[Experimental study on healing of old tear in the avascular portion of menisci in dogs].
32 adult mongrel dogs (male 14, female 18, average weight 16.7 kg) were used to study the healing process of old tear in the avascular portion of the menisci. They were divided into two groups--Control group and repair group which were divided into two subgroups, that is, debrided and undebrided groups. Dogs were sacrificed 2.4 weeks and 4.5 months and one year after repair respectively. Finally results were analysed by means of light and electron microscopy. The following conclusions were drawn: (1) A meniscal tear in the avascular portion would not heal by itself despite some cellular response on the edge of the tear. (2) Tear would not heal after simple vascular access channel procedure. (3)Tear healed with fibrocartilage when vascular access channel with synovial flap implantation was used. Therefore this technique can be considered as an alternative to meniscectomy in the management of torn meniscus clinically. (4) Debridement has a bad effect on the healing process because it makes the tear wider and longer and remove the activated cells. (5) The vascular access channel about one half of the thickness of the peripheral edge of the meniscus does not change the shape of the meniscus.


Sugarcane Aphid (Hemiptera: Aphididae): A New Pest on Sorghum in North America.
In 2013, the sugarcane aphid, Melanaphis sacchari (Zehntner) (Hemiptera: Aphididae), a new invasive pest of sorghum species in North America, was confirmed on sorghum in 4 states and 38 counties in the United States. In 2015, the aphid was reported on sorghum in 17 states and over 400 counties as well as all sorghum-producing regions in Mexico. Ability to overwinter on living annual and perennial hosts in southern sorghum-producing areas and wind-aided movement of alate aphids appear to be the main factors in its impressive geographic spread in North America. Morphological characteristics of the sugarcane aphid include dark tarsi, cornicles, and antennae, allowing easy differentiation from other aphids on the crop. Sugarcane aphid damages sorghum by removing sap and covering plants with honeydew, causing general plant decline and yield loss. Honeydew and sooty mold can disrupt harvesting. The aphid's high reproductive rate on susceptible sorghum hybrids has resulted in reports of yield loss ranging from 10% to greater than 50%. In response, a combination of research-based data and field observations has supported development of state extension identification, scouting, and treatment guides that aid in initiating insecticide applications to prevent yield losses. Highly efficacious insecticides have been identified and when complemented by weekly scouting and use of thresholds, economic loss by sugarcane aphid can be minimized. Some commercial sorghum hybrids are partially resistant to the aphid, and plant breeders have identified other lines with sugarcane aphid resistance. A very diverse community of predators and parasitoids of sugarcane aphid has been identified, and their value to limit sugarcane aphid population growth is under investigation.


Induction of intracellular tau aggregation is promoted by α-synuclein seeds and provides novel insights into the hyperphosphorylation of tau.
Intracytoplasmic proteinaceous inclusions, primarily composed of tau or α-synuclein (α-syn), are predominant pathological features of Alzheimer's disease (AD) and Parkinson's disease (PD), respectively. However, the coexistence of these pathological aggregates is identified in many neurodegenerative disorders, including spectrum disorders of AD and PD. Whereas α-syn can spontaneously polymerize into amyloidogenic fibrils, in vitro, tau polymerization requires an inducing agent. The current study presents a human-derived cellular model, in which recombinant, preformed α-syn fibrils cross-seed intracellular tau to promote the formation of neurofibrillary tangle-like aggregates. These aggregates were hyperphosphorylated, Triton insoluble, and thioflavin-S positive, either comingling with endogenously expressed α-syn aggregates or induced by only exogenously applied recombinant α-syn fibrils. Furthermore, filamentous, amyloidogenic tau took over the cellular soma, displacing the nucleus and isolating or displacing organelles, likely preventing cellular function. Although a significant proportion of wild-type tau formed these cellular inclusions, the P301L mutation in tau increased aggregation propensity resulting from α-syn seeds to over 50% of total tau protein. The role of phosphorylation on the development of these tau aggregates was investigated by coexpressing glycogen synthase kinase 3 β or microtubule-associated protein/microtubule affinity-regulating kinase 2. Expression of either kinase inhibited the formation of α-syn-induced tau aggregates. Analyses of phosphorylation sites suggest that multiple complex factors may be associated with this effect and that Triton-soluble versus Triton-insoluble tau may be independently targeted by kinases. The current work not only provides an exceptional cellular model of tau pathology, but also examines α-syn-induced tau inclusion formation and provides novel insights into hyperphosphorylation observed in disease.


When is fluid in the mastoid cells a worrisome finding?
Mastoiditis is a common clinical entity that is technically present in all cases of otitis media; only a minority of cases actually represents the otolaryngologic emergency of acute coalescent mastoiditis. When reviewing an image with a radiologic diagnosis of mastoiditis, looking for key signs such as destruction of bony septa and considering patient presentation can help distinguish mild mastoiditis from acute coalescent mastoiditis.


Treatment with DHA/EPA ameliorates atopic dermatitis-like skin disease by blocking LTB4 production.
Atopic dermatitis (AD) is caused by both dysregulated immune responses and an impaired skin barrier. Although leukotriene B4 (LTB4) is involved in tissue inflammation that occurs in several disorders, including AD, therapeutic strategies based on LTB4 inhibition have not been explored. Here we demonstrate that progression of an AD-like skin disease in NC/Nga mice is inhibited when docosahexaenoic acid (DHA)/eicosapentaenoic acid (EPA) is administered together with FK506. Treatment with DHA/EPA and FK506 decreases the clinical score of dermatitis in NC/Nga mice and lowers local LTB4 concentrations. The treatment also suppressed the infiltration of T cells, B cells, eosinophils and neutrophils, and promoted reduced serum IgE levels. Secretion of IL-13 and IL-17A in CD4(+) T cells was lower in DHA/EPA- and FK506-treated mice than in mice treated with FK506 alone. The inhibition of disease progression induced by DHA/EPA was reversed by local injection of LTB4, suggesting that the therapeutic effect of DHA/EPA is LTB4-dependent. Our results demonstrate that treatment of AD with DHA/EPA is effective for allergic skin inflammation and acts by suppressing LTB4 production. J. Med. Invest. 63: 187-191, August, 2016.


Establishing technical performance norms for general surgery residents.
Surgery residents are required to achieve performance milestones to advance in their residency. Level-specific, technical performance norms that could be used as milestones, however, do not currently exist. Our aim was to develop level-specific, technical performance norms for general surgery residents on select simulated tasks across multiple institutions. An IRB-approved, prospective, multi-institutional collaborative study with voluntary participation of residents was undertaken at the start of the 2011-2012 academic year. General surgery residents (PGY I-V) from seven institutions were tested on three laparoscopic and five open simulated surgical tasks, and their performance was assessed based on task time and errors. Means and standard deviations of performance for each resident level were calculated and compared. Residents with performance 1 standard deviation below the mean were considered outliers. A total of 147 residents were evaluated. Mean resident age was 28 ± 3 years; 42 % were female; and they had attended 74 different medical schools. Senior residents (PGY III-V) had more clinical and simulator experience than junior residents (PGY I-II) (p < 0.001). Resident performance scores progressively increased in all tasks reaching a plateau at a lower PGY level for open tasks. Depending on the task, 0-18 % of residents were outliers. When surveyed, 66 % of residents agreed that national performance norms for residents should exist. Performance norms were established for select tasks in a representative sample of US surgery residents. Such performance norms allow a more informed assessment of resident skill through comparison to national data and enable the identification of outliers who may benefit from additional training.


Safety and effectiveness of the INCRAFT AAA Stent Graft for endovascular repair of abdominal aortic aneurysms.
This study evaluated the 2-year safety and effectiveness of the European First-in-Human INNOVATION trial for the INCRAFT AAA Stent Graft system (Cordis Corp, Bridgewater, NJ), an ultra-low-profile device for the treatment of abdominal aortic aneurysms. From March 2010 to June 2011, the INNOVATION prospective multicenter trial involving six centers in Europe enrolled and treated 60 asymptomatic patients (95% male; mean age, 74.4 ± 6.9 years) with the INCRAFT System. Main inclusion criteria included proximal aortic neck length of 15 mm or more with a diameter up to 27 mm; distal iliac landing zones with a length greater than 10 mm and a diameter between 9 and 18 mm; and aortic bifurcation >18 mm in diameter and access vessels large enough to accept the 14F outer diameter of the delivery system. The primary end point was technical success at 1 month; 2-year safety end points included the absence of device- or procedure-related major adverse events, absence of type I or III endoleaks, and maintenance of device integrity through 2 years of follow-up. Study oversight was provided by a Data Safety Monitoring Board with event adjudication by a Clinical Events Committee and imaging analysis by an independent core laboratory. Of 60 successfully treated patients, two did not come back for their 1-month evaluation but remained enrolled in the study; 56 were evaluated at 1 year and 52 at 2 years. Of the 58 patients, 56 met the 1-month primary safety and effectiveness end point (97%; 95% confidence interval, 88%-100%). All patients were free from aneurysm enlargement through 2 years. There were no type I or III endoleaks at the 2-year time point. All-cause mortality at 2 years was 11.5%, and no death was device or procedure related. In total, three patients required a postprocedure intervention, two to repair a type I endoleak and one for limb occlusion. Core laboratory evaluation of the postoperative imaging studies documented absence of endograft migration and stent fractures in all patients. The INCRAFT AAA Stent Graft System provides a durable solution for patients with abdominal aortic aneurysms, with a low frequency of device-related events through 2 years of follow-up.


Troponin T isoform expression in humans. A comparison among normal and failing adult heart, fetal heart, and adult and fetal skeletal muscle.
The expression of troponin (Tn) T, a thin-filament regulatory protein, was examined in left ventricular myocardium from normal and from failing adult human hearts. The differences in isoform expression between normal and failing myocardium led us to examine the ontogenic expression of TnT in human striated muscle. Left ventricular samples were obtained from patients with severe heart failure undergoing cardiac transplantation and normal adult organ donors. Fetal muscle was obtained from aborted fetuses after 14-15 weeks of gestation, and adult skeletal muscle was obtained from surgical biopsies. Western blots of normal and failing adult heart proteins demonstrated that two isoforms, TnT1 and TnT2, are expressed in different amounts, with TnT2 being significantly greater in failing hearts (p less than 0.004). Western blots of two-dimensional gels of these proteins resolved two predominant spots of both TnT1 and TnT2 and several minor TnT species. Alkaline phosphatase treatment converted the two major spots of each isoform into the single more basic spots. A comparison of the ATPase activities and the TnT2 percentage of total TnT in individual failing and normal adult hearts demonstrated an inverse and negative relation (r = 0.7, p less than 0.02). In the fetal heart, four TnT isoforms were found, two of which had the same electrophoretic mobilities as the adult cardiac isoforms TnT1 and TnT2. Fetal skeletal muscle expressed two of the four fetal cardiac TnT isoforms, one of which comigrated with adult cardiac TnT1. These cardiac isoforms were expressed in low abundance in fetal skeletal muscle relative to seven fast skeletal muscle TnT isoforms. No cardiac isoforms were present in adult skeletal muscle. Because many etiologies caused heart failure in the transplant patients, we propose that the disease-associated increased expression of the TnT isoform TnT2 is an adaptation to the heart failure state and a partial recapitulation of the fetal expression of cardiac TnT isoforms.


