Emergency Department Patient Experience of Care Survey: Development and Field Test.
The Centers for Medicare & Medicaid Services (CMS) have implemented Consumer Assessment of Healthcare Providers and Systems (CAHPS®) surveys to assess patient experience in a number of settings. Following CAHPS principles, RAND researchers designed and field tested an Emergency Department Patient Experience of Care Survey that consists of three survey instruments for use with adult patients who have visited the emergency department (ED). One instrument is for use with those patients who are discharged to the community following their ED visit; the other two are for use with those patients who are admitted to the hospital from the ED (one for use on its own and one to supplement an existing inpatient survey). The authors conducted a field test of these instruments in 12 hospitals in late 2013 and early 2014 and analyzed the resulting data from 4,101 ED patients. The analyses identified four composite measures (measures composed of responses to multiple survey questions) and ten measures that are each composed of a single survey question. As of September 2014, CMS plans to conduct additional testing on these instruments.


Co-simulation of neuromuscular dynamics and knee mechanics during human walking.
This study introduces a framework for co-simulating neuromuscular dynamics and knee joint mechanics during gait. A knee model was developed that included 17 ligament bundles and a representation of the distributed contact between a femoral component and tibial insert surface. The knee was incorporated into a forward dynamics musculoskeletal model of the lower extremity. A computed muscle control algorithm was then used to modulate the muscle excitations to drive the model to closely track measured hip, knee, and ankle angle trajectories of a subject walking overground with an instrumented knee replacement. The resulting simulations predicted the muscle forces, ligament forces, secondary knee kinematics, and tibiofemoral contact loads. Model-predicted tibiofemoral contact forces were of comparable magnitudes to experimental measurements, with peak medial (1.95 body weight (BW)) and total (2.76 BW) contact forces within 4-17% of measured values. Average root-mean-square errors over a gait cycle were 0.26, 0.42, and 0.51 BW for the medial, lateral, and total contact forces, respectively. The model was subsequently used to predict variations in joint contact pressure that could arise by altering the frontal plane joint alignment. Small variations (±2 deg) in the alignment of the femoral component and tibial insert did not substantially affect the location of contact pressure, but did alter the medio-lateral distribution of load and internal tibia rotation in swing. Thus, the computational framework can be used to virtually assess the coupled influence of both physiological and design factors on in vivo joint mechanics and performance.


Surgical management of clinically early breast cancer.
The management of patients with primary operable or potentially curable breast cancer continues to be controversial. Modified radical mastectomy is still considered the standard approach for patients with this problem, but certain carefully selected patients with smaller tumors are being treated in some centers with a breast-sparing operation, usually with some type of axillary dissection, followed by radical radiation therapy. The manner in which this option is presented to patients varies from the most optimistic (it is proven to be equal to mastectomy) to more guarded appraisals (it is experimental, with long-term results and complications yet to be measured). Lending weight to the latter interpretation is another option open to patients, i.e., to enter randomized prospective trials comparing various locoregional approaches. The approach of the authors to these patients will be discussed.


Role of Fungicide Applications on the Integrated Management of Wheat Stripe Rust.
First described in Europe in 1777, stripe rust (SR) caused by Puccinia striiformis Westend. f. sp. tritici Erikss (Pst) is one of the most important and destructive diseases of wheat worldwide. Until 2000, SR was mainly endemic to cooler regions, but since then, new aggressive strains have emerged, spread intercontinentally, and caused severe epidemics in warmer regions across the world. This has put SR as a disease that poses a threat to the world food security. At present, the preferred strategy for control of SR is the access to wheat cultivars with adequate levels of SR resistance. However, wheat breeding programs are not sufficiently advanced to cope with the recently emerged Pst strains. Under this scenario, foliar fungicide applications have become an important component of SR management, but information on the effects of fungicide applications on SR control and wheat cultivar yield response is scarce. This review seeks to provide an overview of the impact and role of fungicides on SR management. With focus on wheat management in the major wheat-growing regions of the world, the review addresses: (a) the efficacy of different fungicide active ingredients, optimal fungicide timing and number of applications in controlling SR, and (b) the impact of fungicide on wheat grain yield response. Inclusion of fungicides in an integrated crop management approach is discussed.


The effect of different desensitizing agents on initial demineralization of human root dentin.
The aim of this study was to determine the caries-protective effect of 3 different desensitizing agents (Seal & Protect 2.0, D/Sense 2, and Gluma Desensitizer) on root dentin in vitro. The root surfaces of 60 freshly extracted, caries-free human molars were used. After removing the cementum, the teeth were coated with an acid-resistant nail varnish, exposing 2 rectangular windows of 2 X 3 mm each on the root surface. One window served as an untreated control, and the other window was treated with 1 of the desensitizing agents. The specimens were randomly distributed among the following experimental groups: group A, D/Sense 2; group B, Seal & Protect 2.0; and group C, Gluma Desensitizer. Subsequently, all specimens were demineralized for 14 days with acidified gel (HEC, pH 4.8, 37 degrees C). Two dentinal slabs were cut from each window. The slabs were ground to a thickness of 80 microm and immersed in water. The demineralization depth was determined using a polarized light microscope. The nontreated control specimens showed lesions with a mean depth of 84.9 microm (+/- 6.0). In the specimens treated with the desensitizing agents, lesion depth was generally significantly reduced. Statistical analysis revealed significantly lower values for the specimens in group B in comparison with the others. Within the limitations of an in vitro investigation, it can be concluded that the demineralization of the root surface can be hampered by applying the desensitizing agents tested.


Plasticity of amyloid fibrils.
In experiments designed to characterize the basis of amyloid fibril stability through mutational analysis of the Abeta (1-40) molecule, fibrils exhibit consistent, significant structural malleability. In these results, and in other properties, amyloid fibrils appear to more resemble plastic materials generated from synthetic polymers than globular proteins. Thus, like synthetic polymers and plastics, amyloid fibrils exhibit both polymorphism, the ability of one polypeptide to form aggregates of different morphologies, and isomorphism, the ability of different polypeptides to grow into a fibrillar amyloid morphology. This view links amyloid with the prehistorical and 20th century use of proteins as starting materials to make films, fibers, and plastics, and with the classic protein fiber stretching experiments of the Astbury group. Viewing amyloids from the point of view of the polymer chemist may shed new light on a number of issues, such as the role of protofibrils in the mechanism of amyloid formation, the biological potency of fibrils, and the prospects for discovering inhibitors of amyloid fibril formation.


Cancer-associated fibroblasts in pancreatic ductal adenocarcinoma.
Pancreatic ductal adenocarcinoma (PDAC) is a lethal cancer with a prominent extracellular matrix (ECM) deposition and poor prognosis. High levels of ECM proteins derived from tumour cells reduce the efficacy of conventional cancer treatment paradigms and contribute to tumour progression and metastasis. As abundant tumour-promoting cells in the ECM, cancer-associated fibroblasts (CAFs) are promising targets for novel anti-tumour interventions. Nonetheless, related clinical trials are hampered by the lack of specific markers and elusive differences between CAF subtypes. Here, we review the origins and functional diversity of CAFs and show how they create a tumour-promoting milieu, focusing on the crosstalk between CAFs, tumour cells, and immune cells in the tumour microenvironment. Furthermore, relevant clinical advances and potential therapeutic strategies relating to CAFs are discussed.


[Role of chemokines in the vitreous of proliferative diabetic retinopathy].
To investigate the role of chemokines in the pathogenesis of proliferative diabetic retinopathy (PDR). In total, 41 eyes of 38 patients undergoing vitrectomy were divided into two groups; PDR and non-PDR. The PDR group was comprised of 30 eyes, and the non-PDR group of 11 eyes. Vitreous specimens obtained at vitrectomy were centrifuged and separated into supernatants and cellular components. Concentrations of vascular endothelial growth factor(VEGF), interleukin-8 (IL-8), monocyte chemotactic protein-1 (MCP-1), and regulated upon activation, normal T cell expressed and secreted(RANTES) in the supernatants were determined by enzyme-linked immunosorbent assay(ELISA) or chemiluminescence enzyme immunoassay(CLEIA). Expression of VEGF in the cellular components was determined by immunohistochemistry. Vitreous levels of VEGF(p < 0.05), IL8 (p < 0.0001) and MCP-1 (p < 0.05) in the PDR group were significantly higher than in the non-PDR group. However, there was no significant difference in RANTES between the two groups. There was a significant correlation (p < 0.0001, r = 0.84) between vitreous IL-8 and MCP-1 levels in the PDR group. After immunohistochemical staining with antiVEGF monoclonal antibody, VEGF positivity was localized in polymorphonuclear leukocytes and monocytes of the cellular components of PDR vitreous specimens. These results indicate that chemokines are possibly involved in the recruitment of neutrophils and monocytes into the vitreous and that they play a role in the intraocular neovascularization characteristic of PDR.


Prostate cancer incidence and mortality in the Baltic states, Belarus, the Russian Federation and Ukraine.
Prostate cancer incidence varies internationally largely attributable to differences in prostate-specific antigen (PSA) use. The aim of this study was to provide the most recent detailed international epidemiological comparison of prostate cancer incidence and mortality in six north-eastern European countries (Belarus, Estonia, Latvia, Lithuania, the Russian Federation and Ukraine). The number of incident prostate cancer cases was obtained from the countries national cancer registries. Prostate cancer mortality and corresponding population data were extracted from the WHO Mortality Database. Age-specific and age-standardised incidence and mortality rates were calculated (European Standard). The joinpoint regression model was used to provide an average annual percentage change and to detect points in time where significant changes in trends occurred. The observation period was between 13 (Ukraine) and 48 (Estonia) years regarding incidence and around 30 years regarding mortality. The comparison of prostate cancer incidence in six European countries showed almost sixfold differences in the age-adjusted rates in most recent years with highest incidence rates in Lithuania and Estonia. Through the observation period, overall a continuous rise was seen in incidence in all countries and a continuous rise in mortality, with a stabilisation in Estonia and a decrease in Lithuania in recent years. Data limitations included a descriptive design using ecological data. A widespread use of PSA testing seems to be responsible for the changes in the epidemiology of the disease in north-eastern European countries. Substantial variation in the incidence of prostate cancer in the Baltic states suggests the possibility that PSA performance and utilisation spread have had a major influence on observed incidence trends, with a lack of effect on prostate cancer mortality.


Seasonal, extratrigeminal, episodic paroxysmal hemicrania successfully treated with single suboccipital steroid injections.
This case report describes a case of extratrigeminal, episodic paroxysmal hemicrania with a clear seasonal temporal pattern, successfully treated with repeated single suboccipital steroid injections. The pathophysiological and clinical implications of this observation are discussed.


