Hepatitis C virus in autoimmune liver disease in the UK: aetiological agent or artefact?
Hepatitis C virus antibody titres (anti-HCV) were measured in serum from 122 patients with autoimmune liver disease (96 with primary biliary cirrhosis and 26 with autoimmune chronic active hepatitis using three generations of enzyme immunoassay (EIA): first generation--Ortho, EIA1; second generation--Abbott, EIA2; and third generation--Murex, EIA3. Anti-HCV was below the positive cut-off level in all 26 autoimmune chronic active hepatitis patients for all tests, while seropositivity values in primary biliary cirrhosis were 31% (EIA1), 14% (EIA2), and 0% (EIA3). In primary biliary cirrhosis, anti-HCV values as measured by all three tests correlated positively with serum IgG concentrations, serum storage time, and a number of other indices of hepatic dysfunction. Multiple regression analysis showed that anti-HCV values were independently affected by both serum IgG and the length of storage time, although the magnitude of the effects varied between tests. When all three multiple regression models were applied to an extreme clinical example, however, EIA3 was least likely to give a false-positive result. The difference in test performance was emphasised further by examination of anti-HCV values in nine primary biliary cirrhosis patients (confirmed negative by recombinant immunoblot assay 2) in whom serial samples were tested (seven to 14 per patient, stored for one to 138 months). Apparent anti-HCV values (EIA1 and EIA2) increased with increasing serum storage time, but were unchanged when measured by EIA3. A similar pattern was evident in a limited study of five autoimmune chronic active hepatitis patients. As the second generation EIA is in widespread use and confirmatory testing is not always available, the effect of serum storage in addition to hyperglobulinaemia should be considered in the interpretation of positive results in auto immune and in other types of chronic liver disease.


Ruthenium(ii)-catalyzed selective C-H difluoroalkylation of aniline derivatives with pyrimidyl auxiliaries.
A ruthenium-catalyzed alternative para- and meta-difluoroalkylation of anilines is herein reported. The reaction tolerates a broad range of aniline derivatives and provides a convenient approach for accessing the corresponding para/meta-selective difluoroalkylated products. Mechanism studies demonstrated that the initial CAr-H and N-H cycloruthenation is the pivotal step in achieving remote C-H difluoroacetylation.


Isoform-specific O-glycosylation of osteopontin and bone sialoprotein by polypeptide N-acetylgalactosaminyltransferase-1.
Mucin-type O-glycan biosynthesis is regulated by the family of UDP-GalNAc polypeptide:N-acetylgalactosaminlytransfersases (ppGalNAcTs) that catalyzes the first step in the pathway by transferring GalNAc to Ser or Thr residues in a protein from the sugar donor UDP-GalNAc. Because not all Ser/Thr residues are glycosylated, rules must exist that signal which hydroyxamino acids acquire sugar. To date, no universal consensus signal has emerged. Therefore, strategies to deduce the subset of proteins that will be glycosylated by distinct ppGalNAcTs must be developed. Mucin-type O-glycoproteins are present abundantly in bone, where we found multiple ppGalNAcT isoforms, including ppGalNAcT-1, to be highly expressed. Thus, we compared glycoproteins expressed in wild-type and Galnt1-null mice to identify bone-associated proteins that were glycosylated in a ppGalNAcT-1-dependent manner. A reduction in the apparent molecular masses of two SIBLINGs (small integrin binding ligand N-linked glycoproteins), osteopontin (OPN) and bone sialoprotein (BSP) in the Galnt1-null mice relative to those of the wild-type was observed. Several synthetic peptides derived from OPN and BSP sequences were designed to include either known or predicted (in silico) glycosylation sites. In vitro glycosylation assays of these peptides with recombinant ppGalNAcT-1, ppGalNAcT-2, or ppGalNAcT-3 demonstrated that both SIBLINGs contained Thr/Ser residues that were preferentially glycosylated by ppGalNAcT-1. In addition, lysates prepared from wild-type, but not those from Galnt1-null derived osteoblasts, could glycosylate these peptides efficiently, suggesting that OPN and BSP contain sites that are specific for ppGalNAcT-1. Our study presents a novel and systematic approach for identification of isoform-specific substrates of the ppGalNAcT family and suggests ppGalNAcT-1 to be indispensable for O-glycosylation at specific sites of the bone glycoproteins OPN and BSP.


[Children and adolescents with hepatitis C virus infection].
Prevalence of infection with hepatitis C virus (HCV) is lower in children than in adults. The detection rate of anti-HCV antibodies in Western countries is 0.1-0.4% among children and adolescents. Prevalence of serologic response is higher in risk groups. HCV infection in children is usually asymptomatic, most of them have variations in serum levels of alanine aminotransferase (ALA). The laboratory exams for children are the same as those for adults. Histological progression may be faster in children than in adults. In this age group, HCV infection is considered as a special category, in which case it's possible to maintain the patient in observation without antiviral therapy. However, some studies with monotherapy showed that a regime with 1.75-3 MU/m2 of interferon alpha during 6-12 months induces a sustained viral response in 33-56% of the children. Although ribavirin hasn't yet been accepted for pediatric use, there have been several clinical tests in small groups with oral doses of 15 mg/kg a day, combined with interferon, during 12 months. The results are good. Pegylated interferon alpha is not authorized for pediatric use.


[Expression and significance of integrin alpha5, beta1 and E-CD in patients with non-small cell lung carcinoma].
To investigate the relationship between the expressions of integrin alpha5 beta1 and E-CD, and clinicopathological characteristics and prognosis of patients with non-small cell lung carcinoma (NSCLC). The expression of integrin alpha5 beta1 and E-CD were analyzed in 53 NSCLC and 12 control specimens by immunohistochemical assay. The expression of integrin alpha5 beta1 was significantly higher in NSCLC (58.5%) than that in normal lung tissue (16.7%), and also positively related with pathological characteristics (P = 0.021), lymph node metastasis (P = 0.006), and clinical stage (P = 0.002). The 3-year survival rate in NSCLC group was significantly lower than that in control group (22.3% vs 40.6% , P = 0.041). The positive expression of E-CD in NSCLC and control group was 32.1% and 91.7%, respectively, and negatively correlated with pathological characteristics (P = 0.010) and lymph node metastasis (P = 0.002). The 3-year survival rate in control group was 19.9%, lower than that in NSCLC group (41.2%, P > 0.05), but the difference is not significant. The overexpression of integrin alpha5 beta1 may contribute to lymph node metastasis and play an inverse role, while E-CD may be a beneficial prognostic factor in patients with NSCLC.


Reconstitution of human epidermis in vitro is accompanied by transient activation of basal keratinocyte spreading.
Frozen human cadaver skin obtained from the skin bank was thawed and incubated in serum-free medium for 1-2 days, after which the original epidermis could be removed mechanically. Transmission electron microscopic observations showed that the dermal matrix remaining behind contained intact bundles of collagen fibrils but no live cells and that a continuous lamina densa persisted in the basement membrane region. Indirect immunofluorescence analyses demonstrated linear staining of the basement membrane region by antibodies against laminin and type IV collagen and discontinuous staining with antibodies against fibronectin. Scanning electron microscopic observations revealed a normal topographical arrangement of dermal matrix papilla and interspersed crypts on the surface of the matrix. Epidermal cells placed on the dermal matrix attached in 1-2 h and spread by 24 h. After 1 week of culture the epidermis was reconstituted, at which time approximately 30% of the epidermal cells were basal keratinocytes and the remainder were more differentiated keratinocytes. A high degree of differentiation of the reconstituted epidermis was shown by the formation of hemidesmosomes along the basement membrane, the formation of desmosomes characterized by intercellular dense lines, and the presence of a cell layer containing keratohyalin granules. At various times during epidermal reconstitution, cells were harvested and tested in short-term assays for adhesion to fibronectin substrata. During the first several days there was a transient activation of basal keratinocyte spreading analogous to the modulation of keratinocyte spreading that we have observed during epidermal reconstitution in vivo.


Natural Course of Paravalvular Regurgitation After Implantation of the Self-Expanding CoreValve: Insights From Serial TEE Measurements.
This study investigates the natural course of paravalvular regurgitation (PAR) with serial transesophageal echocardiography (TEE) measurements. TEE studies were performed at 30 days and 1 year post TAVI with the self-expanding CoreValve for the treatment of severe aortic valve stenosis in 50 patients. In addition to conventional measurements, PAR perimeter and orifice area were assessed in the cross-sectional short-axis view at the level of the native aortic annulus. At 30 days, PAR was classified as none in 26 patients (52%), mild in 19 patients (38%) and moderate in 5 patients (10%). Between 30 days and 1 year, the number and size of PAR jets decreased and PAR was absent in 30 patients (60%) (P=.58 compared with 30-day results). Paravalvular regurgitation perimeter decreased from 8.2 ± 10.9% to 4.7 ± 7.7% (P<.01), a relative reduction of 43%. Cross-sectional area of regurgitation decreased from 0.22 ± 0.36 cm² to 0.12 ± 0.20 cm² (P=.01), a relative reduction of 45%. This improvement was observed in patients with mild and moderate PAR. No patient without PAR at 30 days developed PAR at 1-year follow-up. PAR perimeter and area as visualized by Color-Doppler TEE in the cross-sectional view decreased by about 45% between 30 days and 1 year post implantation of the self-expanding CoreValve.


The Glidescope system: a clinical assessment of performance.
The Glidescope is a new videolaryngoscope. It has a digital camera incorporated in the blade which displays a view of the vocal cords on a monitor. This allows the placement of a tracheal tube to be visualised. We describe its performance in 50 patients who required orotracheal intubation for elective surgery. Two investigators performed 25 intubations each. Intubation with the Glidescope was successful in 47 of the 50 cases. The three failures occurred early in the series and were attributed to the initial learning curve. The success rate after the first eight patients in each series was 100%. The median (IQR [range]) time to intubation was 40 (30-55 [15-105]) s. The Glidescope provided a grade I view of the glottis in 44 cases and a grade II view in six cases. The view of the larynx was improved in almost half (23) of the cases. The Glidescope improved the view by one grade in 22 and by two grades in one patient. We conclude that the Glidescope is an effective device for tracheal intubation and provides an improved view of the larynx. Further clinical studies are necessary to evaluate its role in airways that are difficult to manage.