Discovery of Novel Caeridins from the Skin Secretion of the Australian White's Tree Frog, Litoria caerulea.
Abundant biologically active peptides have been discovered from frog skin secretions, a rich natural source of bioactive compounds with great potential in drug discovery. In this study, three Caeridin peptides, namely, Caeridin-1, S5-Caeridin-1, and Caeridin-a1, were discovered from the skin secretion of the Australian White's tree frog, Litoria caerulea, for the first time, by means of combining transcriptomic and peptidomic analyses. It also represents the first report on bioactive Caeridins since this family of peptides was initially studied 20 years ago. Chemically synthetic versions of each natural Caeridin demonstrated promising bioactivities either on rat smooth muscles or against microbial growth. Specifically, Caeridin-1 produced contraction of rat bladder smooth muscle, while S5-Caeridin-1 induced relaxation of rat ileum smooth muscle, both at nanomolar concentrations. Moreover, Caeridin-a1 was shown to potently inhibit the growth of the planktonic Gram-positive bacteria Staphylococcus aureus (S. aureus), methicillin-resistant S. aureus (MRSA), and Enterococcus faecalis (E. faecalis), the Gram-negative bacterium, Escherichia coli (E. coli), and the yeast, Candida albicans (C. albicans). The discovery of these Caeridins may induce further intensive and systematic studies of frog skin peptides to promote the discovery of natural templates as lead compounds for drug discovery and therapeutic application.


The biology and pathology of the familial Parkinson's disease protein LRRK2.
Parkinson's disease (PD) is typically a sporadic illness, but the past decade has witnessed the identification of mutations responsible for multiple familial forms of the disease. The proposed functions of some of these genes (e.g., E3 ubiquitin ligase, redox-dependent chaperone) have led to the hypothesis that dysfunction of protein quality control pathways contributes to PD neurodegeneration. However, the key signaling events that act downstream of misfolded proteins to cause cell death remain poorly defined. The discovery of the familial PD kinase leucine-rich repeat kinase 2 (LRRK2) holds great promise for the elucidation of signaling events relevant to PD neurodegeneration. This review will summarize current knowledge of the clinical and cell biological features of LRRK2, the most common inherited cause of Parkinsonism.


The pathophysiology of hyperchloremic metabolic acidosis after urinary diversion through intestinal segments.
The pathophysiology of hyperchloremic metabolic acidosis after urinary diversion through intestinal segments has not been defined. This study employs a canine model in which an ileal segment is interposed between one kidney and the urinary bladder. Comparison of urinary solute excretion rates between the normal and interposed renal units allows quantitation of solute reabsorption and secretion by the ileal segment. Ileal segments reabsorb urinary chloride, potassium, and ammonium. Ammonium is reabsorbed in part as its conjugate free base, ammonia, with the liberated hydrogen ion reabsorbed with chloride or excreted as titratable acid. Inability to excrete acid as ammonium results in depletion of body buffers and a diminished capacity to compensate an additional acid challenge. Bicarbonate is secreted by the ileal segments but not in amounts that are physiologically significant. Impaired renal function predisposes to the development of this syndrome but is not a primary pathophysiologic mechanism.


Nonrashes. 4. Audible signs of cutaneous disease.
There is more than one reason why dermatologists should listen to their patients. Audible signs of cutaneous disease range from the classic, early childhood hoarseness of lipoid proteinosis to a characteristic hoarseness due to syphilitic papules on the epiglottis and arytenoepiglottic folds.


Enhanced scattering and absorption due to the presence of a particle close to an interface.
We study the influence of the presence of an interface on the scattering by a Rayleigh scatterer. The influence of an interface on the spontaneous emission has been known for many years. Here, we study the influence on the extinction cross-section and absorption cross-section. We provide a detailed analysis of interference and near-field effects. We show that the presence of a Rayleigh scatterer may enhance the specular reflection or specular transmission under certain conditions. Finally, we analyze the enhancement of absorption in the bulk in the presence of a small scatterer.


Towards sustainable maize production in the U.S. upper Midwest with interseeded cover crops.
The incorporation of cover crops into the maize (Zea mays L.)-soybean [Glycine max (L.) Merr.] rotation in the U.S. upper Midwest may improve sustainability. Long, cold winters in the region make identifying successful cover crop species and management practices a challenge. Two experiments were conducted in Minnesota, USA from fall 2016 through spring 2019 to examine the effect of cover crops interseeded at four- to six-leaf collar (early-interseeded) and dent to physiological maturity (late-interseeded) on biomass and grain yield of maize. Annual ryegrass (Lolium multiflorum L.) and cereal rye (Secale cereale L.) were evaluated as monocultures and in mixtures with crimson clover (Trifolium incarnatum L.) and forage radish (Raphanus sativus L.). Differences in canopy cover and biomass of late-interseeded cover crops were observed at the southernmost location in 2018. Additional accumulated growing-degree days in fall 2018 did not translate into increased cover crop canopy coverage of late-interseeded cover crops. Differences in cover crop canopy cover and biomass of early-interseeded cover crops were observed by fall frost at all locations in 2017 and at the northernmost location in 2018. Cover crop canopy cover and biomass at termination before planting maize, soil moisture at maize planting as well as maize aboveground biomass and yield were not affected by spring cereal rye regrowth of cover crops late-interseeded the previous year. Similarly, early-interseeded cover crops did not affect maize aboveground biomass or yield. We attribute these results to limited cover crop growth. This highlights the potential of a variety of cover crop strategies interseeded into maize in the U.S. upper Midwest; however, efforts to fine-tuning cover crop management and weather conditions are needed to benefit from such practice.


Regulation of the SOS response in Bacillus subtilis: evidence for a LexA repressor homolog.
The inducible SOS response for DNA repair and mutagenesis in the bacterium Bacillus subtilis resembles the extensively characterized SOS system of Escherichia coli. In this report, we demonstrate that the cellular repressor of the E. coli SOS system, the LexA protein, is specifically cleaved in B. subtilis following exposure of the cells to DNA-damaging treatments that induce the SOS response. The in vivo cleavage of LexA is dependent upon the functions of the E. coli RecA protein homolog in B. subtilis (B. subtilis RecA) and results in the same two cleavage fragments as produced in E. coli cells following the induction of the SOS response. We also show that a mutant form of the E. coli RecA protein (RecA430) can partially substitute for the nonfunctional cellular RecA protein in the B. subtilis recA4 mutant, in a manner consistent with its known activities and deficiencies in E. coli. RecA430 protein, which has impaired repressor cleaving (LexA, UmuD, and bacteriophage lambda cI) functions in E.coli, partially restores genetic exchange to B. subtilis recA4 strains but, unlike wild-type E. coli RecA protein, is not capable of inducing SOS functions (expression of DNA damage-inducible [din::Tn917-lacZ] operons or RecA synthesis) in B. subtilis in response to DNA-damaging agents or those functions that normally accompany the development of physiological competence. Our results provide support for the existence of a cellular repressor in B. subtilis that is functionally homologous to the E. coli LexA repressor and suggest that the mechanism by which B. subtilis RecA protein (like RecA of E. coli) becomes activated to promote the induction of the SOS response is also conserved.


Surface functionalized halloysite nanotubes decorated with silver nanoparticles for enzyme immobilization and biosensing.
Improving enzyme immobilization with high loading capacity and achieving direct electron transfer (DET) between the enzyme and the electrode surface is key to designing highly sensitive enzymatic electrochemical biosensors. Herein, we report a novel approach based on the selective modification of the outer surface of halloysite nanotubes (HNTs) that supports silver nanoparticles (AgNPs) to obtain a hybrid nanocomposite. AgNPs of about 10 nm average size could be uniformly supported on silane-modified HNTs through in situ reduction of Ag+ ions. The resultant nanocomposite shows an excellent support capability for the effective immobilization and electrical wiring of redox enzyme glucose oxidase (GOx). The GOx immobilized HNT/AgNPs were deposited on the glassy carbon electrode (GCE) and utilized for the bioelectrocatalyzed electrochemical detection of glucose. The GOx modified composite electrodes show glucose sensitivity as high as 5.1 μA mM-1 cm-2, which is higher than for the electrodes prepared without surface functionalization.


A mathematical derivation to prove reduced denture retention in v shaped palate.
The impact of oral health on the quality of life of older individuals have significantly increased over the last few decades. Edentulism substantially affect oral and general health and overall quality of life. Complete denture retention is the resistance to displacement of the denture base away from the ridge which is the main source of psychological comfort for the patient. Special retention problems are seen in patients exhibiting highly tapered steep palatal vault. In these cases a metal base or subsequent bench cure reline procedure would be incorporated into the initial treatment plan. In this article, we have derived a mathematical proof for the clinical experience that reduction in the palatal angle, results in lesser denture retention.


Unique structural determinants in the signal peptides of "spontaneously" inserting thylakoid membrane proteins.
A series of thylakoid membrane proteins, including PsbX, PsbY and PsbW, are synthesized with cleavable signal peptides yet inserted using none of the known Sec/SRP/Tat/Oxa1-type insertion machineries. Here, we show that, although superficially similar to Sec-type signal peptides, these thylakoidal signal peptides contain very different determinants. First, we show that basic residues in the N-terminal domain are not important, ruling out electrostatic interactions as an essential element of the insertion mechanism, and implying a fundamentally different targeting mechanism when compared with the structurally similar M13 procoat. Second, we show that acidic residues in the C-domain are essential for the efficient maturation of the PsbX and PsbY-A1 peptides, and that even a single substitution of the -5 Glu by Val in the PsbX signal peptide abolishes maturation in the thylakoid. Processing efficiency is restored to an extent, but not completely, by the highly hydrophilic Asn, implying that this domain is required to be hydrophilic, but preferably negatively charged, in order to present the cleavage site in an optimal manner. We show that substitution of the PsbX C-domain Glu residues by Val leads to a burial of the cleavage site within the bilayer although insertion is unaffected. Finally, we show that substitution of the Glu residues in the lumenal A2 loop of the PsbY polyprotein leads to a block in cleavage on the stromal side of the membrane, and present evidence that the PsbY-A2 signal peptide is required to be relatively hydrophilic and unable to adopt a transmembrane conformation on its own. These data indicate that, rather than being merely additional hydrophobic regions to promote insertion, the signal peptides of these thylakoid proteins are complex domains with uniquely stringent requirements in the C-domain and/or translocated loop regions.