Psychological responses to blood donated by men who have sex with men.
Restrictions previously limiting the ability of men who have sex with men to donate blood are being eased in a number of nations worldwide. In the context of these changes, it is important to determine public perceptions of receiving a transfusion of blood donated by men who have sex with men. In online surveys, 510 (Study 1) and 1,062 (Study 2) heterosexual participants reported attitudes, anxiety, disgust, and gratitude towards potentially receiving a transfusion of blood donated by a homosexual male donor and a heterosexual male donor. In Study 2, half of the participants were reminded of the safety testing carried out on donated blood samples. Negative attitudes, anxiety, disgust, and gratitude were compared between the two donors using t-tests and within-participants indirect effects analysis. Stronger negative attitudes, higher anxiety and disgust, and lower gratitude were reported in relation to a potential transfusion of blood donated by the homosexual male donor relative to the heterosexual male donor (|d|=0.26-0.46). This was the case even when participants were reminded of the safety testing completed on donated blood samples in Study 2. In both studies, the effect of donor sexual orientation on attitudes was explained via heightened anxiety and disgust and attenuated gratitude (b=0.05-0.30). Considering receiving a transfusion of blood donated by a homosexual male donor elicits more negative attitudes, anxiety and disgust, and less positive emotion, relative to blood donated by a heterosexual male donor. These attitudes and emotional reactions are not shifted by a reminder of the safety testing carried out on donated blood samples. In the context of changing restrictions on blood donation by men who have sex with men, these findings highlight a challenge to shift public perception to embrace this cohort of donors.


Compact and user-friendly ultrasound acquisition systems optimized for field recording.
A compact and rugged hardware interface for laptop-based ultrasound recording in the field has been developed. The versatile devices overcome the limitations and drawbacks of previously available solutions. The accompanying recording software enables flexible hard disk recording for both manual and automated operation.


Is L2 paravertebral block comparable to lumbar plexus block for postoperative analgesia after total hip arthroplasty?
Continuous lumbar plexus block (LPB) is a well-accepted technique for regional analgesia after THA. However, many patients experience considerable quadriceps motor weakness with this technique, thus impairing their ability to achieve their physical therapy goals. We asked whether L2 paravertebral block (PVB) provides better postoperative analgesia (defined as decreased postoperative opioid consumption and lower pain scores), better preservation of motor function, and decreased length of hospital stay (LOS) compared to LPB in patients undergoing THA. Sixty patients undergoing minimally invasive THA under standardized spinal anesthesia were enrolled in this randomized controlled study. After exclusions, 53 patients were randomized into the L2 PVB (n = 27) and LPB (n = 26) groups. Patient-controlled analgesia was available for 24 hours. Motor and pain assessments were performed in the recovery room and at the end of 24 hours. LOS was also noted. Postoperative opioid consumption during the first 24 hours was less in the LPB group (mean ± SD: 24 ± 15 mg morphine) than in the L2 PVB group (32 ± 15 mg morphine; p = 0.005); however, postoperative pain scores were not different between groups. Postoperative motor and rehabilitation outcomes and LOS were also similar. Our study demonstrates that use of a LPB results in slightly less morphine consumption but comparable pain scores when compared with continuous L2 PVB. No difference was noted in terms of motor preservation or LOS. Although the difference in morphine consumption was only slightly in favor of the LPB group, the advantage of L2 PVBs noted by previous authors as preservation of motor function, was not seen. At our institute where LPBs have been performed for years, there seems to be no real advantage in switching to L2 PVBs. However, L2 PVB could be a reasonable alternative for operators who are wary of LPBs due to their high potential for complications and/or requiring advanced skills for its placement. But, since L2 PVBs are relatively new, not much is known about their complication profile. We recommend a thorough understanding of both techniques before attempting to place them. Level I, therapeutic study. See Instructions for Authors for a complete description of levels of evidence.


An island in the stream: Australia's place in the cosmopolitan world of Indo-West Pacific freshwater shrimp (Decapoda: Atyidae: Caridina).
Mitochondrial DNA sequences were used to investigate phylogenetic and biogeographic relationships among Australian freshwater shrimp from the genus Caridina H. Milne Edwards, 1837 (Atyidae) and congeners from potential source populations throughout the Indo-West Pacific region. Numerous Australian taxa have close evolutionary relationships with non-Australian taxa from locations throughout the region, indicating a diverse origin of the Australian freshwater fauna. This implies many colonisations to or from Australia over a long period, and thus highlights the surprising adeptness of freshwater shrimp in dispersal across ocean barriers and the unity of much of the region's freshwater biota. Interestingly, a study on Australia's other main genus of atyid shrimp, Paratya Miers, 1882, inferred only a single colonisation. A number of potential species radiations within Australia were also identified. This agrees with patterns detected for a large number of Australian freshwater taxa, and so implies a vicariant explanation due to the development of colder, dryer climates during the late Miocene/early Pliocene.


[Effect of pulse laser UV-radiation on proliferating and resting HeLa tumor cells].
Low-intensity pulse-periodic UV-radiation (lambda = 271.2 nm) within a definite repetition frequency interval (about 10 kHz and 22 kHz) and doses ranging from 0.1 to 10 J/m2 has a selective action on resting HeLa cells. In the same conditions, continuous UV-radiation (lambda = 270 nm) as well as powerful single picosecond pulses (lambda = 266 nm) do not affect the resting cells. The response of proliferating and resting cells to these types of irradiation in different.


A Fast and Validated HPLC Method for Simultaneous Determination of Dopamine, Dobutamine, Phentolamine, Furosemide, and Aminophylline in Infusion Samples and Injection Formulations.
A simple, fast, and validated HPLC method was developed for the simultaneous quantization of five cardiovascular agents: dopamine (DPM), dobutamine (DBM), phentolamine (PTM), furosemide (FSM), and aminophylline (APL) either in infusion samples or in an injection dosage form. The proposed method was achieved with a 150 mm × 4.6 mm, 5.0 μm C18 column, by using a simple linear gradient. Mobile phase A was buffer (50 mM KH2PO4) and mobile Phase B was acetonitrile at a flow rate of 1.0 mL/min. The column temperature was kept at 30°C, and the injection volume was 20 μL. All analytes were separated simultaneously at a retention time (tr) of 3.93, 5.84, 7.06, 8.76, and 9.67 min for DPM, DBM, PTM, FSM, and APL, respectively, with a total run time of less than 15.0 min. The proposed method was validated according to ICH guidelines with respect to accuracy, precision, linearity, limit of detection, limit of quantitation, and robustness. Linearity was obtained over a concentration range of 12.0-240.0, 12.0-240.0, 20.0-200.0, 6.0-240.0, and 10.0-200.0 μg/mL DPM, DBM, PTM, FSM, and APL, respectively. Interday and intraday accuracy and precision data were recorded in the acceptable limits. The new method has successfully been applied for quantification of all five drugs in their injection dosage form, infusion samples, and for evaluation of the stability of investigated drugs in mixtures for endovenous use. The results of the stability study showed that mixtures of DPM, DBM, PTM, FSM, and APL in 5% glucose or 0.9% sodium chloride injection were stable for 48 hours when stored in polypropylene syringes at 25°C.


Cardiac failure detection in 30 minutes: new approach based on gold nanoparticles.
Cardiac failure occurs when heart is unable to pump sufficiently to maintain blood flow to meet the body's needs. The aim of this work is to detect highly expressed genes: follistatin-related protein 1 (FSTL1) in heart failure within 30 minutes, using gold nanoparticles. Gold nanoparticles were prepared by citrate reduction of HAuCl4 3H2O; probe sequence was designed based on the FSTL1 gene region. Preparation of gold nanoprobes (AuNPs) proceeded by treating all the containers with DEPC-treated water, followed by reduction and conjugation. Transmission electron microscopy shows that AuNPs were 10-15 nm in size. The concentration of the nanoprobes was 2.1 nM, and they bind to target. Real-time PCR shows an over-expression of FSTL1 and FSTL3 in heart failure (p < .05). Our data showed that elevated expression of the FSTL1 and FSTL3 is a marker of heart failure as detected within 30 minutes by the synthesised AuNPs; the method is accurate and fast.


Bionomics of Asian Citrus Psyllid (Hemiptera: Liviidae) Associated with Orange Jasmine Hedges in Southeast Central Florida, with Special Reference to Biological Control by Tamarixia radiata.
The Asian citrus psyllid, Diaphorina citri Kuwayama, is an important pest in Florida because it transmits bacteria responsible for citrus huanglongbing disease. In addition to infesting citrus, orange jasmine (Murraya exotica L.) is one of Asian citrus psyllid's preferred host plants and is widely grown as an ornamental hedge. We report on Asian citrus psyllid bionomics over three years at five urban plantings of orange jasmine and on biological control of Asian citrus psyllid by a parasitoid Tamarixia radiata (Waterston). T. radiata had been released in Florida shortly after Asian citrus psyllid was first found, and the parasitoid was known to be established at each planting. Additionally, three new T. radiata haplotypes were released every 3 wk at three plantings during the first study year (one haplotype per planting, over all releases an average of 17 parasitoids per linear meter of hedge); all three haplotypes were released at a fourth planting beginning midway through the study (over all releases, an average combined total of 202 parasitoids per linear meter of hedge). Asian citrus psyllid populations were present year-round at each planting, often at large levels. Such plantings may pose risk to commercial citrus as Asian citrus psyllid reservoirs. Releases of the new haplotypes did not cause any measurable reduction in Asian citrus psyllid population levels during the study, and ironically percentage parasitism was generally highest at a planting where no releases were made. Higher release rates might have been more effective. The probability is discussed that repetitive pruning of orange jasmine reduced the full potential of T. radiata against Asian citrus psyllid in this study.


Absorption of orally supplied natural estrogens correlated with age and somatometric parameters.
The plasma levels of estradiol were studied in 20 postmenopausal women over a period of 8 h following the ingestion of a single dose of 2 mg estradiol valerate and 2 mg micronized estradiol. The determined plasma estradiol concentrations were correlated with somatometric characteristics and age. Twenty-six postcephalic body measurements, the body weight and six skin-fold thickness measurements were taken and the body mass index computed; estradiol serum concentration was measured before and after administration of micronized estradiol and estradiol valerate, respectively. It was shown that estradiol concentrations obtained after estradiol valerate and micronized estradiol ingestion were dependent on the patient's age as well as on the constitutional type. Additionally, we found that there were differences in the reachable estradiol plasma concentrations following the application of the two preparations. Our results justify the individual and differentiated estrogen replacement therapy of the postmenopausal woman.


