The arthrospore-related gene Acaxl2 is involved in cephalosporin C production in industrial Acremonium chrysogenum by the regulatory factors AcFKH1 and CPCR1.
Cephalosporin C (CPC) production is often accompanied by a typical morphological differentiation of Acremonium chrysogenum, involving the fragmentation of its hyphae into arthrospores. The type I integral plasma membrane protein Axl2 is a central component of the bud site selection system (BSSS), which was identified as the regulatory factor involved in the hyphal septation process and arthrospore formation. Using CRISPR/Cas9 technology and homologous recombination (HR), we inserted an egfp donor DNA sequence into the Acaxl2 locus, causing the generation of the deletion strain Ac-ΔAcaxl2:eGFP from Acremonium chrysogenum FC3-5-23, the industrial producer of CPC. The mycelial morphology of the deletion strain Ac-ΔAcaxl2:eGFP was mainly composed of arthrospores with a characteristic diameter of 2-8 µm, which increased from 75% at 48 h to 90% at 72 h post culture and were maintained until the end of the fermentation process. However, the deletion strain showed accelerated production of CPC, and the final titer was 5573 μg/ml, which was nearly three times higher than that of the control strain FC3-5-23. The up-regulation of genes related to the biosynthesis gene cluster in Ac-ΔAcaxl2:eGFP, especially the "late" genes, was one reason why its CPC production was higher than that of the original strain. Furthermore, compared with FC3-5-23, the more significant increase of genes involved in the BSSS (Acbud3 and Acbud4) in Ac-ΔAcaxl2:eGFP in the late stage of fermentation, may be responsible for this increase in arthrospore formation. Similarily, the transcription of the regulatory factors AcFKH1 and CPCR1 were also markedly increased at this time and may be the factors responsible for the regulation of CPC synthesis. These results indicated that Acaxl2 plays an important role in both arthrospore formation and CPC production, strongly implicating these regulatory factors as having pivotal links between mycelial morphology and secondary metabolite production in high-yielding A. chrysogenum. To the opposite, the axl2 gene knockout of wild strain CGMCC 3.3795 did not significantly influence the CPC production, which reflected the complexity of the secondary metabolic process and the differences in the function of axl2 gene in high- and low-yielding strains.


Removal of oxygen demand and nitrogen using different particle-sizes of anthracite coated with nine kinds of LDHs for wastewater treatment.
This paper reports the application of anthracite particles of different sizes and coated with nine kinds of layered double hydroxides (LDHs) varying in M(II)-M(III) cations, as alternative substrates in the simulated vertical-flow constructed wetland columns. Effects of LDHs-coating and particle size of modified anthracites were examined to evaluate their abilities in removing oxygen demand and nitrogen from sewage wastewater. Results showed that LDHs modification effectively enhanced the removal of nitrogen and organics. The removal efficiencies of total nitrogen (TN) , ammonia and chemical oxygen demand (COD) were best improved by 28.5%, 11.9% and 4.1% for the medium particle size (1-3 mm), followed by 9.2%, 5.5% and 13.6% for the large size (3-5 mm), respectively. Only TN removal was improved up to 16.6% for the small particle size (0.5-1 mm). Nitrate tended to accumulate and fluctuate greatly across all the treatments, probably due to the dominancy of aerobic condition in the vertical-flow columns. Overall, MgFe-LDHs was selected as the best-modified coating for anthracite. The results suggested LDHs modification would be one of the promising strategies to provide new-types of highly efficient and lasting wetland substrates.


Hydrocortisone administration for the treatment of refractory hypotension in critically ill newborns.
The purpose of this observation was to evaluate the safety and efficacy of hydrocortisone (HC) for the treatment of refractory hypotension in term and preterm infants. A secondary purpose was to determine the utility of serum cortisol concentrations in predicting the response to treatment. This is a retrospective observational study of 117 infants treated with a standardized HC protocol for refractory hypotension. Refractory hypotension was defined as a mean arterial pressure (MAP) less than the gestational age (GA) despite a total inotrope dose of 20 microg per kg per min. Baseline serum cortisol concentrations were determined prior to treatment with stress dose HC. Treatment with HC increased the MAP at 2, 6, 12 and 24 h after initiation, decreased the total inotrope dose at 6, 12 and 24 h, and was associated with resolution of oliguria. There was no correlation between the pretreatment baseline cortisol concentration and GA, birth weight or the response to treatment. The incidence of grades III to IV intraventricular hemorrhage, periventricular leukomalacia, bacterial or fungal sepsis and spontaneous intestinal perforation (SIP) after HC treatment was similar to institutional historic controls prior to institution of this standardized HC protocol. HC treatment was associated with a rapid resolution of cardiovascular compromise. The incidence of significant side effects was similar to that in previously published reports, including a comparable incidence of SIP. On the basis of our results, measuring baseline serum cortisol concentration to guide the management of refractory hypotension is unwarranted.


A fluorescence polarization assay using oligonucleotide probes for the rapid detection of verotoxin-producing Escherichia coli.
A hybridization assay using fluorescence polarization was combined with the asymmetric polymerase chain reaction (PCR) in a method for the detection of the verotoxin type 2 gene of verotoxin-producing Escherichia coli. Six oligonucleotide probes labeled with FITC were designed and evaluated. One of these gave a detection limit of 10(3) colony forming units per assay, and assay results could be obtained within 5 min after PCR. It appears that the detection limit was restricted mainly by the extent and fidelity of PCR amplification, rather than by the sensitivity of the fluorescence polarization technique, indicating that good probe design facilitates the rapid detection of the PCR product. The fluorescence polarization assay, in conjunction with DNA amplification by PCR, is a powerful and widely applicable method for the rapid and sensitive detection of oligonucleotide sequences.


Factors associated with survival and racing performance of 114 Thoroughbred foals with septic arthritis compared with maternal siblings (2009-2015).
There is little consensus on factors associated with survival in foals with septic arthritis and limited data on long-term racing performance of Thoroughbred foals treated for septic arthritis. A more thorough understanding of short- and long-term outcome is necessary to help inform owners, and subsequently guide treatment. To investigate factors associated with survival, and to analyse racing performance of foals with septic arthritis compared with their maternal siblings. Retrospective cohort and a case-control study. Veterinary clinical records of Thoroughbred foals ≤180 days old that underwent arthroscopic, cannulae or through-and-through needle lavage for the treatment of septic arthritis between 2009 and 2015 were reviewed. Data included signalment, and clinicopathological information. The dam's foaling records were reviewed and the lifetime racing records were obtained for affected foals and two of their maternal siblings. Logistic regression analysis was used to determine factors associated with survival to discharge or racing. Comparisons between treated foals and their maternal siblings were made. Ninety (78%) of 115 foals diagnosed with septic arthritis were discharged alive. Foals <26 days old at the time of admission were five times less likely (P = .003) and foals with concurrent multisystemic disease were six times less likely (P = .02) to be discharged alive. Sixty (67%) foals discharged alive started in ≥1 race, and there was no difference in the proportion of foals that started in a race or racing performance between foals treated for septic arthritis and their maternal siblings. Retrospective study design, limited number of foals with multiple joint involvement and failure to accurately record duration of clinical signs. Foals treated for septic arthritis at the Scone Equine Hospital, New South Wales, Australia had a good prognosis for survival, and for this cohort, foals that survived to discharge had a similar ability to race as their maternal siblings.


Changes in hormone levels in relation to breeding behavior in male bullfrogs (Rana catesbeiana) at the individual and population levels.
Fluctuations in plasma androgen (testosterone and 5 alpha-dihydrotestosterone), corticosterone (B), and luteinizing hormone (LH) of male bullfrogs in central California were measured during the spring mating season. Androgen and LH levels generally increased in the population prior to the initiation of chorusing and establishment of territories, whereas plasma B peaked in a 2-week period at the start of heavy chorusing; this coincided with a transitory, 1 week decline in androgen. Individual males showed fluctuations in plasma androgen and LH levels throughout the breeding season, often within 1 day, but there was no clear correlation between changes in the two hormones. No time of day effect was apparent on any of the hormones. B and androgen levels were significantly but weakly correlated, (r = 0.35) but LH and androgen were not. Although the seasonal trend of increasing androgen corresponded with the start of intense chorusing (and presumably sexual activity) by the population, behavior of individuals and their circulating androgen levels did not correlate. After acquiring territories, males showed no overall trend of increased plasma androgen. Moreover, males that showed no vocal or territorial behavior had significantly higher androgen and lower B levels than calling males. Increased B levels suggest that territorial behavior and especially direct agonistic encounters represent stresses that could have an inhibitory effect on androgen secretion.


Changing horses midstream: the promise and prudence of practice redesign.
An emerging vision for primary care calls for the adoption of information technology and a strong business model to save a dying health care system. The authors are participants in the National Demonstration Project (NDP), a study sponsored by leading organizations in family medicine and directed by a for-profit subsidiary of the American Academy of Family Physicians, TransforMED. The NDP embraces the Future of Family Medicine Report and seeks to test the ability of existing practices to implement its basic tenets. The NDP will conclude in June 2008, but its findings and observations will likely ripple out for years. Our report is a personal reflection that looks beyond the question of whether busy practices and practitioners can change horses midstream. We ask, "Is this primary care, and is this what it needs?"


5-Fluorouracil, folinic acid, epidoxorubicin and cisplatin (FLEP) combination chemotherapy in advanced measurable gastric cancer. A phase II trial of the Spanish Cooperative Group for Gastrointestinal Tumor Therapy (TTD).
Metastatic disease is a common problem in gastric cancer and the development of better chemotherapeutic regimens is a clear priority in gastrointestinal oncology. Ninety consecutive, previously untreated patients with unresectable or measurable metastatic gastric cancer were included in a multicenter phase II trial with a combination of folinic acid (200 mg/m2) and 5-fluorouracil (400 mg/m2) days 1-3, with epidoxorubicin (60 mg/m2) and cisplatin (100 mg/m2) on day 2. A total of 376 courses of FLEP were given, with a median of four courses per patient. Objective responses were observed in 32 (35%) patients (CI at 95%: 25.7%-46.3%). Eight (9%) patients experienced clinical complete remissions. Median time to progression was 25 weeks for the entire group of patients and 38 weeks for responders. Myelosuppression was the primary toxicity. WHO grade 3 leukopenia appeared in 26 patients (29%). Ten presented episodes of febrile neutropenia requiring hospitalization, but no toxic deaths were observed. Grades 3 and 4 thrombocytopenia were seen in 8 and 1 patients, respectively. Median survival time was 8 months for all treated patients and 11 months for responders. The FLEP regimen is an active combination in advanced gastric cancer with moderate toxicity that warrants further testing in a phase III trial.