A combined treatment with melatonin and andrographis promotes autophagy and anticancer activity in colorectal cancer.
Colorectal cancer (CRC) is one of the most frequent malignancies worldwide and remains one of the leading causes of cancer-related deaths in the USA. The high degree of morbidity and mortality associated with this disease is largely due to the inadequate efficacy of current treatments as well the development of chemoresistance. In recent years, several pharmaceutical agents screened from natural products have shown the promise to offer a safe, inexpensive and synergistically multi-targeted treatment option in various cancers. Given the growing evidence of anti-carcinogenic properties of two natural compounds, melatonin (MLT) and andrographis (Andro), we aimed to evaluate their synergistic anticancer effects in CRC. We demonstrate that indeed these two compounds possessed a synergistic anticancer effect in terms of their ability to inhibit cell viability, suppression of colony-formation and induction of apoptosis (P < 0.05). In line with our in vitro findings, we were able to validate this combinatorial anticancer activity in xenograft animal models (P < 0.001) as well as tumor-derived 3D organoids (P < 0.01). RNA-sequencing analysis revealed candidate pathways and genes that mediated antitumor efficacy of MLT and Andro in CRC, among which autophagy pathway and related genes, including NR4A1, CTSL and Atg12, were found to be primarily responsible for the increased anticancer effect by the two natural products. In conclusion, our data reveal a potent and synergistic therapeutic effect of MLT and Andro in the treatment of CRC and provides a rationale for suppressing autophagy in cancer cells as a potential therapeutic strategy for CRC.


If not appendicitis, then what else can it be? A retrospective review of 1492 appendectomies.
To audit the appendectomies at our institute, and summarise atypical pathological results with a discussion of appropriate management. DESIGN. Retrospective study. Regional hospital, Hong Kong. All patients who underwent appendectomy for presumed acute appendicitis from June 2003 to June 2008 were recruited. Incidental appendectomy was excluded. Patient demographics, pathological findings, and surgical outcomes were analysed. RESULTS. The overall negative appendectomy rate was 18.2%. Female patients of reproductive age (11-50 years) conferred an independent risk for a higher negative appendectomy rate than other females (28.7% vs 11.5%; P<0.001). The overall perforation rate was 22.5%; the extremes of age (<11 or >70 years) conferred an independent risk of perforated appendicitis (25.2% vs 16.3%; P=0.002). Preoperative imaging was not associated with a lower negative appendectomy rate or rate for perforated appendicitis (P=0.205 and 0.218, respectively). Multivariate analysis suggested that a preoperative white cell count of less than 13.5 x 10(9) /L was an independent predictor of negative appendectomy (P<0.001); the body temperature and pulse rate of the patients with perforated appendicitis were higher than in those without perforation (P=0.004 and 0.003, respectively). Only 4.0% of the appendectomy specimens contained other appendiceal pathologies. Appendiceal diverticulitis was the most common inflammatory pathology, contributing to 2.7% of all appendectomies, followed by granulomatous appendicitis. In this series there were eight carcinoid tumours, three adenocarcinomas, two mucinous cystadenomas; tubular adenoma, metastatic deposition, mucinous cystadenocarcinoma and pseudomyxoma peritonei each occurred in one patient only. A more focused utilisation of preoperative imaging in females of reproductive age and patients at the extremes of age is suggested. Long-term follow-up should be offered to patients with granulomatous appendicitis and neoplastic appendiceal diseases.


A new approach for anthelmintic discovery for humans.
Natural product-based drug discovery has been deemphasized by the pharmaceutical industry. This situation is discordant with the fact that most people in developing countries rely on traditional medicines derived from local biodiversity for healthcare. Despite economic growth in the past 10 years, Africa remains plagued by parasitic infections, out of reach of eradication. Limited regional funding for drug discovery complicates the situation. Novel models are needed to bring sustainability to local drug discovery programs. This Opinion describes an innovative partnership that promotes local leadership to harness a recombinant yeast-based assay to screen for novel anthelmintic candidates in collections of African natural products. Implementation of this strategy in biodiversity-rich but resource-constrained settings can help build sustainable local capacity for drug discovery.


Same-sex reproduction: medical treatment options and psychosocial considerations.
This review provides an overview of the historical significance of assisted reproduction for gay men and women, discusses current reproductive options for same-sex couples, addresses psychosocial considerations unique to these couples, and reviews the current literature addressing medical and psychosocial aspects of same-sex reproduction. Growing numbers of men and women openly self-identify as gay and lesbian. Accompanying this openness is an increased public acceptance of same-sex relationships and same-sex marriage. The combination of gay/lesbian self-determination and mounting public acceptance of same-sex unions has led these individuals and couples to increasingly seek parenthood through assisted reproduction. Recent studies describe relationship satisfaction in gay couples after assisted reproduction and more positive functioning and less stress associated with parenthood when compared with heterosexual parents. Motivations for parenthood are the same for same-sex couples and heterosexual couples alike. However, achieving the goal of parenthood can be a much greater endeavor medically and psychologically for same-sex couples. Fertility treatment centers increasingly recognize issues unique to gay men and women and are increasingly welcoming.


Host response to bovine ocular squamous cell carcinoma.
Specific humoral and cellular immunologic responses to autologous and heterologous tumors were evaluated in 35 Hereford cows with ocular squamous cell tumors and in 6 healthy cows. Sera from 5 healthy cows and 23 ocular tumor-bearing cows were evaluated for antibody to tumor, using radioimmunoassay, passive hemagglutination, agglutination, and microagglutination assays with various soluble tumor antigen and whole tumor cell preparations. Antibody to tumor was detected in only 2 cows. Using the microagglutination assay, antibody to autologous and heterologous tumor cells was found in the sera of 2 cows inoculated intraocularly with purified, viable autologous tumor cells. Twenty-eight tumor-bearing cows and 3 healthy cows were evaluated for delayed-type hypersensitivity responses, using 5 tumor preparations; positive skin test responses were not observed.


Na+ and K+ concentration and regulation of protein synthesis in L-A9 and Aedes albopictus cells infected with Marituba virus (Bunyaviridae).
Infection of L-A9 cells with Marituba virus produces a severe inhibition of protein synthesis. This inhibition is temporally correlated with an increase in the intracellular Na+ concentration and a decrease in the intracellular K+ concentration. However, in Marituba virus-infected Aedes albopictus cells the intracellular level of Na+ and K+ ions and protein synthesis remained unaltered. Incubation of both cell types at high NaCl concentration facilitated the translation of viral RNA whereas the cellular protein synthesis was inhibited. Using a hypotonic medium, the opposite was found. Results are discussed in terms of a possible involvement of these ions in the viral translational process.


Making More of IT: Enabling Intensive Motor Cognitive Rehabilitation Exercises in Geriatrics Using Information Technology Solutions.
Although the health benefits of physical activity and exercise for older people are well established, a largely sedentary lifestyle still prevails in ageing western societies. Finding new ways to make exercise more accessible and acceptable for older adults must be developed to fully unleash its potential in preventing and weakening age-related physical and cognitive decline. Existing barriers to implement effective exercise-based treatment plans include motivational reservations on both the clinician's and patient's side, but also physical limitations caused by disease or deconditioning. Particularly in the more senior population, debilitating conditions do not allow adherence to currently recommended exercise regimes. A major rethinking of age- and user-adapted exercise is overdue. The high intensities required for physical and mental adaptations must be modifiable and personalized according to the functional status of each patient. Emerging information and communication technologies (ICT) have brought forward a plethora of attractive solutions for smart and adapted exercise, but there remains a vast gap between technological advancement and clinical relevance. Where in the beginning ICT for active ageing mainly focussed on aspects of usability and user experience, the current status of IT as applied in ageing populations noticeably shifted toward new services, applications, and devices that can be offered with the aim to prevent, compensate, care, and/or enhance daily life functioning of senior citizens. In this perspective paper, we aim to summarize the current state of the art in ICT-based interventions aimed at improved motor-cognitive control and make suggestions about how these could be combined with high-intensive interval exercise regimes to make rehabilitation for the impaired older adults more effective, and more fun.


Circulating B Cells With Memory and Antibody-Secreting Phenotypes Are Detectable in Pediatric Kidney Transplant Recipients Before the Development of Antibody-Mediated Rejection.
Development of anti-human leukocyte antigen donor-specific antibodies (DSAs) is associated with antibody-mediated rejection (AMR) and reduced allograft survival in kidney transplant recipients. Whether changes in circulating lymphocytes anticipate DSA or AMR development is unclear. We used time-of-flight mass cytometry to analyze prospectively collected peripheral blood mononuclear cells (PBMC) from pediatric kidney transplant recipients who developed DSA (DSA-positive recipients [DSAPOS], n = 10). PBMC were obtained at 2 months posttransplant, 3 months before DSA development, and at DSA detection. PBMC collected at the same time points posttransplant from recipients who did not develop DSA (DSA-negative recipients [DSANEG], n = 11) were used as controls. DSAPOS and DSANEG recipients had similar baseline characteristics and comparable frequencies of total B and T cells. Within DSAPOS recipients, there was no difference in DSA levels (mean fluorescence intensity [MFI]: 13 687 ± 4159 vs 11 375 ± 1894 in DSAPOSAMR-positive recipients (AMRPOS) vs DSAPOSAMR-negative recipients (AMRNEG), respectively; P = 0.630), C1q binding (5 DSAPOSAMRPOS [100%] vs 4 DSAPOSAMRNEG [80%]; P = 1.000), or C3d binding (3 DSAPOSAMRPOS [60%] vs 1 DSAPOSAMRNEG [20%]; P = 0.520) between patients who developed AMR and those who did not. However, DSAPOS patients who developed AMR (n = 5; 18.0 ± 3.6 mo post-DSA detection) had increased B cells with antibody-secreting (IgD-CD27+CD38+; P = 0.002) and memory (IgD-CD27+CD38-; P = 0.003) phenotypes compared with DSANEG and DSAPOSAMRNEG recipients at DSA detection. Despite the small sample size, our comprehensive phenotypic analyses show that circulating B cells with memory and antibody-secreting phenotypes are present at DSA onset, >1 year before biopsy-proven AMR in pediatric kidney transplant recipients.