Characterization of mass and swelling of hydrogel microstructures using MEMS resonant mass sensor arrays.
The use of hydrogels for biomedical engineering, and for the development of biologically inspired cellular systems at the microscale, is advancing at a rapid pace. Microelectromechanical system (MEMS) resonant mass sensors enable the mass measurement of a range of materials. The integration of hydrogels onto MEMS resonant mass sensors is demonstrated, and these sensors are used to characterize the hydrogel mass and swelling characteristics. The mass values obtained from resonant frequency measurements of poly(ethylene glycol)diacrylate (PEGDA) microstructures match well with the values independently verified through volume measurements. The sensors are also used to measure the influence of fluids of similar and greater density on the mass measurements of microstructures. The data show a size-dependent increase in gel mass when fluid density is increased. Lastly, volume comparisons of bulk hydrogels with a range polymer concentration (5% to 100% (v/v)) show a non-linear swelling trend.


Thickening of the diaphragm: a new computed tomography sign of diaphragm injury.
Computed tomography examination is becoming a standard method of evaluating injury in blunt trauma. This report describes a previously unreported computed tomography sign of diaphragmatic injury. A retrospective review of imaging findings was performed on eight patients with surgically proven traumatic diaphragmatic rupture. Thickening of the diaphragm was present on computed tomography in six of eight patients with surgically proven diaphragm rupture. Two patients demonstrated previously established computed tomography signs of diaphragm rupture and two patients had no findings to suggest diaphragm injury. Diaphragm thickening is associated with diaphragm rupture and may be a useful diagnostic sign in the evaluation of patients with blunt trauma.


A randomized controlled trial comparing the effects of yoga with an active control on ambulatory blood pressure in individuals with prehypertension and stage 1 hypertension.
The purpose of this study was to compare the effects of yoga with an active control (nonaerobic exercise) in individuals with prehypertension and stage 1 hypertension. A randomized clinical trial was performed using two arms: (1) yoga and (2) active control. Primary outcomes were 24-hour day and night ambulatory systolic and diastolic blood pressures. Within-group and between-group analyses were performed using paired t tests and repeated-measures analysis of variance (time × group), respectively. Eighty-four participants enrolled, with 68 participants completing the trial. Within-group analyses found 24-hour diastolic, night diastolic, and mean arterial pressure all significantly reduced in the yoga group (-3.93, -4.7, -4.23 mm Hg, respectively) but no significant within-group changes in the active control group. Direct comparisons of the yoga intervention with the control group found a single blood pressure variable (diastolic night) to be significantly different (P=.038). This study has demonstrated that a yoga intervention can lower blood pressure in patients with mild hypertension. Although this study was not adequately powered to show between-group differences, the size of the yoga-induced blood pressure reduction appears to justify performing a definitive trial of this intervention to test whether it can provide meaningful therapeutic value for the management of hypertension.


Poisonings in Poland reported to the Polish National Health Fund in the years 2009-2011.
Poisonings constitute a significant medical, social and economic problem worldwide. In Poland there is no nationwide registry of poisonings, which results in a lack of accurate epidemiological data. Few publications dealing with the problem are based on data obtained from toxicology units and therefore do not include information about cases treated at emergency departments and other non-toxicology units. We analyzed all admissions due to poisonings reported to the Polish National Health Fund by all hospital units in Poland in the 2009-2011 period. Diagnoses were encoded according to the ICD-10 classification. A total of 254,425 admissions were reported, 85,398 in 2009, 85,230 in 2010 and 83,797 in 2011. The male to female ratios were 1.88, 1.75 and 1.80 respectively. The most frequent causes of admissions were poisonings with ethanol (n = 121,874; 47.9%), carbon monoxide (n = 17,179; 6.8%) and benzodiazepines (n = 10,340; 4.1%). Alcohols were the reason for 104,680 admissions in men (63.2%) and 22,612 admissions in women (25.5%; p < 0.01). Poisonings with pharmaceuticals and other drugs were reported in 34,616 men (20.9%) and 45,238 women (51%; p < 0.01). There were 1680 cases of fatal poisonings in the analyzed period. The hospital mortality due to poisonings increased from 1.1% in 2009 to 1.5% in 2011 (p < 0.01). The mortality in general Intensive Care Units increased from 14.4% in 2009 to 22.3% in 2011 (p < 0.01). The etiology of fatal poisonings was highly dependent on the type of hospital unit. The overall number of admissions due to poisonings decreased slightly during the study period, but they remained a significant cause of morbidity and mortality. Alcohols were the main cause of admissions in the analyzed period. Alcohol intoxications were more frequent in men while poisonings with pharmaceuticals were more frequent in women. Carbon monoxide exposures were a significant cause of morbidity and mortality in the studied period in Poland. A national poison information and toxicovigilance system should be created in Poland, ideally allowing for near real-time monitoring of cases of poisonings.


Historical perspectives on indigenous health in Australia.
In spite of much effort over the past 25 years, the life expectancy of the indigenous people remains nearly 20 years behind the non-Aboriginal white population of Australia. These figures compare unfavourably with the improved life expectancy over the past 25 years of other indigenous peoples, such as the New Zealand Maori and the American Indian populations. By 1990-94, the average Australian indigenous all-cause mortality rate was 1.9 times the Maori rate, 2.4 times the US indigenous rate and 3.15 times the all-Australian rate. The persistence of this discrepancy in Australia is obviously a matter of great concern. There is clearly a gap between available knowledge and its application. Some indication of the possibility of reversal of the current situation is given by a recent report of the beneficial impact of the Homelands Movement on Health Outcomes in Central Australian Aborigines. The study compared the prevalence of obesity, hypertension and diabetes in two groups of Aboriginal adults: those living in homelands versus those living in centralized communities in Central Australia. Baseline studies revealed a lower prevalence of diabetes, hypertension and obesity in the homelands group, compared with those living in centralized communities. They were also less likely to die and less likely to be hospitalized for any cause, particularly infections, injury involving alcohol and other injury. Mean age at death was 58 and 48 years for the residents of homelands and centralized communities, respectively. The benefits were most marked in young adults. It is suggested that the homelands communities have a greater degree of control of their own lives than those living in the centralized communities and this may be an important factor in their improved health status. Improvement in indigenous health should be one of the key issues of reconciliation. Priorities include community control of Aboriginal Health Services under the National Aboriginal Community Controlled Health Organisation (NACCHO), throughout Australia, a greater priority for prevention and public health services (housing, water supply and environmental services) education and economic issues, improved training of indigenous health professionals and increased funding. A national professional organization including NACCHO needs to be established to bridge the big gap between available knowledge and its application for the benefit of the indigenous people of Australia.


Tropism between hepatic and pulmonary metastases in colorectal cancers.
In metastatic colorectal cancers, tumor cells are disseminated prior to surgical resection of the primary tumor but remain dormant until proper colonization mechanisms are activated. To identify the colonization mechanisms of the metastatic tumors, we conducted a pairwise comparison between primary colorectal cancers and metastatic tumors (n=12 pairs), including six hepatic pairs and six pulmonary pairs. The mRNA levels of 224 genes previously reported to be associated with metastasis, cytokines and angiogenesis were quantitatively determined by PCR arrays. Among them, 27 genes were duplicated or triplicated to show consistent expression. Unsupervised hierarchical clustering of the Ct values of metastasis-related genes revealed that liver metastases were indistinguishable from primary colorectal cancers (n=5/6), whereas lung metastases were highly diversified from one another and from the primary tumors (n=6/6). Cytokines and receptor gene expression array data also confirmed the divergence of pulmonary metastases from primary colorectal cancers (n=6/6). Heat map analyses of ΔCt values of the metastasis-related genes identified a 17-gene tropism signature that was sufficient not only to distinguish liver and the lung metastases, but also reconstituted the clustering of primary tumors with the hepatic metastases (n=17/18). In this pilot experiment, pulmonary metastases were significantly diverged from hepatic metastases that were indistinguishable from primary colorectal cancers. Further genomic and clinical studies are in progress to evaluate the potential of the tropism signature as a therapeutic target to inhibit the colonization of metastatic colorectal cancers.


Dental size and shape in the Roman imperial age: two examples from the area of Rome.
Different socioeconomic strata of Roman imperial age are represented by two large dental samples recovered from archaeological excavations near Rome, Italy. Teeth are investigated for crown dimensions and morphological variants. One sample, comprising 1,465 permanent teeth, represents the rural town of Lucus Feroniae (LFR) and is mainly composed of slaves and war veterans. The other, comprising 734 teeth from the Isola Sacra necropolis at Portus Romae (NIS), represents the "middle class" segment of an urban population. Both series show small dental dimensions and fit at the lower end of the trend toward dental reduction in Europe from the Upper Paleolithic to the historical times. The urban sample is less variable metrically and less sexually dimorphic than the rural one. The analysis of discrete crown traits shows absence of rare phenotypic variants in both series. The urban sample is also less variable in this last respect, suggesting that the gene pool of this particular "stratum" of the NIS population was more homogeneous than that of LFR. The occurrence of enamel hypoplasia indicates that metabolic stress during growth and development was similar in LFR and NIS. The overall set of available data is evaluated in the light of the history of the two Roman sites and the composition of each population.


Unveiling the additive-assisted oriented growth of perovskite crystallite for high performance light-emitting diodes.
Solution-processed metal halide perovskites have been recognized as one of the most promising semiconductors, with applications in light-emitting diodes (LEDs), solar cells and lasers. Various additives have been widely used in perovskite precursor solutions, aiming to improve the formed perovskite film quality through passivating defects and controlling the crystallinity. The additive's role of defect passivation has been intensively investigated, while a deep understanding of how additives influence the crystallization process of perovskites is lacking. Here, we reveal a general additive-assisted crystal formation pathway for FAPbI3 perovskite with vertical orientation, by tracking the chemical interaction in the precursor solution and crystallographic evolution during the film formation process. The resulting understanding motivates us to use a new additive with multi-functional groups, 2-(2-(2-Aminoethoxy)ethoxy)acetic acid, which can facilitate the orientated growth of perovskite and passivate defects, leading to perovskite layer with high crystallinity and low defect density and thereby record-high performance NIR perovskite LEDs (~800 nm emission peak, a peak external quantum efficiency of 22.2% with enhanced stability).


