the remainder from free AA in the maternal serum. We also demonstrated that embryonic methionine was derived primarily from the digestion of maternal serum proteins but that more of the methionine was diverted from the synthesis of embryonic proteins, supporting the view that it has important functions other than the synthesis of proteins. All these studies focus on the role of the YS in human development and whether human YS dysfunction may play a role in the pathogenesis of congenital malformations. Further studies on the delivery of AA to the embryo are warranted to determine whether certain AA are in short supply in maternal serum and place the embryo at risk if nutritional alterations in the maternal environment occurs. Furthermore, the YS may be an organ whose role might offer opportunities for pregnancy control.


Plant invasion is associated with higher plant-soil nutrient concentrations in nutrient-poor environments.
Plant invasion is an emerging driver of global change worldwide. We aimed to disentangle its impacts on plant-soil nutrient concentrations. We conducted a meta-analysis of 215 peer-reviewed articles and 1233 observations. Invasive plant species had globally higher N and P concentrations in photosynthetic tissues but not in foliar litter, in comparison with their native competitors. Invasive plants were also associated with higher soil C and N stocks and N, P, and K availabilities. The differences in N and P concentrations in photosynthetic tissues and in soil total C and N, soil N, P, and K availabilities between invasive and native species decreased when the environment was richer in nutrient resources. The results thus suggested higher nutrient resorption efficiencies in invasive than in native species in nutrient-poor environments. There were differences in soil total N concentrations but not in total P concentrations, indicating that the differences associated to invasive plants were related with biological processes, not with geochemical processes. The results suggest that invasiveness is not only a driver of changes in ecosystem species composition but that it is also associated with significant changes in plant-soil elemental composition and stoichiometry.


Apoptosis induced by selenomethionine and methioninase is superoxide mediated and p53 dependent in human prostate cancer cells.
Selenomethionine (SeMet) is the chemical form or major component of selenium used for cancer chemoprevention in several clinical trials. However, evidence from experimental studies indicates that SeMet has weaker anticancer effects than most other forms of selenium. Recent studies showed that the anticancer activity of SeMet can be enhanced by methioninase (METase), indicating that SeMet metabolites are responsible for its anticancer activity. In the present study, we showed that wild-type p53-expressing LNCaP human prostate cancer cells were more sensitive to cotreatment with SeMet and METase than p53-null PC3 human prostate cancer cells. SeMet and METase cotreatment significantly increased levels of superoxide and apoptosis in LNCaP cells. Cotreatment with SeMet and METase resulted in increased levels of phosphorylated p53 (Ser15), total p53, Bax, and p21(Waf1) proteins. LNCaP cells treated with SeMet and METase also showed p53 translocation to mitochondria, decreased mitochondrial membrane potential, cytochrome c release into the cytosol, and activation of caspase-9. The effects of SeMet and METase were suppressed by pretreatment with a synthetic superoxide dismutase mimic or by knockdown of p53 via RNA interference. Reexpression of wild-type p53 in PC3 cells resulted in increases in superoxide production, apoptosis, and caspase-9 activity and a decrease in mitochondrial membrane potential following cotreatment with SeMet and METase. Our study shows that apoptosis induced by SeMet plus METase is superoxide mediated and p53 dependent via mitochondrial pathway(s). These results suggest that superoxide and p53 may play a role in cancer chemoprevention by selenium.


Aging Does Not Enhance Social Contagion Effect.
Studies on the social contagion of memory show that it is possible to create false memories from the wrong responses from other people without requiring their physical presence. The current study examined age differences between false memories via the modified social contagion paradigm. Twenty older and twenty younger adults were shown six household scenes and were exposed to the erroneous memory reports of an implied confederate who was not physically present. The presentation time of the scenes and the typicality of the contagion items were manipulated. The participants watched each scene individually and then took turns giving their recall responses with the responses belonging to a fictional participant provided by written cards. The results in a final individual recall test indicated a significant contagion effect in both groups of participants. Additionally, an effect of the typicality of the contagion items was observed, such that the more typical items produced more contagion than the less typical items. In relation to true recall, the older adults remembered significantly fewer items from the scenes than the younger ones and obtained a lower score in the word list subtest of the Weschler Memory Scale. Although the older group had an episodic memory deficit, they were not more susceptible to being affected by the wrong responses of other people than younger group.


Immunotherapy in hematologic malignancies.
The management of hematologic malignancies has traditionally relied on chemotherapy regimens, many of which are still in use today. However, with advancements in the knowledge of tumour pathophysiology, therapies are continually evolving. Monoclonal antibodies against specific targets on tumour cells are now widely used to treat hematologic malignancies, either in combination with chemotherapy or as single agents. Rituximab, a monoclonal antibody against the CD20 antigen, is a good example of successful monoclonal antibody therapy that has improved outcomes for patients with B cell non-Hodgkin lymphomas. Monoclonal antibodies are now being used against the immune checkpoints that function to inhibit T cell activation and subsequent tumour eradication by those cytotoxic T cells. Such therapies enhance T cell-mediated tumour eradication and are widely successful in treating patients with solid tumours such as malignant melanoma. Now, they are slowly finding their place in the management of hematologic neoplasms. Even though, currently, immune checkpoint inhibitors are used for relapsed or refractory hematologic neoplasms, trials are ongoing to evaluate their role in frontline treatment. Our review focuses on the current use of immunotherapies in various hematologic malignancies.