High-Speed In Situ Observation System for Sonoporation of Cells With Size- and Position-Controlled Microbubbles.
A high-speed in situ microscopic observation system developed for basic studies on mechanisms of sonoporation is introduced in this paper. The main part of the system is an inverted-type fluorescence microscope, and a high-speed camera of 20 MHz in a maximum framing rate was used to visualize the dynamics of cavitation bubbles that causes a sonoporation effect. Differential interference contrast and fluorescence techniques were used for sensitive visualization of cell changes during sonoporation. The system is also equipped with optical tweezers that can move a microbubble of several microns in size by using a donut-shaped light beam. In situ microscopic observation of sonoporation was carried out using a cell with a size- and position-controlled microbubble. The experimental results showed that the ability of cells to repair sonoporation-induced damage depends on their membrane tension, indicating the usefulness of the observation system as a basic tool for the investigation of sonoporation phenomena.


Cytoplasmic induction of changes in the ultrastructure of the Acetabularia nucleus and perinuclear cytoplasm.
The ultrasturcture of the cell nucleus and perinuclear cytoplasm in Acetabularia is quite different in young and old cells. When a nucleus of an old cell was implanted into the cytoplasm of a young cell the nucleus assumed the morphology typical of a young cell within less than 10 days. The cytoplasm of an old cell was able to induce the reverse change in an implanted nucleus from a young cell. This cytoplasmic induction of nuclear ageing appeared to proceed more rapidly than rejuvenation.


[Continuing education and matrix support: training, experience, and practices of health professionals in the Centers for the Support of Family Health and the supported teams].
: This study aimed to understand and analyze how processes of continuing education are experienced by professionals in the Centers for the Support of Family Health (NASF) and family health teams. The aim was also to understand how matrix support as a strategy for continuing education was incorporated into the work of these professionals. This was a multiple case study with a qualitative approach in six municipalities in southeast Bahia State, Brazil, which had type-I NASF. Semi-structured interviews were held with 43 NASF professionals and 40 with physicians and nurses from the supported teams. The interviews were categorized and analyzed using thematic content analysis. The results showed that the activities in continuing education are insufficient and inadequate, with a supply of sporadic training developed with a traditional teaching methodology. Continuing education had not been institutionally consolidated in the six municipalities. Training in matrix support and the work process in the NASF proved weak for both groups, which interfered in the support function and management of care. The study revealed the limited action of NASF as promotors of continuing education for the supported teams.


Glycolipid-dependent and lectin-driven transcytosis in mouse enterocytes.
Glycoproteins and glycolipids at the plasma membrane contribute to a range of functions from growth factor signaling to cell adhesion and migration. Glycoconjugates undergo endocytic trafficking. According to the glycolipid-lectin (GL-Lect) hypothesis, the construction of tubular endocytic pits is driven in a glycosphingolipid-dependent manner by sugar-binding proteins of the galectin family. Here, we provide evidence for a function of the GL-Lect mechanism in transcytosis across enterocytes in the mouse intestine. We show that galectin-3 (Gal3) and its newly identified binding partner lactotransferrin are transported in a glycosphingolipid-dependent manner from the apical to the basolateral membrane. Transcytosis of lactotransferrin is perturbed in Gal3 knockout mice and can be rescued by exogenous Gal3. Inside enterocytes, Gal3 is localized to hallmark structures of the GL-Lect mechanism, termed clathrin-independent carriers. These data pioneer the existence of GL-Lect endocytosis in vivo and strongly suggest that polarized trafficking across the intestinal barrier relies on this mechanism.


Predicting cisplatin-induced acute kidney injury by urinary neutrophil gelatinase-associated lipocalin excretion: a pilot prospective case-control study.
Nephrotoxicity is the major limitation to cisplatin therapy for solid tumors. We aimed at evaluating whether early increases in serum/urine levels of neutrophil gelatinase-associated lipocalin (NGAL), a biomarker of tubular damage and regeneration, predict cisplatin-induced acute kidney injury (AKI). We compared changes in serum creatinine and serum/urine NGAL levels (ELISA assay) at 1 and 4 h and 1, 2, 3, 7 and 15 days after cisplatin infusion (70-80 mg/m(2)) versus baseline in 12 consecutive cancer patients (cases) with AKI (>25% serum creatinine increase vs. baseline) and 12 consecutive controls without AKI. Baseline characteristics and posttreatment serum NGAL levels were similar in both groups. Urinary NGAL levels increased significantly more in cases than in controls at 1, 2, 3 and 15 days after cisplatin. The NGAL increase preceded AKI by 4.5 days and the NGAL increase at day 2 after cisplatin independently predicted AKI (p < 0.05). Six cases with residual kidney dysfunction at 15 days showed a trend to earlier and higher increase in urinary NGAL levels compared to cases with renal function recovery. An early increase in urinary NGAL excretion may help in identifying patients at risk of cisplatin-induced AKI who might benefit from innovative treatments to prevent cisplatin nephrotoxicity.


Applying the COM-B model to patient-reported barriers to medication adherence in pediatric acute lymphoblastic leukemia.
Adherence to oral chemotherapy, including 6-mercaptopurine (6-MP), is suboptimal in pediatric acute lymphoblastic leukemia (ALL), which is associated with increased risk of relapse. Study objectives were to examine self-reported adherence to 6-MP and related barriers to adherence, mapped to the capability, opportunity, motivation, and behavior (COM-B) model for behavior change. Forty-nine parents (median, 39 years old; 76% females) and 15 patients (median, 17 years old, 20% females) completed the study survey. Suboptimal adherence was reported in 43% of parents and 73% of patients. Most parents and patients (80% and 90%, respectively) reported ≥1 adherence barrier. Parents reported difficulty helping their child meet others with ALL (43%), contacting community organizations (39%), and meeting other parents (37%). Patients reported difficulty finding out what their medications are (40%), finding out what 6-MP does (47%), and meeting other patients (40%). Using the COM-B, we found that parents and patients endorsed barriers in multiple components, especially physical (55%, 67%) and social opportunity (56%, 47%), highlighting that barriers to adherence may be multifaceted. Our results suggest that parents and patients with ALL face various prevalent barriers to medication adherence and provide insight into the development of behavioral interventions focused on promoting adherence, which is essential to prevent relapse and optimize health outcomes in ALL.


[Falciparum malaria in Oslo and Akerhus].
Imported falciparum malaria in an increasingly frequent health problem in many areas in which it is not endemic. Complications are commonly seen, and reported case-fatality rates may exceed 3%. The study is a medical chart-based retrospective study of all cases of falciparum malaria diagnosed in Oslo and Akershus counties, south-eastern Norway, 1988-1997. We identified 232 diagnosed cases; of these, records were available for 222 cases (95%). The incidence rate almost quadrupled during the study period. The two largest groups were immigrants visiting their country of origin (35%) and Norwegian tourists (29%). 95% of the cases were infected in Sub-Saharan Africa. There were no fatal cases, and only eight cases (3.6%) developed complicated falciparum malaria. In a statistical analysis, the following factors were found to be significantly associated with complicated disease: higher age, noncompliance to recommended chemoprophylaxis in assumed non-immune subjects, prolonged doctor's delay and prolonged diagnostic delay. The study suggests that complications in imported falciparum malaria may largely be prevented by a high rate of chemoprophylaxis compliance in non-immune travellers and a high awareness of this possibility among physicians evaluating febrile travellers from endemic areas.


Structure and function of pulmonary surfactant proteins.
In summary, the isolation and characterization of three surfactant proteins have considerably changed our understanding of the nature of pulmonary surfactant and its metabolism. The isolation of the cDNAs and genes encoding the proteins and the elucidation of their structure now makes possible the generation of surfactant proteins for further study and for therapy of surfactant deficient states. Artificial surfactant consisting of appropriately modified human proteins can be produced by recombinant DNA technology. Mixed with appropriate synthetic phospholipids, these proteins may be useful for the treatment of infants with hyaline membrane disease. Significant progress has been made in identifying and characterizing these three more abundant proteins; however, many questions regarding the functions of numerous other proteins present in surfactant (whether produced by Type II cells, Type I cell, Clara cell, or others) remain unanswered. Knowledge of the factors controlling developmental expression of surfactant phospholipids and protein as well as the molecular basis of the interactions among the surfactant proteins and phospholipids may lead to new strategies for therapy of hyaline membrane disease. Major questions regarding the site and nature of posttranslational modification of the surfactant proteins, their sites of assembly with lipids, and their precise roles in the metabolism of surfactant in vitro remain to be clarified. It is hoped that answers to these questions will facilitate the identification and design of appropriate therapy of infants and adults with pulmonary disease associated with surfactant deficiency.


Movement and Mobility: A Concept Analysis.
This article provides an analysis of the concepts of movement and mobility within the context of the International Classification of Functioning, Disability, and Health (ICF) for patients' functioning, disability, and health. The methodology developed by Walker and Avant was used to clarify definitions, components, and relationships relevant to the 2 concepts and to the elements of the ICF framework. Definitions and the relationship between concepts are key information that clinicians and researchers need to measure the correct concept when they are assessing the effectiveness of interventions in nursing practice. Concept analysis findings are grounded by the notion that movement occurs when the body causes its own displacement and is explained by the basic principles of physics, human anatomy, and physiology. Mobility is then distinct because it is affected by the environment that the individual is in and can be assisted by any type of mobility aid. Mobility does not need to be generated by the individual's muscles but does need to be controlled by the individual who is mobile. An individual's mobility in his or her environment is important to his or her well-being and needs to be understood in relationship to his or her movements.