Reviving the guardian of the genome: Small molecule activators of p53.
The tumor suppressor p53 is one of the most important proteins for protection of genomic stability and cancer prevention. Cancers often inactivate it by either mutating its gene or disabling its function. Thus, activating p53 becomes an attractive approach for the development of molecule-based anti-cancer therapy. The past decade and half have witnessed tremendous progress in this area. This essay offers readers with a grand review on this progress with updated information about small molecule activators of p53 either still at bench work or in clinical trials.


Heterozygous loss-of-function mutation in Odd-skipped related 1 (Osr1) is associated with vesicoureteric reflux, duplex systems, and hydronephrosis.
Odd-skipped related 1 (Osr1) is a transcriptional repressor that plays critical roles in maintaining the mesenchymal stem cell population within the developing kidney. Here, we report that newborn pups with a heterozygous null mutation in Osr1 exhibit a 21% incidence of vesicoureteric reflux and have hydronephrosis and urinary tract duplications. Newborn pups have a short intravesical ureter, resulting in a less competent ureterovesical junction which arises from a delay in urinary tract development. We describe a new domain of Osr1 expression in the ureteral mesenchyme and within the developing bladder in the mouse. OSR1 was sequenced in 186 children with primary vesicoureteric reflux, and 17 have single nucleotide polymorphisms. Fifteen children have a common synonymous variant, rs12329305, one child has a rare nonsynonymous variant, rs3440471, and one child has a rare 5'-UTR variant, rs45535040 The impact of these SNPs is not clear; therefore, the role of OSR1 in human disease remains to be elucidated. Osr1 is a candidate gene implicated in the pathogenesis of vesicoureteric reflux and congenital abnormalities of the kidney and urinary tract in mice.


Brain lesions manifesting as psychiatric disorders: eight cases.
Sometimes patients with organic brain lesions in neurologically silent brain areas might present only with psychiatric symptoms, such as depression, anxiety disorders, schizophrenia, anorexia nervosa, or cognitive dysfunction. This study presents eight cases of patients with brain lesions (four cases of meningiomas, one case of intracerebral cysts, one case of anaplastic oligodendroglioma, one case of multiform glioblastoma, and one case of occlusive hydrocephalus) who, for a significant period of time, were diagnosed and treated for psychiatric disorders (three cases of Alzheimer's disease, two cases of schizoaffective disorder, one case of schizophrenia, one case of depression, and one case of organic emotional lability disorder). When neurologic symptoms developed, they underwent neuroimaging studies and organic brain lesions were diagnosed. Further treatment required neurosurgical interventions. These cases show that brain tumors can be neurologically silent for a sufficient period of time and manifest as psychiatric disorders. Therefore, neuroimaging studies are needed when atypical changes in mental status or neurologic symptoms and signs develop.


A conserved structure and function of the YidC homologous protein Slr1471 from Synechocystis sp. PCC 6803.
In this article, we show that the orf slr1471 from Synechocystis sp. PCC 6803 codes for a functional member of the YidC/Alb3/Oxa1 protein family, and the encoded protein has a transmembrane topology with a common core structure. Using specific antibodies raised against the Synechocystis YidC homologous protein, we further show that the Synechocystis YidC protein appears to be predominantly localized in the cyanobacterial cytoplasmic membrane. The impact of the described findings for synthesis of membrane proteins and for protein sorting within cyanobacterial cells is discussed.


Substance misuse among health care workers: national survey of occupational physicians.
National Health Service (NHS) occupational health departments assist in the identification and assessment of substance-use disorders among health care workers (HCWs) and are involved in the management of an individual's return to work after treatment. To determine the experience and training of NHS occupational health physicians (OHPs) in identifying substance misuse among HCWs. A national, cross-sectional, postal-based questionnaire was administered to the Association of National Health Occupational Physicians membership. A total of 145/224 (65%) OHPs (55% male), with a mean age of 49 years (SD ± 9.1; range 28-76), who had worked in the NHS on average for 9.6 years took part. The majority of respondents were consultant grade (59%). Since taking up their NHS post, 26% had received no training in substance misuse. Of those who had undergone formal training, the mean duration received was 2.8 days for drugs and 3.5 days for alcohol. OHPs reported that they did not feel sufficiently trained in this area. Most (65%) did not routinely include standardized screening tools or deliver 'brief interventions' (78%), although many reported that they would routinely ask about substance use when there was no clear indication of use (42%). The majority did not feel they were adequately supported (54%) in this work, nor did they have adequate resources for these patients within their organization (68%). OHPs see HCWs with substance-use problems as part of their work, but the support provided is likely limited by insufficient training and inadequate support.


Origins of lineage-specific elements via gene duplication, relocation, and regional rearrangement in Neurospora crassa.
The origin of new genes has long been a central interest of evolutionary biologists. However, their novelty means that they evade reconstruction by the classical tools of evolutionary modelling. This evasion of deep ancestral investigation necessitates intensive study of model species within well-sampled, recently diversified, clades. One such clade is the model genus Neurospora, members of which lack recent gene duplications. Several Neurospora species are comprehensively characterized organisms apt for studying the evolution of lineage-specific genes (LSGs). Using gene synteny, we documented that 78% of Neurospora LSG clusters are located adjacent to the telomeres featuring extensive tracts of non-coding DNA and duplicated genes. Here, we report several instances of LSGs that are likely from regional rearrangements and potentially from gene rebirth. To broadly investigate the functions of LSGs, we assembled transcriptomics data from 68 experimental data points and identified co-regulatory modules using Weighted Gene Correlation Network Analysis, revealing that LSGs are widely but peripherally involved in known regulatory machinery for diverse functions. The ancestral status of the LSG mas-1, a gene with roles in cell-wall integrity and cellular sensitivity to antifungal toxins, was investigated in detail alongside its genomic neighbours, indicating that it arose from an ancient lysophospholipase precursor that is ubiquitous in lineages of the Sordariomycetes. Our discoveries illuminate a "rummage region" in the N. crassa genome that enables the formation of new genes and functions to arise via gene duplication and relocation, followed by fast mutation and recombination facilitated by sequence repeats and unconstrained non-coding sequences.


The effects of cigarette smoking on T cell subsets. A population-based survey of healthy caucasians.
To investigate the influence of cigarette smoking on mononuclear cell subsets, we determined T cell, B cell, monocyte, and HLA-DR+ subsets in a population-based, stratified, random sample of healthy Caucasians using monoclonal antibodies and flow cytometry. The study population consisted of 282 subjects 20 to 69 yr of age, including 108 smokers and 174 nonsmokers. Multivariate analysis techniques were used to assess the influence of cigarette smoking status after controlling for the effects of age and gender. Cigarette smoking was associated with a nonspecific increase in the leukocyte count involving all major cell types (smokers: 8.50 +/- 0.15 versus nonsmokers: 7.33 +/- 0.12 cells/mm3; p less than or equal to 0.0001). In addition, cigarette smokers had a selective increase in CD4+ cells (helper-inducer T cells) compared with nonsmokers (55.3 +/- 0.8 versus 52.2 +/- 0.6% of lymphoid cells; p = 0.002), resulting in a statistically significant increase in the CD4+/CD8+ (helper/suppressor) ratio (2.42 +/- 0.1 versus 2.13 +/- 0.16; p = 0.02). There was no significant difference between smokers and nonsmokers in the level of CD3+ cells (total T cells: 76.8 +/- 0.7 versus 76.1 +/- 0.5; p = 0.5), CD8+ cells (suppressor-cytotoxic T cells: 25.7 +/- 0.8 versus 27.0 +/- 0.5%; p = 0.1), CD19+ cells (B cells) (10.7 +/- 0.4 versus 10.0 +/- 0.3%; p = 0.2), CD14+ cells (monocytes) (18.0 +/- 0.6 versus 17.0 +/- 0.4%; p = 0.2), or HLA-DR+ cells (14.5 +/- 0.5 versus 14.0 +/- 0.4%; p = 0.4).(ABSTRACT TRUNCATED AT 250 WORDS)


Functional characterization and modulation of cytokine production by CD8+ T cells from human immunodeficiency virus-infected individuals.
CD8+ T-cell clones were generated from peripheral blood mononuclear cells (PBMC) of three human immunodeficiency virus (HIV)-seronegative individuals and six HIV-seropositive individuals and assessed for their cytokine secretion profile, cytolytic potential, and chemokine production. While the great majority of CD8+ T-cell clones generated from HIV-seronegative individuals produced interferon (IFN)-gamma, but not interleukin-4 (IL-4), that is a type 1 cytotoxic (Tc1) profile, high numbers of CD8+ T-cell clones generated from HIV-seropositive individuals produced IL-4 in addition to IFN-gamma or IL-4 alone, thus showing a type 0 cytotoxic (Tc0)- or a type 2 cytotoxic (Tc2) profile, respectively. Tc0/Tc2 cells displayed lower cytolytic activity than Tc1 cells, including a reduced ability to lyse autologous targets pulsed with HIV or HIV peptides. By contrast, the production of chemokines RANTES and macrophage inflammatory protein-1alpha was comparable in Tc1, Tc0, and Tc2 clones irrespective of whether they were derived from HIV-seronegative or HIV-seropositive individuals. When CD8+ T-cell clones were generated from PBMC cultures of HIV-seronegative individuals conditioned with IL-4 plus an anti-IL-12 antibody (Ab), a shift towards the Tc0/Tc2-like profile was observed. Conversely, the addition to PBMC cultures of IL-12 plus an anti-IL-4 Ab shifted the differentiation of CD8+ T cells from HIV-infected individuals towards the Tc1-like profile, whereas IL-12 or anti-IL-4 Ab alone had a lower Tc1-promoting effect. Irradiated PBMC from HIV-infected individuals, used as feeder cells, shifted the differentiation of CD8+ T cells from a healthy HIV-seronegative individual towards the Tc0/Tc2-like profile. On the other hand, a shift towards the Tcl-like profile was noted in CD8+ T-cell clones generated from the skin specimens of two HIV-seropositive patients with Kaposi's sarcoma, successfully treated with IFN-alpha, in comparison to CD8+ clones generated from the same skin areas before treatment. The IFN-alpha-induced Tc1 shift could be prevented by the incubation of skin-infiltrating CD8+ T cells with IL-4 before cloning. Taken together, these data indicate that both defective production of IL-12 and abnormal IL-4 production in bulk PBMC populations of HIV-infected individuals may contribute to the development of high numbers of CD8+ T-cell clones showing a Tc0/Tc2-like phenotype and reduced cytolytic potential against HIV itself. They also suggest that the cytokine profile of CD8+ T-cell clones can be modulated by cytokines (or anticytokine Ab) both in vitro and in vivo.