O6-methylguanine-DNA methyltransferase activity in the human lung persists with advancing age.
O6-methylguanine-DNA methyltransferase (MGMT) plays an important role in the repair of DNA lesions induced by alkylating carcinogens in a wide range of animals. To determine the relationship between DNA repair activity and tumor susceptibility with advancing age the activity of MGMT was measured in normal lung tissue extracts from 66 patients with primary or metastatic lung cancer obtained at surgery. The age of the patients whose lung MGMT activity was measured ranged from 40 to 79 years (males) and from 40 to 80 years (females), and the values varied over 10-fold in both sexes (33 males and 33 females). Our results indicate that the MGMT enzyme protein is expressed at appreciable levels throughout life, and on age-associated rapid decline is not a feature in the human lung. Thus, any age-associated increase in lung cancer incidence cannot be ascribed to any change in MGMT activity. In order to assess other possible factors which might exert a modulating influence we examined the MGMT activity in relation to sex, tumor type and smoking. However, in none of these cases was any significant correlation evident.


Fasciocutaneous Propeller Flap Based on Perforating Branch of Ulnar Artery for Soft Tissue Reconstruction of the Hand and Wrist.
A skin defect of the hand and wrist is a common manifestation in industrial crushing injuries, traffic accidents or after excision of tumors. We reconstructed a skin defect in the ulnar aspect of the hand and wrist with a perforator-based propeller flap from the ulnar artery. The aims of our study are to evaluate the utility and effectiveness of this flap and to discuss the advantages and disadvantages of the flap in hand and wrist reconstruction with a review of the literature. Between April 2011 and November 2016, five cases of skin defect were reconstructed with a perforator-based propeller flap from the ulnar artery. There were four males and one female. The age of patients ranged from 36 to 73 years. Skin defect sites were on the dorso-ulnar side of the hand in three cases and palmar-ulnar side of the wrist in two cases. The size of the skin defect ranged from 4 × 3 cm to 8 × 5 cm. We evaluated the viability of the flap, postoperative complication and patient's satisfaction. There was no failure of flap in all cases. The size of the flap ranged from 4 × 4 cm to 12 × 4 cm. One patient, who had a burn scar contracture, presented with limited active and passive motion of the wrist after the operation. The other patients had no complications postoperatively. Cosmetic results of the surgery were excellent in one patient, good in three patients, and fair in one patient. The fasciocutaneous propeller flap based on a perforating branch of the ulnar artery is a reliable treatment option for the ulnar side skin defect of the hand and wrist.


Bioaccessibility, solid phase distribution, and speciation of Sb in soils and in digestive fluids.
Antimony (Sb) is a pollutant found in lead-contaminated soils and occurs mainly in oxidation states Sb(III) or Sb(V), this latter being the less toxic forms to organisms. In human health risk assessment, soil ingestion is one of a major exposure route in which Human bioavailability is not considered. The objectives of this paper were to establish a link between Sb speciation in soils and its bioaccessibility considering the possible evolution of the speciation in the digestive tract. Four soils were sampled around a former lead-extraction site. Total Sb content on these soils ranged between 26mgkg(-1) and 1150mgkg(-1). Sb bioaccessibility in the soils was measured using an in vitro Bioaccessibility Group Research Europe (BARGE) digestion test. A sequential extraction procedure and physical analyses were carried out on these samples to estimate Sb speciation in soils. Differential pulse anodic stripping voltammetry (DPASV) was also used to study the evolution of Sb speciation during the digestion process. For the four samples, Sb bioaccessibility was low, ranging between 1.5% and 12% of total Sb content. Sequential extractions and physical analysis on the most contaminated soil showed that Sb was mostly occurring in the iron oxide and sulphide phases and in the pentavalent form. No change of this speciation was occurring during the digestion process. Determination of the Sb speciation in the soil matrix is thus an accurate way to demonstrate that this metalloïd can be considered as a non major issue in the overall risk characterisation if it occurs under the pentavalent form.


Embracing human genetics: a primer for developmental biologists.
Understanding the etiology of congenital disorders requires interdisciplinary research and close collaborations between clinicians, geneticists and developmental biologists. The pace of gene discovery has quickened due to advances in sequencing technology, resulting in a wealth of publicly available sequence data but also a gap between gene discovery and crucial mechanistic insights provided by studies in model systems. In this Spotlight, I highlight the opportunities for developmental biologists to engage with human geneticists and genetic resources to advance the study of congenital disorders.