Decreased bombesin peptide response to cigarette smoking in schizophrenia.
Schizophrenic patients are extremely heavy tobacco smokers. However, a lower incidence of lung cancer in schizophrenic patients has been observed in comparison to other heavy smokers. Nicotine increases the proliferation of pulmonary neuroendocrine tissue, causing the release of a bombesin-like peptide. Thus, bombesin-like peptide levels in urine may be an indicator of precancerous, cigarette-induced lung damage. Bombesin-like peptide levels of 10 schizophrenic smokers and 11 schizophrenic nonsmokers were compared to those of nonschizophrenic subjects matched for age and pack-years of smoking. The nonschizophrenic smokers showed the expected increase in urinary bombesin-like peptide levels, as compared to nonschizophrenic nonsmokers. Schizophrenic patients had lower bombesin-like peptide levels independent of smoking effects. The mechanism of the difference in bombesin-like peptide levels between schizophrenic patients and nonschizophrenic subjects is unknown, but one possibility involves alteration in the alpha 7-nicotinic acetylcholine receptor, which mediates the growth of some neuroendocrine cell lines in vitro.


Intraoperative 5-aminolevulinic acid-induced photodynamic diagnosis of metastatic brain tumors with histopathological analysis.
Fluorescence-guided surgery using 5-aminolevulinic acid (5-ALA) is a promising real-time navigation method in the surgical resection of malignant gliomas. In order to determine whether this method is applicable to metastatic brain tumors, we evaluated the usefulness of intraoperative fluorescence patterns and histopathological features in patients with metastatic brain tumors. We retrospectively reviewed the cases of 16 patients with metastatic brain tumors who underwent intraoperative 5-ALA fluorescence-guided resection. Patients were given 20 mg/kg of 5-ALA orally 2 h prior to the surgery. High-powered excitation illumination and a low-pass filter (420, 450, or 500 nm) were used to visualize the fluorescence of protoporphyrin IX (PpIX), the 5-ALA metabolite. We evaluated the relationships between the fluorescence and histopathological findings in both tumoral and peritumoral brain tissue. Tumoral PpIX fluorescence was seen in only 5 patients (31%); in the remaining 11 patients (69%), there was no fluorescence in the tumor bulk itself. In 14 patients (86%), vague fluorescence was seen in peritumoral brain tissue, at a thickness of 2-6 mm. The histopathological examination found cancer cell invasion of adjacent brain tissue in 75% of patients (12/16), at a mean ± SD depth of 1.4 ± 1.0 mm (range 0.2-3.4 mm) from the microscopic border of the tumor. There was a moderate correlation between vague fluorescence in adjacent brain tissue and the depth of cancer cell invasion (P = 0.004). Peritumoral fluorescence may be a good intraoperative indicator of tumor extent, preceding more complete microscopic gross total resection. Institutional Review Board of Osaka Medical College No. 42, registered February 17, 1998, and No. 300, registered April 1, 2008. They were retrospectively registered.


Thermal Ablation for Intrahepatic Cholangiocarcinoma in Cirrhosis: Safety and Efficacy in Non-Surgical Patients.
To assess the effectiveness, safety, and overall survival (OS) of thermal ablation as upfront treatment of intrahepatic colangiocarcinoma (ICC) in patients with cirrhosis. This was a retrospective analysis of all biopsy-confirmed ICC in cirrhotic patients treated in the authors' unit from 2001 to 2017. Baseline characteristics, ablation procedures, and complications were recorded, and time to recurrence (TTR) and OS were calculated. Twenty-seven patients were identified. Seventy percent had Child-Pugh A disease, and most had clinically significant portal hypertension. Median tumor size was 21 mm. Twenty-one cases were uninodular, and 10 were single ≤ 2 cm. Complete radiologic necrosis was achieved in 25 cases (92.6%). Median OS was 30.6 months (95% confidence interval [CI], 22.6-46.5), and recurrence was detected in 21 cases (77.8%) with a TTR of 10.1 months (95% CI, 7.7-20.9). In those patients with single ≤ 2-cm ICC, the OS was 94.5 months (95% CI, 11.7-not reached). Differences in OS were statistically significant between patients with single ICC ≤ 2 cm and patients with single ICC > 2 cm (P = .04) and between patients with single ICC > 2 cm and patients with multinodular ICC (P = .02). Only 1 patient had a treatment-related complication. Thermal ablation is a safe and effective treatment for ICC in patients with cirrhosis who are not candidates for surgery. The OS is similar to that reported in surgical series, but the initial treatment success is hampered by a high rate of tumor recurrence. Encouraging long-term survival after thermal ablation is achieved in patients with single ≤ 2-cm ICC.


Silencing of lncRNA PVT1 by miR-214 inhibits the oncogenic GDF15 signaling and suppresses hepatocarcinogenesis.
The prognosis for hepatocellular carcinoma (HCC) is dismal. Long noncoding RNA PVT1 has been linked to malignancies and might be a deleterious therapy target. However, the key events controlling its expression in HCC remain undetermined. Here, we address how PVT1 is fine-regulated and its downstream signaling in hepatoma cells. Interestingly, we found that c-Myc and P53 could divergently regulate PVT1 transcription. Oncoprotein c-Myc enhances PVT1 expression, whereas P53 suppresses its expression. We also identified miR-214 as a crucial, negative regulator of PVT1. Consistently, high miR-214 levels were significantly correlated with diminished PVT1 expression in HCC specimens. Silencing of PVT1 by ectopic miR-214 or siRNAs markedly inhibited viability and invasion of HCC cells. In opposition, inhibition of endogenous miR-214 promoted PVT1 expression and enhanced cell proliferation. Notably, oncogenic GDF15 is a potential downstream target of the miR-214-PVT1 signaling. Collectively, our results show that the c-Myc/P53/miR-214-PVT1-GDF15 axis is implicated in HCC development, shedding light on the mechanistic actions of PVT1 and representing potential targets for HCC clinical intervention.


Hydatid disease of the ribs: An exceptional location.
Hydatid disease is a zoonosis caused by larval stages of cestodes belonging to the genus Echinococcus. The rib location is exceptional. It presents a real diagnostic and therapeutic challenge. To describe the clinical, serological and radiological features and surgical management of rib hydatidosis. This is a retrospective study conducted over 4 years, on five cases of rib hydatidosis. We analysed the clinical and radiological presentations and the adopted therapeutic procedure. The average age of our patients was 44 years, without gender predominance. The clinical signs were dominated by the presence of an immobile swelling of fluid consistency without inflammatory sign, accompanied by moderate and intermittent localised pain. The laboratory assessment was nonspecific. The radiological assessment, including chest X-ray and thoracic computed tomography, with and without contrast, was essential in order to assess the extent of the lesion. Thoracic magnetic resonance imaging was requested in one case because of suspicions of a spinal extension. All of our patients underwent a rib excision accompanied by medical treatment of albendazole 24 hours after the surgery. The follow-up ranged from 1 year to 4 years and did not show any recurrence. Hydatidosis of the ribs is an exceptional location of hydatid disease. The diagnosis was based on radiology and intraoperative exploration. The treatment remained essentially surgical by rib excision with anthelmintic drugs to prevent recurrence.


Nuclear factor-kappaB contributes to infarction after permanent focal ischemia.
Activation of transcription factor nuclear factor-kappaB (NF-kappaB) may induce expression of either proinflammatory/apoptotic genes or antiapoptotic genes. Because a considerable number of middle cerebral artery occlusions (MCAOs) in humans are not associated with reperfusion during the first 24 hours, the role of NF-kappaB after permanent MCAO (pMCAO) was investigated. Mice transgenic for a NF-kappaB-driven beta-globin reporter were exposed to pMCAO, and the expression of the reporter gene was quantified with real-time polymerase chain reaction. Mice lacking the p50 subunit of NF-kappaB and wild-type controls were exposed to pMCAO with or without treatment with pyrrolidinedithiocarbamate (PDTC), an NF-kappaB inhibitor. Brain sections of human stroke patients were immunostained for the activated NF-kappaB. pMCAO increased NF-kappaB transcriptional activity to 260% (36.9+/-4.5 compared with 14.4+/-2.6; n=10; P<0.01) in the brain; this NF-kappaB activation was completely blocked by PDTC (17.2+/-2.6; n=9; P<0.05). In p50-/- mice, pMCAO resulted in 41% (18+/-3.2 mm3; n=12) smaller infarcts compared with wild-type controls (32.9+/-3.8 mm3; n=9; P<0.05), which was comparable to the protection achieved with PDTC in wild-type mice (19.6+/-4.2 mm3; n=8). Pro-DTC, a PDTC analogue that does not cross the blood-brain barrier, had no effect, even though Pro-DTC and PDTC were equally protective in vitro. During the first 2 days of human stroke, NF-kappaB was activated in neurons in the penumbral areas. NF-kappaB is induced in neurons during human stroke, and activation of NF-kappaB in the brain may contribute to infarction in pMCAO.


Early dosing and efficacy of triptans in acute migraine treatment: the TEMPO study.
To compare the effectiveness of early or late triptan intake in the treatment of acute migraine attacks. The TEMPO study was a French prospective, multicentre, two-phase study conducted in neurological practice. Two-hundred-and-ten migraine patients who were regular triptan users were enrolled. In the first phase, patients treated three attacks as they usually did. In the second phase, those who initially practiced late dosing ( ≥ 1 hour after headache onset) were instructed to change to early dosing ( < 1 hour). A total of 144 patients completed the first phase. Seventy-nine patients constituted the 'early dosers' group and 65 patients the 'late dosers' group. In this phase, early dosing produced higher rates (n = 38; 52.8%) of freedom from pain at 2 hours in at least two of three attacks compared with late dosing (n = 19; 30.2%; p < 0.01). In the second phase, switching from late to early dosing following the physician's instruction (n = 42 patients) also improved the rates of freedom from pain at 2 hours (from 38.1% (n = 16) to 53.7% (n = 22); p < 0.05). This suggests that advising patients on the importance of early triptan intake after headache onset may help improve the efficacy of acute migraine treatments.