The cardiac ventricular defibrillation threshold: inherent limitations in its application and interpretation.
A quantity termed the "threshold" has been used to describe the results of electrical ventricular defibrillation studies, with the implication of a clear distinction between ineffective and effective shock intensities. Although several definitions of the threshold have been suggested, and various methods have been used to quantify it, no comparison of the accuracies of the various methods could be found in the literature. This article, after presenting a method of applying basic probability theory to an assumed distribution relating probability of successful defibrillation to current amplitude, uses the method to examine several popular algorithms for defibrillation-threshold determination. The results show that where a sharp transition exists from ineffective to effective current amplitudes, most algorithms yield fairly good results. Where that transition is gradual (as it appears to be in all of the published reports examined), the algorithms are shown to be inadequate.


The conserved U.G pair in the 5' splice site duplex of a group I intron is required in the first but not the second step of self-splicing.
Group I self-splicing introns have a 5' splice site duplex (P1) that contains a single conserved base pair (U.G). The U is the last nucleotide of the 5' exon, and the G is part of the internal guide sequence within the intron. Using site-specific mutagenesis and analysis of the rate and accuracy of splicing of the Tetrahymena thermophila group I intron, we found that both the U and the G of the U.G pair are important for the first step of self-splicing (attack of GTP at the 5' splice site). Mutation of the U to a purine activated cryptic 5' splice sites in which a U.G pair was restored; this result emphasizes the preference for a U.G at the splice site. Nevertheless, some splicing persisted at the normal site after introduction of a purine, suggesting that position within the P1 helix is another determinant of 5' splice site choice. When the U was changed to a C, the accuracy of splicing was not affected, but the Km for GTP was increased by a factor of 15 and the catalytic rate constant was decreased by a factor of 7. Substitution of U.A, U.U, G.G, or A.G for the conserved U.G decreased the rate of splicing by an even greater amount. In contrast, mutation of the conserved G enhanced the second step of splicing, as evidenced by a trans-splicing assay. Furthermore, a free 5' exon ending in A or C instead of the conserved U underwent efficient ligation. Thus, unlike the remainder of the P1 helix, which functions in both the first and second steps of self-splicing, the conserved U.G appears to be important only for the first step.


Causes of poor outcome after cataract surgery in Satkhira district, Bangladesh.
Recent data have raised concerns about visual outcome following cataract surgery. The aim of this study was to assess the frequency and causes of poor and borderline outcome after cataract surgery in a population-based case series in Satkhira district, Bangladesh. A population-based case series was conducted within a population-based cluster survey of people aged over 50 years in Satkhira district where 4868 people underwent visual acuity (VA) screening. Eyes operated for cataract with VA<6/18 were examined in detail by an ophthalmologist, including a full history and dilated fundoscopy, to determine the cause of the visual outcome. Cataract surgery was performed on 213 eyes. Outcome was good (VA>6/18) for 128 eyes (60.1%), borderline (VA<6/18 to 6/60) for 35 eyes (16.4%), and poor (VA<6/60) for 50 eyes (23.5%) with available correction. Borderline and poor outcomes were most commonly due to lack of spectacles (25.8%), poor selection (33.8%), or surgical complications (30.6%). Surgical sequelae, namely posterior capsule opacification, was a less common cause of poor or borderline outcome (9.7%). Quality of surgical outcomes is of concern in Satkhira district. Increased emphasis on selection of subjects for surgery, provision of spectacles, and monitoring of surgery may improve outcomes.


Evaluation of two neck ultrasound measurements as predictors of difficult direct laryngoscopy: A prospective observational study.
Unpredictable difficult laryngoscopy remains a challenge for anaesthesiologists, especially if difficult ventilation occurs during standard laryngoscopy. Accurate airway assessment should always be performed, but the common clinical screening tests have shown low sensitivity and specificity with a limited predictive value. Ultrasound-based airway assessment has been proposed recently as a useful, simple, noninvasive bedside tool as an adjunct to clinical methods, but to date, few studies are available about the potential role of ultrasound in difficult airway evaluation, and these are mostly limited to specific groups of patients. The aim of this study was to determine the correlation between the sonographic measurements of anterior cervical soft tissues thickness and Cormack-Lehane grade view at direct laryngoscopy in patients with normal clinical screening tests. Prospective, single blinded, observational study. Operating theatre of a teaching hospital from May 2017 to September 2017. A total of 301 patients at least 18 years of age undergoing elective surgery under general anaesthesia with tracheal intubation were included in the study. Pre-operative evaluation was performed before surgery, demographic variables were collected and clinical screening tests to predict a difficult airway were performed. Patients with predicted difficult intubation were excluded. A 10 to 13-MHz linear ultrasound transducer was placed in the transverse plane and the thickness of the anterior cervical soft tissues was measured at two levels [thyrohyoid membrane (pre-epiglottic space) and vocal cords (laryngeal inlet)] with the patient's head in a neutral position. At each level, the distance from the skin in the median axis and the surrounding area was measured. The laryngoscopic view was graded by a different anaesthetist with more than 5 years of experience with direct laryngoscopy, blinded to the ultrasound assessments. The 'pre-epiglottic space thickness' at the level of thyrohyoid membrane was measured as the median distance from skin to epiglottis (mDSE) and the pre-epiglottic area was calculated; the mDSE cut-off value of 2.54 cm (sensitivity 82%, specificity 91%) and the pre-epiglottic area cut-off value of 5.04 cm (sensitivity 85%, specificity 88%) were the best predictors of a Cormack-Lehane grade at least 2b at direct laryngoscopy and of difficult intubation. The cut-off value of mDSE showed greater sensitivity in female patients (94 vs. 86%) and greater specificity in male patients (92 vs. 83%). No correlation was found between difficult laryngoscopy and ultrasound assessments at the level of the vocal cords. Airways ultrasounds might be considered as a predictor of restricted/difficult laryngoscopy and unpredicted difficult intubation. ClinicalTrials.gov: NCT03547193.


Raeder's syndrome [corrected]: paratrigeminal paralysis of the oculopupillary sympathetic system.
Raeder described five patients with mixed features of trigeminal nerve pathology and oculosympathetic impairment, with or without other cranial nerve lesions. This constellation of clinical features drew the original author's attention to the paratrigeminal region as a likely site for the causative lesion in this syndrome. An analysis of the anatomy of the oculosympathetic innervation supports the view that a restricted lesion in the middle cranial fossa might cause the syndrome of trigeminal nerve involvement, neuralgic pain or sensory change, with ptosis or miosis, or both, but no anhidrosis. Such a paratrigeminal oculosympathetic syndrome (POSS) usefully reminds clinicians to pursue vigorously possible lesions of the middle cranial fossa with careful, and possibly repeated, imaging studies. Attaching the eponym Raeder's syndrome or Raeder's paratrigeminal neuralgia to this syndrome adds nothing valuable to the anatomical description (POSS), which might be preferred for clarity.


Trimethoprim sulphamoxole in the treatment of chancroid. Comparison of two single dose treatment regimens with a five day regimen.
In a prospective blinded study, 135 men with genital ulcers culture positive for Haemophilus ducreyi, were randomized to one of three regimens. Two single dose regimens, either the combination of sulphamoxole 3200 mg/trimethoprim 640 mg or trimethoprim 700 mg alone were compared to a five day regimen of sulphamoxole 800 mg/trimethoprim 160 mg twice daily. All 31 treated with a five day regimen of trimethoprim sulphamoxole healed without further treatment. Of 27 patients treated with the single dose sulphamoxole/trimethoprim regimen, only 21 were cured and of 34 treated with trimethoprim alone, 25 responded. Antibacterial susceptibilities were performed on 31 H. ducreyi isolates. The laboratory susceptibility of these strains to trimethoprim correlated with the clinical response to the single agent. Trimethoprim alone in a dose of 700 mg or the combination of sulphamoxole (3200 mg) and trimethoprim (640 mg) is not satisfactory for the single dose treatment of genital ulcer disease. However, when prescribed for five days, sulphamoxole/trimethoprim is effective and compares favourably with other treatment regimens.


Risk factors for inguinal hernia in middle-aged and elderly men: results from the Rotterdam Study.
Prospective data on risk factors and the incidence of inguinal hernia are sparse, especially in an elderly population. The aim of this study was to determine the incidence of and risk factors for inguinal hernia. We analyzed data from the Rotterdam Study, a prospective cohort study that observed the general population aged ≥45 years of Ommoord, a district in Rotterdam, from baseline (1990) over a period of >20 years. Diagnoses of inguinal hernia were obtained from hospital discharge records and records from general practitioners. Multivariate regression analysis was performed to determine risk factors for inguinal hernia development. Among 5,780 men, with a total of 50,802 person-years, who did not have a hernia at baseline, 416 cases of inguinal hernia (7.2%) occurred. The 20-year cumulative incidence was 14%. Age-adjusted hazard ratio (HR) for inguinal hernia for men relative to women was 12.4 (95% CI, 9.5-16.3; P < .001). On multivariate analysis, the risk of inguinal hernia increased with advancing age (HR per 1-year increase in age, 1.03; 95% CI, 1.02-1.04; P < .001). Participants with a body mass index (BMI) of 25-30 kg/m2 had an HR of 0.72 (95% CI, 0.58-0.89; P = .003) compared with a BMI of <25; a BMI of >30 had an associated HR of 0.63 (95% CI, 0.42-0.94; P = .025). Inguinal hernia is common in the middle-aged and elderly male population and its incidence increases with advancing age. Overweight or obese patients have a lesser risk of developing an inguinal hernia.


Observations on massive retrieved human allografts.
Radiographic and histological studies of sixteen massive retrieved human allografts were carried out after the allografts had been in situ for four to sixty-five months. The studies demonstrated that union between the allograft and the host took place slowly at cortical-cortical junctions by the formation of an external callus derived from the cortex of the host, and it took place more rapidly at cancellous-cancellous junctions by internal callus advancing from the host into the allograft. Internal repair took place very slowly, was confined to the superficial surface and the ends of the graft, and had involved only 20 per cent of the graft by five years. The deep unrepaired portions of the graft retained their architecture, and where bone cement had been used to fix a prosthetic stem or an intramedullary rod to the allograft, there was no evidence of resorption of bone or loosening of the device. Soft tissues of the host became attached to the graft by deposition of a thin seam of new bone on the surface of the graft. A previous fracture of two grafts had healed before the time of retrieval. Analysis of the articular cartilage revealed no evidence that any chondrocytes had survived, even when the graft had been cryoprotected before it was preserved by freezing. The necrotic cartilage functioned well for as long as five years, and as it degenerated, it was covered by a pannus of fibrovascular reparative tissue. Two allografts that had been removed because of rejection were surrounded by an envelope of chronic inflammatory tissue that prevented union, adherence of soft tissue, and internal repair. Internal repair was more advanced about sites of fracture and adjacent to recurrent tumors than in other portions of the graft. These findings suggest that large frozen allografts in humans are osteoconductive rather than osteoinductive.