Primary presentation and clinical course of pediatric and adolescent patients with differentiated thyroid carcinoma after radioiodine therapy.
Differentiated thyroid carcinoma (DTC) in childhood and during adolescence is extremely rare. Pediatric DTC commonly presents with advanced disease at diagnosis including a high prevalence of cervical lymph node metastases and pulmonary metastases. Studies in children with DTC are limited. Therefore, we aimed to evaluate the initial presentation, effectiveness of radioiodine therapy (RIT), and long-term outcome of prepubertal in comparison to pubertal/postpubertal patients. Eighty-five pediatric and young patients aged 6.4 to 21.9 years with histopathologically confirmed DTC were retrospectively included. They all underwent total thyroidectomy followed by RIT. Initial presentation and outcome of prepubertal and pubertal/postpubertal patients were compared 1 year after RIT, during follow-up, and at the last visit of follow-up. Prepubertal patients presented with significantly higher T and M stages. One year after RIT, 42/81 (52%) patients still presented with evidence of disease (ED). During follow-up of a median of 7.9 years, prepubertal patients were less often in complete remission (58% vs. 82% in pubertal patients). At the last visit of follow-up, 19/80 (24%) patients still had ED without statistical differences between the two groups (42% prepubertal vs. 18% pubertal/postpubertal, p-value 0.06). None of our patients died disease-related over the observed period. Prepubertal children with DTC presented with a more advanced tumor stage at the initial presentation. During follow-up, they present more often with ED. However, at the end of our study, we did not observe statistically relevant differences in patient outcomes between the prepubertal and pubertal/postpubertal groups.


Hyperglycemic Stress Impairs the Stemness Capacity of Kidney Stem Cells in Rats.
The incidence of acute kidney injury in patients with diabetes is significantly higher than that of patients without diabetes, and may be associated with the poor stemness capacity of kidney stem cells (KSCs) and limited recovery of injured renal tubules. To investigate the effects of hyperglycemic stress on KSC stemness, KSCs were isolated from the rat renal papilla and analyzed for their self-renewal and differentiation abilities. Our results showed that isolated KSCs expressed the mesenchymal stem cell markers N-cadherin, Nestin, CD133, CD29, CD90, and CD73. Moreover, KSCs co-cultured with hypoxia-injured renal tubular epithelial cell (RTECs) induced the expression of the mature epithelial cell marker CK18, suggesting that the KSCs could differentiate into RTECs in vitro. However, KSC proliferation, differentiation ability and tolerance to hypoxia were decreased in high-glucose cultures. Taken together, these results suggest the high-glucose microenvironment can damage the reparative ability of KSCs. It may result in a decreased of recovery capability of renal tubules from injury.


Predicting potential lncRNA biomarkers for lung cancer and neuroblastoma based on an ensemble of a deep neural network and LightGBM.
Introduction: Lung cancer is one of the most frequent neoplasms worldwide with approximately 2.2 million new cases and 1.8 million deaths each year. The expression levels of programmed death ligand-1 (PDL1) demonstrate a complex association with lung cancer. Neuroblastoma is a high-risk malignant tumor and is mainly involved in childhood patients. Identification of new biomarkers for these two diseases can significantly promote their diagnosis and therapy. However, in vivo experiments to discover potential biomarkers are costly and laborious. Consequently, artificial intelligence technologies, especially machine learning methods, provide a powerful avenue to find new biomarkers for various diseases. Methods: We developed a machine learning-based method named LDAenDL to detect potential long noncoding RNA (lncRNA) biomarkers for lung cancer and neuroblastoma using an ensemble of a deep neural network and LightGBM. LDAenDL first computes the Gaussian kernel similarity and functional similarity of lncRNAs and the Gaussian kernel similarity and semantic similarity of diseases to obtain their similar networks. Next, LDAenDL combines a graph convolutional network, graph attention network, and convolutional neural network to learn the biological features of the lncRNAs and diseases based on their similarity networks. Third, these features are concatenated and fed to an ensemble model composed of a deep neural network and LightGBM to find new lncRNA-disease associations (LDAs). Finally, the proposed LDAenDL method is applied to identify possible lncRNA biomarkers associated with lung cancer and neuroblastoma. Results: The experimental results show that LDAenDL computed the best AUCs of 0.8701, 107 0.8953, and 0.9110 under cross-validation on lncRNAs, diseases, and lncRNA-disease pairs on Dataset 1, respectively, and 0.9490, 0.9157, and 0.9708 on Dataset 2, respectively. Furthermore, AUPRs of 0.8903, 0.9061, and 0.9166 under three cross-validations were obtained on Dataset 1, and 0.9582, 0.9122, and 0.9743 on Dataset 2. The results demonstrate that LDAenDL significantly outperformed the other four classical LDA prediction methods (i.e., SDLDA, LDNFSGB, IPCAF, and LDASR). Case studies demonstrate that CCDC26 and IFNG-AS1 may be new biomarkers of lung cancer, SNHG3 may associate with PDL1 for lung cancer, and HOTAIR and BDNF-AS may be potential biomarkers of neuroblastoma. Conclusion: We hope that the proposed LDAenDL method can help the development of targeted therapies for these two diseases.


Characterization of proanthocyanidins from Parkia biglobosa (Jacq.) G. Don. (Fabaceae) by Flow Injection Analysis-Electrospray Ionization Ion Trap Tandem Mass Spectrometry and Liquid Chromatography/Electrospray Ionization Mass Spectrometry.
The present study investigates the chemical composition of the African plant Parkia biglobosa (Fabaceae) roots and barks by Liquid Chromatography-Electrospray Ionization and Direct Injection Tandem Mass Spectrometry analysis. Mass spectral data indicated that B-type oligomers are present, namely procyanidins and prodelphinidins, with their gallate and glucuronide derivatives, some of them in different isomeric forms. The analysis evidenced the presence of up to 40 proanthocyanidins, some of which are reported for the first time. In this study, the antiradical activity of extracts of roots and barks from Parkia biglobosa was evaluated using DPPH method and they showed satisfactory activities.