Patch dynamics ofGlaucopsyche lygdamus (Lycaenidae): correlations between butterfly density and host species diversity.
Glaucopsyche lygdamus egg densities were surveyed over a 2000-m section of Gold Creek and at 30 different isolated patches in the Gold Basin drainage in Colorado. Host plant numbers and diversity were quantified, as well as other variables potentially influencing butterfly population size, such as patch size and isolation. Egg densities correlated significantly only with measures of host species diversity. Patches consisting of a single host species, no matter how large, did not support high butterfly densities, but patches of multiple, equitably distributed host species did. The most likely explantation, in light of oviposition preference and larval performance data accumulated for this butterfly species, is that host species diversity is necessary for the persistence ofG. lygdamus populations, because alternative host species buffer population losses during poor or unusual years. The dependence of both ovipositing butterflies and developing larvae on the ephemeral, young, host plant flowers make the butterfly especially vulnerable to year-to-year variation in host plant availability and quality.


Comparative pathology of canine hereditary nephropathies: an interpretive review.
Hereditary nephropathies in dogs represent multiple complex clinicopathological entities which cause renal failure in juvenile, adolescent, and young adult dogs. To date, nephropathies believed to have a genetic basis have been described in 11 breeds. These disorders represent a variety of developmental, degenerative, and metabolic defects. Many canine hereditary nephropathies are analogous to childhood nephropathies in man. Review of the veterinary literature, comparative pathology, and avenues of future research are discussed.


Synthesis, structural characteristics, DNA binding properties and cytotoxicity studies of a series of Ru(III) complexes.
Four related ruthenium(III) complexes, with the formula mer-[RuCl(3)(dmso)(N-N)] (dmso=dimethyl sulfoxide; N-N=2,2'-bipyridine (1), 1,10-phenantroline (2), dipyrido[3,2-f:2',3'-h]quinoxaline (3) and dipyrido[3,2-a:2',3'-c]phenazine (4)), have been reported. Complexes 3 and 4 are newly synthesized and characterized by X-ray diffraction. The hydrolysis process of 1-4 has been studied by UV-vis measurement, and it has been found that the extension of the N-N ligands can increase the stability of the complexes. The binding of these complexes with DNA has been investigated by plasmid cleavage assay, competitive binding with ethidium bromide (EB), DNA melting experiments and viscosity measurements. The DNA binding affinity is increased with the extension of the planar area of the N-N ligands, and complex 4 shows an intercalative mode of interaction with DNA. The in vitro anticancer activities of these compounds are moderate on the five human cancer cell lines screened.


Successful dietary control of tyrosinemia II.
A Japanese girl, 2 years, 8 months of age, with palmoplantar keratosis and dendritic corneal opacities, showed increased tyrosine levels in the plasma, urine, and cerebrospinal fluid. The mental and physical growth was not retarded. The hepatorenal functions were within normal limits. Electron microscopically, the epidermal keratinocytes showed increased tonofibrils and no structures suggestive of tyrosine crystals. Cytosol and mitochondrial tyrosine aminotransferase (TAT) activities of the liver were greatly decreased, while p-hydroxyphenyl pyruvate oxidase (p-HPPO) activity was not decreased. The plasma tyrosine levels were controlled for 3 years with low phenylalanine-tyrosine diet.


Herpes Simplex Virus Cervicitis Mimicking Preterm Premature Rupture of Membranes.
The diagnosis of preterm premature rupture of membranes (PROM) is based on pooling, ferning, and Nitrazine tests; definitive diagnosis is made with a blue dye test. A 21-year-old woman, gravida 1 para 0, at 25 5/7 weeks of gestation was admitted for preterm PROM with positive findings of pooling, Nitrazine, and ferning. Her cervix was bluish with white plaques. Amniotic fluid volume was normal. On hospital day 8, her discharge ceased; examination was negative for pooling, Nitrazine, and ferning. A blue dye tampon test was negative. A Pap test result from her hospitalization returned consistent with herpes infection. The diagnosis of preterm PROM should be constantly reevaluated in the setting of a normal amniotic fluid volume.


Efficient Semitransparent Solar Cells with High NIR Responsiveness Enabled by a Small-Bandgap Electron Acceptor.
Inspired by the remarkable promotion of power conversion efficiency (PCE), commercial applications of organic photovoltaics (OPVs) can be foreseen in near future. One of the most promising applications is semitransparent (ST) solar cells that can be utilized in value-added applications such as energy-harvesting windows. However, the single-junction STOPVs utilizing fullerene acceptors show relatively low PCEs of 4%-6% due to the limited sunlight absorption because it is a dilemma that more photons need to be harvested in UV-vis-near-infrared (NIR) region to generate high photocurrent, which leads to the significant reduction of device transparency. This study describes the development of a new small-bandgap electron-acceptor material ATT-2, which shows a strong NIR absorption between 600 and 940 nm with an Egopt of 1.32 eV. By combining with PTB7-Th, the as-cast OPVs yield PCEs of up to 9.58% with a fill factor of 0.63, an open-circuit voltage of 0.73 V, and a very high short-circuit current of 20.75 mA cm-2 . Owing to the favorable complementary absorption of low-bangap PTB7-Th and small-bandgap ATT-2 in NIR region, the proof-of-concept STOPVs show the highest PCE of 7.7% so far reported for single-junction STOPVs with a high transparency of 37%.


Loss of CD28 on Peripheral T Cells Decreases the Risk for Early Acute Rejection after Kidney Transplantation.
End-stage renal disease patients have a dysfunctional, prematurely aged peripheral T-cell system. Here we hypothesized that the degree of premature T-cell ageing before kidney transplantation predicts the risk for early acute allograft rejection (EAR). 222 living donor kidney transplant recipients were prospectively analyzed. EAR was defined as biopsy proven acute allograft rejection within 3 months after kidney transplantation. The differentiation status of circulating T cells, the relative telomere length and the number of CD31+ naive T cells were determined as T-cell ageing parameters. Of the 222 patients analyzed, 30 (14%) developed an EAR. The donor age and the historical panel reactive antibody score were significantly higher (p = 0.024 and p = 0.039 respectively) and the number of related donor kidney transplantation was significantly lower (p = 0.018) in the EAR group. EAR-patients showed lower CD4+CD28null T-cell numbers (p<0.01) and the same trend was observed for CD8+CD28null T-cell numbers (p = 0.08). No differences regarding the other ageing parameters were found. A multivariate Cox regression analysis showed that higher CD4+CD28null T-cell numbers was associated with a lower risk for EAR (HR: 0.65, p = 0.028). In vitro, a significant lower percentage of alloreactive T cells was observed within CD28null T cells (p<0.001). Immunological ageing-related expansion of highly differentiated CD28null T cells is associated with a lower risk for EAR.


Genetic epistasis regulates amyloid deposition in resilient aging.
The brain-derived neurotrophic factor (BDNF) interacts with important genetic Alzheimer's disease (AD) risk factors. Specifically, variants within the SORL1 gene determine BDNF's ability to reduce amyloid β (Aβ) in vitro. We sought to test whether functional BDNF variation interacts with SORL1 genotypes to influence expression and downstream AD-related processes in humans. We analyzed postmortem brain RNA sequencing and neuropathological data for 441 subjects from the Religious Orders Study/Memory and Aging Project and molecular and structural neuroimaging data for 1285 subjects from the Alzheimer's Disease Neuroimaging Initiative. We found one SORL1 RNA transcript strongly regulated by SORL1-BDNF interactions in elderly without pathological AD and showing stronger associations with diffuse than neuritic Aβ plaques. The same SORL1-BDNF interactions also significantly influenced Aβ load as measured with [18F]Florbetapir positron emission tomography. Our results bridge the gap between risk and resilience factors for AD, demonstrating interdependent roles of established SORL1 and BDNF functional genotypes.


Bronchial asthma due to allergy to pancreatic extract: a hazard in the treatment of cystic fibrosis.
The mother of two children suffering from cystic fibrosis developed marked allergic symptoms affecting the eyes, nose and bronchi and severe bronchial asthma ensued. The cause of this allergy proved to be the powder released on opening the capsules of pancreatic extract which she provided for her children at each mealtime. The risk of pancreatic extract to those handling it, and possibly also to patients suffering from cystic fibrosis, is emphasized.


Immunohistochemical analysis of mechanoreceptors in the human posterior cruciate ligament: a demonstration of its proprioceptive role and clinical relevance.
Although long-term studies report successful results with total knee arthroplasty (TKA), performed with or without posterior cruciate ligament (PCL) retention, controversy exists as to which is preferable in regard to patient outcome and satisfaction. The possible proprioceptive role of the PCL may account for a more normal feeling of the arthroplasty. Although the PCL has been examined using various histological techniques, immunohistochemical techniques are the most sensitive for neural elements. Therefore an immunohistochemical study was designed to determine the patterns of innervation, the morphological types of the proprioceptors, and their immunohistochemical profile. During TKA, samples were obtained from 22 osteoarthritic PCLs and subjected to immunohistochemical analysis with mouse monoclonal antibodies against neurofilament protein (NFP), S100 protein (S100P), epithelial membrane antigen (EMA), and vimentin (all present in neuromechanoreceptors). Three normal PCLs from cadaveric specimens were also obtained and analyzed for comparison. Five types of sensory corpuscles were observed in both the normal and the arthritic PCLs: simple lamellar, Pacini-like, Ruffini, Krause-like, and morphologically unclassified. Their structure included a central axon, inner core, and capsule in lamellar and Pacini corpuscles and variable intracorpuscular axons and periaxonal cells in the Ruffini and Krause-like corpuscles. The immunohistochemical profile showed the central axon to have NFP immunoreactivity, periaxonal cells to have S100P and vimentin immunoreactivity, and the capsule to have EMA and vimentin immunoreactivity. Nerve fibers and free nerve endings displayed NFP and S100P immunoreactivity. The immunohistochemical profile of the PCL sensory corpuscles is almost identical to that of cutaneous sensory corpuscles. Some prior histological studies of the PCL reported Golgi-like mechanoreceptors, and others found encapsulated corpuscles but no Golgi-like structures. This report determined the innervation of the PCL by the more sensitive immunohistochemical means, revealing four major types of encapsulated mechanoreceptors. The plentiful and varied types of encapsulated mechanoreceptors found in even the arthritic PCL suggests a rich proprioceptive role. It is controversial as to whether preservation of the PCL at TKA improves postoperative proprioception. Our findings tend to support those clinical reports of improved proprioception after PCL-retaining versus PCL-substituting TKAs. The presence of many and varied types of mechanoreceptors may account for the improved stair climbing reported in patients with PCL-retaining TKA and may contribute to patient satisfaction and a more normal feeling after TKA.