Nonrandom chromosomal change (trisomy 11) in murine plasmacytomas induced by an ABL-MYC retrovirus.
Trisomy of chromosome 11 (Ts11) is the second most frequent nonrandom chromosomal change in murine plasmacytomas (PCTs). The frequency of Ts11 is significantly higher in PCTs induced in pristane-conditioned mice infected by Abelson-murine leukemia virus (52%) compared to those induced by pristane alone (8.1%). Although the significance of Ts11 in mouse plasmacytomagenesis is not clearly understood it is hypothesized that a gene or genes located on chromosome (Chr) 11 may specifically promote the development of PCTs in which both oncogenes, c-myc and v-abl, are abundantly expressed. To test this assumption we induced PCTs by three highly effective plasmacytomagenic retroviruses: ABL-MYC, J3V1, and RIM. Nearly 90% of PCTs that arose in BALB/c, (BALB/c x DBA/2N)F1, BALB/c-nu/nu, and 5-month-old SCID mice infected with ABL-MYC virus were trisomic for Chr 11. In contrast, < 10% of PCTs induced by J3V1 or RIM retroviral constructs encompassing either v-myc and v-raf or c-myc and v-Ha-ras oncogenes, respectively, contained Ts11. We have also investigated whether the entire Chr 11 or any particular subregion is preferentially duplicated in the process of ABL-MYC plasmacytomagenesis. By inducing PCTs in F1 heterozygous mice that are carriers of reciprocal translocations involving Chr 11 we found that the duplicated chromosomal region is located distal to the T4Dn breakpoint (11B5 band) on the telomeric segment of Chr 11. The regular duplication of this chromosomal segment strongly suggests the presence of a gene or genes whose amplification is of critical importance for v-abl associated murine plasmacytomagenesis.


The monovalent ionophore monensin maintains the nuclear localization of the human stress protein hsp28 during heat shock recovery.
In HeLa cells exposed to supra-optimal temperatures, the alpha-crystallin-related stress protein hsp28 is reversibly redistributed inside the nucleus and increases its level of phosphorylation and aggregation. Here, I show that, at normal temperature after a heat stress, the sodium ionophore monensin maintains the nuclear localization of hsp28 without impairing the dephosphorylation of this protein. This phenomenon is not due to a prolongation, by monensin, of the synthesis of the heat-shock proteins after the heat stress. In contrast, the potassium ionophore nonactin induces only a weak alteration in the hsp28 locale, while the calcium ionophore A23187 and the uncoupler of oxidative phosphorylation FCCP have no effect. Following the removal of monensin 15 h after the heat stress, a further incubation of the cells for at least 36 h is necessary in order to observe a redistribution of hsp28 into the cytoplasm. A large fraction of hsp28 is then observed as dense excretion granules. In control cells kept at normal temperature, monensin, like nonactin, A23187 and FCCP, does not induce the redistribution of hsp28 inside the nucleus. Taken together, these results suggest that the disruption of the Na+ active transport by monensin probably inhibits the redistribution of hsp28 in the cytoplasm after heat shock.


Toxoplasma gondii Infection in Alpine Red Deer (Cervus elaphus): Its Spread and Effects on Fertility [corrected].
In contrast to the depth of knowledge on the pathological effects of parasitism in domestic animals, the impact of the vast majority of parasites on wildlife hosts is poorly understood and, besides, information from domestics is rarely usable to disclose the parasites' impact on free-ranging populations' dynamics. Here we use Toxoplasmosis as a study model since, until now, the infection process and the protozoan's effects in natural conditions has received little attention. We analysed 81 sera from red deer (Cervus elaphus) sampled in Italian Alps and through generalized linear models we evaluated (1) the epidemiological factors influencing T. gondii infection dynamics; (2) its impact on female fertility [corrected]. High seroprevalence of T. gondii infection was recorded in yearling (1 year-old; prevalence = 52.4%) and adult (>2 year-old; prevalence = 51.3%) red deer, while calves (<1 year-old) did not contract the infection suggesting horizontal transmission as the main route of infection. The stable prevalence between yearlings and adults and the higher serological titres of younger individuals lead to two alternative infection processes suggesting a difference between age classes or in acquiring the infection or in responding to the pathogen. No associations between T. gondii serological titres and pregnancy status was observed indicating no direct effect on the probability of being pregnant; nevertheless a relation between females' higher serological titres and lower foetal development emerged, suggesting potential effects of the parasite infection on deer reproduction. The results demonstrate high seroprevalence of T. gondii infection in free-ranging red deer and, furthermore, the effect on foetal development suggests the potential impact of the parasite on red deer fertility and thus on its population dynamics.


Epidermal growth factor activates nuclear factor-kappaB in human proximal tubule cells.
The promotion of cell survival and regeneration in acute renal failure (ARF) is important for the restitution of renal function. Epidermal growth factor (EGF) has been implicated in the regulation of cell proliferation. We provide evidence for a direct link between EGF, nuclear factor-kappaB (NF-kappaB), and cell cycle regulation (cyclin D1). EGF was found to stimulate NF-kappaB-dependent gene transcription and DNA binding. In addition, EGF stimulated cyclin D1 promoter activity as well as cyclin D1 expression. Moreover, inhibition of NF-kappaB caused a pronounced reduction of EGF-induced cyclin D1 promoter activity. Furthermore, both EGF-mediated NF-kappaB activation and cyclin D1 expression were inhibited by coexpression of super IkappaB. Taken together, these data identify NF-kappaB and cyclin D1 as downstream targets of EGF and establish a molecular link between stimulation of EGF via activation of NF-kappaB and cyclin D1 expression in human proximal tubular cells.


Can a two-hour lecture by a pharmacist improve the quality of prescriptions in a pediatric hospital? A retrospective cohort study.
A high rate of prescription errors exists in pediatric teaching hospitals, especially during initial training. To determine the effectiveness of a two-hour lecture by a pharmacist on rates of prescription errors and quality of prescriptions. A two-hour lecture led by a pharmacist was provided to 11 junior pediatric residents (PGY-1) as part of a one-month immersion program. A control group included 15 residents without the intervention. We reviewed charts to analyze the first 50 prescriptions of each resident. Data were collected from 1300 prescriptions involving 451 patients, 550 in the intervention group and 750 in the control group. The rate of prescription errors in the intervention group was 9.6% compared to 11.3% in the control group (p=0.32), affecting 106 patients. Statistically significant differences between both groups were prescriptions with unwritten doses (p=0.01) and errors involving overdosing (p=0.04). We identified many errors as well as issues surrounding quality of prescriptions. We found a 10.6% prescription error rate. This two-hour lecture seems insufficient to reduce prescription errors among junior pediatric residents. This study highlights the most frequent types of errors and prescription quality issues that should be targeted by future educational interventions.


An idealized polyhedral model and geometric structure for silicon nanotubes.
In this paper, we introduce an idealized model of silicon nanotubes comprising an exact polyhedral geometric structure for single-walled silicon nanotubes. The silicon nanotubes considered here are assumed to be formed by sp(3) hybridization and thus the nanotube lattice is assumed to comprise only squares or skew rhombi. Beginning with the three postulates that all bond lengths are equal, all adjacent bond angles are equal, and all atoms are equidistant from a common axis of symmetry, we derive exact formulae for the geometric parameters such as radii, bond angles and unit cell length. We present asymptotic expansions for these quantities to the first two orders of magnitude. Because of the faceted nature of the polyhedral model we may determine a perceived inner radius for the nanotube, from which an expression for the wall thickness emerges. We also describe the geometric properties of some ultra-small silicon nanotubes. Finally, the values of the diameters for the polyhedral model are compared with results obtained from molecular dynamics simulations and some limited numerical calculations are undertaken to confirm the meta-stability of the proposed structures.


Neural Processing and Production of Gesture in Children and Adolescents With Autism Spectrum Disorder.
Individuals with autism spectrum disorder (ASD) demonstrate impairments in non-verbal communication, including gesturing and imitation deficits. Reduced sensitivity to biological motion (BM) in ASD may impair processing of dynamic social cues like gestures, which in turn may impede encoding and subsequent performance of these actions. Using both an fMRI task involving observation of action gestures and a charade style paradigm assessing gesture performance, this study examined the brain-behavior relationships between neural activity during gesture processing, gesturing abilities and social symptomology in a group of children and adolescents with and without ASD. Compared to typically developing (TD) controls, participants with ASD showed atypical sensitivity to movement in right posterior superior temporal sulcus (pSTS), a region implicated in action processing, and had poorer overall gesture performance with specific deficits in hand posture. The TD group showed associations between neural activity, gesture performance and social skills, that were weak or non-significant in the ASD group. These findings suggest that those with ASD demonstrate abnormalities in both processing and production of gestures and may reflect dysfunction in the mechanism underlying perception-action coupling resulting in atypical development of social and communicative skills.


Vascular endothelial growth factor isoforms display distinct activities in promoting tumor angiogenesis at different anatomic sites.
The gene for the major angiogenic factor, vascular endothelial growth factor (VEGF), encodes several spliced isoforms. We reported previously that overexpression of two VEGF isoforms, VEGF(121) and VEGF(165), by human glioma U87 MG cells induced tumor-associated intracerebral hemorrhage, whereas expression of a third form, VEGF(189), did not cause vessel rupture. Here, we test whether these VEGF isoforms have distinct activities for enhancing vascularization and growth of gliomas in mice. U87 MG cells that overexpressed VEGF(165) or VEGF(189) grew more rapidly than the parental cells in both s.c. and intracranial (i.c.) locations. However, cells that overexpressed VEGF(121) only showed enhancement of i.c. tumor growth but had a minimal effect on s.c. glioma progression. At both anatomical sties, VEGF(165) and VEGF(189) strongly augmented neovascularization, whereas VEGF(121) only increased vessel density in brain tumors. In each type of glioma, expression of VEGF receptors -1 and -2 largely phenocopied the tumor vasculature, because increased VEGF/VEGF receptor-activated microvessel densities were strongly correlated with the angiogenicity and tumorigenicity elicited by the VEGF isoforms at both anatomical sites. One notable difference between the sites was the expression of vitronectin, a prototypic ligand of alpha(v)beta(3) and alpha(v)beta(5) integrins, detected in i.c. but not in s.c., gliomas. Endothelial cell migration stimulated by VEGF(121) was potentiated by vitronectin to a greater extent than that stimulated by VEGF(165). This data demonstrates that VEGF isoforms have distinct activities at different anatomical sites and suggest that the microenvironment of different tissues affects the function of VEGF isoforms.