Making the AGREE tool more user-friendly: the feasibility of a user guide based on Boolean operators.
Rationale, aims and objectives The Appraisal of Guidelines Research and Evaluation (AGREE) instrument is a generic tool for assessing guideline quality. This feasibility study aimed to reduce the ambiguity and subjectivity associated with AGREE item scoring, and to augment the tool's capacity to differentiate between good- and poor-quality guidelines. Methods A literature review was conducted to ascertain what AGREE instrument adjustments had been reported to date. The AGREE User Guide was then modified by: 1 constructing a detailed set of instructions, or dictionary, using Boolean operators, and 2 overlaying seven criteria to categorize guideline quality. The feasibility of the Boolean-based dictionary was tested by three appraisers using three randomly selected guidelines on low back pain management. The dictionary was then revised and re-tested. Results Of the 52 published studies identified, 14% had modified the instrument by adding or deleting items and 35% had adopted strategies, such as using a consensus approach, to overcome inconsistencies and ensure identical item scoring among appraisers. For the feasibility test, Pearson correlation coefficients ranged from 0.27 to 0.81. Revision and re-testing of the dictionary increased the level of agreement (range 0.41 to 0.94). Application of the revised dictionary not only decreased the variability of the domain scores, but also reduced the tool's reliability among inexperienced appraisers. Conclusion Appraisers found the Boolean-based AGREE User Guide easier to use than the original, which improved their confidence in the tool. Good reliability was achieved in the feasibility test, but the reliability and validity of some of the changes will require further evaluation.


Aspects of neurodegeneration in the canine brain.
The process of neurodegeneration displays some common morphological characteristics, most of which are jointly observed in the brains of most mammalian species. In the canine brain, neurodegeneration is frequently typified by an extensive beta-amyloid (A beta) deposition (mainly of the C-terminal A beta1-42 form) within the neurones and at the synaptic regions, in the early stages of the process. These deposits subsequently appear to give rise to the formation of senile plaques of the diffuse (non-beta-sheet) subtype, which tend to develop spontaneously but rarely proceed to form neuritic plaques. Additional features accompanying neurodegeneration include accumulations of the "aging pigment," lipofuscin, intraneuronal changes in the cytoskeleton, vascular changes in the cerebrum, cortical cerebral atrophy, enlargement of the ventricles and increased concentration of oxidative stress markers, many of which are perceived as cardinal features of extensive dysfunction in the protein turnover network. The involvement of ubiquitin is discrete but consistent in many of these molecular structures and seems to account for some critical aspects of the associated neuropathology. Irrespective of these, though, the degenerated canine brain seems to be devoid of neurofibrillary tangle formation, a manifestation commonly observed in the brain of both aged (cognitively normal) and Alzheimer-affected human subjects. The fact that canines exhibit clear symptoms of an age-related cognitive decline pertains to the concept of A beta playing a central role in age-related cognitive dysfunction and neurodegeneration.


Median polish for quality assurance of a PET scanner.
Sensitivity data for the PC 4600, a multiplanar positron emission tomograph system, were obtained over a period of several months during installation and routine clinical operation. These data were analyzed using the exploratory data analysis techniques of median polish (MP), box and whisker plots, and coded residuals. These techniques proved to be useful in spotting trends and identifying problems. Median polish had advantages over traditional percent difference techniques under some conditions because it allows separate study of more than one effect and is particularly resistant to the influence of outliers. The other exploratory data analysis techniques used are of value in interpreting the results of the MP procedure. The methods presented have direct application to any quantitative multiplanar emission tomographic imaging quality assurance program.


In vitro analysis of the penetration of methylene blue dye in human radicular dentin using different methods of impregnation.
The purpose of the present study was to verity whether different conditions of contact with a dye solution would result in different levels of the impregnation by the marker agent in human radicular dentin. The root canals of 60 upper human canines were immersed in methylene blue dye (0.5%, pH 7.0) for 24 h under six different conditions: passive immersion; pre-agitation for 10 min using an endodontic ultrasound; pre-agitation for 10 min using a cleaning ultrasound; 25-mmHg vacuum for 10 min followed by passive immersion; 30-mmHg and 650-mmHg vacuum for 24 h. The roots were longitudinally sectioned and dye impregnation was assessed by a trained and calibrated examiner, using the Q 550 IW image analyser (Leica Qwin, Cambridge, England). The measurements were analysed with anova and Tukey test. The results revealed no significant difference among the different methods for dye impregnation. In the cervical third, impregnation with 30 mmHg was significantly more effective than with endodontic ultrasound or 25 mmHg. At the apical third, no differences were observed between any of the dye impregnation methods. The comparison of different thirds revealed a statistically significant difference between the cervical and apical and the middle and apical thirds, but not between the cervical and middle thirds.