Live birth in male de novo Kallmann syndrome after cross-generational genetic sequencing.
To present the first case proposing the use of preimplantation genetic testing for monogeneic disorders for Kallmann syndrome, providing comprehensive care in the genomic era of precision medicine. Gonadotropin therapy was used for spermatogenesis, followed by in vitro fertilization by intracytoplasmic sperm injection and embryo transfer. Cross-generational targeted next-generation sequencing was then done for genes known to cause Kallmann syndrome. A heterozygous mutation at codon 102 of the FGFR1 gene was found in the patient, but the father was found to have the same mutation yet is unaffected by Kallmann syndrome. Since no causative mutation was found, a de novo or sporadic mutation was suspected as the cause of Kallmann syndrome in this case. Comprehensive care must be available for male Kallmann syndrome patients, as treatment should not stop at spermatogenesis, but continue with genetic counseling due to possible inheritance.


24 h Hydration profile of collegiate soccer players training once versus twice per day in the heat.
This study examined 24-h hydration parameters among collegiate, male soccer players (n = 17) during twice (X2) and once (X1) per day practice schedules in the heat. Urine specific gravity (USG) and body mass were measured before morning practices, afternoon practice (X2)/team meeting (X1), and the next morning practices. Fluid intake, sweat losses, and urinary losses were assessed during each 24-h window. Pre-practice body mass or USG did not differ among the timepoints. Sweat losses differed among all practices (p < 0.05) and averaged approximately 2.181±0.693 (X2AM) 1.710±0.474 (X2PM), and 3.361±0.956 L (X1AM), but players averaged replacing >50% of sweat losses with fluid intake every practice. Fluid intake during and between practices from practice 1 to the afternoon practice for X2 resulted in a positive fluid balance for X2 (+0.446±0.916 L). However, higher sweat loss during the initial morning practice and lower relative fluid intake prior to the afternoon team meeting the following morning resulted in a negative fluid balance (-0.304±0.675 L; p < 0.05: Cohen's d = 0.94) over the same time period for X1. By the start of the next morning practice sessions, both X1 (+0.664±1.051 L) and X2 (+0.446±0.916 L) were in positive fluid balance, respectively. Ample fluid consumption opportunities, scaled down practice intensities during X2, and potentially intentional greater relative fluid intake during X2 training resulted in no difference in fluid shift versus an X1 schedule before the start of practices. The majority of players maintained fluid balance drinking ad libitum regardless of practice schedule.


Extracellular proton alters the divalent cation binding affinity in a cyclic nucleotide-gated channel pore.
Extracellular protons in the range of 10(-9) to 10(-5) M effectively suppressed Na+ current (K(1/2) = 10(-6.1)) through the bovine retinal guanosine 3',5'-cyclic mononucleotide-gated ion channel expressed in Xenopus oocytes. The reduction of channel current was mediated by a single glutamate residue (Glu363) within the pore-forming region of the channel, also involved in extracellular divalent cation binding. Increasing the concentration of extracellular proton decreased the binding affinity of the extracellular divalent cation (e.g. Sr2+) and the large difference of binding affinity previously observed between the wild-type and E363D mutant channel disappeared. These results indicate that the permeation characteristics of cyclic nucleotide-gated ion channel can be altered by extracellular pH through a single acidic residue in the channel conduction pathway.


Survival rates and outcomes of pregnancies with prenatal diagnosis of trisomy 18: A 16-year experience from a public hospital in South Africa.
Many studies, largely from high-income countries (HIC), have reported outcomes in babies with trisomy 18 (T18), with a paucity of data from Africa. Knowledge of outcomes is important in counselling women prenatally diagnosed with T18. We aimed to review all prenatally diagnosed cases of T18 between January 2006 and December 2021. Demographic data, diagnosis, gestation and outcome data were obtained from the Astraia® database and patient files. We included 88 pregnant women of whom 46 terminated their pregnancies (30 beyond 24 weeks' gestation). Three underwent foeticides, one had a caesarean section for maternal obstetric reasons and 26 underwent inductions of labour without foetal monitoring. Four neonates were live born but none lived >8 h. In those who continued their pregnancies, the mean gestation at delivery was 34.8 weeks, 14 (33%) were live births and only 5 survived for >24 h with none surviving to 1 year of life. In our cohort, infants with T18 had lower live birth rates and shorter survival than in the current literature from HIC. This may be due to the implementation of non-aggressive intrapartum care and comfort care for the neonates. This has implications for counselling in our setting.


Improved determination of plasmid copy number using quantitative real-time PCR for monitoring fermentation processes.
Recombinant protein production in Escherichia coli cells is a complex process, where among other parameters, plasmid copy number, structural and segregational stability of plasmid have an important impact on the success of productivity. It was recognised that a method for accurate and rapid quantification of plasmid copy number is necessary for optimization and better understanding of this process. Lately, qPCR is becoming the method of choice for this purpose. In the presented work, an improved qPCR method adopted for PCN determination in various fermentation processes was developed. To avoid experimental errors arising from irreproducible DNA isolation, whole cells, treated by heating at 95 degrees C for 10 minutes prior to storage at -20 degrees C, were used as a template source. Relative quantification, taking into account different amplification efficiencies of amplicons for chromosome and plasmid, was used in the PCN calculation. The best reproducibility was achieved when the efficiency estimated for specific amplicon, obtained within one run, was averaged. It was demonstrated that the quantification range of 2 log units (100 to 10000 bacteria per well) enable quantification in each time point during fermentation. The method was applied to study PCN variation in fermentation at 25 degrees C and the correlation between PCN and protein accumulation was established. Using whole cells as a template source and relative quantification considering different PCR amplification efficiencies are significant improvements of the qPCR method for PCN determination. Due to the approaches used, the method is suitable for PCN determination in fermentation processes using various media and conditions.


Rapid venous access using saphenous vein cutdown at the ankle.
Injured adults can usually be treated adequately with peripheral intravenous lines. However, in hypotensive patients, alternative techniques such as venous cutdown may be necessary. There are no adult studies that document the success rate or time required to complete this procedure in the emergent situation. During a 1-year period, 73 cutdowns were attempted on 56 patients. Sixty-two of these attempts resulted in venous cannulation with a free flow of fluid (84.9%). The average time required for placement was 4.9 minutes. First-year residents had a significantly lower success rate (70%) than emergency department staff (89%) or second- through fifth-year surgical residents (94%). One patient who lived developed a local cellulitis. There were no other complications. In the hands of an experienced operator, saphenous vein cutdown at the ankle is a reliable method of rapidly gaining venous access in the adult patient. There are few immediate or late complications if the catheter is removed within 24 hours.


Loss of Brca1 and Trp53 in adult mouse mammary ductal epithelium results in development of hormone receptor-positive or hormone receptor-negative tumors, depending on inactivation of Rb family proteins.
Breast cancer is a heterogenous disease with several histological and molecular subtypes. Models that represent these subtypes are essential for translational research aimed at improving clinical strategy for targeted therapeutics. Different combinations of genetic aberrations (Brca1 and Trp53 loss, and inhibition of proteins of the Rb family) were induced in the mammary gland by injection of adenovirus expressing Cre recombinase into the mammary ducts of adult genetically engineered mice. Mammary tumors with different genetic aberrations were classified into molecular subtypes based on expression of molecular markers and RNAseq analysis. In vitro potency assays and Western blots were used to examine their drug sensitivities. Induction of Brca1 and Trp53 loss in mammary ductal epithelium resulted in development of basal-like hormone receptor (HR)-negative mammary tumors. Inhibition of Rb and Trp53 loss or the combination of Rb, Trp53 and Brca1 aberrations resulted in development of luminal ductal carcinoma positive for ER, PR, and Her2 expression. HR positivity in tumors with Rb, Trp53 and Brca1 aberrations indicated that functionality of the Rb pathway rather than Brca1 status affected HR status in these models. Mammary tumor gene expression profiles recapitulated human basal-like or luminal B breast cancer signatures, but HR-positive luminal cancer models were endocrine resistant and exhibited upregulation of PI3K signaling and sensitivity to this pathway inhibition. Furthermore, both tumor subtypes were resistant to CDK4/6 inhibition. Examination of molecular expression profiles and drug sensitivities of tumors indicate that these breast cancer models can be utilized as a translational platform for evaluation of targeted combinations to improve chemotherapeutic response in patients that no longer respond to hormone therapy or that are resistant to CDK4/6 inhibition.


The Development of Neuroimaging Biomarkers for Cognitive Decline in Sickle Cell Disease.
Sickle cell disease (SCD) is complicated by neurologic complications including vasculopathy, hemorrhagic or ischemic overt stroke, silent cerebral infarcts and cognitive dysfunction. Patients with SCD, even in the absence of vasculopathy or stroke, have experience cognitive dysfunction that progresses with age. Transcranial Doppler ultrasound and structural brain MRI are currently used for primary and secondary stroke prevention, but laboratory or imaging biomarkers do not currently exist that are specific to the risk of cognitive dysfunction in patients with SCD. Recent investigations have used advanced MR sequences assessing cerebral hemodynamics, white matter microstructure and functional connectivity to better understand the pathophysiology of cognitive decline in SCD, with the long-term goal of developing neuroimaging biomarkers to be used in risk prediction algorithms and to assess the efficacy of treatment options for patients with SCD.


Transcription of the human adenovirus E1a gene in Saccharomyces cerevisiae.
The early region 1a (E1a) and its flanking sequences of human adenovirus type 5 (Ad5) have been cloned in the yeast-Escherichia coli shuttle vector YEp13 and transferred into the yeast Saccharomyces cerevisiae. The E1a-specific RNAs were produced in the transformed yeast cells. The 5' ends of these transcripts were capped but were lacking 10 to 45 nucleotides from the 5' end of the proper E1a mRNA. These transcripts terminated approx. 1000 nucleotides downstream from the proper 3' end. No splicing of the E1a-specific RNA could be detected in the yeast cells.


Tympanometry and isolated fracture of the stapes.
Temporal bone fractures are frequently associated with ossicular dislocations or fractures, most commonly involving the incus. To our knowledge, isolated fracture of the posterior crus of the stapes has not been previously reported. A 20-year-old man consulted for persistent left hypoacusis several months after a head injury. Initial computed tomography of the temporal bone showed a simple temporal bone fracture with no other associated abnormalities. The diagnosis of stapes fracture was suggested by increased compliance on tympanometry, leading to a second thin-section temporal bone computed tomography, which suggested a fracture of the posterior crus of the stapes. Endaural surgical exploration confirmed the diagnosis and allowed placement of ionomer cement in the posterior crus. The possibility of ossicular dislocation or fracture must be considered in patients with persistent conductive hearing loss associated with increased compliance on tympanometry, even when computed tomography of the temporal bone does not show dislocation of the ossicular chain or ossicular fracture. Hearing rehabilitation can be performed by hearing aid or surgical reconstruction of the ossicles.