Bunyavirus requirement for endosomal K+ reveals new roles of cellular ion channels during infection.
In order to multiply and cause disease a virus must transport its genome from outside the cell into the cytosol, most commonly achieved through the endocytic network. Endosomes transport virus particles to specific cellular destinations and viruses exploit the changing environment of maturing endocytic vesicles as triggers to mediate genome release. Previously we demonstrated that several bunyaviruses, which comprise the largest family of negative sense RNA viruses, require the activity of cellular potassium (K+) channels to cause productive infection. Specifically, we demonstrated a surprising role for K+ channels during virus endosomal trafficking. In this study, we have used the prototype bunyavirus, Bunyamwera virus (BUNV), as a tool to understand why K+ channels are required for progression of these viruses through the endocytic network. We report three major findings: First, the production of a dual fluorescently labelled bunyavirus to visualize virus trafficking in live cells. Second, we show that BUNV traffics through endosomes containing high [K+] and that these K+ ions influence the infectivity of virions. Third, we show that K+ channel inhibition can alter the distribution of K+ across the endosomal system and arrest virus trafficking in endosomes. These data suggest high endosomal [K+] is a critical cue that is required for virus infection, and is controlled by cellular K+ channels resident within the endosome network. This highlights cellular K+ channels as druggable targets to impede virus entry, infection and disease.


Pyoderma gangrenosum: a systematic review.
Pyoderma gangrenosum (PG) is a rare, chronic neutrophilic dermatosis of unknown etiology. The world wide incidence is estimated to be around 3-10 cases per million population per year. In 50-70% of cases inflammatory bowel diseases, hematological malignancies or rheumatologic disorders are associated to PG. Although the etiology is uncertain, the dysregulation of the immune system appears to be implied. Pathergy is the most important triggering factor of PG. Indeed, 20-30% of patients report the onset of PG following trivial trauma. Four main variants of PG have been described, namely classic, pustular, bullous, and vegetative forms. The classic form of PG is characterized by ulcers with a raised, undermined, inflammatory border. Intense pain is generally associated to PG. The diagnosis is mainly clinical and of exclusion. The differential diagnosis should take into account infections, vascular disorders and malignancies. The clinical course can be explosive and rapidly progressive or indolent and gradually progressive. Often patients develop only one episode and the overall prognosis is good but extremely influenced by the underlying disorders. Local therapy, mainly with topic steroids is used for mild to moderate lesions. For severe forms of PG a systemic therapy with glucocorticoids and/or other drugs such as tacrolimus, cyclosporine, etc. is needed. This paper is a systematic review of literature on PG.


A comparative study of the structural organization of spheres derived from the adult human subventricular zone and glioblastoma biopsies.
Sphere forming assays have been useful to enrich for stem like cells in a range of tumors. The robustness of this system contrasts the difficulties in defining a stem cell population based on cell surface markers. We have undertaken a study to describe the cellular and organizational composition of tumorspheres, directly comparing these to neurospheres derived from the adult human subventricular zone (SVZ). Primary cell cultures from brain tumors were found to contain variable fractions of cells positive for tumor stem cell markers (CD133 (2-93%)/SSEA1 (3-15%)/CXCR4 (1-72%)). All cultures produced tumors upon xenografting. Tumorspheres contained a heterogeneous population of cells, but were structurally organized with stem cell markers present at the core of spheres, with markers of more mature glial progenitors and astrocytes at more peripheral location. Ultrastructural studies showed that tumorspheres contained a higher fraction of electron dense cells in the core than the periphery (36% and 19%, respectively). Neurospheres also contained a heterogeneous cell population, but did not have an organization similar to tumorspheres. Although tumorspheres clearly display irregular and neoplastic cells, they establish an organized structure with an outward gradient of differentiation. We suggest that this organization is central in maintaining the tumor stem cell pool.


New Models to Predict the Acute and Chronic Toxicities of Representative Species of the Main Trophic Levels of Aquatic Environments.
To assess the impact of chemicals on an aquatic environment, toxicological data for three trophic levels are needed to address the chronic and acute toxicities. The use of non-testing methods, such as predictive computational models, was proposed to avoid or reduce the need for animal models and speed up the process when there are many substances to be tested. We developed predictive models for Raphidocelis subcapitata, Daphnia magna, and fish for acute and chronic toxicities. The random forest machine learning approach gave the best results. The models gave good statistical quality for all endpoints. These models are freely available for use as individual models in the VEGA platform and for prioritization in JANUS software.


Humid tropical rain forest has expanded into eucalypt forest and savanna over the last 50 years.
Tropical rain forest expansion and savanna woody vegetation thickening appear to be a global trend, but there remains uncertainty about whether there is a common set of global drivers. Using geographic information techniques, we analyzed aerial photography of five areas in the humid tropics of northeastern Queensland, Australia, taken in the 1950s and 2008, to determine if changes in rain forest extent match those reported for the Australian monsoon tropics using similar techniques. Mapping of the 1950s aerial photography showed that of the combined study area (64,430 ha), 63% was classified as eucalypt forests/woodland and 37% as rain forest. Our mapping revealed that although most boundaries remained stable, there was a net increase of 732 ha of the original rain forest area over the study period, and negligible conversion of rain forest to eucalypt forest/woodland. Statistical modeling, controlling for spatial autocorrelation, indicated distance from preexisting rain forest as the strongest determinant of rain forest expansion. Margin extension had a mean rate across the five sites of 0.6 m per decade. Expansion was greater in tall open forest types but also occurred in shorter, more flammable woodland vegetation types. No correlations were detected with other local variables (aspect, elevation, geology, topography, drainage). Using a geographically weighted mean rate of rain forest margin extension across the whole region, we predict that over 25% of tall open forest (a forest type of high conservation significance) would still remain after 2000 years of rain forest expansion. This slow replacement is due to the convoluted nature of the rain forest boundary and the irregular shape of the tall open forest patches. Our analyses point to the increased concentration of atmospheric CO(2) as the most likely global driver of indiscriminate rain forest expansion occurring in northeastern Australia, by increasing tree growth and thereby overriding the effects of fire disturbance.


Guillain-Barré and Miller Fisher syndromes occurring with tumor necrosis factor alpha antagonist therapy.
Diverse neurologic syndromes have been described in association with tumor necrosis factor alpha (TNFalpha) antagonist therapy for inflammatory arthritides and Crohn's disease. The objective of this study was to review the occurrence and clinical features of Guillain-Barré syndrome and its variant, the Miller Fisher syndrome, during TNFalpha antagonist therapy. The postmarketing database of the US Food and Drug Administration (FDA) was searched, following our experience with a patient with rheumatoid arthritis in whom the Miller Fisher syndrome variant of the Guillain-Barré syndrome developed while he was receiving infliximab therapy. Our index patient had a neurologic illness defined initially by ataxia and dysarthria, which fluctuated in relation to each subsequent infliximab infusion and, after 6 months, culminated in areflexic flaccid quadriplegia. In addition, 15 patients in whom Guillain-Barré syndrome developed following TNFalpha antagonist therapy were identified from the FDA database. Guillain-Barré syndrome developed following infliximab therapy in 9 patients, following etanercept therapy in 5 patients, and following adalimumab therapy in 1 patient. Among the 13 patients for whom followup data were available, 1 patient experienced no resolution, 9 patients had partial resolution, and 3 patients had complete resolution of Guillain-Barré syndrome following therapy. An association of Guillain-Barré syndrome with TNFalpha antagonist therapy is supported by the worsening of neurologic symptoms that occurred in our index patient following each infusion of infliximab, and by the temporal association of this syndrome with TNFalpha antagonist therapy in 15 other patients. An acute or subacute demyelinating polyneuropathy should be considered a potential adverse effect of TNFalpha antagonist therapy.


Uptake and elimination of ionizable organic chemicals at fish gills: I. Model formulation, parameterization, and behavior.
A mechanistic model for the uptake and elimination of ionizable organic chemicals at fish gills is presented. This model is a modification of a previous model for nonionizable organic chemicals that addressed the transport of chemical to and from gill surfaces in water and blood, diffusion of chemical across epithelial cells, and binding of chemical to components in water and blood. For ionizable chemicals, three additional processes are included. First, excretory products alter the pH at gill surfaces, affecting the relative amounts of neutral and ionized molecules compared with that in the bulk exposure water. Second, ionized molecules support chemical flux to and from epithelial cell membranes and help maintain high diffusion gradients of neutral molecules across these membranes, thereby contributing to uptake and elimination even if the membranes are impermeable to ionized molecules. Third, membrane barriers are not completely impermeable to ionized molecules, and even limited permeability can have appreciable effects on chemical flux. Approaches for model parameterization are discussed. Model-predicted relationships of uptake and elimination rates to exposure water pH, alkalinity, and chemical properties are presented and discussed in terms of model processes. The model is shown to predict important features of reported effects of pH on uptake rates of weak organic acids.


Fatal intoxications associated with the designer opioid AH-7921.
AH-7921 (3,4-dichloro-N-[(1-dimethylamino)cyclohexylmethyl]benzamide) is a designer opioid with ∼80% of morphine's µ-agonist activity. Over a 6-month period, we encountered nine deaths where AH-7921 was involved and detected in blood from the deceased. Shortly after the last death, on August 1 2013, AH-7921 was scheduled as a narcotic and largely disappeared from the illicit market in Sweden. AH-7921 was measured by a selective liquid chromatography-MS-MS method and the concentrations of AH-7921 ranged from 0.03 to 0.99 µg/g blood. Six of our cases had other drugs of abuse on board and most had other medications such as benzodiazepines, antidepressants and analgesics. However, the other medicinal drugs encountered were present in postmortem therapeutic concentrations and unlikely to have contributed to death. In addition to the parent compound, we identified six possible metabolites where two N-demethylated dominated and four mono-hydroxylated were found in trace amounts in the blood. In conclusion, deaths with AH-7921 seem to occur both at low and high concentrations, probably a result of different tolerance to the drug. Hence, it is reasonable to assume that no sharp dividing line exists between lethal and non-lethal concentrations. Further, poly-drug use did not seem to be a major contributing factor for the fatal outcome.


More is Better: English Language Statistics are Biased Toward Addition.
We have evolved to become who we are, at least in part, due to our general drive to create new things and ideas. When seeking to improve our creations, ideas, or situations, we systematically overlook opportunities to perform subtractive changes. For example, when tasked with giving feedback on an academic paper, reviewers will tend to suggest additional explanations and analyses rather than delete existing ones. Here, we show that this addition bias is systematically reflected in English language statistics along several distinct dimensions. First, we show that words associated with an increase in quantity or number (e.g., add, addition, more, most) are more frequent than words associated with a decrease in quantity or number (e.g., subtract, subtraction, less, least). Second, we show that in binomial expressions, addition-related words are mentioned first, that is, add and subtract rather than subtract and add. Third, we show that the distributional semantics of verbs of change, such as to improve and to transform, overlap more with the distributional semantics of add/increase than subtract/decrease, which suggests that change verbs are implicitly biased toward addition. Fourth, addition-related words have more positive connotations than subtraction-related words. Fifth, we demonstrate that state-of-the-art large language models, such as the Generative Pre-trained Transformer (GPT-3), are also biased toward addition. We discuss the implications of our results for research on cognitive biases and decision-making.