Ins and outs of enteral access. Part 1: short-term enteral access.
Proper selection of the type of enteral access in a given clinical situation is important to maximize the benefit to the patient and minimize the discomfort to the patient, the morbidity and mortality, and the cost. This decision is based on whether or not the patient needs short-term (<4-6 weeks) or long-term enteral nutrition support. This article focuses on short-term access including nasogastric (NG) and nasoenteric (NE) tubes and reviews each of these enteral access tubes along with their indications, contraindications, advantages, and disadvantages.


High-speed 3D shape measurement using a rotary mechanical projector.
In this paper, a fast rotary mechanical projector (RMP) is designed and manufactured for high-speed 3D shape measurement. Compared with the common high-speed projectors, RMP has a good performance in high-speed projection, which can obtain high quality projected fringes with shorter camera exposure time by using the error diffusion binary coding method and chrome plating technology. The magnitude, acceptability of systemic projection error is analyzed and quantified in detail. For the quantified error, the probability distribution function (PDF) algorithm is introduced to correct the error. Corrected projection error is reduced to more than one third of the original error. Subsequently, a monocular measurement system composed of the RMP and a single camera is constructed. The combination of the RMP device and PDF algorithm ensure the accuracy of a corresponding 3D shape measurement system. Experiments have demonstrated that the proposed solution has a good performance for the 3D measurement of high-speed scenes.


The transcription factor IRF4 determines the anti-tumor immunity of CD8+ T cells.
Understanding the factors that regulate T cell infiltration and functional states in solid tumors is crucial for advancing cancer immunotherapies. Here, we discovered that the expression of interferon regulatory factor 4 (IRF4) was a critical T cell intrinsic requirement for effective anti-tumor immunity. Mice with T-cell-specific ablation of IRF4 showed significantly reduced T cell tumor infiltration and function, resulting in accelerated growth of subcutaneous syngeneic tumors and allowing the growth of allogeneic tumors. Additionally, engineered overexpression of IRF4 in anti-tumor CD8+ T cells that were adoptively transferred significantly promoted their tumor infiltration and transition from a naive/memory-like cell state into effector T cell states. As a result, IRF4-engineered anti-tumor T cells exhibited significantly improved anti-tumor efficacy, and inhibited tumor growth either alone or in combination with PD-L1 blockade. These findings identify IRF4 as a crucial cell-intrinsic driver of T cell infiltration and function in tumors, emphasizing the potential of IRF4-engineering as an immunotherapeutic approach.


Can We StoP Worrying about Long-term PPIs and Gastric Cancer Risk?
Proton pump inhibitors (PPI) are a cornerstone of management for many digestive diseases. While chronic PPI use induces physiologic changes including gastric acid suppression and hypergastrinemia, existing data are conflicting on whether this impacts the risk of gastric cancer among PPI users. Sassano and colleagues utilized pooled case-control data from five studies in the Stomach cancer Pooling (StoP) Project to investigate the association between PPI use and histologically confirmed gastric cancer. Short-term PPI use (6 months) was associated with increased risk of gastric cancer, but no association was found between long-term PPI use (3 years or more) and gastric cancer. Although the authors relied on patient-reported PPI use data, and data related to Helicobacter pylori infection and eradication rates were missing, no histologic gastric cancer subtypes in this international case-control study were associated with any PPI use. Currently reported findings provide patients and clinicians with reassuring observations that long-term PPI use does not significantly increase gastric cancer risk. The relationship identified among short-term PPI users may reflect reverse causality. Our understanding will be furthered by additional assessment of potential confounders, including comorbid conditions, PPI metabolism, and social determinants of health. See related article by Sassano et al., p. 1174.


Single amino acids changes in the signal receptor domain of XylR resulted in mutants that stimulate transcription in the absence of effectors.
The XylR protein positively controls expression from the Pseudomonas putida TOL plasmid sigma 54-dependent "upper" pathway operon promoter (Pu) and the xylS gene promoter (Ps), in response to the presence of aromatic effectors. Two mutant XylR regulators able to stimulate transcription from Pu and Ps in the absence of effectors were isolated. These mutants exhibited single point mutations, namely Asp135-->Asn and Pro85-->Ser. Both mutations are located in the amino termini domain of XylR, which is thought to be responsible for interactions with effectors. The effector profile of XylRP85S was similar to that of wild-type XylR protein; however, XylRD135N exhibited an altered pattern of effector recognition: with m-nitrotoluene it stimulated transcription from the Pu promoter above the high basal level, whereas this nitroarene inhibited the wild-type regulator. Previous work (Delgado, A., and Ramos, J.L. (1994) J. Biol. Chem. 269, 8059-8062) showed that residue 172 was involved in effector interactions, as mutant XylRE172K also recognized m-nitrotoluene. However, double mutant XylR135N/E172K did not stimulate transcription in the absence of effector, but retained the ability to stimulate transcription with m-nitrotoluene. Transcription mediated by XylRD135N and XylRP85S from Pu::lacZ was analyzed in detail. Like the wild-type regulator, XylRD135N and XylRP85S required sigma 54 for full transcription activation, but in contrast with the wild-type regulator, XylRD135N, but not XylRP85S, stimulated transcription from Pu in the absence of the integration host factor protein. XylRD135N, also in contrast with XylR and XylRP85S, mediated transcription from a mutant Pu promoter that lacked one of the upstream regulator binding sites (delta UAS1), but not when both upstream regulator binding sites were deleted. The level of autoregulation of XylRD135N was at least 2-fold higher than that found with the wild-type XylR regulator and the mutant XylRP85S.


Impalement injuries of the oral cavity in children: a case report and survey of the literature.
Impalement injuries of the soft palate and oropharynx are common in children, especially those of preschool age. These injuries are particularly common in toddlers, given their propensity to fall easily while carrying objects in their mouths. Although most of these injuries do not have lasting sequelae, some can have devastating neurological complications, and consequently, careful assessment of the patient during the early stages of trauma is imperative. Close follow-up for up to 72 h, and parental counselling and instruction should be considered as part of their immediate care. A case of toothbrush impalement in a 4-year-old child is presented. The foreign body was noted to be lodged in the cheek. After careful triaging, history taking and appropriate imaging, surgical removal of this brush was carried out uneventfully under general anaesthesia. No further complications were noted postoperatively, and because of the type of injury, the patient was discharged within 24 h and reviewed as an outpatient. A survey of the literature confirms that most injuries of this kind can be treated conservatively, and in many cases, without surgical intervention. However, careful assessment by an experienced clinician is necessary to rule out other complications. The present paper discusses the prevalence, management and complications associated with impalement injuries of the oral cavity in children.


Civilian Penetrating Gunshot Injury to the Neurocranium in Enugu.
Civilian penetrating gunshot injuries to the neurocranium are no longer uncommon in Nigeria. Such injuries are however poorly reported. They are associated with poor outcome and, at close range, are frequently fatal, especially when inflicted by high-velocity weapons. Prompt transfer to neurosurgical service and urgent intervention may improve outcome in those that are not mortally wounded. Fifty-two patients with civilian penetrating gunshot wounds seen over a 10-year period (2004-2014) at the University of Nigeria Teaching Hospital and Memfys Hospital for Neurosurgery Enugu were reviewed retrospectively, and their data were analyzed to evaluate factors that impacted on outcome. Only patients with clinical and imaging evidence of cranial gunshot injuries who reached hospital alive were included in the study. The overall mortality and Glasgow outcome score were analyzed. Fifty-two patients with isolated civilian penetrating gunshot wounds were identified (M:F = 7.7:1); mean (standard deviation) age was 32.8 (11.9) years. There was a high correlation (0.983) between the sex of the patients and the outcome. The overall mortality was 30.8%, whereas the mortality for patients with postresuscitation Glasgow coma scale (GCS) score ≤8 was 57%, as against 12.9% in those in whom postresuscitation GCS was >8; meaning that 87.1% of patients in whom postresuscitation GCS was >8 survived. Thirty-one patients (59.6%) had papillary abnormalities. Majority of patients with monohemispheric lesions survived while all those with diencephalic, transventricular, and posterior fossa involvement had 100% mortality. Admitting GCS and bullet trajectory were predictive of outcome.


Isoleucine synthesis by Clostridium sporogenes from propionate or alpha-methylbutyrate.
Preliminary studies demonstrated that Clostridium sporogenes synthesized isoleucine by a pathway not involving threonine or threonine dehydratase. Radiotracer experiments with cells grown in a defined carbohydrate-free medium showed that radioactivity from [U-14C]serine, [3-14C]pyruvate, [14C]NaHCO3 and [1-], [2-] and [3-14C]propionate was incorporated into isoleucine. Conversely, there was no detectable incorporation of 14C into isoleucine during growth with [U-14C]glutamate, [U-14C]threonine, [U-14C]valine, [U-14C]leucine or [U-14C]methionine. Crude extracts of the bacteria grown in a minimal medium contained levels of alpha-acetohydroxyacid synthase activities comparable to those in Escherichia coli K12 grown in minimal medium. Stepwise degradation of isoleucine obtained from C. sporogenes grown in the presence of specifically-labelled precursors indicated that C. sporogenes can make isoleucine via the reductive carboxylation of propionate to yield alpha-oxobutyrate, which is metabolized to isoleucine in the classical fashion. Isoleucine was also formed by C. sporogenes via the reductive carboxylation of alpha-methylbutyrate to alpha-oxo-beta-methylvalerate.


Long-term outcome of microvascular decompression for hemifacial spasm.
To investigate the long term outcomes of microvascular decompression (MVD) for hemifacial spasm (HFS) and to identify any prognostic factors. A retrospective analysis of 189 consecutive patients with typical HFS who underwent MVD. Multiple logistic regression analysis of variables at various time points including at least immediate time point and one at no less than six years was performed. Short-term follow-up showed a cure rate of 91%, including 51 cases of delayed resolution (27%). At two years or more information was available in 148 (out of 189) cases of patients. 101 cases (68% - of 148 cases) had complete recovery, 28 cases (19%) achieved a partial though worthwhile recovery, so that the effective rate of symptoms relief at six years was 87%. Complications were found (66/189, 34.92%) and cured within the follow-up period (cure rate of 100%). In both the univariate and multivariate analyses, the postoperative findings of clinical outcomes showed that preoperative illness duration, compressive pattern, the intraoperative indentation of the root exit zone (REZ) of the facial nerve and intraoperative AMR disappearance were negative predictors and age considered to be positive, which significantly predicted the clinical outcome of patients following MVD. MVD may be a safe and effective strategy for HFS patients in view of relatively higher cure rates and lower complication risks within follow-up. Besides, patients' age, duration of disease, intraoperative indentation of the REZ of the facial nerve, and disappearance of AMR were the major influential variables may be useful for the prediction of prognosis in the patients underwent MVD.