BioTile, a Perl based tool for the identification of differentially enriched regions in tiling microarray data.
Genome-wide tiling array experiments are increasingly used for the analysis of DNA methylation. Because DNA methylation patterns are tissue and cell type specific, the detection of differentially methylated regions (DMRs) with small effect size is a necessary feature of tiling microarray 'peak' finding algorithms, as cellular heterogeneity within a studied tissue may lead to a dilution of the phenotypically relevant effects. Additionally, the ability to detect short length DMRs is necessary as biologically relevant signal may occur in focused regions throughout the genome. We present a free open-source Perl application, Binding Intensity Only Tile array analysis or "BioTile", for the identification of differentially enriched regions (DERs) in tiling array data. The application of BioTile to non-smoothed data allows for the identification of shorter length and smaller effect-size DERs, while correcting for probe specific variation by inversely weighting on probe variance through a permutation corrected meta-analysis procedure employed at identified regions. BioTile exhibits higher power to identify significant DERs of low effect size and across shorter genomic stretches as compared to other peak finding algorithms, while not sacrificing power to detect longer DERs. BioTile represents an easy to use analysis option applicable to multiple microarray platforms, allowing for its integration into the analysis workflow of array data analysis.


Cancer incidence in a cohort of Swedish chimney sweeps, 1958-2006.
We examined cancer incidence in an expanded cohort of Swedish chimney sweeps. We added male chimney sweep trade union members (1981-2006) to an earlier cohort (employed 1918-1980) and linked them to nationwide registers of cancer, causes of deaths, and total population. The total cohort (n = 6320) was followed from 1958 through 2006. We estimated standardized incidence ratios (SIRs) using the male Swedish population as reference. We estimated exposure as years of employment and analyzed for exposure-response associations by Poisson regression. A total of 813 primary cancers were observed versus 626 expected (SIR = 1.30; 95% confidence interval = 1.21, 1.39). As in a previous follow-up, SIRs were significantly increased for cancer of the esophagus, liver, lung, bladder, and all hematopoietic cancer. New findings included significantly elevated SIRs for cancer of the colon, pleura, adenocarcinoma of the lung, and at unspecified sites. Total cancer and bladder cancer demonstrated positive exposure-response associations. Exposure to soot and asbestos are likely causes of the observed cancer excesses, with contributions from adverse lifestyle factors. Preventive actions to control work exposures and promote healthier lifestyles are an important priority.


When "humanitarianism" becomes "development": the politics of international aid in Syria's Palestinian refugee camps.
In recent years, the United Nations Relief and Works Agency for Palestine Refugees in the Near East (UNRWA) has attempted to go beyond its role as a provider of relief and basic services in Palestinian refugee camps and emphasize its role as a development agency. In this article, I focus on the Neirab Rehabilitation Project, an UNRWA-sponsored development project taking place in the Palestinian refugee camps of Ein el Tal and Neirab in northern Syria. I argue that UNRWA's role as a relief-centered humanitarian organization highlights the everyday suffering of Palestinian refugees, suffering that has become embedded in refugees' political claims. I show that UNRWA's emphasis on "development" in the refugee camps is forcing Palestinian refugees in Ein el Tal and Neirab to reassess the political narrative through which they have understood their relationship with UNRWA.


LpxC inhibitors: a patent review (2010-2016).
The Zn2+-dependent deacetylase LpxC is an essential enzyme of lipid A biosynthesis in Gram-negative bacteria and a promising target for the development of antibiotics selectively combating Gram-negative pathogens. Researchers from industry and academia have synthesized structurally diverse LpxC inhibitors, exhibiting different LpxC inhibitory and antibacterial activities. Areas covered: A brief introduction into the structure and function of LpxC, showing its suitability as antibacterial target, along with the structures of several reported LpxC inhibitors, is given. The article reviews patents (reported between 2010 and 2016) and related research publications on novel small-molecule LpxC inhibitors. Emphasis is placed on structure-activity relationships within the reported series of LpxC inhibitors. Expert opinion: The performed analysis of patents revealed that the current search for novel LpxC inhibitors is focused on small molecules, sharing common structural features like a Zn2+-chelating group as well as a highly lipophilic side-chain. However, despite the promising preclinical data of many of the reported compounds, besides the recently withdrawn clinical candidate ACHN-975, no other LpxC inhibitor has entered clinical trials. The lack of clinical candidates might be related with undesired effects caused by the common structural elements of the LpxC inhibitors.


Sugar Dating, Perceptions of Power, and Condom Use: Comparing the Sexual Health Risk Behaviours of Sugar Dating to Non-Sugar Dating Women.
Sugar dating is a form of dating typically characterized by wealthier, older men providing financial support to younger, less financially secure women in exchange for companionship and sexual intimacy. The goals of the current study were to (1) quantitatively assess the sexual practices of sugar dating women in their arrangements with sugar daddies, including time spent on sexual activity, average number of current partners, and relative perceptions of relationship power, (2) examine how perceptions of power within arrangements relate to condom use with sugar daddies, and (3) compare samples of sugar dating and non-sugar dating women on both condom use consistency by partner type and rates of STI testing and diagnoses. Overall, condom use for all women was highest with casual sexual partners and lowest with romantic partners, with sugar dating women's condom use with sugar daddies in between. Consistent with social exchange theory, perception of power within sugar dating arrangements predicted condom use with sugar daddies, such that women who felt they held more power reported more consistent condom use. Further, sugar dating women were twice as likely to have been diagnosed with an STI but were more than six times as likely to have been tested for STIs.