The II Brazilian Guidelines for the pharmacological treatment of patients hospitalized with COVID-19 Joint Guidelines of the Associação Brasileira de Medicina de Emergência, Associação de Medicina Intensiva Brasileira, Associação Médica Brasileira, Sociedade Brasileira de Angiologia e Cirurgia Vascular, Sociedade Brasileira de Infectologia, Sociedade Brasileira de Pneumologia e Tisiologia and Sociedade Brasileira de Reumatologia.
To update the recommendations to support decisions regarding the pharmacological treatment of patients hospitalized with COVID-19 in Brazil. Experts, including representatives of the Ministry of Health and methodologists, created this guideline. The method used for the rapid development of guidelines was based on the adoption and/or adaptation of existing international guidelines (GRADE ADOLOPMENT) and supported by the e-COVID-19 RecMap platform. The quality of the evidence and the preparation of the recommendations followed the GRADE method. Twenty-one recommendations were generated, including strong recommendations for the use of corticosteroids in patients using supplemental oxygen and conditional recommendations for the use of tocilizumab and baricitinib for patients on supplemental oxygen or on noninvasive ventilation and anticoagulants to prevent thromboembolism. Due to suspension of use authorization, it was not possible to make recommendations regarding the use of casirivimab + imdevimab. Strong recommendations against the use of azithromycin in patients without suspected bacterial infection, hydroxychloroquine, convalescent plasma, colchicine, and lopinavir + ritonavir and conditional recommendations against the use of ivermectin and remdesivir were made. New recommendations for the treatment of hospitalized patients with COVID-19 were generated, such as those for tocilizumab and baricitinib. Corticosteroids and prophylaxis for thromboembolism are still recommended, the latter with conditional recommendation. Several drugs were considered ineffective and should not be used to provide the best treatment according to the principles of evidence-based medicine and to promote resource economy.


Membrane interactions between adjacent mucols secretion granules.
In primate goblet cells, the membranes of adjacent mucous granules from contact areas which appear as extensive pentalaminar fusion sites in thin sections. In freeze-fracture replicas, the same membrane areas are smooth, except for a few 6-8-nm particles which adhere to the E face. These protein-poor membrane interaction sites are relatively long-lived, and it is proposed that further stimulus may be required to trigger membrane fission.


Symptoms, signs, and functional disability in adult spondylolisthesis.
A cross-sectional clinical study. To determine whether there are specific symptoms, signs, and functional disability associated with adult spondylolisthesis. In spite of the common occurrence of adult spondylolisthesis, the symptoms, signs, and disability associated with it have not been analyzed in a large, well-defined group of patients. The symptoms, signs, and disability of 111 consecutive patients with adult spondylolisthesis, before randomized treatment with fusion or physiotherapy, were compared with those of 39 patients with nonspecific low back pain before lumbar fusion. The patients completed a questionnaire covering clinical history and symptoms and submitted a pain drawing. The signs were documented. Functional disability and pain were quantified by 12-function and 2-pain visual analog scales, respectively. Sixty-two percent of the patients reported low back pain as well as sciatica, 7% sciatica only, and 31% low back pain only. Specific signs were infrequent. A positive straight leg raising test result in 12% and an L5 sensory disturbance in 13% were the most common. The symptoms were similar in patients with spondylolisthesis and chronic low back pain, but the chronic low back pain group reported more functional disability. Patients with a nonorganic pain drawing (widespread, nonspecific pain) were more often blue collar workers; were more often and longer on sick leave; and reported reduced mental condition, sexual function, functional ability, and more pain than patients with an organic pain drawing (localized, specific pain). The clinical pattern and functional disability in adult spondylolisthesis and in low back pain of nonspecific origin are similar. Sciatica in adult spondylolisthesis is typically not associated with a positive straight leg raising test result.


Recent Trends in Oral Anticoagulant Use and Post-Discharge Complications Among Atrial Fibrillation Patients with Acute Myocardial Infarction.
Atrial fibrillation (AF) is a common complication of acute myocardial infarction (AMI).The CHA2DS2VAScand CHADS2risk scoresare used to identifypatients with AF at risk for strokeand to guide oral anticoagulants (OAC) use, including patients with AMI. However, the epidemiology of AF, further stratifiedaccording to patients' risk of stroke, has not been wellcharacterized among those hospitalized for AMI. We examined trends in the frequency of AF, rates of discharge OAC use, and post-discharge outcomes among 6,627 residents of the Worcester, Massachusetts area who survived hospitalization for AMI at 11 medical centers between 1997 and 2011. A total of 1,050AMI patients had AF (16%) andthe majority (91%)had a CHA2DS2VAScscore >2.AF rates were highest among patients in the highest stroke risk group.In comparison to patients without AF, patients with AMI and AF in the highest stroke risk category had higher rates of post-discharge complications, including higher 30-day re-hospitalization [27 % vs. 17 %], 30-day post-discharge death [10 % vs. 5%], and 1-year post-discharge death [46 % vs. 18 %] (p < 0.001 for all). Notably, fewerthan half of guideline-eligible AF patientsreceived an OACprescription at discharge. Usage rates for other evidence-based therapiessuch as statins and beta-blockers,lagged in comparison to AMI patients free from AF. Our findings highlight the need to enhance efforts towards stroke prevention among AMI survivors with AF.


Cisplatin plus gemcitabine or vinorelbine for elderly patients with advanced non small-cell lung cancer: the MILES-2P studies.
Two phase I/II trials were done to evaluate the feasibility of cisplatin combined with gemcitabine or vinorelbine in elderly patients with advanced non-small-cell lung cancer (NSCLC). Patients with advanced NSCLC who were older than 70 years of age and who had a performance status of 0 to 1 were eligible. Cisplatin was given on day 1 (a starting dose of 50 mg/m2 with increasing increments of 10 mg/m2 at each level) and gemcitabine (1,000 mg/m2) or vinorelbine (25 mg/m2) on days 1 and 8. Cycles were repeated every 21 days. A two-stage flexible optimal design was applied in the phase II study, and unacceptable toxicity was the primary end point. Overall, 159 patients were enrolled: 38 in phase I and 121 in phase II studies. Cisplatin was feasible at 60 mg/m2 with gemcitabine and at 40 mg/m2 with vinorelbine. With the former combination, 50 of 60 (83.3%) patients were treated without unacceptable toxicity; objective responses were reported in 26 of 60 patients (43.5%; 95% CI, 30.6 to 56.8); median progression-free and overall survivals were 25.3 and 43.6 weeks, respectively. With the latter combination, 50 (82.0%) of 61 patients were treated without unacceptable toxicity; objective responses were reported in 22 of 61 patients (36.1%; 95% CI, 24.2 to 49.4); median progression-free and overall survivals were 21.1 and 33.1 weeks, respectively. Both cisplatin (60 mg/m2) plus gemcitabine and cisplatin (40 mg/m2) plus vinorelbine are feasible and active in the treatment of elderly patients with advanced NSCLC. The former combination, which provides a higher dose of cisplatin, deserves comparison versus single-agent chemotherapy in this setting of patients.


Climate forcing of an emerging pathogenic fungus across a montane multi-host community.
Changes in the timings of seasonality as a result of anthropogenic climate change are predicted to occur over the coming decades. While this is expected to have widespread impacts on the dynamics of infectious disease through environmental forcing, empirical data are lacking. Here, we investigated whether seasonality, specifically the timing of spring ice-thaw, affected susceptibility to infection by the emerging pathogenic fungus Batrachochytrium dendrobatidis (Bd) across a montane community of amphibians that are suffering declines and extirpations as a consequence of this infection. We found a robust temporal association between the timing of the spring thaw and Bd infection in two host species, where we show that an early onset of spring forced high prevalences of infection. A third highly susceptible species (the midwife toad, Alytes obstetricans) maintained a high prevalence of infection independent of time of spring thaw. Our data show that perennially overwintering midwife toad larvae may act as a year-round reservoir of infection with variation in time of spring thaw determining the extent to which infection spills over into sympatric species. We used future temperature projections based on global climate models to demonstrate that the timing of spring thaw in this region will advance markedly by the 2050s, indicating that climate change will further force the severity of infection. Our findings on the effect of annual variability on multi-host infection dynamics show that the community-level impact of fungal infectious disease on biodiversity will need to be re-evaluated in the face of climate change.This article is part of the themed issue 'Tackling emerging fungal threats to animal health, food security and ecosystem resilience'.


Vaccine refusal, mandatory immunization, and the risks of vaccine-preventable diseases.
Vaccines are among the most effective prevention tools available to clinicians. However, the success of an immunization program depends on high rates of acceptance and coverage. There is evidence of an increase in vaccine refusal in the United States and of geographic clustering of refusals that results in outbreaks. Children with exemptions from school immunization requirements (a measure of vaccine refusal) are at increased risk for measles and pertussis and can infect others who are too young to be vaccinated, cannot be vaccinated for medical reasons, or were vaccinated but did not have a sufficient immunologic response. Clinicians can play a crucial role in parental decision making. Health care providers are cited as the most frequent source of immunization information by parents, including parents of unvaccinated children. Although some clinicians have discontinued or have considered discontinuing their provider relationship with patients who refuse vaccines, the American Academy of Pediatrics Committee on Bioethics advises against this and recommends that clinicians address vaccine refusal by respectfully listening to parental concerns and discussing the risks of nonvaccination.


Association of adiponectin with carotid arteriosclerosis in predialysis chronic kidney disease.
Adiponectin is an adipocyte-derived protein with antiatherogenic properties. Chronic kidney disease (CKD) is one of the risk factors for cardiovascular disease. We investigated the potential association between adiponectin and carotid arteriosclerosis in patients with predialysis CKD. We enrolled 95 CKD patients without dialysis and 81 non-CKD patients. Intima-media thickness (IMT) and plaque score (PS) in the common carotid artery were measured using an ultrasound system. Carotid arteriosclerosis was defined as IMT >1.2 mm and/or PS >5.0 mm. The prevalence of CKD was independently associated with carotid arteriosclerosis after adjustment for other risk factors. Higher adiponectin levels were observed in CKD patients compared with non-CKD patients. Adiponectin levels were not independently correlated with the presence of carotid arteriosclerosis in all subjects. To evaluate the association between adiponectin and carotid arteriosclerosis among a CKD population, we divided the CKD patients into 2 groups according to a cutoff level of adiponectin determined by ROC analysis. The prevalence of carotid arteriosclerosis was significantly higher in the low-adiponectin group than in the high-adiponectin group among CKD patients. After adjusting for other risk factors, low levels of adiponectin were independently correlated with carotid arteriosclerosis in CKD patients. Our data document that adiponectin is associated with increased risk of carotid atherosclerosis in a predialysis CKD population.