Clinical pharmacokinetic/pharmacodynamic and physiologically based pharmacokinetic modeling in new drug development: the capecitabine experience.
Preclinical studies, along with Phase I, II, and III clinical trials demonstrate the pharmacokinetics, pharmacodynamics, safety and efficacy of a new drug under well controlled circumstances in relatively homogeneous populations. However, these types of studies generally do not answer important questions about variability in specific factors that predict pharmacokinetic and pharmacodynamic (PKPD) activity, in turn affecting safety and efficacy. Semi-physiological and clinical PKPD modeling and simulation offer the possibility of utilizing data obtained in the laboratory and the clinic to make accurate characterizations and predictions of PKPD activity in the target population, based on variability in predictive factors. Capecitabine is an orally administered pro-drug of 5-fluorouracil (5-FU), designed to exploit tissue-specific differences in metabolic enzyme activities in order to enhance efficacy and safety. It undergoes extensive metabolism in multiple physiologic compartments, and presents particular challenges for predicting pharmacokinetic and pharmacodynamic activity in humans. Clinical and physiologically based pharmacokinetic (PBPK) and pharmacodynamic models were developed to characterize the activity of capecitabine and its metabolites, and the clinical consequences under varying physiological conditions such as creatinine clearance or activity of key metabolic enzymes. The results of the modeling investigations were consistent with capecitabine's rational design as a triple pro-drug of 5-FU. This paper reviews and discusses the PKPD and PBPK modeling approaches used in capecitabine development to provide a more thorough understanding of what the key predictors of its PBPK activity are, and how variability in these predictors may affect its PKPD, and ultimately, clinical outcomes.


Expression of the autonomous parvovirus H1 genome: evidence for a single transcriptional unit and multiple spliced polyadenylated transcripts.
We identified viral transcripts in parvovirus H1-infected rodent cells using the S1 nuclease mapping technique of Berk and Sharp (1977, 1978). The most abundant viral transcript, present in both nucleus and cytoplasm, is approximately 2.8 kb long and represents about 56% of the viral genome. Less abundant viral transcripts of 3.0, 1.45 and 1.30 kb, and possibly other minor viral transcripts, are also detected in nuclear and cytoplasmic fractions. In contrast, a prominent 4.7 kb viral transcript which corresponds to 95% of the viral DNA is found only in the nucleus; this finding suggests that the parvovirus genome may function as a single transcription unit. Virus-infected cells pretreated with cycloheximide accumulate all these viral transcripts. Analyses of RNA-DNA hybrids (isolated from neutral agarose gels) by electrophoresis on alkaline agarose gels indicate that the 4.7, 3.0 and 2.8 kb viral transcripts are "spliced" RNAs. The nuclear-specific 4.7 kb transcript appears to be encoded by two noncontiguous DNA segments of 2.2 and 2.6 kb. The 3.0 and 2.8 kb transcripts are apparently encoded by.a 2.6 kb segment of DNA and one or more much smaller noncontiguous DNA segments, one of which is approximately 170 nucleotides long.


Proposal for diagnostic criteria for IgG4-related kidney disease.
IgG4-related disease has attracted wide attention recently. It is characterized by a high level of serum IgG4 and dense infiltration of IgG4-positive plasma cells into multiple organs, with the kidney being one representative target. Although several sets of diagnostic criteria for autoimmune pancreatitis (AIP) are available and renal lesion is recognized as an extra-pancreatic manifestation of AIP, it is difficult to differentiate IgG4-related tubulointerstitial nephritis (TIN) without AIP from other types of TIN. To clarify the entity of IgG4-related kidney disease (IgG4-RKD) and support in-depth studies, the Japanese Society of Nephrology has established a working group to prepare diagnostic criteria for IgG4-RKD. The working group analyzed 41 patients with IgG4-RKD, and collected the following data to devise a diagnostic algorithm and diagnostic criteria for IgG4-RKD: clinical features including extra-renal organ involvement, urinalysis and serological features including serum IgG4 levels, imaging findings demonstrated by computed tomography (CT), renal histology with IgG4 immunostaining, and response to steroid therapy. The conditions for criteria are as follows. (1) Presence of some kidney damage, as manifested by abnormal urinalysis or urine marker(s) and/or decreased kidney function with either elevated serum IgG level, hypocomplementemia, or elevated serum IgE level. (2) Kidney imaging studies showing abnormal renal imaging findings, i.e., multiple low density lesions on enhanced CT, diffuse kidney enlargement, hypovascular solitary mass in the kidney, and hypertrophic lesion of the renal pelvic wall without irregularity of the renal pelvic surface. (3) Serum IgG4 level exceeding 135 mg/dl. (4) Renal histology showing two abnormal findings: (a) dense lymphoplasmacytic infiltration with infiltrating IgG4-positive plasma cells >10/high power field (HPF) and/or ratio of IgG4-positive plasma cells/IgG positive plasma cells >40%. (b) Characteristic 'storiform' fibrosis surrounding nests of lymphocytes and/or plasma cells. (5) Extra-renal histology showing dense lymphoplasmacytic infiltration with infiltrating IgG4-positive plasma cells >10/HPF and/or ratio of IgG4-positive plasma cells/IgG-positive plasma cells >40%. The diagnosis is classified into 3 stages of definite, probable and possible according to the combinations of the above conditions. Thirty-nine cases (95.1%) were diagnosed with IgG4-RKD according to the criteria. The provisional criteria and algorithm appear to be useful for clarifying the entity of IgG4-RKD and seeking underlying IgG4-RKD cases; however, further experience is needed to confirm the validity of these criteria.


Synthesis and characterization of a paramagnetic sialic acid conjugate as probe for magnetic resonance applications.
Magnetic Resonance Imaging (MRI) using paramagnetic systems as contrast agents is receiving increased attention as diagnostic tool in the clinic. At the same time, NMR of paramagnetic systems can also be applied in biochemical fields; for example, the use of Paramagnetic Relaxation Enhancement (PRE) allows structure refinement and the analysis of transient dynamic processes involved in macromolecular complex formation. Herein we report the synthesis and computational characterization of a new DOTA-like sialic acid conjugate, which can be used both in MRI and PRE applications when coordinated to a suitable paramagnetic metal.


Isoquinoline Kelch-like ECH-Associated Protein 1-Nuclear Factor (Erythroid-Derived 2)-like 2 (KEAP1-NRF2) Inhibitors with High Metabolic Stability.
Pharmacological activation of NRF2 (nuclear factor erythroid 2-related factor 2) arises from blocking the interaction of NRF2 with its negative regulator, KEAP1 (Kelch-like ECH-associated protein 1). We previously reported an isoquinoline-based NRF2 activator, but this compound showed negative logD7.4 and a -2 charge at physiological pH, which may have limited its membrane permeability. In this work, we report potent, metabolically stable analogs that result from replacing a carboxymethyl group at the 4-position with a fluoroalkyl group.


The presence of platelet-activating factor binding sites in human myometrium and their role in uterine contraction.
It has recently been suggested that platelet-activating factor is involved in a number of processes associated with reproductive biology. We have demonstrated a critical role for this autacoid in the initiation and maintenance of parturition both in animals and man. In the present investigation, it has been demonstrated that platelet-activating factor affects contraction in strips of human myometrium at concentrations as low as 10(-10) mol/L. A platelet-activating factor receptor has been identified and characterized in human myometrium. Platelet-activating factor acts to cause an increase in intracellular Ca2+ concentration of isolated myometrial smooth muscle cells in culture and an increase in the phosphorylation of the 20 kd light chain of myosin in a concentration-dependent fashion. The presence of platelet-activating factor receptors in human myometrium, the observed increase in Ca2+ concentration and phosphorylation of myosin light chain in the presence of platelet-activating factor provide further support for the importance of this autacoid in the initiation of parturition.


Familial clustering of schizophrenia, bipolar disorder, and major depressive disorder.
To investigate familial clustering of schizophrenia, bipolar disorder, and major depressive disorder. Combining data from a psychiatric case registry and Statistics Netherlands provided information on 4,673 affected probands and 18,692 matched population controls. Probands with schizophrenia had relative risks (RRs) for having a sibling with schizophrenia of 3.77 (95% confidence interval (CI): 2.60-5.46) and with bipolar disorder of 1.79 (95% CI: 0.64-4.96) as compared with a reference proband. Probands affected with bipolar disorder have an RR of 6.51 (95% CI: 2.60-16.29) for having a sibling with bipolar disorder and of 1.71 (95% CI: 0.71-4.14) for having a sibling with schizophrenia as compared with a reference proband. Probands affected with major depressive disorder also have increased risk for having a sibling with schizophrenia (RR: 2.04, 95% CI: 1.54-2.72) as compared with a reference proband, which was similar to the risk for having a sibling with major depressive disorder (RR: 1.91, 95% CI: 1.63-2.24) or bipolar disorder (RR: 2.06, 95% CI: 1.18-3.60). Our findings suggest, as previous studies have, that risk across schizophrenia and bipolar disorder is considerably lower (twofold) than within diagnostic entities, whereas for major depressive disorder risk is similar within and across diagnostic entities.


Science, marketing and wishful thinking in quantitative proteomics.
In a recent editorial (J. Proteome Res. 2007, 6, 1633) and elsewhere questions have been raised regarding the lack of attention paid to good analytical practice with respect to the reporting of quantitative results in proteomics. Using those comments as a starting point, several issues are discussed that relate to the challenges involved in achieving adequate sampling with MS-based methods in order to generate valid data for large-scale studies. The discussion touches on the relationships that connect sampling depth and the power to detect protein abundance change, conflict of interest, and strategies to overcome bureaucratic obstacles that impede the use of peer-to-peer technologies for transfer and storage of large data files generated in such experiments.