Fast activation of dihydropyridine-sensitive calcium channels of skeletal muscle. Multiple pathways of channel gating.
Dihydropyridine (DHP) receptors of the transverse tubule membrane play two roles in excitation-contraction coupling in skeletal muscle: (a) they function as the voltage sensor which undergoes fast transition to control release of calcium from sarcoplasmic reticulum, and (b) they provide the conducting unit of a slowly activating L-type calcium channel. To understand this dual function of the DHP receptor, we studied the effect of depolarizing conditioning pulse on the activation kinetics of the skeletal muscle DHP-sensitive calcium channels reconstituted into lipid bilayer membranes. Activation of the incorporated calcium channel was imposed by depolarizing test pulses from a holding potential of -80 mV. The gating kinetics of the channel was studied with ensemble averages of repeated episodes. Based on a first latency analysis, two distinct classes of channel openings occurred after depolarization: most had delayed latencies, distributed with a mode of 70 ms (slow gating); a small number of openings had short first latencies, < 12 ms (fast gating). A depolarizing conditioning pulse to +20 mV placed 200 ms before the test pulse (-10 mV), led to a significant increase in the activation rate of the ensemble averaged-current; the time constant of activation went from tau m = 110 ms (reference) to tau m = 45 ms after conditioning. This enhanced activation by the conditioning pulse was due to the increase in frequency of fast open events, which was a steep function of the intermediate voltage and the interval between the conditioning pulse and the test pulse. Additional analysis demonstrated that fast gating is the property of the same individual channels that normally gate slowly and that the channels adopt this property after a sojourn in the open state. The rapid secondary activation seen after depolarizing prepulses is not compatible with a linear activation model for the calcium channel, but is highly consistent with a cyclical model. A six-state cyclical model is proposed for the DHP-sensitive Ca channel, which pictures the normal pathway of activation of the calcium channel as two voltage-dependent steps in sequence, plus a voltage-independent step which is rate limiting. The model reproduced well the fast and slow gating models of the calcium channel, and the effects of conditioning pulses. It is possible that the voltage-sensitive gating transitions of the DHP receptor, which occur early in the calcium channel activation sequence, could underlie the role of the voltage sensor and yield the rapid excitation-contraction coupling in skeletal muscle, through either electrostatic or allosteric linkage to the ryanodine receptors/calcium release channels.


Variants in the ASB10 Gene Are Associated with Primary Open Angle Glaucoma.
Recently nonsynonymous coding variants in the ankyrin repeats and suppressor of cytokine signaling box-containing protein 10 (ASB10) gene were found to be associated with primary open angle glaucoma (POAG) in cohorts from Oregon and Germany, but this finding was not confirmed in an independent cohort from Iowa. The aim of the current study was to assess the role of ASB10 gene variants in Pakistani glaucoma patients. Sanger sequencing of the coding exons and splice junctions of the ASB10 gene was performed in 30 probands of multiplex POAG families, 208 sporadic POAG patients and 151 healthy controls from Pakistan. Genotypic associations of individual variants with POAG were analyzed with the Fisher's exact or Chi-square test. In total 24 variants were identified in POAG probands and sporadic patients, including 11 novel variants and 13 known variants. 13 of the variants were nonsynonymous, 6 were synonymous, and 5 were intronic. Three nonsynonymous variants (p.Arg49Cys, p.Arg237Gly, p.Arg453Cys) identified in the probands were not segregating in the respective families. This is not surprising since glaucoma is a multifactorial disease, and multiple factors are likely to be involved in the disease manifestation in these families. However a nonsynonymous variant, p.Arg453Cys (rs3800791), was found in 6 sporadic POAG patients but not in controls, suggesting that it infers increased risk for the disease. In addition, one synonymous variant was found to be associated with sporadic POAG: p.Ala290Ala and the association of the variant with POAG remained significant after correction for multiple testing (uncorrected p-value 0.002, corrected p-value 0.047). The cumulative burden of rare, nonsynonymous variants was significantly higher in sporadic POAG patients compared to control individuals (p-value 0.000006). Variants in ASB10 were found to be significantly associated with sporadic POAG in the Pakistani population. This supports previous findings that sequence variants in the ASB10 gene may act as a risk factor for glaucoma.


What can flies tell us about copper homeostasis?
Copper (Cu) is an essential redox active metal that is potentially toxic in excess. Multicellular organisms acquire Cu from the diet and must regulate uptake, storage, distribution and export of Cu at both the cellular and organismal levels. Systemic Cu deficiency can be fatal, as seen in Menkes disease patients. Conversely Cu toxicity occurs in patients with Wilson disease. Cu dyshomeostasis has also been implicated in neurodegenerative disorders such as Alzheimer's disease. Over the last decade, the fly Drosophila melanogaster has become an important model organism for the elucidation of eukaryotic Cu regulatory mechanisms. Gene discovery approaches with Drosophila have identified novel genes with conserved protein functions relevant to Cu homeostasis in humans. This review focuses on our current understanding of Cu uptake, distribution and export in Drosophila and the implications for mammals.


[Changes in the renin-angiotensin-aldosterone system in elderly patients with chronic ischemic heart disease. 4. The renin-angiotensin-aldosterone system in elderly patients with ischemic heart disease and cardiovascular insufficiency].
The elderly chronic ischemic heart disease (IHK) patients with cardiac failure show a higher activation of the renin-angiotensin-aldosterone system compared to the younger patients. It was noted functional activity of the renin-angiotensin-aldosterone system increases with a progress of the disease (decompensation). Changes occur not only in the basal level of plasma reninactivity and circulating aldosterone concentration, but also the 24 hour rhythm to the side of an increased hormonal level during the evening hours, evidencing thus for disadaption of the renin-angiotensin-aldosterone system and its decreased reliability under conditions of habitual life activity. Administration of the converting enzyme inhibitor, Captopril, has confirmed a pathogenetic role of the renin-angiotensin-aldosterone system in the development of cardiac failure syndrome in the chronic IHK patients as well as verified a new approach in the treatment of this pathology.


The Fracture Risk Assessment Tool Probability and Trabecular Bone Score Mediate the Relationship between Sphingosine 1-phosphate Levels and Fracture Risk.
The sphingosine 1-phosphate (S1P) concentration is a potential biomarker of osteoporotic fracture and is associated with both the fracture risk assessment tool (FRAX) probability and trabecular bone score (TBS), which are well-known predictors of fracture. We sought to estimate the effect of the S1P concentration on fracture risk using the FRAX probability and TBS as mediators. Plasma S1P concentrations, FRAX variables, and TBSs were measured in 66 postmenopausal women with fractures and 273 postmenopausal women without fractures. Associations between S1P concentration, FRAX probability, TBS, and fracture risk were analyzed using correlation, logistic regression, and mediation analyses. Subjects in the highest S1P concentration tertile had a higher fracture risk (odds ratio [OR], 5.09; 95% confidence interval [CI], 2.22-11.67) than those in the lowest S1P concentration tertile before adjustment. Subjects in the highest FRAX probability tertile had a higher fracture risk (OR, 14.59; 95% CI, 5.01-42.53) than those in the lowest FRAX probability tertile before adjustment. Subjects in the lowest TBS tertile had a higher fracture risk (OR, 4.76; 95% CI, 2.28-9.93) than those in the highest TBS tertile before adjustment. After adjustment for FRAX probability and TBS, the highest S1P concentration tertile was still associated with a higher fracture risk (OR, 3.13; 95% CI, 1.28-7.66). The FRAX probability and TBS accounted for 32.6% and 21.7%, respectively, of the relationship between the S1P concentration and fracture risk. The relationship between the circulating S1P concentration and fracture risk was partly mediated by the FRAX probability, bone microarchitecture, and other factors.


Network-based approaches elucidate differences within APOBEC and clock-like signatures in breast cancer.
Studies of cancer mutations have typically focused on identifying cancer driving mutations that confer growth advantage to cancer cells. However, cancer genomes accumulate a large number of passenger somatic mutations resulting from various endogenous and exogenous causes, including normal DNA damage and repair processes or cancer-related aberrations of DNA maintenance machinery as well as mutations triggered by carcinogenic exposures. Different mutagenic processes often produce characteristic mutational patterns called mutational signatures. Identifying mutagenic processes underlying mutational signatures shaping a cancer genome is an important step towards understanding tumorigenesis. To investigate the genetic aberrations associated with mutational signatures, we took a network-based approach considering mutational signatures as cancer phenotypes. Specifically, our analysis aims to answer the following two complementary questions: (i) what are functional pathways whose gene expression activities correlate with the strengths of mutational signatures, and (ii) are there pathways whose genetic alterations might have led to specific mutational signatures? To identify mutated pathways, we adopted a recently developed optimization method based on integer linear programming. Analyzing a breast cancer dataset, we identified pathways associated with mutational signatures on both expression and mutation levels. Our analysis captured important differences in the etiology of the APOBEC-related signatures and the two clock-like signatures. In particular, it revealed that clustered and dispersed APOBEC mutations may be caused by different mutagenic processes. In addition, our analysis elucidated differences between two age-related signatures-one of the signatures is correlated with the expression of cell cycle genes while the other has no such correlation but shows patterns consistent with the exposure to environmental/external processes. This work investigated, for the first time, a network-level association of mutational signatures and dysregulated pathways. The identified pathways and subnetworks provide novel insights into mutagenic processes that the cancer genomes might have undergone and important clues for developing personalized drug therapies.


PoreGlow: A split green fluorescent protein-based system for rapid detection of Listeria monocytogenes.
Listeria monocytogenes, a severe foodborne pathogen causing severe diseases underscores the necessity for the development of a detection system with high specificity, sensitivity and utility. Herein, the PoreGlow system, based on split green fluorescent protein (GFP), was developed and assessed for the fast and accurate detection of L. monocytogenes. Split GFP-encapsulated liposomes were optimized for targeted analysis. The system utilizes listeriolysin O (LLO), a toxin produced by L. monocytogenes that enlarges the pores split GFP-encapsulated liposomes, to detect L. monocytogenes by measuring the fluorescent signal generated when the encapsulated GFP is released and reacted with the externally added fragment of the split GFP. The system exhibited a limit of detection of 0.17 μg/ml for LLO toxin and 10 CFU/mL for L. monocytogenes with high sensitivity and specificity and no cross-reactivity with other bacteria. The PoreGlow system is practical, rapid, and does not require sample pre-treatment, making it a promising tool for the early detection of L. monocytogenes in food products, which is crucial for preventing outbreaks and protecting public health.