Proliferation Drives Aging-Related Functional Decline in a Subpopulation of the Hematopoietic Stem Cell Compartment.
Aging of the hematopoietic stem cell (HSC) compartment is characterized by lineage bias and reduced stem cell function, the molecular basis of which is largely unknown. Using single-cell transcriptomics, we identified a distinct subpopulation of old HSCs carrying a p53 signature indicative of stem cell decline alongside pro-proliferative JAK/STAT signaling. To investigate the relationship between JAK/STAT and p53 signaling, we challenged HSCs with a constitutively active form of JAK2 (V617F) and observed an expansion of the p53-positive subpopulation in old mice. Our results reveal cellular heterogeneity in the onset of HSC aging and implicate a role for JAK2V617F-driven proliferation in the p53-mediated functional decline of old HSCs.


Effects of thermocycling on the degree of cure of two lingual retainer composites.
The aim of this study was to evaluate the effects of thermocycling on the degree of cure (DC) and water sorption behavior of two lingual retainer composites. A total of 50 composite specimens, 5 mm diameter and 2 mm height, were prepared using Light Cure Retainer (LCR) (Reliance) and Transbond Lingual Retainer (TLR) (3M Unitek). After 40-second curing with a halogen light and after 24-hour water storage, the composite specimens were subjected to different thermocycling regimes. Absorbance peaks to monitor the DC were recorded using Fourier transform infrared (FTIR) spectroscopy, while water sorption was calculated in microg/mm(3). On DC, significant differences among the different thermocycling regimes were observed only for the TLR specimens (p<0.05). On water sorption behavior, an increase in the number of thermal cycles resulted in increased water sorption for both composites, but the statistical differences in these groups were not significant. On the effect of thermocycling up to 20,000 cycles, present findings showed that LCR was less affected than TLR. Further, LCR exhibited higher DC and lower water sorption values than TLR after thermal cycling.


REPETITIVE CYCLES OF BIOLUMINESCENCE AND SPAWNING IN THE POLYCHAETE, ODONTOSYLLIS PHOSPHOREA.
Spawning by large numbers of the marine polychaete, Odontosyllis phosphorea, occurred at fortnightly intervals. The animals appeared at the surface of the water shortly after sunset and luminesced and spawned for approximately 30 minutes. The spawning was correlated with the monthly lunar and tidal cycles and lasted from June through October.


[Study of the Tangut Prescription Sanxiandan unearthed in Heishuicheng].
Prescriptions (ИHB.NO.4384) in the10th Volume of Heishuicheng Manuscripts Collected in Russia covered 9 pages. The 4(th) and the 7(th) pages recorded a prescription with no name and composed of Radix Aconiti, Atractylodes lan and Foeniculumvulgare. By checking Chinese Medical books, the name of this prescription was probably Sanxiandan and it may have been recorded in Chenshi Jingyanfang for the first time. Chenshi Jingyanfang is not extant now but its narrative style, which is different from that of the Tangut prescription, could be speculated upon from other literature that mentioned it. So those Tangut prescriptions could have been compiled according to some Chinese Medical books after Chunyou Periods.


Microscale multilayer cocultures for biomimetic blood vessels.
Living tissues have complex and well-organized microstructures. Although microtechnology has been used to create in vivo-like cell microstructures in vitro, most available microscale systems are two-dimensional, and few three-dimensional (3D) systems have been explored. This article demonstrates a 3D hierarchical biomimetic multilayer microsystem created by a generally applicable technique. The technique employs layer-by-layer microfluidics to build layers of cells and biopolymers in microchannels, allowing controlled patterning of cells and their microenvironments in the x, y, and z-dimension. As a prototype, a multilayer system was created using three vascular cell types within heterogeneous types of biopolymers to mimic the structure and composition of a blood vessel wall. The effects of matrix composition and multilayer configurations on 3D cell-cell interactions and cell biology were revealed. Cell migration in the z-dimension, matrix remodeling, intercellular adhesion molecule expression and actin organization were examined under different 3D coculture conditions. A more biomimetic coculture was found to reproduce a more stable structure and in vivo-like function. This approach provides a method to fabricate microscale hierarchical "neotissues" with 3D configurations of matrix materials and multiple cell types, and an in vitro cell coculture model to understand 3D processes of cell-cell and cell-matrix interactions.


[Study of 164 episodes of infectious endocarditis in drug addicts: comparison of HIV positive and negative patients].
Intravenous drug-addicts (IVDA) are the most important risk group for acquiring HIV and developing infective endocarditis (IE) in Spain. The clinical and evolutive features of a large series of IE in HIV-infected IVDA are reported and compared with those observed in a smaller group of patients without HIV infection. A retrospective analysis was made of 164 episodes of IE from 136 patients diagnosed in a Service of Infectious Diseases in Madrid (1986-1992). IE was defined according to the modified Von Reyn criteria and only the probable and definite IE episodes were evaluated. A total of 143 IE episodes occurred in 117 IVDA infected with HIV (group A) and 21 episodes in 19 HIV-negative patients (group B). Most of seropositive patients were asymptomatic carriers of HIV (54%) and only 22% had AIDS. The IE onset was acute for 97% patients in group A and 81% in group B, with fever and respiratory symptoms as main complaints. Chest X-ray was normal in 19% of cases in group A and in 28% in group B. Septic embolisms were observed in 56% and 41% of patients in group A and B, respectively. The vegetation originated mainly on the tricuspid valve and Staphylococcus aureus was recovered from most blood-cultures. The mortality rate was similar in both groups, 6% and 5% in groups A and B, respectively. Most IVDA with IE were HIV-positive patients in this series. IE is usually reported in the early stages of HIV infection and apparently its presence has no influence on the clinical course of IE.


Conjugated polymers/semiconductor nanocrystals hybrid materials--preparation, electrical transport properties and applications.
This critical review discusses specific preparation and characterization methods applied to hybrid materials consisting of π-conjugated polymers (or oligomers) and semiconductor nanocrystals. These materials are of great importance in the quickly growing field of hybrid organic/inorganic electronics since they can serve as active components of photovoltaic cells, light emitting diodes, photodetectors and other devices. The electronic energy levels of the organic and inorganic components of the hybrid can be tuned individually and thin hybrid films can be processed using low cost solution based techniques. However, the interface between the hybrid components and the morphology of the hybrid directly influences the generation, separation and transport of charge carriers and those parameters are not easy to control. Therefore a large variety of different approaches for assembling the building blocks--conjugated polymers and semiconductor nanocrystals--has been developed. They range from their simple blending through various grafting procedures to methods exploiting specific non-covalent interactions between both components, induced by their tailor-made functionalization. In the first part of this review, we discuss the preparation of the building blocks (nanocrystals and polymers) and the strategies for their assembly into hybrid materials' thin films. In the second part, we focus on the charge carriers' generation and their transport within the hybrids. Finally, we summarize the performances of solar cells using conjugated polymer/semiconductor nanocrystals hybrids and give perspectives for future developments.


WNT antagonists exhibit unique combinatorial antitumor activity with taxanes by potentiating mitotic cell death.
The WNT pathway mediates intercellular signaling that regulates cell fate in both normal development and cancer. It is widely appreciated that the WNT pathway is frequently dysregulated in human cancers through a variety of genetic and epigenetic mechanisms. Targets in the WNT pathway are being extensively pursued for the development of new anticancer therapies, and we have advanced two WNT antagonists for clinical development: vantictumab (anti-FZD) and ipafricept (FZD8-Fc). We examined the antitumor efficacy of these WNT antagonists in combination with various chemotherapies in a large set of patient-derived xenograft models. In responsive models, WNT blockade led to profound synergy with taxanes such as paclitaxel, and the combination activity with taxanes was consistently more effective than with other classes of chemotherapy. Taxane monotherapy increased the frequency of cells with active WNT signaling. This selection of WNT-active chemotherapy-resistant tumorigenic cells was prevented by WNT-antagonizing biologics and required sequential dosing of the WNT antagonist followed by the taxane. The WNT antagonists potentiated paclitaxel-mediated mitotic blockade and promoted widespread mitotic cell death. By blocking WNT/β-catenin signaling before mitotic blockade by paclitaxel, we found that this treatment effectively sensitizes cancer stem cells to taxanes. This combination strategy and treatment regimen has been incorporated into ongoing clinical testing for vantictumab and ipafricept.


Real world outcomes in patients with neuroendocrine tumor receiving peptide receptor radionucleotide therapy.
177Lu-Dotatate (Lu-177), a form of peptide receptor radionuclide therapy (PRRT), was approved by Food and Drug Administration (FDA) for the treatment of somatostatin-receptor-positive neuroendocrine tumors (NETs) in 2018. Clinical trials prior to the FDA approval of Lu-177 showed favorable outcomes but there is limited published real world outcomes data. This study aims to describe and analyze real world outcomes of patients with NET who received Lu-177. After obtaining institutional review board approval, retrospective evaluation was performed to analyze the efficacy of Lu-177 for somatostatin receptor-positive gastro-entero-pancreatic NETs (GEP-NETs) patients at the University of Kansas Cancer Center between June 2018 and September 2021. This study aims to determine the response rate to the treatment of the entire cohort and subgroups. A total of 65 patients received Lu-177 of which 58 completed treatment. The 58 patients had a median age of 61.5 years, 24 females and 34 males, 86% Caucasian and 12% black. The origins of NETs were primarily small bowel (n = 24) and pancreatic (n = 14). Pathology showed grades 1 (n = 21), 2 (n = 25), and 3 (n = 4) and were primarily well-differentiated tumors (n = 4). Among the cohort, 52 patients had imaging to assess response with 14 (26.9%) patients with partial response (PR), 31 (59.6%) with stable disease (SD), and 7 (13.5%) with progressive disease (PD). In a subset analysis, patients with non-functional disease (n = 29) had higher rates of PR 42.3% (compared to 11.5%, P = 0.0147) and higher disease control rate of 96% (compared to 78%, P = 0.042) than patients with functional disease (n = 29). Patients with non-functional disease had a lower PD of 3.85% (compared to 23%, P = 0.0147) than those with functional disease. This real world outcomes analysis of NETs treated with Lu-177 shows improved PR when compared to the initial clinical trials and is promising for patients. In addition, patients with non-functional tumors were found to have a statistically significant improved response rate which has not been described in the literature before. If these study findings are validated in a larger cohort they may guide patient selection for Lu-177 therapy in the future.