The influence of crank rate on peak oxygen consumption during arm crank ergometry.
The aim of this study was to assess the influence of three imposed crank rates on the attainment of peak oxygen consumption (VO2peak) and other physiological responses during incremental arm crank ergometry. Twenty physically active, although non-specifically trained, males volunteered for the study. They completed an exercise protocol using an electrically braked arm ergometer (Lode Angio, Groningen, Netherlands) at crank rates of 60, 70 and 80 rev x min(-1). The order of tests was randomized and they were separated by at least 2 days. Peak VO2 was significantly higher (P < 0.05) at 70 and 80 rev x min(-1) than at 60 rev x min(-1). Peak ventilation volume increased as a function of crank rate and was higher (P < 0.05) at 80 than at 60 rev x min(-1). Peak heart rate was higher (P < 0.05) at 70 and 80 rev x min(-1) than at 60 rev x min(-1). Furthermore, 70 and 80 rev x min(-1) resulted in an extended test time compared with 60 rev x min(-1). The greater physiological responses observed during the tests at the two faster crank rates might have been the result of a postponement of acute localized neuromuscular fatigue, allowing for more work to be completed. We recommend, therefore, that an imposed crank rate between 70 and 80 rev x min(-1) should be used to elicit VO2peak and other physiological responses in arm crank ergometry.


Plasticity of the myelination genomic fabric.
This study aimed to quantify the influence of the astrocyte proximity on myelination genomic fabric (MYE) of oligodendrocytes, defined as the most interconnected and stably expressed gene web responsible for myelination. Such quantitation is important to evaluate whether astrocyte signaling may contribute to demyelination when impaired and remyelination when properly restored. For this, we compared changes in the gene expression profiles of immortalized precursor oligodendrocytes (Oli-neu), stimulated to differentiate by the proximity of nontouching astrocytes or treatment with db-cAMP. In a previous paper, we reported that the astrocyte proximity upregulated or turned-on a large number of myelination genes and substantially enriched the Ca(2+)-signaling and cytokine receptor regulatory networks of MYE in Oli-neu cells. Here, we introduce the "transcriptomic distance" to evaluate fabric remodeling and "pair-wise relevance" to identify the most influential gene pairs. Together with the prominence gene analysis used to select and rank the fabric genes, these novel analytical tools provide a comprehensively quantitative view of the physio/pathological transformations of the transcriptomic programs of myelinating cells. Applied to our data, the analyses revealed not only that the astrocyte neighborhood is a substantially more powerful regulator of myelination than the differentiating treatment but also the molecular mechanisms of the two differentiating paradigms are different. By inducing a profound remodeling of MYE and regulatory transcriptomic networks, the astrocyte-oligodendrocyte intercommunication may be considered as a major player in both pathophysiology and therapy of neurodegenerative diseases related to myelination.


Forebrain lesions and retention of imprinting in domestic chicks.
Newly-hatched domestic chicks were reared separately with one of two types of imprinting object hanging in their cages. On the 5th day discrimination tests demonstrated strong preferences for the familiar object. On the 6th day the chicks were subjected to bilateral aspiration lesions of either the anterior or the lateral forebrain and their imprinting objects removed from the cage. Discrimination tests were made on the 9th and 12th days, the imprinting objects replaced, and a final relearning test given on the 14th day. The anterior lesion group was slightly impaired in the retention and relearning tests while the lateral lesion group was drastically affected, being significantly worse than the anterior group and with choices only just significantly above random expectation in the retention tests and no different from random choice in the relearning test. It was concluded that a lateral forebrain region is necessary for retention and relearning of a previously acquired imprinted preference.


Development of locomotion in the rat: the significance of early movements.
The development of the nervous system is determined by an interaction between genetic and epigenetic factors. We investigated the possible role of proprioceptive afferent input in the development of locomotion in the rat. Kinematic analysis of locomotion in normal rats reveals a marked transition from immature overground locomotion into the adult pattern around the 15th postnatal day. Around this age, the timing of EMG activity in the tibialis anterior and the gastrocnemius muscle in the hindpaw gains accuracy and the intensity of the EMG increases. In the soleus muscle we observed an increased regularity in the shape of individual motor unit potentials. Neuroanatomical research revealed the occurrence of dendrite bundles in the motoneuronal pool of the soleus muscle pool from the 16th day as the result of a reorganisation. Immobilisation of one hindlimb in an extended position by casts around the leg from the first till the 20th postnatal day does not interfere with the development of dendrite bundles in the soleus motoneuronal pool. However, we demonstrated long lasting abnormalities in the timing of the EMG activation patterns in the gastrocnemius and the tibialis anterior muscles. It is conceivable that the experimental interference with afferent feed back is the causative factor of these functional abnormalities.


Clinicopathological Features and Prognosis of Resected Pancreatic Ductal Adenocarcinoma Patients with Claudin-18 Overexpression.
Claudin-18.2 (CLDN18.2) is specifically expressed in pancreatic precancerous lesions and pancreatic ductal adenocarcinoma (PDAC). We assessed the clinical characteristics of patients with CLDN18.2-overexpressing pancreatic cancer to identify patients who might benefit from CLDN18-targeted treatment. A total of 130 patients with surgically resected PDAC were investigated for the immunohistochemical expression of claudin-18 (CLDN18). The CLDN18 staining intensities (0-3+) and relative proportion of positive tumor cells were analyzed by two independent raters. Tumors positive for CLDN18 expression were defined as ≥80% of tumor cells with 2+ or 3+ staining intensity in a CLDN18 immunohistochemical assay. Positive CLDN18 expression was present in 41/130 (31.5%) surgically resected PDACs and the relative proportion of positive tumor cells and the staining intensity were directly correlated (p < 0.001). Positive CLDN18 expression was significantly associated with well-differentiated tumors (p < 0.001) and less regional node involvement (p = 0.045). The positive CLDN18-expressing group showed no statistical difference in median overall survival (17.4 months vs. 20.6 months, p = 0.770) compared to the negative CLDN18-expressing group. Distant nodal metastasis was more frequent in the positive CLDN18-expressing group (p = 0.011). CLDN18 is frequently expressed in PDAC, and high CLDN18-expressing PDACs showed some different clinicopathologic characteristics. High CLDN18 expression was not associated with prognosis in patients with surgically resected PDAC.


[Holistic treatment of endometriosis].
The pain phenomena caused by endometriosis are manifold. In addition to nociceptive pain there is also a nociplastic reaction with central sensitization. Atypical symptoms, such as acyclic lower abdominal pain, radiating pain, nonspecific bladder and intestinal complaints or even depression increasingly occur in addition to the classical cyclic complaints, such as severe dysmenorrhea, cyclic lower abdominal pain, dyspareunia, dysuria and dyschezia. Due to the diffuse range of symptoms, affected patients often consult not just gynecologists but also specialists from other disciplines (internal medicine, gastroenterology, orthopedics, pain therapy, psychology etc.). The complexity of endometriosis is presented. The resulting approaches to multimodal interdisciplinary holistic treatment are described. Interdisciplinary concepts should be involved in the optimal treatment of endometriosis patients along with hormonal and surgical treatment, mostly under the supervision of a gynecologist and pain management, dietary changes, psychological support and physiotherapeutic management should also be included. This article provides an overview of possible treatment strategies for chronic symptomatic endometriosis. Based on multimodal treatment strategies and regarding the complex pathophysiological alterations of this disease, the complex complaints that significantly impair the quality of life of endometriosis patients can be greatly improved.


A method for following patients with retrievable inferior vena cava filters: results and lessons learned from the first 1,100 patients.
Patients who have undergone implantation of a retrievable inferior vena cava (IVC) filter require continued follow-up to have the device removed when clinically appropriate and in a timely fashion to avoid potential long-term filter-related complications. The efficacy of a method for patient follow-up was evaluated based on a retrospective review of a single-institutional retrievable IVC filter experience. Patients with retrievable IVC filters were tracked via a prospectively collected database designed specifically for patient follow-up. Follow-up consisted of periodic review of the electronic medical record. Patients were contacted by mail (at regular intervals one or more times) when removal of the filter was deemed appropriate. A retrospective review of the ultimate fate of the first 1,127 retrievable IVC filters placed at a single institution was performed. Retrieval rates were compared with those seen in the initial experience, during which no structured follow-up was performed. Of 1,127 filters placed, 658 (58.4%) were removed. Filter removal or declaration of the device as permanent was achieved in 860 patients (76.3%). Filter removal, declaration of the device as permanent, or establishment of the need for continued follow-up was achieved in 941 patients (83.5%). Only 186 patients (16.5%) were lost to follow-up. The follow-up method described in the present study resulted in a statistically significant difference (P < .001) in the likelihood of a patient returning for IVC filter removal compared with a lack of follow-up (59% vs 24%).


Carcinogenicity of nickel subsulfide in Fischer rats and Syrian hamsters after administration by various routes.
In an endeavor to expand the variety of experimental models for study of nickel carcinogenesis, nickel subsulfide (Ni3S2) was administered to rodents by five previously untested routes. In two groups of Syrian hamsters, Ni3S2 induced multiple sarcomas at the sites of single im injections (5 or 10 mg of Ni3S2). In contrast, Ni3S2 did not induce any malignant tumors of the cheek pouches, oral cavity or gastrointestinal tract, despite multiple local applications to the cheek pouches of several groups of hamsters in total dosages as large as 1.1 g of Ni3S2. In a group of Fischer rats, single intratesticular injections of Ni3S2 (10 mg) induced many testicular sarcomas. In contrast, no malignant tumors developed in two groups of rats that received single injections into the submaxillary gland (2.5 mg of Ni3S2) or into the liver (5 mg of Ni3S2 via the portal venous system).


Versatile perfusion chamber for high-magnification microscopy.
The large number of culture or perfusion chambers reflects the necessity of researchers to develop them from time to time because of specific biologic or optical requirements. The paper described a new versatile perfusion chamber, which can be machined at low cost and with a very rapid assembly procedure. An additional interesting feature is the lock system with an annular threaded screw that fits the angle of the objective, thus increasing the field of observation.


Three-Component Coupling of Aromatic Aldehydes, 1-Morpholino-2-nitroalkenes, and 3-Aminoazoles via Boron Trifluoride Etherate Catalysis: Reaction Pathway and Features of the Formation of Intermediates.
4,7-Dihydro-6-nitro-7-Ar-5-R-azolo[1,5-a]pyrimidines were obtained by the multicomponent reaction of aminoazoles, morpholino-nitroalkenes, and aromatic aldehydes in the catalysis of boron trifluoride etherate. The optimal reaction conditions were determined, and the formation of the target regioisomer was demonstrated. The pathway for multicomponent transformation, including the formation of azolyl-nitroalkene, was determined. Morpholino-nitroalkenes were assumed to convert into the corresponding nitroalkynes during catalysis of boron trifluoride etherate. For a multicomponent reaction with 4-nitrobenzaldehyde, conditions have been proposed that exclude the formation of a side regioisomer.