Butt lengths differ by area deprivation level: a field study to explore intensive smoking.
We collected cigarette butts in a range of residential areas, to assess differences in the length of unburnt tobacco in the butts, and in proportions of roll-your-own (RYO) cigarettes. Two high, two medium, and two low deprivation areas, as classified by deciles of the New Zealand Deprivation Index, were selected for the Wellington region. Collected butts were systematically classified and measured. A mixed model of analysis, treating location clusters nested within deprivation level areas as a random effect, was used to assess differences in mean length of unburnt tobacco in the butts. A total of 6,262 cigarette butts and separate filters were collected, of which 3,509 (56.0%) were measurable manufactured cigarette butts, 1,069 were unmeasurable manufactured butts, 1,450 were RYO butts, and 236 were RYO filters. The RYO butts were not measured because of the extent of their degradation. The unburnt tobacco lengths in manufactured cigarette butts were significantly shorter in the most deprived areas, relative to the least deprived areas (p = .035). Deformed manufactured cigarette butts (i.e., that potentially were stubbed out) showed the same pattern (p = .011 between the most and least deprived areas). We found no significant difference between deprivation areas in the proportion of RYO material found. The shorter mean unburnt tobacco length in the most deprived areas is consistent with more intensive smoking among smokers in those areas. This finding is consistent with other evidence of increased price sensitivity among poorer smokers, and with basic economic theory. Further evidence on observed smoking behavior in the field is necessary to better interpret these preliminary findings.


Evidence from lesion studies for epileptogenic and non-epileptogenic neurotoxic interactions between kainic acid and excitatory innervation.
The toxicity of kainic acid toward rat hippocampal neurons depends on the presence of specific excitatory afferents. Acute destruction of the critical pathway essentially abolishes the neurotoxicity of intraventricular kainic acid, but some or all hippocampal neurons continue to be destroyed by locally-injected kainic acid until the critical pathway(s) degenerates. These results support the view that kainic acid destroys hippocampal neurons in two ways: (1) by initiating a lethal status epilepticus; and (2) by interacting with certain pathways independently of on-going electrical activity within those pathways.


Comparison between interventional versus medical therapy in patients with rheumatic mitral valve stenosis in Tanzania.
Rheumatic heart disease remains the most common cardiovascular disease in children and young adults. The outcome of interventional versus medical therapy on the long term is not fully elucidated yet. This study provides contemporary data on the clinical profile, treatment and follow up of patients with rheumatic mitral stenosis (MS) in Tanzania. Patients' medical information, investigations and treatment data were recorded in this prospective cohort study. They were followed up for 6-24 months to determine the long-term outcome. Interventional therapy was defined as a combination of surgery and percutaneous balloon mitral valvuloplasty. Kaplan-Meier curves and Cox proportional hazards model were used in analyses. p-Value < 0.05 was considered statistically significant. We enrolled 290 consecutive patients. Interventions were done in half of the patients. Median follow up was 23.5 months. Mortality was higher in the medical than interventional treatment (10.4% vs. 4%, log-rank p = 0.001). Median age was 36 years, females (68.3%) and low income (55.5%). Multivalvular disease was found in 116 (40%) patients, atrial fibrillation (31.4%), stroke/transient ischaemic attack (18.9%) and heart failure class III-IV (44.1%). Median (IQR) duration of disease was 3 (4) years, secondary prophylaxis (27.7%) and oral anticoagulants use (62.3%). In multivariable analysis, the risk of death among patients on medical was 3.07 times higher than those on interventional treatment (crude HR 3.07, 95% CI 1.43-6.56, p = 0.004), 2.44 times higher among patients with arrhythmias versus without arrhythmias (crude HR 2.44, 95% CI 1.19-4.49, p = 0.015) and 2.13 times higher among patients with multivalvular than single valve disease (crude HR 2.13, 95% CI 1.09-4.16, p = 0.026). Intervention is carrying low mortality compared to medical treatment. Arrhythmias and multivalvular disease are associated with a high mortality. Rheumatic MS is more prevalent in young people, females and individuals with low income. There is a late hospital presentation and a low use of both secondary prophylactic antibiotics and anticoagulants.


Methicillin-resistant Staphylococcus aureus infection in the hand.
Methicillin-resistant Staphylococcus aureus has become an increasingly common cause of skin and soft-issue infections. The purpose of this study was to review and summarize the most recent literature on methicillin-resistant S. aureus infections of the hand. The English-language literature related to methicillin-resistant S. aureus infections of the hand was reviewed, and information related to the history, epidemiology, clinical presentation, and treatment of methicillin-resistant S. aureus infections of the hand was summarized. Community-acquired methicillin-resistant S. aureus hand infections have increased rapidly in prevalence over the past two decades. The prevalence varies markedly around the world, but in the United States, community-acquired methicillin-resistant S. aureus now causes the majority of purulent hand infections. Purulent hand infections should be treated with surgical drainage. When antibiotics are indicated, empiric coverage for community-acquired methicillin-resistant S. aureus should be provided if local prevalence rates exceed 10 to 15 percent.


Metabolomic analysis of platelets during storage: a comparison between apheresis- and buffy coat-derived platelet concentrates.
Platelet concentrates (PCs) can be prepared using three methods: platelet (PLT)-rich plasma, apheresis, and buffy coat. The aim of this study was to obtain a comprehensive data set that describes metabolism of buffy coat-derived PLTs during storage and to compare it with a previously published parallel data set obtained for apheresis-derived PLTs. During storage we measured more than 150 variables in 8 PLT units, prepared by the buffy coat method. Samples were collected at seven different time points resulting in a data set containing more than 8000 measurements. This data set was obtained by combining a series of standard quality control assays to monitor the quality of stored PLTs and a deep coverage metabolomics study using liquid chromatography coupled with mass spectrometry. Stored PLTs showed a distinct metabolic transition occurring 4 days after their collection. The transition was evident in PLT produced by both production methods. Apheresis-derived PLTs showed a clearer phenotype of PLT activation during early days of storage. The activated phenotype of apheresis PLTs was accompanied by a higher metabolic activity, especially related to glycolysis and the tricarboxylic acid cycle. Moreover, the extent of the activation differed between bags resulting in interbag variability in the storage lesion of apheresis-prepared PLTs. This may be related to donor-related polymorphism. This study demonstrated two discrete metabolic phenotypes in stored PLTs prepared with both apheresis and buffy coat methods. PLT activation occurs during the first metabolic phenotype and might lead to a low reproducibility of the apheresis PCs.


Effects of Liraglutide Versus Placebo on Cardiovascular Events in Patients With Type 2 Diabetes Mellitus and Chronic Kidney Disease.
LEADER trial (Liraglutide Effect and Action in Diabetes: Evaluation of CV Outcome Results) results demonstrated cardiovascular benefits for patients with type 2 diabetes mellitus at high cardiovascular risk on standard of care randomized to liraglutide versus placebo. The effect of glucagon-like peptide-1 receptor agonist liraglutide on cardiovascular events and all-cause mortality in patients with type 2 diabetes mellitus and chronic kidney disease is unknown. Liraglutide's treatment effects in patients with and without kidney disease were analyzed post hoc. Patients were randomized (1:1) to liraglutide or placebo, both in addition to standard of care. These analyses assessed outcomes stratified by baseline estimated glomerular filtration rate (eGFR; <60 versus ≥60 mL/min/1.73 m2) and baseline albuminuria. The primary outcome (composite of cardiovascular death, nonfatal myocardial infarction, or nonfatal stroke) and secondary outcomes, including all-cause mortality and individual components of the primary composite outcome, were analyzed using Cox regression. Overall, 2158 and 7182 patients had baseline eGFR <60 or ≥60 mL/min/1.73 m2, respectively. In patients with eGFR <60 mL/min/1.73 m2, risk reduction for the primary composite cardiovascular outcome with liraglutide was greater (hazard ratio [HR], 0.69; 95% CI, 0.57-0.85) versus those with eGFR ≥60 mL/min/1.73 m2 (HR, 0.94; 95% CI, 0.83-1.07; interaction P=0.01). There was no consistent effect modification with liraglutide across finer eGFR subgroups (interaction P=0.13) and when analyzing eGFR as a continuous variable (interaction P=0.61). Risk reductions in those with eGFR <60 versus ≥60 mL/min/1.73 m2 were as follows: for nonfatal myocardial infarction, HR, 0.74; 95% CI, 0.55-0.99 versus HR, 0.93; 95% CI, 0.77-1.13; for nonfatal stroke, HR, 0.51; 95% CI, 0.33-0.80 versus HR, 1.07; 95% CI, 0.84-1.37; for cardiovascular death, HR, 0.67; 95% CI, 0.50-0.90 versus HR, 0.84; 95% CI, 0.67-1.05; for all-cause mortality, HR, 0.74; 95% CI, 0.60-0.92 versus HR, 0.90; 95% CI, 0.75-1.07. Risk reduction for the primary composite cardiovascular outcome was not different for those with versus without baseline albuminuria (HR, 0.83; 95% CI, 0.71-0.97; and HR, 0.92; 95% CI, 0.79-1.07, respectively; interaction P=0.36). Liraglutide added to standard of care reduced the risk for major cardiovascular events and all-cause mortality in patients with type 2 diabetes mellitus and chronic kidney disease. These results appear to apply across the chronic kidney disease spectrum enrolled. URL: https://www.clinicaltrials.gov/ . Unique identifier: NCT01179048.


Ascorbic acid and the common cold. Evaluation of its efficacy and toxicity.
We reviewed the clinical data relating to the efficacy and safety of pharmacologic doses of ascorbic acid in the prevention and treatment of the common cold. Although one study tentatively supports the hypothesis that such doses of ascorbic acid may be efficacious, a second study by the same group did not confirm the significant findings, and no clear, reproducible pattern of efficacy has emerged from the review of all the evidence. Similarly, there is currently little adequate evidence on either the presence or the absence of serious adverse reactions to such doses of ascorbic acid, although many such reactions have been hypothesized. The unrestricted use of ascorbic acid for these purposes cannot be advocated on the basis of the evidence currently available.


Streptococcus pyogenes cervical intertrigo with secondary bacteremia.
We report a 12-month-old infant girl with cervical intertrigo caused by Streptococcus pyogenes. This form of intertrigo has been reported in only five children, but it is clearly underestimated. It should be suspected for well-demarcated beefy-red lesions of the neck not responding to antifungal therapy. A rapid streptococcal antigen test of a lesion specimen is a useful diagnostic tool. Our patient was notable for the development of S. pyogenes bacteremia, a complication that has not been previously associated with this condition.