Acute childhood hemiplegia associated with chickenpox and elevated anticardiolipin antibody.
Chickenpox is a common infection of childhood. Central nervous system complications, such as cerebellar ataxia and meningoencephalitis, are estimated to occur in less than 1% of cases of chickenpox. Although transverse myelitis and hemiplegia have also, although rarely, been reported, hemiplegia associated with chickenpox and elevated anticardiolipin antibody has not been reported in the literature. We report the case of a 2.5-year-old boy who developed a right hemiplegia 3 weeks after the onset of a primary varicella infection. The serum level of anticardiolipin antibody IgG was markedly elevated. Brain magnetic resonance imaging (MRI) revealed an infarction involving the left globus pallidus, caudate nucleus, and posterior leg of the internal capsule. Laboratory studies ruled out all known causes of stroke. Neurologic signs and symptoms slowly improved with supportive treatment. Varicella infection should be considered one of the possible causes of acute ischemic strokes in children. Usually, two or more risk factors are detected in children with ischemic strokes, and we suggest that all possible causes of strokes be investigated in children with ischemic strokes.


Effects of 3-aminobenzamide on the induction of morphologic transformation by diverse compounds in Balb/3T3 cells in vitro.
When cells were exposed simultaneously for a 24-h period to the poly(ADP-ribose) synthetase inhibitor 3-aminobenzamide (3AB) (1 or 3 mM) plus aflatoxin B1 (AfB1), no increase in toxicity and limited enhancement of transformation frequency (less than 2 X) was observed. Similarly, simultaneous treatment of cell with 3AB plus methylcholanthrene (MCA) had limited effects, slightly decreasing both toxicity and transformation. In contrast, simultaneous treatment with non-toxic doses of 3AB together with the alkylating agents N-methyl-N'-N-nitro-nitrosoguanidine (MNNG) or ethyl methanesulfonate (EMS) resulted in substantial enhancement of the toxicity and transforming effects of both short-chain alkylating agents. When EMS and varying doses of 3AB (0.1-3 mM) were administered simultaneously for 24 h, increasing levels of 3AB were found to cause a dose-dependent enhancement in toxicity and transformation. To explore the relationship of MNNG- and 3AB-induced effects, two further experiments were performed. First, cells were treated with MNNG plus 3AB for varying lengths of time (4, 24, 72 h). Although exposure for as little as 4 h enhanced toxicity and transformation, these effects were even more profound following 24 or 72 h exposure. Second, cells were exposed to 3AB for varing times prior to or after MNNG exposure. Under these conditions the addition of 3AB up to 6 h post MNNG exposure caused profound enhancement of toxicity and transformation, whereas addition of 3AB 24 h post exposure had minimal effects. Thus the co-carcinogenic effect of 3AB is agent-specific, time-specific and dose-dependent.


Exploring the distribution of genetic markers of pharmacogenomics relevance in Brazilian and Mexican populations.
Studies of pharmacogenomics-related traits are increasingly being performed to identify loci that affect either drug response or susceptibility to adverse drug reactions. However, the effect of the polymorphisms can differ in magnitude or be absent depending on the population being assessed. We used the Affymetrix Drug Metabolizing Enzymes and Transporters (DMET) Plus array to characterize the distribution of polymorphisms of pharmacogenetics and pharmacogenomics (PGx) relevance in two samples from the most populous Latin American countries, Brazil and Mexico. The sample from Brazil included 268 individuals from the southeastern state of Rio de Janeiro, and was stratified into census categories. The sample from Mexico comprised 45 Native American Zapotecas and 224 self-identified Mestizo individuals from 5 states located in geographically distant regions in Mexico. We evaluated the admixture proportions in the Brazilian and Mexican samples using a panel of Ancestry Informative Markers extracted from the DMET array, which was validated with genome-wide data. A substantial variation in ancestral proportions across census categories in Brazil, and geographic regions in Mexico was identified. We evaluated the extent of genetic differentiation (measured as FST values) of the genetic markers of the DMET Plus array between the relevant parental populations. Although the average levels of genetic differentiation are low, there is a long tail of markers showing large frequency differences, including markers located in genes belonging to the Cytochrome P450, Solute Carrier (SLC) and UDP-glucuronyltransferase (UGT) families as well as other genes of PGx relevance such as ABCC8, ADH1A, CHST3, PON1, PPARD, PPARG, and VKORC1. We show how differences in admixture history may have an important impact in the distribution of allele and genotype frequencies at the population level.


Lactobacillus paracasei BEJ01 prevents immunotoxic effects during chronic zearalenone exposure in Balb/c mice.
Zearalenone (ZEN) is an estrogenic mycotoxin produced by numerous Fusarium species in pre- or post-harvest cereals. ZEN displays a potent estrogenicity in livestock and also causes severe immunological problems. The aims of this study were to isolate a new ZEN-degrading micro-organism for biological detoxification, to examine its ability to degrade ZEN in liquid medium, and to evaluate its potential for in vivo preventitive effects against ZEN (as would occur with contaminated feed)-induced immunomodulation in mice. Lactobacillus paracasei BEJ01 (LP) isolated from Tunisian artisanal butter was found to display significant binding ability to ZEN in phosphate-buffered saline (i.e. 96.6%) within 24 h of incubation. The in vivo study was conducted using Balb/c mice that received either vehicle (control), LP only (at 2 × 10(9 )cfu/l, ∼2 mg/kg BW), ZEN alone (at 40 mg/kg BW), or ZEN + LP daily for 15 d. Compared to control mice, ZEN treatment led to significantly decreased body weight gains and decrements in all immune parameters assessed. The addition of LP to ZEN strongly reduced the adverse effects of ZEN on each parameter. In fact, mice receiving ZEN + LP co-treatment displayed no significant differences in the assayed parameters as compared to the control mice. The exposures to the bacteria alone had no adverse effects in the mice. From these data, we conclude that LP bacteria could be beneficial in human and animals for protection against immunotoxicity from ZEN at high levels and during chronic exposures.


Serine biosynthesis in human hair follicles by the phosphorylated pathway: follicular 3-phosphoglycerate dehydrogenase.
The phosphorylated pathway of serine biosynthesis was demonstrated in human hair bulbs and sheaths by the formation of phosphoserine and serine from (14C)3-phosphoglyceric acid. The initial and rate limiting enzyme of the pathway, 3-phosphoglycerate dehydrogenase (3-PGDH) was demonstrated by enzyme determinations in human and rat hair follicles, human epidermis, and chicken epidermis. Follicular 3-PGDH was characterized using a sensitive fluorometric assay with NADH as a co-substrate. Monovalent cations (Na+, K+, Li+, or NH4+) were necessary for full enzyme activity. p-Hydroxymercuribenzoate inhibited activity, and activity was 3 times higher with NADH as a co-substrate than with NADPH. The apparent Km for the substrate hydroxyphosphopyruvic acid was 32.8 muM, and the apparent Km for NADH 4.8 muM similar to the Kms for other mammalian 3-PGDHs. Enzyme activity was not altered by parenteral corticosteroids, a high carbohydrate diet, low protein diet, or starvation. Enzyme activity decreased over the first 12 days of life in newborn rats. The phosphorylated pathway of serine synthesis provides a potential nondietary and nonhepatic source of serine, glycine, and their products in keratinizing tissues.


[Cortisone anxiety in chronic obstructive respiratory tract diseases: a patient survey].
Patients with chronic-obstructive lung diseases often develop unrealistic anxiety about the side-effects of cortisone therapy. Distorted descriptions in mass-media are the reason for the anxiety. The relations between cortisone anxiety and other variables of the subjective and objective situation will be investigated within a model of health psychology. About 10% of the 66 patients completing a questionnaire express attitudes, which could be interpreted as cortisone anxiety. The patients with cortisone anxiety differ in other characteristics as well.


The assessment of evoked potential contrast thresholds using real-time retrieval.
Electrophysiologic contrast sensitivity functions (CSF) have been estimated using lock-in amplifier signal retrieval of the visually evoked response (VER). These CSFs were compared with CSFs obtained psychophysically using the same stimulus conditions. The two measures of contrast sensitivity behave similarly in response to variations of temporal and spatial frequency. The major advantage of using real-time retrieval is speed. Threshold for a single spatiotemporal condition can be estimated in as little as 20 sec, making the application of electrophysiologic contrast sensitivity testing feasible for clinical populations.


The linear chromosome of the plant-pathogenic mycoplasma 'Candidatus Phytoplasma mali'.
Phytoplasmas are insect-transmitted, uncultivable bacterial plant pathogens that cause diseases in hundreds of economically important plants. They represent a monophyletic group within the class Mollicutes (trivial name mycoplasmas) and are characterized by a small genome with a low GC content, and the lack of a firm cell wall. All mycoplasmas, including strains of 'Candidatus (Ca.) Phytoplasma asteris' and 'Ca. P. australiense', examined so far have circular chromosomes, as is the case for almost all walled bacteria. Our work has shown that 'Ca. Phytoplasma mali', the causative agent of apple proliferation disease, has a linear chromosome. Linear chromosomes were also identified in the closely related provisional species 'Ca. P. pyri' and 'Ca. P. prunorum'. The chromosome of 'Ca. P. mali' strain AT is 601,943 bp in size and has a GC content of 21.4%. The chromosome is further characterized by large terminal inverted repeats and covalently closed hairpin ends. Analysis of the protein-coding genes revealed that glycolysis, the major energy-yielding pathway supposed for 'Ca. P. asteris', is incomplete in 'Ca. P. mali'. Due to the apparent lack of other metabolic pathways present in mycoplasmas, it is proposed that maltose and malate are utilized as carbon and energy sources. However, complete ATP-yielding pathways were not identified. 'Ca. P. mali' also differs from 'Ca. P. asteris' by a smaller genome, a lower GC content, a lower number of paralogous genes, fewer insertions of potential mobile DNA elements, and a strongly reduced number of ABC transporters for amino acids. In contrast, 'Ca. P. mali' has an extended set of genes for homologous recombination, excision repair and SOS response than 'Ca. P. asteris'. The small linear chromosome with large terminal inverted repeats and covalently closed hairpin ends, the extremely low GC content and the limited metabolic capabilities reflect unique features of 'Ca. P. mali', not only within phytoplasmas, but all mycoplasmas. It is expected that the genome information obtained here will contribute to a better understanding of the reduced metabolism of phytoplasmas, their fastidious nutrition requirements that prevented axenic cultivation, and the mechanisms involved in pathogenicity.