Foot and ankle reconstruction with vertically designed deep inferior epigastric perforator flap.
The deep inferior epigastric perforator (DIEP) flap is one of the most commonly utilized flaps of reconstructive surgery. Although the horizontal flap design is the most commonly used, this flap can be vertically designed to avoid drawbacks such as excessive tissue dissection, relatively reduced flap perfusion, and scarification of the contralateral flap opportunity. The aim of this report is to present our case series for foot and ankle reconstruction with vertical designed DIEP flap. The free vertically designed DIEP flaps (VDIEP) were used in eight patients (7 male, 1 female) whose age is in a range of 20-66 years for soft tissue reconstructions in the ankle and foot region over a five-year period. The range of defects' size was from 8 × 5 cm to 15 × 7 cm and the causes were electrical burn, trauma and diabetic foot infections. Flap dimensions varied from 10 × 6 cm to 17 × 9 cm. All the flaps had two or more perforators, and all flaps survived completely. There were no early or late complications. We followed up the patients for 10 months in average. We observed no functional problems, especially in main motions of foot and ankle like eversion, inversion, flexion or extension except one patient. Donor site scars were acceptable in all patients. The VDIEP flap may be an option for selected lower extremity soft tissue reconstructions, and it may be an alternative to classically designed abdominal flaps.


The ICF as a conceptual platform to specify and discuss health and health-related concepts.
The World Health Organization's International Classification of Functioning, Disability and Health (ICF) has provided a new foundation for our understanding of health, functioning, and disability. As a content-valid, comprehensive and universally applicable health classification, it serves as a platform to clarify and specify health-related concepts that are frequently used in the medical literature. The health concepts to which we refer are: well-being, health status, quality of life (QoL) and health-related quality of life (HRQoL). The aim of this paper is to use the ICF as a conceptual platform to specify and discuss health-related concepts. The ICF entities health and health-related domains and functioning will be used as starting point to reach the objective of the paper. Health domains refer to domains intrinsic to the person as a physiological and psychological entity, such as mental functions, seeing functions, and mobility. Health-related domains are not part of a person's health but are so closely related that a description of a person's lived experience of health would be incomplete without them. Examples of health-related domains are work, education, and social activities. Functioning refers to all health and health-related domains within the ICF. Well-being is made up of health, health-related, and non-health-related domains, such as autonomy and integrity. Health state is a health profile that results from collecting together health domains. Functioning states is a profile that results from collecting both health and health-related domains. Health status is a summary measure of health state. Functioning status is a summary measure of functioning state. QoL is the individual's perceptions of how the life is going in health, health-related, and non-health domains. HRQoL is the individual's perceptions of how the life is going in health and health-related domains. "HRQoL is to QoL as functioning is to well-being". The ICF represents a standardized and international basis for the operationalization of health based on its health domains. It refers to the more restricted concepts of health state and health status. The ICF is also the basis for the operationalization of functioning based on all health and health-related domains contained therein. The authors argue that functioning is an operationalization of health from a broader perspective. It refers to an operational concept of health in terms of a set of health domains ('under the skin') and health-related domains ('outside the skin') that consider the individual person not only as a biological but also as a social entity. Health from this perspective refers to the broader notion of functioning state and functioning status. Nevertheless, the ICF provides more than a basis for the operationalization of health and functioning. The ICF also contains contextual factors.


A Fourier approach to pulse pile-up in photon-counting x-ray detectors.
An analytic Fourier approach to predict the expected number of counts registered in a photon-counting detector subject to pulse pile-up for arbitrary photon flux, detector response function, and pulse-shape is presented. The analysis provides a complete forward model for energy-sensitive, photon-counting x-ray detectors for spectral computed tomography. The formalism of the stochastic theory of the expected frequency of level crossings of shot noise processes is applied to the pulse pile-up effect and build on a recently published analytic Fourier representation of the level crossing frequency of shot noise processes with piece-wise continuous kernels with jumps. The general analytic result is validated by a Monte Carlo simulation for pulses of the form g(t) = e(-t/τ) (t > 0) and a Gaussian detector response function. The Monte Carlo simulations are in excellent agreement with the analytic predictions of photon counts within the numerical accuracy of the calculations. The phenomenon of pulse pile-up is identified with the level-crossing problem of shot noise processes and an exact, analytic formula for the expected number of counts in energy-sensitive, photon-counting x-ray detectors for arbitrary photon flux, response function, and pulse-shapes is derived. The framework serves as a theoretical foundation for future works on pulse pile-up.


Single-leg forward hopping exposures adversely affect knee joint health among persons with unilateral lower limb loss: A predictive model.
Single-leg hopping is an atypical, yet convenient, method of ambulation for individuals who have sustained unilateral lower limb-loss. Hopping is generally discouraged by therapists but many patients report hopping, and the potential deleterious effects of frequent hopping on knee joint health remains unclear. Mechanical fatigue due to repeated exposures to increased or abnormal loading on the intact limb is thought to be a primary contributor to the high prevalence of knee osteoarthritis among individuals with unilateral lower limb amputation. We aimed to compare knee joint mechanics between single-leg hopping and walking at self-selected paces among individuals with unilateral lower limb-loss, and estimated the associated probability of knee cartilage failure. Thirty-two males with traumatic unilateral lower limb-loss (22 transtibial, 10 transfemoral) hopped and walked at a self-selected pace along a 15-m walkway. Peak knee moments were input to a phenomenological model of cartilage fatigue to estimate the damage and long-term failure probability of the medial knee cartilage when hopping vs. walking. We estimate that each hop accumulates as much damage as at least 8 strides of walking (p < 0.001), and each meter of hopping accumulates as much damage as at least 12 m of walking (p < 0.001). The 30-year failure probability of the medial knee cartilage exceeded a "coin-flip" chance (50%) when performing more than 197 hops per day. Although a convenient mode of ambulation for persons with unilateral lower limb-loss, to mitigate risk for knee osteoarthritis it is advisable to minimize exposure to single-leg forward hopping.


Prenatal sonography for the detection of fetal anomalies: results of a prospective study and comparison with prior series.
Reported rates of sonographic detection of fetal anomaly vary widely. The purpose of our study was to determine how well we were detecting fetal anomaly in our region and to compare our results with published series using standardized criteria. Over a 2-year period, we compared the reports of sonographic studies done in 7880 pregnancies at 16-20 weeks' gestation with all pregnancy outcomes established by medical records and pathologic results of the infants, whether born alive or dead. Fetal anomaly was defined as any structural anomaly except for those specifically excluded. We compared our results, expressed as percentage of anomalies found and anomalies found per 1000 pregnancies screened, with those of published studies, modifying published figures where necessary to compensate for our defined exclusion criteria. At postnatal examination, we found 157 anomalies in 144 babies who had prenatal sonography at 16-20 weeks' gestation. We detected 93 anomalies (60%) in 84 fetuses, leading to termination of pregnancy in 42 cases (50% of anomalous fetuses). The prevalence of fetal anomaly in our population was 19.8 anomalies per 1000 pregnancies screened. We detected 11.8 anomalies per 1000 screened, leading to termination of 5.3 pregnancies per 1000 screened. Our success rate for fetal anomaly detection ranged from 92% for CNS anomalies to 31% for cardiac anomalies and 25% for craniofacial anomalies. There were fewer babies born with a major malformation in the group whose anomalies were detected prenatally (21%) than in the group whose anomalies were not detected (58%). By comparison, in prior series, prevalence of anomalies varied from 5.67 to 25.95 per-11000 screened; anomaly detection varied from 16.6% to 74.4%; detection rate varied from 3.08 to 11.03 per 1000 screened. There was no correlation between the percentage and per 1000 detection rates. Thus, the RADIUS and Helsinki trials had similar detection rates per 1000, but more than a twofold difference in percentage detection rate. Similarly, the percentage success rate in studies could be the same, but the detection rate per 1000 different threshold. There was a similar lack of correlation for rates of pregnancy termination. All series except for the RADIUS study reported a reduction in the rate of adverse outcome if the anomalies were detected prenatally. Our rate of detection of fetal anomaly is satisfactory for most organ systems but remains poor for cardiac, skeletal, and craniofacial anomalies. The relative ranking of the success of fetal anomaly detection in prior studies depends on whether the detection rate is reported as a percentage or as the number of anomalies detected per 1000 screened. Our results compare favorably with those reported in other series.


Type IV fimbrial subunit protein ApfA contributes to protection against porcine pleuropneumonia.
Porcine pleuropneumonia caused by Actinobacillus pleuropneumoniae accounts for serious economic losses in the pig farming industry worldwide. We examined here the immunogenicity and protective efficacy of the recombinant type IV fimbrial subunit protein ApfA as a single antigen vaccine against pleuropneumonia, or as a component of a multi-antigen preparation comprising five other recombinant antigens derived from key virulence factors of A. pleuropneumoniae (ApxIA, ApxIIA, ApxIIIA, ApxIVA and TbpB). Immunization of pigs with recombinant ApfA alone induced high levels of specific serum antibodies and provided partial protection against challenge with the heterologous A. pleuropneumoniae serotype 9 strain. This protection was higher than that engendered by vaccination with rApxIVA or rTbpB alone and similar to that observed after immunization with the tri-antigen combination of rApxIA, rApxIIA and rApxIIIA. In addition, rApfA improved the vaccination potential of the penta-antigen mixture of rApxIA, rApxIIA, rApxIIIA, rApxIVA and rTbpB proteins, where the hexa-antigen vaccine containing rApfA conferred a high level of protection on pigs against the disease. Moreover, when rApfA was used for vaccination alone or in combination with other antigens, such immunization reduced the number of pigs colonized with the challenge strain. These results indicate that ApfA could be a valuable component of an efficient subunit vaccine for the prevention of porcine pleuropneumonia.


Age and 5HT 2-receptor blockade with ketanserin in essential hypertension.
Platelet derived serotonin (5HT) contributes to blood pressure elevation and the development of thromboembolic complications. Among the pathophysiological mechanisms involved in these vascular events, derangements in 5HT kinetics and exaggerated platelet response to 5HT may be part of the major triggering factors. An age-dependent attenuation platelet 5HT kinetics was revealed in normotensive control subjects but not in patients with essential hypertension. In older hypertensive patients, particularly in men, platelet 5HT uptake was decreased. In parallel, platelet reactivity was increased with advancing age as shown by a greater 5HT induced platelet aggregation and higher plasma concentration of beta-thromboglobulin. Antihypertensive treatment with the 5HT2-receptor antagonist ketanserin attenuated platelet 5HT turnover and inhibited 5HT induced platelet aggregation; the latter was more pronounced in older patients. The clinical efficacy and tolerability of ketanserin 20-40 mg twice daily given as mono- or combination therapy was evaluated in 188 patients with mild to moderate essential hypertension for a period of 12 weeks. A greater fraction of patients greater than or equal to 60 years achieved diastolic target pressure of less than or equal to 95 mgHg. Complaints related to the central nervous system or the peripheral circulation were greatly reduced in patients older than 60 years. In older patients, over and above the antihypertensive effect, 5HT2-receptor blockade may play an important role in the prevention of thromboembolic complications by inhibition of 5HT induced platelet activation.