miR-618 rs2682818 C>A polymorphism decreases Hirschsprung disease risk in Chinese children.
MicroRNAs (miRNAs) are endogenous non-coding small RNAs that play an important role in the development of many malignant tumors. In addition, recent studies have reported that single nucleotide polymorphisms (SNPs) located in the miRNA functional region was inextricably linked to tumor susceptibility. In the present study, we investigated the susceptibility between miR-618 rs2682818 C>A and Hirschsprung disease (HSCR) in the Southern Chinese population (1470 patients and 1473 controls). Odds ratios (ORs) and 95% confidence intervals (CIs) were used for estimating the strength of interrelation between them. We found that the CA/AA genotypes of miR-618 rs2682818 were associated with a decreased risk of HSCR when compared with the CC genotype (OR = 0.84, 95% CI = 0.72-0.99, P=0.032). Based on the stratified analysis of HSCR subtypes, the rs2682818 CA/AA genotypes were able to significantly lessen the risk of HSCR compared with CC genotype in patients with long-segment HSCR (adjusted OR = 0.70, 95% CI = 0.52-0.93, P=0.013). In conclusion, our results indicated that the miR-618 rs2682818 C>A polymorphism was associated with a reduced risk of HSCR in Chinese children, especially in patients with long-segment HSCR (L-HSCR) subtype.


Development of a separate tank with an electrolysis-dependent bacteria controlling system for the long term storage of potable water.
For the long term storage of tap water, we developed a separate type of tank (5 m3) equipped with an electrolysis system to control bacterial growth. The electrolysis conditions using 20A direct current and a water flow rate of 10 L/min were capable of producing available chlorine (AC) at the rate of 5-8mg/min and raising the AC level of the stored tap water by about 0.2 mg/kg within 20-30 min The electrolyzed tap water with 0.2 mg/kg AC showed a capability per ml of killing 10(5)-10(6) cfu of bacteria such as Escherichia coli and Pseudomonas aeruginosa within 15 sec. A 6-month trial operation of the storage system with an automatic electrolysis control to keep AC level ranging 0.2-0.4 mg/kg demonstrated that the system worked well for the stored tap water in suppressing bacterial growth as well as in keeping good potable quality with reference to the 46 parameters specified for Japanese tap water. Actually, the electrolysis treatment was administered intermittently with an interval of about two weeks. Thus we believe the developed system has good potential to secure a potable water supply not only in the occasion of emergencies but also in countries having problems in the supply of safe drinking water.


[Structural transitions of serum proteins in solutions during high- and low-temperature dehydration].
Infrared and ultraviolet spectroscopy, potentiometric titration, and refractometry, as well as analysis of lysine, tryptophan, and tyrosine availability, and enzymic hydrolysis were used to establish that high-temperature and low-temperature dehydration of solutions of serum protein is accompanied by a rearrangement of its molecular structure, resulting in changes in the number of polar radicals and hydrophobic bonds, intermolecular and intramolecular interactions, and the formation of a new stable conformation. Comparison of the two methods of dehydration showed that cryoconcentration resulted in a greater accessibility of the protein molecule to chemical agents and proteolytic enzymes, whereas heating had an opposite effect.


Identification of microRNAs in corpus luteum of pregnancy in buffalo (Bubalus bubalis) by deep sequencing.
This study was aimed to identify miRNAs of corpus luteum (CL) in buffaloes during pregnancy. For this study, CL (n=2) were collected from gravid uteri of buffalo and RNA was isolated. Following this, the purity and integrity of RNA was checked and used for deep sequencing using Illumina Hiseq 2500 platform. The reads' quality was checked prior to in silico analyses viz. identification of conserved, novel and target of miRNAs. In this study, out of identified miRNAs (3018), 3013 were known and 5 were novel miRNAs on alignment with reference genomes. In addition, prediction of putative target genes for identified abundant miRNAs revealed several genes viz. HOX, KLF4, NCOR2, CDKN2Z, MAPK7, COX2, PPARA, PTEN, ASS3A, ELK1, CASP3, BCL211, MCL1, CCND2, Cyclin A2 and CDC25A during early pregnancy in buffalo. These predicted target genes have been associated with various cellular house-keeping processes including apoptosis. In conclusion, this study reports the identification of conserved and novel microRNAs (miRNAs) in CL during pregnancy in buffalo by deep sequencing.


Incidence and characteristics of facial nerve stimulation in children with cochlear implants.
Electrical stimulation from a cochlear implant can spread beyond the auditory nerve. The aims of this study were to accurately measure facial nerve stimulation in pediatric implant users and to determine the characteristics and incidence of this unwanted activity. Part A consisted of a prospective study of a randomized sample of 44 pediatric implant users. Part B consisted of a retrospective analysis of 121 children with previously recorded electrically evoked auditory brainstem responses (EABR). Responses were evoked by 3 electrodes along the implant array in three groups of children: 1) postmeningitic, 2) abnormal cochlea, and 3) neither. Intraoperative measures were obtained under anesthesia; all other recordings were completed in awake children. Intraoperative recordings revealed large nonauditory responses in a number of channels, including the midline EABR. Under paralysis, these responses disappeared, and clear EABRs were recorded. Similarly, prospective postoperative electromyographic (EMG) responses from the facial nerve were found in more than 59% (26 of 44) of experienced implant users (Nucleus 24): 31% of postmeningitic children (4 of 13), 80% of those with abnormal cochlea (8 of 10), and 66% of those with neither (14 of 21). Retrospective analysis of previously recorded postoperative EABRs demonstrated facial nerve stimulation in 35% (42 of 121). In most cases, facial nerve stimulation occurred when levels were perceptually loud but comfortable. 1) Facial nerve potentials can be recorded using EMG in a large proportion of cochlear implant users at high levels of stimulation. 2) The EABR can be obscured in the presence of facial nerve stimulation and care should be taken to distinguish it from the EMG response, particularly when auditory brainstem activity is in question. 3) Use of surface EMG provides an additional objective measure to ensure the safe and comfortable use of cochlear implants.