Emerged HA and NA mutants of the pandemic influenza H1N1 viruses with increasing epidemiological significance in Taipei and Kaohsiung, Taiwan, 2009-10.
The 2009 influenza pandemic provided an opportunity to observe dynamic changes of the hemagglutinin (HA) and neuraminidase (NA) of pH1N1 strains that spread in two metropolitan areas--Taipei and Kaohsiung. We observed cumulative increases of amino acid substitutions of both HA and NA that were higher in the post-peak than in the pre-peak period of the epidemic. About 14.94% and 3.44% of 174 isolates had one and two amino acids changes, respective, in the four antigenic sites. One unique adaptive mutation of HA2 (E374K) was first detected three weeks before the epidemic peak. This mutation evolved through the epidemic, and finally emerged as the major circulated strain, with significantly higher frequency in the post-peak period than in the pre-peak (64.65% vs 9.28%, p<0.0001). E374K persisted until ten months post-nationwide vaccination without further antigenic changes (e.g. prior to the highest selective pressure). In public health measures, the epidemic peaked at seven weeks after oseltamivir treatment was initiated. The emerging E374K mutants spread before the first peak of school class suspension, extended their survival in high-density population areas before vaccination, dominated in the second wave of class suspension, and were fixed as herd immunity developed. The tempo-spatial spreading of E374K mutants was more concentrated during the post-peak (p = 0.000004) in seven districts with higher spatial clusters (p<0.001). This is the first study examining viral changes during the naïve phase of a pandemic of influenza through integrated virological/serological/clinical surveillance, tempo-spatial analysis, and intervention policies. The vaccination increased the percentage of E374K mutants (22.86% vs 72.34%, p<0.001) and significantly elevated the frequency of mutations in Sa antigenic site (2.36% vs 23.40%, p<0.001). Future pre-vaccination public health efforts should monitor amino acids of HA and NA of pandemic influenza viruses isolated at exponential and peak phases in areas with high cluster cases.


Template-stripped nanoaperture tweezer integrated with optical fiber.
We demonstrate an optical trapping technique that integrates the light guiding of an optical fiber with the field localization of a nanoaperture in a gold film. A key innovation of our technique is to use template-stripping for easy planar fabrication without the need for nanofabrication on the tip itself. As a proof of principle, we demonstrate the trapping of 20 nm and 30 nm polystyrene nanoparticles in solution, as observed by a jump in the transmitted laser intensity through the aperture. We use the finite difference time domain technique to simulate this intensity jump with the addition of a nanoparticle in the aperture, showing reasonable agreement with the experimental data. This simple nano-aperture optical fiber tip eliminates the need for a microscope setup while allowing for trapping nanoparticles, so it is anticipated to have applications in biology (e.g. viruses), biophysics (e.g. protein interactions), physics (e.g. quantum emitters), and chemistry (e.g. colloidal particles).


The melanoma epidemic. Is increased surveillance the solution or the problem?
Melanoma has a very bad reputation, and rightfully so. Until relatively recently, the prognosis for melanoma was dismal and, very reasonably, early detection has been proposed as a viable approach to preventing melanoma deaths. On the surface, this simple approach appears to have been a stellar success, with melanoma survival rates improving markedly. There is no question that there has been a dramatic increase in the number of people with a diagnosis of melanoma. On careful scrutiny, the numbers reflected in the increased incidence may not add up. Mortality rates and the diagnosis rate for thick tumors ( > 1.5 mm thick) have not gone up commensurate with the growing rate of diagnosis. The obvious explanation for this discrepancy is that we are doing a great job! However, this explanation must assume that an overwhelming percentage of those at risk for melanoma are in a position to benefit from our service. It is very unlikely that this is the case. It is not clear whether the increase in the number of melanoma cases diagnosed reflects an actual increase in real disease. Could this increase be in part a product of aggressive surveillance resulting in the identification of "atypical" pigmented skin tumors of limited or nonexistent potential for malignant behavior? Could the dramatic improvement in the prognosis of melanoma simply be a consequence of removing biologically benign pigmented tumors that are inadvertently classified as malignant?


Metformin acts on the gut-brain axis to ameliorate antipsychotic-induced metabolic dysfunction.
Antipsychotic-induced metabolic dysfunction (AIMD) is an intractable clinical challenge worldwide. The situation is becoming more critical as second-generation antipsychotics (SGAs), to a great extent, have replaced the role of first-generation antipsychotics in managing major psychiatric disorders. Although the exact mechanisms for developing AIMD is intricate, emerging evidence has indicated the involvement of the microbiota-gut-brain axis in AIMD. SGAs treatment may change the diversity and compositions of intestinal flora (e.g., decreased abundance of Bacteroidetes and Akkermansia muciniphila, and increased Firmicutes). Short-chain fatty acids and other metabolites derived from gut microbiota, on the one hand, can regulate the activity of intestinal endocrine cells and their secretion of satiety hormones (e.g., glucagon-like peptide 1, peptide YY, cholecystokinin and ghrelin); on the other hand, can activate the vagus nerve or transport into the brain to exert a central modulation of foraging behaviors via binding to neuropeptide receptors. Interestingly, metformin, a classical antidiabetic agent, is capable of alleviating AIMD possibly by regulating the microbiota-gut-brain axis. That is, metformin can not only partially reverse the alterations of gut microbial communities due to SGAs treatment, but also play a positive role in rectifying the disturbances of peripheral and central satiety-related neuropeptides. Current evidence has indicated a promising role for metformin on ameliorating AMID, but further verifications in well-designed clinical trials are still warranted.


Mutational bias in penguin microsatellite DNA.
Analysis of nucleotide sequence variation at a microsatellite DNA locus revealed extensive size homoplasy of alleles in Adélie penguins (Pygoscelis adeliae). Variation in the flanking regions at this locus allowed discrimination between mechanisms proposed for length changes in microsatellite DNA alleles. We further examined the structure of alleles for the same microsatellite DNA locus across 11 additional species of penguin (Spheniscidae) by mapping allele sequences onto an independent penguin phylogeny. Our analysis indicated that the repeat motifs appear to have evolved independently on several occasions. We observed sequence instability in the region bordering the repeat tract with a transversional bias predominating. We propose that this bias results from inaccurate DNA replication owing to the sequence context of this repeat tract. Because we show that regions flanking repeat sequences exhibit this mutational bias, this cautions against the use of such regions for phylogeny reconstruction.


Systemic but not central administration of tumor necrosis factor-alpha attenuates LPS-induced fever in rats.
The purpose of this study was to test the hypothesis that tumor necrosis factor-alpha (TNF) limits fever induced by lipopolysaccharide (LPS) in rats and to determine whether such antipyretic action of this cytokine is outside or inside the central nervous system (CNS). The CNS effects on LPS-induced fever were tested by injecting a subpyrogenic amount (0.20 microgram) of human recombinant TNF (hrTNF) intracerebroventricularly or by slowly infusing into the anterior hypothalamus an amount previously measured in this brain region during LPS fever (0.24 U in 0.13 microliter of artificial cerebrospinal fluid/min). The peripheral effects of this cytokine on LPS fever were tested by injecting 1 micrograms/kg of hrTNF intraperitoneally or by intraperitoneal administration of 300 micrograms/kg of the hrTNF soluble receptor p80 (hrTNFsr). The core temperature (measured by biotelemetry) during LPS fever was not significantly affected by administration of hrTNF intracerebroventricularly or intrahypothalamically. An intraperitoneal injection of hrTNF (1 microgram/kg) had a significant antipyretic effect on febrile response to LPS (mean temperature 2-8 h after injections was 37.28 +/- 0.12 degrees C in rats injected with hrTNF and LPS vs. 38.73 +/- 0.04 degrees C in rats injected with saline and LPS; analysis of variance among groups, P = 0.0001; Fisher's protected least significant difference, P < 0.05). When rats were injected intraperitoneally with hrTNFsr, the febrile response to LPS was enhanced (analysis of variance among groups, P = 0.0001; Fisher's protected least significant difference, P < 0.05). These results support the hypothesis that TNF acts to limit the magnitude of LPS-induced fever and that this action occurs outside the CNS.


Identification of two proteins induced by exposure of the pathogenic fungus Candida glabrata to fluconazole.
Candida glabrata is an increasingly important cause of opportunistic fungal infection of humans and appears to be intrinsically resistant to the triazole antifungal fluconazole. However, the mechanisms responsible for reduced susceptibility to azole drugs are not understood. Fluconazole exposure rapidly induced expression of a 169-kDa protein band in plasma membrane fractions of C. glabrata cells. Mass spectrometry of trypsin-digested peptide fragments showed that the induced protein band comprised the ATP binding cassette-type drug efflux transporter CgCdr1p. CgCdr1p was also functionally overexpressed in S. cerevisiae and similarly identified by mass spectrometry. A 61-kDa protein band in the plasma membrane fraction from C. glabrata was also induced by fluconazole exposure. Mass spectrometric peptide fingerprinting identified this band as lanosterol 14alpha-demethylase, the enzyme in the ergosterol biosynthesis pathway targeted by fluconazole. The rapid induction of a multidrug efflux pump and/or overproduction of lanosterol 14alpha-demethylase are mechanisms that could make C. glabrata appear intrinsically resistant to fluconazole. Mass spectrometric fingerprint analysis of SDS-PAGE separated plasma membrane fractions combined with heterologous hyper-expression provides a convenient method for protein identification and functional evaluation of induced proteins, even in an organism where the genome sequence database is incomplete.