A functional comparison of animal anterior cruciate ligament models to the human anterior cruciate ligament.
Many investigators have used animal models to clarify the role of the human anterior cruciate ligament (ACL). Because none of these models are anatomically and biomechanically identical to the human ACL, there exists a need for an objective comparison of these models. To do this, we used a universal force-moment sensor to measure and compare the in situ forces, including magnitude and direction, of the ACL and the anteromedial (AM) and posterolateral (PL) bundles of human, pig, goat, and sheep knees. An Instron was used to apply 50 and 100 N anterior tibial loads at 90 degrees of knee flexion, while a universal force-moment sensor was used to measure the forces applied by the ACL to the tibia, the in situ force of the ACL. We found significant differences between the magnitude of force experienced by the goat and sheep ACL and AM and PL bundles when compared with the human ACL and AM and PL bundles. Also, the direction of the in situ force in the ACL and AM bundles of the goat and sheep were different from the human. The pig knee differed from the human only in the magnitude and direction of the in situ force in the PL bundle in response under anterior tibial loading. A tally of the significant differences between the animal models and the human knees indicates that goat and sheep knees may have limitations in modeling the human ACL, while the pig knee may be the preferred model for experimental studies.


Postgraduate education and staff development in Spain.
The evolution of the number of hospital pharmacists and pharmacy services in Spanish hospitals is presented and compared with different European countries. The reasons and regulations that have motivated this improvement also are explained. The program for residency training in accredited hospital pharmacies is discussed. In Spain, three years of residency are necessary to become a hospital pharmacy specialist. The Spanish Society of Hospital Pharmacy (AEFH) developed workshops, following the criteria of the World Health Organization, in order to produce an education guidelines document for residency training programs. These educational objectives are presented and the lack of support from Spanish schools of pharmacy is emphasized.


Yeast-based heterologous production of the Colletochlorin family of fungal secondary metabolites.
Transcriptomic studies have revealed that fungal pathogens of plants activate the expression of numerous biosynthetic gene clusters (BGC) exclusively when in presence of a living host plant. The identification and structural elucidation of the corresponding secondary metabolites remain challenging. The aim was to develop a polycistronic system for heterologous expression of fungal BGCs in Saccharomyces cerevisiae. Here we adapted a polycistronic vector for efficient, seamless and cost-effective cloning of biosynthetic genes using in vivo assembly (also called transformation-assisted recombination) directly in Escherichia coli followed by heterologous expression in S. cerevisiae. Two vectors were generated with different auto-inducible yeast promoters and selection markers. The effectiveness of these vectors was validated with fluorescent proteins. As a proof-of-principle, we applied our approach to the Colletochlorin family of molecules. These polyketide secondary metabolites were known from the phytopathogenic fungus Colletotrichum higginsianum but had never been linked to their biosynthetic genes. Considering the requirement for a halogenase, and by applying comparative genomics, we identified a BGC putatively involved in the biosynthesis of Colletochlorins in C. higginsianum. Following the expression of those genes in S. cerevisiae, we could identify the presence of the precursor Orsellinic acid, Colletochlorins and their non-chlorinated counterparts, the Colletorins. In conclusion, the polycistronic vectors described herein were adapted for the host S. cerevisiae and allowed to link the Colletochlorin compound family to their corresponding biosynthetic genes. This system will now enable the production and purification of infection-specific secondary metabolites of fungal phytopathogens. More widely, this system could be applied to any fungal BGC of interest.


[Initial cellular damage in the developing rat brain caused by cytotoxicity of ethylnitrosourea].
Initial cellular degeneration in developing rat fetal brains were induced during the early cytogenetic period of neuroblasts by cytotoxic action of ethylnitrosourea (ENU). This was transplacentally administered. After a single intravenous administration of 60 mg/kg body weight ENU to 16th day-pregnant rats, each fetus was removed surgically from the uteri every hour for 24 hours and every day for 9 days. These were fixed by the transcardial perfusion method to observe them with light and electron microscopes. From 4 hours after the ENU-treatment, the degenerative alteration of cells, characterized ultrastructurally by cytoplasmic condensation and nuclear pyknosis, developed selectively in the matrix cell layer of the fetal cerebrum where active cell division was carried out to produce neuroblasts. Cellular degeneration was first noticed at the zone of DNA synthesis and then all the matrix cell layer including the mitotic phase. Furthermore, the cell processes of matrix cells, or so-called radical fibers, were also recognized to be in the same degenerative processes. These affected cells increased in number and became arranged in columns along the radial fibers through the process of time, and successively migrated to the migrating zone through the outer part of the matrix cell layer. At 10 hours after treatment, many cells were observed in the migrating zone and a few cells were noticed in the matrix cell layer. The degenerated neuroblasts reached the cortical plate about 3 hours after the development of initial degeneration. In contrast, the neuroblasts in the cerebral cortical plate which had already been formed at the time of ENU administration were not affected by the ENU and persisted for the following developments.(ABSTRACT TRUNCATED AT 250 WORDS)


Suicide Attempts Among Pregnant and Postpartum Women in Japan: A Nationwide Retrospective Cohort Study.
To describe the demographic and clinical characteristics of pregnant and postpartum women in Japan with psychoneurological disorders who attempt suicide. A nationwide retrospective cohort study was conducted using the Diagnosis Procedure Combination database, a national database on acute-care inpatients in Japan. All pregnant and postpartum women who had psychoneurological disorders (ICD-10 codes: O993, F530, F531) and were admitted to participating hospitals from January 2016 to March 2018 were identified. Among eligible patients, the prevalence of suicide attempts, risk factors for suicidal behaviors, maternal outcomes, and other characteristics were investigated. Among the 3,286 eligible patients (3,026 pregnant women and 260 postpartum women), 22 pregnant women and 16 postpartum women had attempted suicide. The prevalence of suicide attempts was significantly higher among postpartum women (6.2%) than among pregnant women (0.7%; P < .001). Postpartum patients were more likely to be aged 30 years or older and to have depression. Wrist cutting was the main method of suicide attempt among pregnant patients, whereas hanging was the main method among postpartum patients. Three postpartum patients died during hospitalization. Postpartum patients were more likely to be in critical condition and to use fatal suicide attempt methods compared with pregnant women.


Curitiba acute ischemic stroke protocol: a university hospital and EMS initiative in a large Brazilian city.
Few healthcare centers in Brazil perform thrombolytic therapy for acute ischemic stroke (AIS) patients. The aim of this study was to describe an interinstitutional protocol for the rapid identification and thrombolytic treatment of AIS patients at a public health hospital in a large Brazilian city. Emergency medical services (EMS) personnel evaluated 433 patients with possible stroke during a six-month period. After a standard checklist, patients with suspected AIS and symptoms onset of less than two hours were evaluated at our University Hospital (UH). Sixty-five (15%) patients met the checklist criteria and had a symptom onset of less than two hours, but only 50 (11%) patients were evaluated at the UH. Among them, 35 (70%) patients had ischemic stroke, 10 (20%) had hemorrhagic stroke, and 5 (10%) had other diagnoses. Of the 35 ischemic stroke patients, 15 (43%) underwent IV thrombolysis. The present study demonstrated that trained EMS workers could help to improve the rate of thrombolytic treatment in large Brazilian cities. Permanent training programs for EMS and hospital staff, with quality control and correct identification of AIS patients, should be implemented to increase appropriate thrombolytic therapy rates in Brazil.


Evolving strategies in mechanobiology to more effectively treat damaged musculoskeletal tissues.
In this paper, we had four primary objectives. (1) We reviewed a brief history of the Lissner award and the individual for whom it is named, H.R. Lissner. We examined the type (musculoskeletal, cardiovascular, and other) and scale (organism to molecular) of research performed by prior Lissner awardees using a hierarchical paradigm adopted at the 2007 Biomechanics Summit of the US National Committee on Biomechanics. (2) We compared the research conducted by the Lissner award winners working in the musculoskeletal (MS) field with the evolution of our MS research and showed similar trends in scale over the past 35 years. (3) We discussed our evolving mechanobiology strategies for treating musculoskeletal injuries by accounting for clinical, biomechanical, and biological considerations. These strategies included studies to determine the function of the anterior cruciate ligament and its graft replacements as well as novel methods to enhance soft tissue healing using tissue engineering, functional tissue engineering, and, more recently, fundamental tissue engineering approaches. (4) We concluded with thoughts about future directions, suggesting grand challenges still facing bioengineers as well as the immense opportunities for young investigators working in musculoskeletal research. Hopefully, these retrospective and prospective analyses will be useful as the ASME Bioengineering Division charts future research directions.


Multi-probe relaxation dispersion measurements increase sensitivity to protein dynamics.
Carr-Purcell-Meiboom-Gill (CPMG) relaxation dispersion measurements are a valuable tool for the characterization of structural transitions on the micro-millisecond timescale. While the measurement of (15)N relaxation dispersion is now routine, the measurements with alternative nuclei remain limited. Here we report (15)N as well as (1)H R2 relaxation dispersion measurements of the N23PP/S148A "dynamic knockout" mutant of dihydrofolate reductase. The (1)H dispersion measurements are complementary to (15)N data as many additional residues are observed to have dispersive behavior for the (1)H nucleus. Simultaneous fitting of the dispersion profiles for the two nuclei increases the accuracy of exchange parameters determined for individual residues and clustered groups of residues. The different sensitivity of the two nuclei to changes in backbone torsional angles, ring currents, and hydrogen bonding effects provides important insights into the nature of the structural changes that take place during the exchange process. We observe clear evidence of direct and indirect hydrogen bond effects for the (15)N and (1)H chemical shift changes in the active-site, modulation of ring current shielding in the CD-loop and backbone torsional changes in a cluster of residues associated with the C-terminus. This work demonstrates the power of combined (1)H and (15)N probes for the study of backbone dynamics on the micro-millisecond timescale though the analysis of chemical shift changes.


[Therapeutic advances in monoclonal gammopathy-associated renal diseases].
During the last ten years, significant advances have been made in the field of monoclonal gammopathy-associated renal diseases. These have consisted in a better recognition of their pathological and clinical significations and correlation. Furthermore, therapeutic advances have allowed improving the prognosis of B-cell monoclonal proliferations and of the related renal complications. In the peculiar case of the multiple myeloma with severe renal failure requiring dialysis, one can now expect better survival and chance of recovering renal function.