Discovery of a novel melanin concentrating hormone receptor 1 (MCHR1) antagonist with reduced hERG inhibition.
An initial SAR study resulted in the identification of the novel, potent MCHR1 antagonist 2. After further profiling, compound 2 was discovered to be a potent inhibitor of the hERG potassium channel, which prevented its further development. Additional optimization of this structure resulted in the discovery of the potent MCHR1 antagonist 11 with a dramatically reduced hERG liability. The decrease in hERG activity was confirmed by several in vivo preclinical cardiovascular studies examining QT prolongation. This compound demonstrated good selectivity for MCHR1 and possessed good pharmacokinetic properties across preclinical species. Compound 11 was also efficacious in reducing body weight in two in vivo mouse models. This compound was selected for clinical evaluation and was given the code AMG 076.


Alternative and non-hormonal treatments to symptoms of menopause.
Ovarian function can start to decline a few years before the eventual cessation of the menstrual cycle. The average age of menopause in the UK is 51 years, and it is a retrospective diagnosis after 12 months of amenorrhoea. Women can experience many symptoms such as vasomotor and vulvovaginal symptoms that impact them physically, psychologically, sexually and thus their overall wellbeing. Women may have medical contraindications to hormonal therapy or may prefer non-hormonal or alternative treatments. This review looks at the evidence, efficacy, and safety of a range of complementary or alternative treatments and non-hormonal pharmacological treatments for the treatment of vasomotor symptoms and vulvovaginal atrophy of menopause.


Multivariate framework for the assessment of key forcing to Lake Malawi level variations in non-stationary frequency analysis.
Lake Malawi in south eastern Africa is a very important freshwater system for the socio-economic development of the riparian countries and communities. The lake has however experienced considerable recession in the levels in recent years. Consequently, frequency analyses of the lake levels premised on time-invariance (or stationarity) in the parameters of the underlying probability distribution functions (pdfs) can no longer be assumed. In this study, the role of hydroclimate forcing factors (rainfall, lake evaporation, and inflowing discharge) and low frequency climate variability indicators (e.g., El Nino Southern Oscillation-ENSO and the Indian Ocean Dipole Mode-IODM) on lake level variations is investigated using a monthly mean lake level dataset from 1899 to 2017. Non-stationarity in the lake levels was tested and confirmed using the Mann-Kendall trend test (α = 0.05 level) for the first moment and the F test for the second moment (α = 0.05 level). Change points in the series were identified using the Mann-Whitney-Pettit test. The study also compared stationary and non-stationary lake level frequency during 1961 to 2004, the common period where data were available for all the forcing factors considered. Annual maximum series (AMS) and peak over threshold (POT) analysis were conducted by fitting various candidate extreme value distributions (EVD) and parameter fitting methods. The Akaike information criteria (AIC), Bayesian information criteria (BIC), deviance information criteria (DIC), and likelihood ratios (RL) served as model evaluation criteria. Under stationary conditions, the AMS when fitted to the generalized extreme value (GEV) distribution with maximum likelihood estimation (MLE) was found to be superior to POT analysis. For the non-stationary models, open water evaporation as a covariate of the lake levels with the GEV and MLE was found to have the most influence on the lake level variations as compared with rainfall, discharge, and the low frequency climatic forcing. The results are very critical in flood zoning especially with various planned infrastructural developments around the lakeshore.


Regulation of the expression of genes encoding types I, II, and III collagen during chick embryonic development.
During the embryonic development of the chicken, stimulation of production of collagen-enriched tissue such as bone matrix, cartilage matrix, and skin dermis occurs between day 7 and day 15. We have examined the levels of the RNAs encoding the interstitial collagens (types I, II, and III) to determine if this developmental progress is associated with increased accumulation of collagen RNA. Using cell-free translation and DNA:RNA hybridization techniques, we have detected a coordinate enhancement of approximately 10-fold in the steady state levels of whole embryo RNAs encoding the alpha 1(I), alpha 2(I), alpha 1(II), and alpha 1(III) collagens between 5 and 10 days of chick embryonic development. The developmental pattern of expression of these collagen genes in whole embryos is in marked contrast to that of two noncollagenous proteins, cellular fibronectin and beta-actin, whose RNA levels were not found to change dramatically during early chicken development. In addition, we have observed that at least 3 of the 4 collagen genes examined were expressed at low levels beginning between 1-2 days. Both 5'- and 3'-specific alpha 2(I) collagen gene DNA probes hybridized to early stage and late stage chick embryo RNAs of identical sizes. DNase I-hypersensitive sites have been detected near the 5' end of the alpha 2(I) collagen gene in chromatin isolated from both 2-day and 5-day embryos, representing developmental time points well before and at the threshold of the onset of enhanced collagen RNA synthesis, respectively. These results suggest that the same gene is expressed in early and late chick embryos to yield alpha 2(I) collagen RNAs of similar structure.


Computer simulations: tools for population and evolutionary genetics.
Computer simulations are excellent tools for understanding the evolutionary and genetic consequences of complex processes whose interactions cannot be analytically predicted. Simulations have traditionally been used in population genetics by a fairly small community with programming expertise, but the recent availability of dozens of sophisticated, customizable software packages for simulation now makes simulation an accessible option for researchers in many fields. The in silico genetic data produced by simulations, along with greater availability of population-genomics data, are transforming genetic epidemiology, anthropology, evolutionary and population genetics and conservation. In this Review of the state-of-the-art of simulation software, we identify applications of simulations, evaluate simulator capabilities, provide a guide for their use and summarize future directions.


Evidence for an essential histidine in carboxypeptidase Y. Reaction with the chloromethyl ketone derivative of benzyloxycarbonyl-L-phenylalanine.
The possible role of histidine residues in the catalytic function of carboxypeptidase Y from bakers' yeast has been investigated using site-specific reagents. Among the reagents tested, benzyloxy-L-phenylalanylchloromethane (Z-PheCH2Cl) was the most powerful inhibitor of the enzyme. It irreversibly inactivated both the peptidase and esterase activities with an apparent second order rate constant of 3.8 M-minus 1 S-minus 1; the D isomer caused essentially no effect on either activity. Inhibition by L-Z-PheCH2Cl, the reaction retarded by certain competitive inhibitors of the enzyme. Using radioactive L-Z-PheCH2Cl, the reaction with the enzyme was shown to be essentially stoichiometric. Diisopropylphosphorofluoridate (iPr2PF)-inactivated enzyme failed to react with Z-PheCH2Cl, and conversely, the Z-PheCH2Cl-inhibited enzyme failed to react with radioactive iPr2PF. Amino acid analyses of the Z-PheCH2Cl-inactivated enzyme revealed the loss of essentially 1 residue, with a concomitant yield of a 0.62 residue of N-t-carboxymethylhistidine. Since carboxypeptidase Y has a reactive serine at its active center, we concluded from these results that the mechanism involves a charge-relay system in the hydrolysis of peptide and ester substrates, as in chymotrypsin. An -SH group of carboxypeptidase Y was not affected during the reaction with L-Z-PheCH2Cl. The generic name "serine carboxypeptidase" has been proposed for carboxypeptidase Y and for the iPr2PF-sensitive carboxypeptidases from plants, molds, and animal tissues, in order to distinguish them from "metal carboxypeptidase" to which carboxypeptidase A (EC 184.108.40.206) and B (EC 184.108.40.206) belong.


Genetic influences on epilepsy outcomes: A whole-exome sequencing and health care records data linkage study.
This study was undertaken to develop a novel pathway linking genetic data with routinely collected data for people with epilepsy, and to analyze the influence of rare, deleterious genetic variants on epilepsy outcomes. We linked whole-exome sequencing (WES) data with routinely collected primary and secondary care data and natural language processing (NLP)-derived seizure frequency information for people with epilepsy within the Secure Anonymised Information Linkage Databank. The study participants were adults who had consented to participate in the Swansea Neurology Biobank, Wales, between 2016 and 2018. DNA sequencing was carried out as part of the Epi25 collaboration. For each individual, we calculated the total number and cumulative burden of rare and predicted deleterious genetic variants and the total of rare and deleterious variants in epilepsy and drug metabolism genes. We compared these measures with the following outcomes: (1) no unscheduled hospital admissions versus unscheduled admissions for epilepsy, (2) antiseizure medication (ASM) monotherapy versus polytherapy, and (3) at least 1 year of seizure freedom versus <1 year of seizure freedom. We linked genetic data for 107 individuals with epilepsy (52% female) to electronic health records. Twenty-six percent had unscheduled hospital admissions, and 70% were prescribed ASM polytherapy. Seizure frequency information was linked for 100 individuals, and 10 were seizure-free. There was no significant difference between the outcome groups in terms of the exome-wide and gene-based burden of rare and deleterious genetic variants. We successfully uploaded, annotated, and linked genetic sequence data and NLP-derived seizure frequency data to anonymized health care records in this proof-of-concept study. We did not detect a genetic influence on real-world epilepsy outcomes, but our study was limited by a small sample size. Future studies will require larger (WES) data to establish genetic variant contribution to epilepsy outcomes.


Monolithically, widely tunable quantum cascade lasers based on a heterogeneous active region design.
Quantum cascade lasers (QCLs) have become important laser sources for accessing the mid-infrared (mid-IR) spectral range, achieving watt-level continuous wave operation in a compact package at room temperature. However, up to now, wavelength tuning, which is desirable for most applications, has relied on external cavity feedback or exhibited a limited monolithic tuning range. Here we demonstrate a widely tunable QCL source over the 6.2 to 9.1 μm wavelength range with a single emitting aperture by integrating an eight-laser sampled grating distributed feedback laser array with an on-chip beam combiner. The laser gain medium is based on a five-core heterogeneous QCL wafer. A compact tunable laser system was built to drive the individual lasers within the array and produce any desired wavelength within the available spectral range. A rapid, broadband spectral measurement (520 cm(-1)) of methane using the tunable laser source shows excellent agreement to a measurement made using a standard low-speed infrared spectrometer. This monolithic, widely tunable laser technology is compact, with no moving parts, and will open new opportunities for MIR spectroscopy and chemical sensing.