Effective specialist or jack of all trades? Experimental evolution of a crop pest in fluctuating and stable environments.
Understanding pest evolution in agricultural systems is crucial for developing effective and innovative pest control strategies. Types of cultivation, such as crop monocultures versus polycultures or crop rotation, may act as a selective pressure on pests' capability to exploit the host's resources. In this study, we examined the herbivorous mite Aceria tosichella (commonly known as wheat curl mite), a widespread wheat pest, to understand how fluctuating versus stable environments influence its niche breadth and ability to utilize different host plant species. We subjected a wheat-bred mite population to replicated experimental evolution in a single-host environment (either wheat or barley), or in an alternation between these two plant species every three mite generations. Next, we tested the fitness of these evolving populations on wheat, barley, and on two other plant species not encountered during experimental evolution, namely rye and smooth brome. Our results revealed that the niche breadth of A. tosichella evolved in response to the level of environmental variability. The fluctuating environment expanded the niche breadth by increasing the mite's ability to utilize different plant species, including novel ones. Such an environment may thus promote flexible host-use generalist phenotypes. However, the niche expansion resulted in some costs expressed as reduced performances on both wheat and barley as compared to specialists. Stable host environments led to specialized phenotypes. The population that evolved in a constant environment consisting of barley increased its fitness on barley without the cost of utilizing wheat. However, the population evolving on wheat did not significantly increase its fitness on wheat, but decreased its performance on barley. Altogether, our results indicated that, depending on the degree of environmental heterogeneity, agricultural systems create different conditions that influence pests' niche breadth evolution, which may in turn affect the ability of pests to persist in such systems.


Cone and rod dysfunction in fundus albipunctatus with RDH5 mutation: an electrophysiological study.
A prior study showed that some patients with fundus albipunctatus (FA) have severely reduced full-field cone ERGs. The purpose of this study was to investigate the frequency of cone dysfunction and to determine the cause of the reduced full-field cone ERGs in patients with FA and whether the rod system is affected in patients with FA. Sixteen consecutive patients with FA (from 1993 to 2003; eight males, eight females; mean age, 25.4 years) with an RDH5 gene mutation were studied. The amplitudes and implicit times of the standard cone ERGs in the patients with FA were compared to those obtained from normal subjects (n = 55). The a-waves of cone ERGs were also elicited by a bright flash and were fitted to a mathematical model of the a-wave. Rod ERG responses were elicited by dim blue flashes after 3 hours of dark adaptation. The amplitude of the b-wave of the cone ERG in the FA group varied considerably from within the normal limits to markedly decreased. Six of 16 patients with FA had b-wave amplitudes that were smaller than the lowest limit of the control subjects. The degree of cone dysfunction tended to be more severe in older patients. The analysis of the cone a-wave demonstrated that R(m) (maximal response amplitude) in the patients with FA with reduced standard cone ERGs was significantly smaller than that in control subjects. Rod ERGs were also reduced in the patients with FA who had reduced cone ERGs. In patients with FA, 38% had extensive cone dysfunction. The reduced full-field cone ERGs were mainly due to the loss of cone photoreceptors, and the rod system was also affected in some patients.


[Cor triatriatum diagnosed in a 55-year-old man].
Cor triatriatum is a rare congenital anomaly in adults. The left atrium is devised by a fibro-muscular membrane. It is usually detected in infancy or early childhood, although some cases remain undetected until adult life. As a consequence of trans-membrane flow obstruction, the clinical features often mimic mitral stenosis. We describe an adult patient, in whom this anomaly was diagnosed in 55 year of life.


SenseLab: new developments in disseminating neuroscience information.
This article presents the latest developments in neuroscience information dissemination through the SenseLab suite of databases: NeuronDB, CellPropDB, ORDB, OdorDB, OdorMapDB, ModelDB and BrainPharm. These databases include information related to: (i) neuronal membrane properties and neuronal models, and (ii) genetics, genomics, proteomics and imaging studies of the olfactory system. We describe here: the new features for each database, the evolution of SenseLab's unifying database architecture and instances of SenseLab database interoperation with other neuroscience online resources.


BQ323636.1, a Novel Splice Variant to NCOR2, as a Predictor for Tamoxifen-Resistant Breast Cancer.
Purpose: Adjuvant tamoxifen treatment revolutionized the management of estrogen receptor (ER)-positive breast cancers to prevent cancer recurrence; however, drug resistance compromises its clinical efficacy. The mechanisms underlying tamoxifen resistance are not fully understood, and no robust biomarker is available to reliably predict those who will be resistant. Here, we study BQ323636.1, a novel splice variant of the NCOR2 gene, and evaluate its efficacy in predicting tamoxifen resistance in patients with breast cancer.Experimental Design: A monoclonal anti-BQ323636.1 antibody that specifically recognizes the unique epitope of this splice variant was generated for in vitro mechanistic studies and for in vivo analysis by immunohistochemistry on tissue microarrays of two independent cohorts of 358 patients with more than 10 years clinical follow-up data, who had ER-positive primary breast cancer and received adjuvant tamoxifen treatment. An orthotopic mouse model was also used.Results: Overexpression of BQ323636.1 conferred resistance to tamoxifen in both in vitro and in an orthotopic mouse model. Mechanistically, coimmunoprecipitation showed BQ323636.1 could bind to NCOR2 and inhibit the formation of corepressor complex for the suppression of ER signaling. Nuclear BQ3232636.1 overexpression in patients samples was significantly associated with tamoxifen resistance (P = 1.79 × 10-6, sensitivity 52.9%, specificity 72.0%). In tamoxifen-treated patients, nuclear BQ323636.1 overexpression was significantly correlated with cancer metastasis and disease relapse. Nuclear BQ323636.1 was also significantly associated with poorer overall survival (P = 1.13 × 10-4) and disease-specific survival (P = 4.02 × 10-5).Conclusions: These findings demonstrate that BQ323636.1 can be a reliable biomarker to predict tamoxifen resistance in patients with ER-positive breast cancer. Clin Cancer Res; 24(15); 3681-91. ©2018 AACRSee related commentary by Jordan, p. 3480.


Effects of extracellular pH, CO2, and HCO3- on ketogenesis in perfused rat liver.
Effects of extracellular pH, CO2, and HCO3- on ketone body production from octanoate were studied in perfused livers from fasted rats. pH was adjusted to 7.1-7.5 by varying perfusate [HCO3-] and [CO2], where brackets denote concentration. At constant 25 mM [HCO3-], total production of beta-hydroxybutyrate (beta-OHB) + acetoacetate (AcAc) was constant from pH 7.1 to 7.5. However, the [beta-OHB]/[AcAc] ratio decreased from 1.60 to 1.00 when pH decreased from 7.3 to 7.1; there was no change at pH 7.4. At constant [CO2], decreasing pH from 7.4 to 7.1 did not alter either total ketogenesis or the [beta-OHB]/[AcAc] ratio. This suggests that high [CO2] rather than low pH was responsible for the alteration in the redox ratio. At constant pH of 7.4, variations in [HCO3-] between 15 and 25 mM did not influence total ketogenesis or the [beta-OHB]/[AcAc] ratio. However, increasing [HCO3-] from 25 to 35 mM decreased the [beta-OHB]/[AcAc] ratio from 1.76 to approximately 1, again without affecting total ketogenesis. At constant 1.75 mM [CO2], increasing [HCO3-] from 25 to 35 mM also reduced the [beta-OHB]/[AcAc] ratio from 1.63 to approximately 1, suggesting that the effect of high [HCO3-] on this redox ratio can be ascribed to HCO3- itself. It is concluded that high [CO2] or [HCO3-] decreases the mitochondrial [NADH]/[NAD+] ratio in hepatocytes, resulting in a decreased [beta-OHB]/[AcAc] ratio.


Transient high thyroid stimulating hormone and hypothyroidism incidence during follow up of subclinical hypothyroidism.
Objectives. Given the high prevalence of subclinical hypothyroidism (SCH), defined as high thyroid stimulating hormone (TSH) and normal free thyroxine (FT4), and uncertainty on treatment, one of the major challenges in clinical practice is whether to initiate the treatment for SCH or to keep the patients under surveillance. There is no published study that has identified predictors of short-term changes in thyroid status amongst patients with mild elevation of TSH (4.5-10 mIU/L). Subjects and Results. A cohort study was conducted on patients with SCH detected through a general population screening program, who were followed for six months. This project identified factors predicting progression to hypothyroid status, persistent SCH and transient cases. A total of 656 participants joined the study (431 controls and 225 were patients with SCH). A part of participants (12.2%) developed biochemical hypothyroidism during the follow-up, while 73.8% of the subjects became euthyroid and the remained ones (13.4%) stayed in the SCH status. The incidence of overt hypothyroidism for participants with TSH above 6.9 mIU/L was 36.7%, with incidence of 42.3% for females. Anti-thyroid peroxidase antibodies (TPO) positivity is an important predictor of development of hypothyroidism; however, it could be also positive due to transient thyroiditis. Conclusions. It can be concluded that females with TSH above 6.9 mIU/L, particularly those with free triiodothyronine (FT3) and FT4 in the lower half of the reference range, are more likely to develop biochemical hypothyroidism. Therefore, it is recommended to give them a trial of levothyroxine replacement. It is also recommended to repeat TSH after six months for male subjects and participants with baseline TSH equal or less than 6.9 mIU/L.


Adaptation and Initial Validation of Minimum Data Set (MDS) Mortality Risk Index to MDS Version 3.0.
To evaluate the predictive validity of an adapted version of the Minimum Data Set (MDS) Mortality Risk Index-Revised (MMRI-R) based on MDS version 3.0 assessment items (MMRI-v3) and to compare the predictive validity of the MMRI-v3 with that of a single MDS item indicating limited life expectancy (LLE). Retrospective, cross-sectional study of MDS assessments. Other data sources included the Veterans Affairs (VA) Residential History File and Vital Status File. VA nursing homes (NHs). Veterans aged 65 and older newly admitted to VA NHs between July 1, 2012, and September 30, 2015. The dependent variable was death within 6 months of admission date. Independent variables included MDS items used to calculate MMRI-v3 scores (renal failure, chronic heart failure, sex, age, dehydration, cancer, unintentional weight loss, shortness of breath, activity of daily living scale, poor appetite, acute change in mental status) and the MDS item indicating LLE. The predictive ability of the MMRI-v3 for 6-month mortality (c-statistic 0.81) is as good as that of the original MMRI-R (c-statistic 0.76). Scores generated using the MMRI-v3 had greater predictive ability than that of the single MDS indicator for LLE (c-statistic 0.76); using the 2 together resulted in greater predictive ability (c-statistic 0.86). The MMRI-v3 is a useful tool in research and clinical practice that accurately predicts 6-month mortality in veterans residing in Veterans Affairs NHs. Identification of residents with LLE has great utility for studying palliative care interventions and may be helpful in guiding allocation of these services in clinical practice. J Am Geriatr Soc 66:2353-2359, 2018.