Bowel dysfunction in patients with cauda equina lesions.
Despite their serious sequels on bowel function, lesions of the cauda equina have not been previously systematically studied in larger patient populations. This was the aim of the present report. From the registrars of a diagnostic and rehabilitation centres 67 patients with clinical, electrodiagnostic and radiological findings supportive of the cauda equina lesions were recruited. The Slovene versions of the standard questionnaires for anal incontinence and constipation were used. The responses were scored, impairments categorized and previous treatments noted. Neurological examination, electromyography (EMG) of lumbo-sacral myotomes, quantitative anal sphincter EMG and electromyographic evaluation of the sacral reflex were performed. Severe anal incontinence/constipation was reported by 18%/0%, moderate by 36%/33%, and slight by 28%/43% of our patients. Twenty-one per cent of patients wore pads continuously and 14% occasionally. More than half of the patients (60%) reported changes in their lifestyle due to anal incontinence. No patient had completely normal findings on neurological examination. Perianal sensory loss correlated (P < 0.05) with anal incontinence and gender with constipation (women > men). Only two patients had received medical attention for bowel dysfunction. Study thus demonstrated significant bowel impairment in patients with lesions of the cauda equina, which has received insufficient medical attention.


Noncoordinated expression of luteal cell messenger ribonucleic acids during human chorionic gonadotropin stimulation of the primate corpus luteum.
In nonfertile cycles, the absolute steroidogenic capacity of the primate corpus luteum, as reflected in the expression of messenger RNA (mRNA) for the progesterone biosynthetic enzymes cytochrome P450 cholesterol side-chain cleavage (P450SCC) and 3 beta-hydroxysteroid dehydrogenase (3 beta-HSD), progressively declines until luteal regression. Despite this progressive loss in luteal cell function, the elaboration of CG by the implanted blastocyst is able to prolong the functional lifespan of the corpus luteum. It was the purpose of this study to investigate the relationship between aging of the primate corpus luteum and the cellular mechanisms by which the decline in luteal cell function is arrested by CG. Corpora lutea were obtained from cynomolgus monkeys on days 11 or 16 of the luteal phase after a 7-day treatment period with increasing doses of human CG (hCG) given intramuscularly beginning on days 5 or 10. Corpora lutea were also obtained from control animals on days 5, 10, 11, and 16 of the luteal phase. Human CG treatment significantly (P < 0.05) elevated both serum progesterone and estradiol levels throughout the treatment period; however, progesterone production in animals treated with hCG late in the luteal phase (days 10-16) steadily declined after the third treatment day. Expression of mRNA for P450SCC and 3 beta-HSD was markedly stimulated (P < 0.05) by hCG treatment early in the luteal phase. However, 3 beta-HSD message levels in corpora lutea from animals treated with hCG on days 10-16 were not different from those of day-16 control corpora lutea, whereas P450SCC mRNA was only minimally stimulated. There was a dramatic (P < 0.05) increase in mRNA levels for the aromatase enzyme and low density lipoprotein receptor in animals given hCG in both the early and the late luteal phase. In conclusion, there appears to be a differential responsiveness of the primate corpus luteum to hCG stimulation dependent upon luteal age. The loss in responsiveness to hCG in terms of maintenance of mRNA levels is reflective of the inability of the late luteal phase corpus luteum for continued progesterone biosynthesis in the face of heightened luteotropic stimulation.


Cardiac-specific overexpression of catalase prolongs lifespan and attenuates ageing-induced cardiomyocyte contractile dysfunction and protein damage.
1. Oxidative stress plays a role in senescence-associated organ deterioration. This is supported by the beneficial effects of anti-oxidants against ageing-related organ damage, although their role in cardiac ageing has not been elucidated. 2. The aim of the present study was to examine the impact of cardiac-specific overexpression of catalase, an enzyme for H(2)O(2) detoxification, on cardiac contractile function and protein damage in young (3-4 months) and old (26-28 months) male mice. Lifespan was analysed using the Kaplan-Meier survival curve. Cardiomyocyte contractile indices at various stimulus frequencies (0.1-5.0 Hz) were analysed, including peak shortening (PS), time to 90% PS, time to 90% relengthening (TR(90)) and maximal velocity of shortening/relengthening (+/-dL/dt). Protein damage was assessed using protein carbonyl formation. Catalase transgenic mice showed longer lifespan than wild-type FVB mice. The catalase transgene itself did not alter bodyweight or organ weight, or myocyte function. Ageing depressed +/-dL/dt and prolonged TR(90), but had no effect on other indices in FVB mice. Increased frequency triggered decreases in PS amplitude were exaggerated in aged FVB myocytes. Interestingly, ageing-induced mechanical defects were significantly attenuated in myocytes from catalase mice. Protein carbonyl formation was elevated in aged FVB compared with young FVB mice, which was significantly diminished in catalase mice. The proteomes of the myocardium of young or old FVB and catalase mice were compared using two-dimensional gel electrophoresis and mass spectrometry. Six proteins with differential expression between young and old FVB groups were tentatively identified, some of which were reversed by catalase. 3. In summary, the present data suggest that catalase protects cardiomyocytes from ageing-induced contractile defects and protein damage.