Conjunctival Icterus - An Important but Neglected Sign of Clinically Relevant Hyperbilirubinemia in Jaundiced Neonates.
Conjunctival icterus is a largely neglected physical sign that may be helpful in identifying neonates with clinically relevant hyperbilirubinemia by practitioners in the hospital and outpatient clinic or parents at home. A recent NICU based study reported that conjunctival icterus is often a sign of significant (TSB ≥ 17 mg/dl) hyperbilirubinemia and TSB levels ≥ 76th-95th percentile on the Bhutani nomogram. In contrast, others report that conjunctival icterus, although frequently present at high TSB levels, may also be detected at lower TSB concentrations; suggesting instead that its absence may help to rule out significant hyperbilirubinemia. The current review details the nature of conjunctival icterus and presents new data on its clinical occurrence in relation to TSB levels that re-affirm its correlation with elevated TSB concentrations and use to trigger TSB measurement in the jaundiced neonate.


Acromegaly and Cancer: An Update.
Acromegaly is a chronic and rare disease. The diagnosis usually takes several years. Multiple comorbidities are associated with acromegaly. Long-term exposure to growth factors may lead to complications such as the development of benign or malignant tumors. However, the association between acromegaly and cancer remains a matter of debate due to multiple limitations in epidemiological data. There is controversy between acromegaly and mortality, but evidence shows a significant improvement in mortality rates with disease control and careful management of comorbidities. Older age, increased growth hormone levels (GH) at last follow-up, higher insulin-like growth factor-1 (IGF-1) levels at diagnosis, malignancy and radiotherapy were proposed as independent predictors of mortality. In this review we summarize the current state of knowledge in this field. Incidence of different cancer types is described. Rigorous surveillance of endocrine diseases may contribute to increased tumor detection. Personalized screening should probably be recommended.


A graphical method of estimating fatal radiation-induced cancers using the BEIR V method.
A method is presented that permits health physicists to estimate the risks of radiation-induced cancers in populations of males or females exposed to whole-body external irradiation at any age. Graphics are presented that may be used for either acute or chronic exposures. There is good agreement for chronic exposures when the cancer estimates determined here are compared with those prescribed in BEIR V. There are differences between the two when considering some acute exposures. In such circumstances, the method given here generally gives conservative results.


Listening preference for the native language compared to an unfamiliar language in hearing and hearing-impaired infants after cochlear implantation.
The goal of this study was to investigate the preference for the native language compared with an unfamiliar language in normally hearing (NH) and hearing-impaired infants with cochlear implants (CIs). Preference for the native language is an important step in the process of language acquisition because it helps infants to attend to the important signals of their environment. This preference has been shown very early in the infants' life. In the case of infants with CI, it is of interest to determine whether they show similar perceptual biases as NH infants despite their impaired auditory system and impoverished input provided by the CI device. Nine hearing-impaired infants with CI (most with 1-2 mo of CI use) and 19 NH infants were tested on their preference for their native language (Hebrew child-directed speech) compared with a nonnative language (English child-directed speech). The central fixation preference procedure was used in which listening times were measured via orientation responses of the infant to visual stimuli. 1) Normal hearing and infants with CI had perceptual bias for their native language (Hebrew) when compared with a nonnative language (English). 2) Infants with CI have shorter attention to speech stimuli compared with NH. The findings of the present study are the first to show that hearing-impaired infants with CI bring the same perceptual biases to the task of language learning as NH infants. These have important implications on understanding the process by which infants with CI acquire language via the CI device.


A Spatial Analysis of Land Use and Network Effects on Frequency and Severity of Cyclist-Motorist Crashes in the Copenhagen Region.
Urban and transport planners worldwide have recently designed and implemented policies for increasing the number of cyclists. Although cycling is on the rise even in car-oriented cities and regions, the fear of being involved in a crash is still the main obstacle to further increases in cycling market shares. The current study proposes the first joint model of frequency and severity of cyclist-motorist collisions with the aim of unraveling the factors contributing to both the probability of being involved in a crash and, conditional on the crash occurrence, experiencing a severe injury outcome. A multivariate Poisson-lognormal model with correlated autoregressive priors was estimated on a sample of 5,349 cyclist-motorist crashes that occurred in the Copenhagen region between 2009 and 2013. The model considered the links of the road network in the region as the unit of observation, controlled for traffic exposure of nonmotorized and motorized transport modes, evaluated the effect of infrastructure and land use, and accounted for heterogeneity and spatial correlation across links. Results confirmed the existence of the phenomenon of safety in numbers and added to the narrative by emphasizing that the most severe crashes are the ones most benefiting from an increase in the number of cyclists. In addition, results argued that the construction of Copenhagen-style bicycle paths would significantly contribute to increasing safety, especially in suburban areas where the speed differential between cyclists and motorists is greater. Last, results illustrated a need for thinking about cycling safety in intersection design and reflecting on the importance of spatial and aspatial correlation both within and between injury categories. The findings from this study illustrated how encouraging cycling would increase safety in relation to the phenomenon of safety in numbers and how, in turn, increasing safety would convince more people to cycle. In addition, they suggested how the design of bicycle infrastructure should not only consider bicycle lanes but in particular focus on bicycle paths where the number of conflicts and the stress for sharing the road are highly reduced and how thinking about road design should extend to the general level and include a discourse about safer intersections. Last, attention should be given to the road design in the city center and to traffic management, because clearly safer traffic implies more cyclists and, in turn, more cyclists imply fewer cars and less congestion.


MMN and attention: competition for deviance detection.
We addressed the question of whether the mismatch negativity (MMN) event-related potential reflects an attention-independent process. Previous studies have shown that the MMN response to intensity deviation was significantly reduced or even abolished when attention was highly focused on a concurrent sound channel, whereas no conclusive evidence of attentional sensitivity has been obtained for frequency deviation. We tested a new hypothesis suggesting that competition between detection of identical deviations in attended and unattended channels and the biasing of this competition induced by the subject's task account for the observed MMN effects. In a fast-paced dichotic paradigm, we set up competition for frequency MMN and removed it for intensity MMN. We found that frequency MMN was now abolished in the unattended channel, whereas the amplitude of the intensity MMN was unaffected. These results support the competition hypothesis and suggest that selective attention in and of itself does not affect the MMN. Top-down processes can determine what information reaches the deviance-detection process when changes in multiple channels vie for the same MMN resource and one of the competing changes is relevant for the subject's task.


An antifibrin monoclonal antibody useful in immunoscintigraphic detection of thrombi.
Balb/c mice were immunized with human plasmin-generated fibrinogen degradation product Y. Spleen cells were fused with P3X63-Ag8.653 myeloma cells. A clone (Y22) was found that produces monoclonal antibodies (MoAbs) with a strong reactivity with human fibrin and only a weak reactivity with fibrinogen in an enzyme immunoassay (EIA). Y22 also reacts with fibrin of rabbits, rats, sheep, and dogs. The antibodies are of the IgG1 kappa-type and appear to be directed against a conformation-dependent epitope in the D-domain of fibrin. Experiments with 99mTc-labeled Y22 in vitro show that Y22 binds rapidly to forming clots. 99mTc-Y22 also binds to preformed plasma clots in a plasma milieu, even in the presence of high concentrations of heparin. Clot localization experiments in rabbits and rats confirm the high fibrin specificity and the potential of 99mTc-Y22 for thrombus imaging in vivo.


Clinical study of alar anatomy and surgery of the alar base.
To analyze and quantify specific aspects of alar base anatomy and to identify anatomic configurations that may be correlated with specific surgical manipulations. Analysis in a population of patients presenting for aesthetic nasal surgery. Facial plastic surgery practice. The photographic slides of 120 white patients who presented for consultation were reviewed. On the base view of photographic slides, observations were made on the width of the alar base, recurvature of the alar base, thickness of the alar lobule, thickness of the alar wall, and flare of the alar wall. On the lateral view, observations were made on the vertical insertion of the ala on the face (cephalic, normal, or caudal), contour of alar rim in profile (gentle S-shape or straight), size of alar lobule (small, normal, or large), and alar-columellar relationship, with special attention to the presence of alar hooding. A spreadsheet program was used to analyze the incidence of each configuration and any association between various features of the alar base. Anatomic diversity exists that requires a careful individual analysis for each patient. A planned surgical intervention must fit the patient's unique anatomy. Distinctive configurations of recurvature, vertical insertion, and other aspects of the alar base were observed, with special implications for the surgeon's approach. Anatomic diversity requires a thorough preoperative examination followed by selection of an approach that addresses the specific anatomic findings. The choice of the best alar reduction and sculpture technique ultimately relies on a precise analysis of the anatomic configuration of the patient's alar base.


Endoscopic management of eroded prosthesis in vertical banded gastroplasty patients.
One of the major complications of applying a prosthetic device to facilitate gastric partitioning has been intraluminal erosion of the prosthesis. Removing an eroded gastric band is fraught with difficulty secondary to the extensive inflammatory response around the proximal stomach and left lobe of the liver. As a result, bariatric clinicians have attempted to apply endoscopic technology to facilitate removal of eroded gastric prostheses. Our study reports on our experience of applying endoscopic scissor transection to remove eroded gastric prostheses in a large tertiary-care medical center. A retrospective chart review was conducted to identify all adult (>18 years old) patients managed endoscopically for removal of eroded prosthesis post bariatric surgery at the Baylor College of Medicine Comprehensive Bariatric Surgery Center and the University of Texas-Houston Bariatric Surgery Program. Nine patients, mean body mass index (BMI) 39.5 +/- 7.3 kg/m(2), were managed endoscopically to remove eroded gastric prosthesis post vertical banded gastroplasty. All patients (9/9) reported symptoms consistent with a proximal gastric outlet obstruction (dysphagia, nausea, vomiting). Inadequate weight loss was reported by 67% (6/9) of patients. The mean time for endoscopic band removal was 28.0 +/- 8.8 min. All eroded bands were evident at the end of the gastric pouch and were visualized with either a normal or retroflexed endoscopic view. All nine patients were discharged the same day of their procedure. Mean follow-up of the patients was 9.7 weeks (1-30 weeks). Three patients underwent revisional bariatric surgery following the band removal. Our study demonstrates that endoscopic scissor transaction is a safe and effective modality in removing eroded gastric prostheses after vertical banded gastroplasty and avoids the pitfalls associated with removing the eroded bands surgically. It also allows the patient and surgeon to make an insightful decision regarding a revisional bariatric surgical procedure, without being contingent on addressing the eroded gastric band.