Anthropometry-based generation of personalized and population-specific human airway models.
Understanding aerosol deposition in the human lung is of great significance in pulmonary toxicology and inhalation pharmacology. Adverse effects of inhaled environmental aerosols and pharmacological efficacy of inhaled therapeutics are dependent on aerosol properties as well as person-specific respiratory tract anatomy and physiology. Anatomical geometry and physiological function of human airways depend on age, gender, weight, fitness, health, and disease status. Tools for the generation of the population- and subject-specific virtual airway anatomical geometry based on anthropometric data and physiological vitals are invaluable in respiratory diagnostics, personalized pulmonary pharmacology, and model-based management of chronic respiratory diseases. Here we present a novel protocol and software framework for the generation of subject-specific airways based on anthropometric measurements of the subject's body, using the anatomical input, and the conventional spirometry, providing the functional (physiological) data. This model can be used for subject-specific simulations of respiration physiology, gas exchange, and aerosol inhalation and deposition.


A Novel Fungal Lipase With Methanol Tolerance and Preference for Macaw Palm Oil.
Macaw palm is a highly oil-producing plant, which presents high contents of free fatty acids, being a promising feedstock for biofuel production. The current chemical routes are costly and complex, involving highly harsh industrial conditions. Enzymatic processing is a potential alternative; however, it is hampered by the scarce knowledge on biocatalysts adapted to this acidic feedstock. This work describes a novel lipase isolated from the thermophilic fungus Rasamsonia emersonii (ReLip), which tolerates extreme conditions such as the presence of methanol, high temperatures, and acidic medium. Among the tested feedstocks, the enzyme showed the highest preference for macaw palm oil, producing a hydrolyzate with a final free fatty acid content of 92%. Crystallographic studies revealed a closed conformation of the helical amphipathic lid that typically undergoes conformational changes in a mechanism of interfacial activation. Such conformation of the lid is stabilized by a salt bridge, not observed in other structurally characterized homologs, which is likely involved in the tolerance to organic solvents. Moreover, the lack of conservation of the aromatic cluster IxxWxxxxxF in the lid of ReLip with the natural mutation of the phenylalanine by an alanine might be correlated with the preference of short acyl chains, although preserving catalytic activity on insoluble substrates. In addition, the presence of five acidic amino acids in the lid of ReLip, a rare property reported in other lipases, may have contributed to its ability to tolerate and be effective in acidic environments. Therefore, our work describes a new fungal biocatalyst capable of efficiently hydrolyzing macaw oil, an attractive feedstock for the production of "drop-in" biofuels, with high desirable feature for industrial conditions such as thermal and methanol tolerance, and optimum acidic pH. Moreover, the crystallographic structure was elucidated, providing a structural basis for the enzyme substrate preference and tolerance to organic solvents.


(HOMOGRAFTS IN THYMECTOMIZED, IRRADIATED MICE: RESPONSES TO PRIMARY AND SECONDARY SKIN GRAFTS.)
Adult thymectomized, irradiated mice restored with syngeneic bone marrow exhibit an accelerated response toward second-set skin homografts, although the response to the primary grafts is impaired. Such mice have a long-lasting impairment (up to 223 days) of the primary response to allogeneic grafts (but not to xenogeneic grafts) and show the greatest deficiency of response toward grafts sharing the same H2 locus as the host. Thymectomy in the adult mouse is ineffective in altering a state of homograft sensitivity induced prior to irradiation.


Effect of patient age on increasing morbidity and mortality following urogynecologic surgery.
The purpose of this study was to estimate the effect of age on the risk of in-hospital mortality and morbidity following urogynecologic surgery and to compare risks associated with obliterative versus reconstructive procedures for prolapse in elderly women. We conducted a retrospective cohort study utilizing data from 1998 to 2002 from the Nationwide Inpatient Sample. Multivariable logistic regression was performed to obtain odds ratios estimating the effect of age on risk of death and complications, adjusting for comorbidities and demographic factors. There were 264,340 women in our study population. Increasing age was associated with higher mortality risks per 1000 women (< 60 years, 0.1; 60-69 years, 0.5; 70-79 years, 0.9; > or = 80 years, 2.8; P < .01) and higher complication risks per 1000 women (< 60 years, 140; 60-69 years, 130; 70-79 years, 160; > or = 80 years, 200; P < .01). Using multivariable logistic regression, increasing age was associated with an increased risk of death (60-69 years, odds ratio [OR] 3.4 [95% CI 1.7-6.9]; 70-79 years, OR 4.9 [95% CI 2.2-10.9]; > or = 80 years, OR 13.6 [95% CI 5.9-31.4]), compared with women < 60 years. The risk of peri-operative complications was also higher in elderly women 80 years of age and older (OR 1.4 [95% CI 1.3-1.5]) compared with younger women. Elderly women 80 years and over who underwent obliterative procedures had a lower risk of complication compared with those who underwent reconstructive procedures for prolapse (17.0% vs 24.7%, P < .01). Although the absolute risk of death is low, elderly women have a higher risk of mortality and morbidity following urogynecologic surgery.


Clinical prediction of fall risk and white matter abnormalities: a diffusion tensor imaging study.
The Tinetti scale is a simple clinical tool designed to predict risk of falling by focusing on gait and stance impairment in elderly persons. Gait impairment is also associated with white matter (WM) abnormalities. To test the hypothesis that elderly subjects at risk for falling, as determined by the Tinetti scale, have specific patterns of WM abnormalities on diffusion tensor imaging. Community-based cohort of 125 homebound elderly individuals. Diffusion tensor imaging scans were analyzed using tract-based spatial statistics analysis to determine the location of WM abnormalities in subjects with Tinetti scale scores of 25 or higher (without risk of falls) and lower than 25 (with risk of falls).Multivariate linear least squares correlation analysis was performed to determine the association between Tinetti scale scores and local fractional anisotropy values on each skeletal voxel controlling for possible confounders. In subjects with risk of falls (Tinetti scale score <25), clusters of abnormal WM were seen in the medial frontal and parietal subcortical pathways, genu and splenium of corpus callosum, posterior cingulum, prefrontal and orbitofrontal pathways, and longitudinal pathways that connect frontal-parietal-temporal lobes. Among these abnormalities, those in medial frontal and parietal subcortical pathways correlated with Mini-Mental State Examination scores, while the other locations were unrelated to these scores. Elderly individuals at risk for falls as determined by the Tinetti scale have WM abnormalities in specific locations on diffusion tensor imaging, some of which correlate with cognitive function scores.


Human T-cell leukemia virus type I Tax transactivates human interleukin 8 gene through acting concurrently on AP-1 and nuclear factor-kappaB-like sites.
The transactivator protein, Tax, from the human T-cell leukemia virus type I (HTLV-I) transactivates both viral and cellular genes. Previously, we had shown that interleukin 8 (IL-8) is constitutively expressed in HTLV-I-infected cells and in cells transiently expressing Tax. We show here that the IL-8 promoter is Tax responsive in Jurkat T cells. Furthermore, using several deletion and mutated plasmids of the 5'-flanking regulatory region of the IL-8 gene linked to the luciferase gene as a reporter and mutant tax gene expression vectors, we have established that both AP-1 at -126 to -120 and nuclear factor (NF)-kappaB-like cis-element at -80 to -71 are essential and sufficient for the induction of the IL-8 gene by HTLV-I Tax. In addition, overexpression of the dominant-negative mutants of NF-kappaB inhibitor molecules, IkappaBalpha and IkappaBbeta, abolished the Tax-induced activation of IL-8 gene. Gel mobility shift assays detected proteins specifically binding to the AP-1 and NF-kappaB-like sites in Tax-expressing T-cell lines infected with HTLV-I. Similarly, the nuclear translocation of proteins specifically bound to these two motifs was shown in JPX-9 cells, a subclone of Jurkat cells, carrying the Tax sequences under the control of an inducible promoter. Taken together, these results suggest that the cooperation of transcription factors NF-kappaB and AP-1 is essential for transactivation of IL-8 gene by HTLV-I Tax.


Artificial oligopeptide scaffolds for stoichiometric metal binding.
Two artificial peptides with pendant pyridine or bipyridine ligands have been synthesized and incorporated into oligomeric strands that are analogous to peptide nucleic acid. Spectrophotometric titrations with Cu(2+) and Fe(2+) show that the oligomers bind stoichiometric quantities of transition metals based on the number of pendant ligands. The identities of the titration products are confirmed by high resolution mass spectrometry. In the case of the bipyridine tripeptides, the titration stoichiometry and mass spectra indicate that the metal ions form interstrand cross-links between two oligopeptides, creating duplex structures linked exclusively by metal ions. Calculated molecular structures of the metalated oligopeptides and duplexes indicate that the peptide backbone acts as a scaffold for the directed assembly of metal ions. Electron paramagnetic resonance spectroscopy of the Cu-containing molecules have varying degrees of electronic interaction based on their charge and supramolecular structure. Cyclic voltammetry of the Fe(2+)- and Cu(2+)-linked bpy oligopeptide duplexes shows that they possess unique electrochemical signatures based on the redox reactivity of the metal complex.


The Role of Headache in the Classification and Management of Hypertensive Disorders in Pregnancy.
Hypertensive disorders of pregnancy remain among the leading causes of maternal morbidity and mortality. The onset of headaches in patients with hypertensive disorders of pregnancy has been considered as a premonitory symptom for eclampsia and other adverse maternal outcomes. Headaches are very common symptoms during pregnancy and the postpartum period with a reported incidence of 39%; however, headache is absent in 30-50% of women before the onset of eclampsia and is a poor predictor of eclampsia and adverse maternal outcomes. If included in the definition of cerebral or visual disturbances, headache may be considered a symptom of preeclampsia, a diagnostic feature of preeclampsia with severe features, a premonitory symptom of eclampsia, and an indication for delivery. Inclusion of this nonspecific symptom in the diagnosis and management of hypertensive disorders of pregnancy in the absence of an evidence basis may lead to unintended consequences including excessive testing, visits to outpatient offices or emergency departments, additional hospitalization, and iatrogenic preterm delivery without proven benefit. If a cerebral disturbance such as severe or persistent headache presents for the first time during pregnancy or postpartum, an evaluation should be performed that considers a broad differential diagnosis, including but not limited to hypertensive disorders of pregnancy, and the diagnostic evaluation is similar to that in nonpregnant adults. This commentary draws attention to the implications of considering the cerebral disturbance of headache as a symptom that portends adverse pregnancy outcome in the current recommendations for diagnosing and managing hypertensive disorders of pregnancy.


[Improved access to the superficial femoral artery in PTA using a Tuohy cannula].
Description of a new needle well suited for problem cases in percutaneous transluminal angioplasty of the superficial femoral artery. This needle facilitates probing of the vessel in antegrade puncture and makes sliding off into the deep femoral artery less likely.