The Chlamydia pneumoniae Tarp Ortholog CPn0572 Stabilizes Host F-Actin by Displacement of Cofilin.
Pathogenic Chlamydia species force entry into human cells via specific adhesin-receptor interactions and subsequently secrete effector proteins into the host cytoplasm, which in turn modulate host-cell processes to promote infection. One such effector, the C. trachomatis Tarp factor, nucleates actin polymerization in vitro. Here we show that its C. pneumoniae ortholog, CPn0572, associates with actin patches upon bacterial invasion. GFP-CPn0572 ectopically expressed in yeast and human cells co-localizes with actin patches and distinctly aberrantly thickened and extended actin cables. A 59-aa DUF 1547 (DUF) domain, which overlaps with the minimal actin-binding and protein oligomerization fragment required for actin nucleation in other Tarp orthologs, is responsible for the aberrant actin phenotype in yeast. Interestingly, GFP-CPn0572 in human cells associated with and led to the formation of non-actin microfilaments. This phenotype is strongly enhanced in human cells expressing the GFP-tagged DUF deletion variant (GFP-ΔDUF). Finally ectopic CPn0572 expression in yeast and in-vitro actin filament binding assays, demonstrated that CPn0572 stabilizes pre-assembled F-actin by displacing and/or inhibiting binding of the actin-severing protein cofilin. Remarkably, the DUF domain suffices to displace cofilin from F actin. Thus, in addition to its actin-nucleating activities, the C. pneumoniae CPn0572 also stabilizes preformed host actin filaments.


Factor XIII mediates adhesion of platelets to endothelial cells through alpha(v)beta(3) and glycoprotein IIb/IIIa integrins.
Coagulation factor XIII (FXIII) is a transglutaminase that catalyzes crosslink formation in fibrin clots. Endothelial cells (EC) were demonstrated to bind FXIII via their alpha(v)beta3 integrin receptor. FXIII was also shown to bind platelet glycoprotein IIb/IIIa receptor. In the present study, we analyzed if FXIII can mediate platelet-EC interaction. Both FXIII and activated FXIII (FXIIIa) bound to EC monolayers; this binding was enhanced by the addition of Mn2+ and was inhibited by the monoclonal antibody L609 against alpha(v)beta3 integrin. Normal washed platelets also bound surface-immobilized or soluble FXIII and FXIIIa, and the binding was GPIIb/IIIa dependent. The effect of FXIII concentrate (Fibrogammin-P) treatment on the interaction of ECs with platelets from six FXIII-deficient patients was studied. Patients' platelets were radiolabeled with 3H-Adenine, washed, resuspended in autologous plasma and allowed to adhere to immortalized EC line EAhy926. Adhesion of platelets from FXIII-deficient patients to ECs increased 1.7+/-0.4-fold (P=.01) following intravenous infusion of FXIII concentrate. Similarly, addition of 1 U/ml of FXIII concentrate to the patients' PRP in vitro increased the adhesion 1.8+/-0.5-fold (P=.008). Preincubation of the EC monolayers with increasing concentrations of either FXIII or FXIIIa augmented the adhesion of normal washed platelets to ECs in a dose-dependent manner. At 10 U/ml of EC-bound FXIII or FXIIIa, platelet adhesion enhanced 1.7+/-0.25-fold (P=.03) and 2.5+/-0.5-fold (P=.02), respectively. The increase in platelet adhesion was completely abolished by pretreatment of ECs with the anti-alpha(v)beta3 antibody L609 or by preincubation of the platelets with the GPIIb/IIIa inhibitor Abciximab. Taken together, our data indicate that FXIII mediates the interaction of platelets with ECs by bridging between endothelial alpha(v)beta3 and platelet GPIIb/IIIa integrins. This interaction may be relevant for tissue remodeling and wound repair after vascular injury in FXIII-deficient patients.


"One plate/three-reporter" assay format for the detection and validation of yeast two-hybrid interactions.
We describe a novel assay format for the Gal4-based yeast two-hybrid-system, in which the readout from three different reporter genes is measured sequentially in a single microplate. Activation of the URA3, MEL1, and lacZ reporters in response to a protein-protein interaction is monitored by measuring sequentially: (i) growth in medium lacking uracil, (ii) alpha-galactosidase activity, and (iii) beta-galactosidase. The data thus generated permit elimination of many false positive signals and provide a preliminary measurement of reporter activation-strength that may be confirmed by further analysis. The assay procedure is inexpensive and requires few liquid-handling steps. It is appropriate for automated high-throughput interaction mating assays, validation of putative interactor strains and hybrid-protein self-activator tests.