Effectiveness of a nutraceutical supplement containing highly standardized perilla and ginger extracts in patients with functional dyspepsia.
In Western countries functional dyspepsia (FD) has a prevalence of 10-20% among adults and although many drugs are currently available for use within clinical practice, FD remains an important challenge for physicians. Recently, food supplements that are ginger-based, along with other botanicals, have been proposed to be a possible natural alternative to pharmaceutical drugs to empirically counteract the symptoms of FD. We have therefore retrospectively analyzed the efficacy and safety profiles of a nutraceutical containing, in addition to a highly standardized ginger root extract, a multi-fractionated botanical obtained from Perilla frutescens leaf containing an innovative bouquet of compounds, including hydrophilic polyphenols and the lipophilic terpenoid perilla ketone. The results of our single-group study, obtained from patients with a diagnosis of FD who were treated with the perilla/ginger nutraceutical, demonstrated a good efficacy profile, with a significant reduction observed in nearly all evaluated symptoms (epigastric pain, heartburn, gastric reflux, nausea, borborygmi, early satiety, diarrhea/constipation) starting from the first week of treatment that was further improved after 2 weeks. The treatment was well tolerated with very mild side effects (flatulence, meteorism, gastric burning, difficulty in falling asleep) lasting 3-4 days, which disappeared without stopping the treatment. Despite all the limitations of our pragmatic study, we believe that the perilla and ginger supplement we have used can be considered a valid tool for an empirical approach to treating patients with FD, especially when a non-conventional drug treatment is preferable to the patient and considered suitable by the physician.


Review of the pharmacology, clinical efficacy, and safety of azelastine hydrochloride.
Rhinitis is one of the most common diseases in the general population. Although it is not a life-threatening condition, rhinitis can cause significant discomfort and, therefore, negatively impact quality of life. Several treatment options are available; however, optimal relief of symptoms is difficult to achieve for most patients. Azelastine hydrochloride (Astelin) nasal spray is the only prescription intranasal antihistamine available in the USA, and is approved for treating symptoms of both seasonal allergic rhinitis and nonallergic vasomotor rhinitis. Oral formulations of azelastine are available outside the USA for use in seasonal and perennial allergic rhinitis, asthma and urticaria. Azelastine hydrochloride has demonstrated a favorable safety profile during approximately 20 years of clinical use.


Isolation, analysis, and expression of two genes from Thermoanaerobacterium sp. strain JW/SL YS485: a beta-xylosidase and a novel acetyl xylan esterase with cephalosporin C deacetylase activity.
The genes encoding acetyl xylan esterase 1 (axe1) and a beta-xylosidase (xylB) have been cloned and sequenced from Thermoanaerobacterium sp. strain JW/SL YS485. axe1 is located 22 nucleotides 3' of the xylB sequence. The identity of axe1 was confirmed by comparison of the deduced amino acid sequence to peptide sequence analysis data from purified acetyl xylan esterase 1. The xylB gene was identified by expression cloning and by sequence homology to known beta-xylosidases. Plasmids which independently expressed either acetyl xylan esterase 1 (pAct1BK) or beta-xylosidase (pXylo-1.1) were constructed in Escherichia coli. Plasmid pXylAct-1 contained both genes joined at a unique EcoRI site and expressed both activities. Substrate specificity, pH, and temperature optima were determined for partially purified recombinant acetyl xylan esterase 1 and for crude recombinant beta-xylosidase. Similarity searches showed that the axe1 and xylB genes were homologs of the ORF-1 and xynB genes, respectively, isolated from Thermoanaerobacterium saccharolyticum. Although the deduced sequence of the axe1 product had no significant amino acid sequence similarity to any reported acetyl xylan esterase sequence, it did have strong similarity to cephalosporin C deacetylase from Bacillus subtilis. Recombinant acetyl xylan esterase 1 was found to have thermostable deacetylase activity towards a number of acetylated substrates, including cephalosporin C and 7-aminocephalosporanic acid.


Gene expression-based classification of nonseminomatous male germ cell tumors.
Male adult germ cell tumors (GCTs) comprise two major histologic groups: seminomas and nonseminomas. Nonseminomatous GCTs (NSGCTs) can be further divided into embryonal carcinoma (EC), teratoma (T), yolk sac tumor (YS), and choriocarcinoma (CC) on the basis of the lineage differentiation that they exhibit. NSGCTs frequently present as mixed tumors consisting of two or more histological subtypes, often limiting correlative studies of clinical and molecular features to histology. We sought to develop a molecular classifier that could predict the predominant histologic subtype within mixed NSGCT tumor samples. The expression profiles of 84 NSGCTs (42 pure and 42 mixed) and normal age-matched testes were obtained using Affymetrix microarrays. Using prediction analysis for microarrays, we identified 146 transcripts that classified the histology of pure NSGCTs samples with 93% accuracy. When applied to mixed NSGCTs, the classifier predicted a histology that was consistent with one of the reported components in 93% of cases. Among the predictive transcripts were CGB (high in CC), LCN2 (high in T), BMP2 (high in YS), and POU5F1 (high in EC). Thus, the expression-based classifier accurately assigned a single predominant histology to mixed NSGCTs, and identified transcripts differentially expressed between histologic components with relevance to NSGCT differentiation.


Colocated Mental Health/Developmental Care.
The inadequacy of mental health and developmental services for children is a widely recognized and growing problem. Although a variety of solutions have been proposed, none has been generally successful or feasible. This research describes models of colocation that have evolved in primary care settings in Massachusetts and reports on pediatricians' and their colocated colleagues' impressions of their benefits and challenges. Pediatricians in 18 practices that included a colocated mental health/developmental specialist (MH/DS) were identified through a survey administered through the state American Academy of Pediatrics Chapter, and interviewed. Practices varied widely in the professional expertise/training and roles of the MH/DSs, communication among providers, and financial arrangements. The majority of pediatricians and MH/DSs reported being pleased with their colocated arrangements, despite the costs rarely being supported by billing revenues. This study suggests that further development of such systems hold promise to meet the growing need for accessible pediatric mental and developmental health care.


Ventilation/perfusion lung scintigraphy: what is still needed? A review considering technetium-99m-labeled macro-aggregates of albumin.
Lung perfusion scintigraphy (LPS) with technetium-99m-labeled macro-aggregates of albumin (Tc-99m-MAA) is well established in the diagnostic of pulmonary embolism (PE). In the last decade, it was shown that single-photon emission computer tomography (SPECT) acquisition of LPS overcame static scintigraphy. Furthermore, there are rare indications for LPS, such as preoperative quantification of regional lung function prior to lung resection or transplantation, optimization of lung cancer radiation therapy, quantification of right-left shunt, planning of intra-arterial chemotherapy, and several rare indications in pediatrics. Moreover, LPS with Tc-99m-MAA is a safe method with low radiation exposure. PE can also be diagnosed by spiral computer tomography (CT), ultrasound, magnetic resonance angiography, or pulmonary angiography (PA, former gold standard). The present review considers all these methods, especially spiral CT, and compares them with LPS with respect to sensitivity and specificity and gives an overview of established and newer publications. It shows that LPS with Tc-99m-MAA represents a diagnostic method of continuing value for PE. In comparison with spiral CT and/or PA, LPS is not to be defeated as mentioned also by the most actual Prospective Investigation of Pulmonary Embolism Diagnosis (PIOPED) II reports. This applies in particular to chronic or recurring embolisms, whereas currently spiral CT may be of greater value for major or life-threatening embolisms. At present, LPS cannot be replaced by other methods in some applications, such as pediatrics or in the quantification of regional pulmonary function in a preoperative context or prior to radiation therapy. LPS still has a place in the diagnostics of PE and is irreplaceable in several rare indications as described earlier.


Polyamine oxidase activity and polyamine levels in human colorectal cancer and in normal surrounding mucosa.
Polyamine oxidase (PAO) is an enzyme involved in the interconversion pathway of polyamines, compounds required for cell proliferation and differentiation. As the role of PAO in tumor growth is unclear, and no data about PAO activity in human colorectal carcinoma are available, our aim was to investigate PAO activity and polyamine levels in this kind of tumor. Polyamine levels and PAO activity were detected in 30 neoplastic colorectal samples and surrounding mucosa by HPLC. Free and N1-acetylated polyamine levels were higher in the neoplastic tissue than surrounding mucosa of the same patient. On the contrary, PAO activity was significantly lower in the neoplastic tissue than surrounding mucosa. It seems that PAO activity does not play an important role in the increased free polyamine levels in human colorectal carcinoma. Instead, the low PAO activity observed in our study let us to hypothesize that polyamine analogues can have an antitumoral effect on colorectal carcinoma.


Amino acid transport and rubidium-ion uptake in monolayer cultures of hepatocytes from neonatal rats.
Amino acid and K(+) transport during development has been investigated in hepatocyte monolayer cultures with either alpha-amino[1-(14)C]isobutyrate or (86)Rb(+) used as a tracer for K(+). Parenchymal cells from neo- and post-natal rat livers have been isolated by an improved non-perfusion technique [Bellemann, Gebhardt & Mecke (1977)Anal.Biochem.81, 408-415], and the resulting hepatocyte suspensions purified from non-hepatocytes before inoculation. In the presence of Na(+) (Na(+)-dependent component), the rates of amino acid uptake in neonatal hepatocytes were markedly enhanced compared with cells from 30-day-old rats. When Na(+) was replaced by choline (Na(+)-independent component) the accumulation of alpha-aminoisobutyrate was decreased and it was not affected by the age of the animals. Kinetic analysis of Na(+)-dependent alpha-aminoisobutyrate transport revealed the existence of a high-affinity low-K(m) component (K(m)0.91mm) with a V(max.) of 2.44nmol/mg of protein per 4min, which later declined gradually with progressive development. Rates of Rb(+) transport were concomitantly enhanced in neonatal hepatocytes and thereafter declined with postnatal age. The increased Rb(+) influx was effectively inhibited by ouabain and reflected elevated activity of the electrogenic Na(+)/K(+)-pump during early stages of development. Kinetic evaluation of the enhanced rates of Rb(+) uptake indicates multiple and co-operative binding sites of the enzyme involved in the Rb(+) uptake, and the transport system is positively co-operative (the Hill coefficient h is >1.0). In short, amino acid transport in neonatal rat hepatocytes is increased as a result of an existing low-K(m) component for the Na(+)-dependent alpha-aminoisobutyrate uptake, which endows the hepatocytes with a high capability for concentrating amino acids at low ambient values. The concomitant enhancement of K(+) transport reflects changes in the electrochemical gradient for Na(+) across the hepatocellular membrane and, along with this, presumably alterations in the membrane potential; the latter might be the driving force for the enhanced alpha-aminoisobutyrate transport in the alanine-preferring system during postnatal age.