Automated video analysis system reveals distinct diurnal behaviors in C57BL/6 and C3H/HeN mice.
Advances in rodent behavior dissection using automated video recording and analysis allows detailed phenotyping. This study compared and contrasted 15 diurnal behaviors recorded continuously using an automated behavioral analysis system for a period of 14 days under a 14/10 light/dark cycle in single housed C3H/HeN (C3H) or C57BL/6 (C57) male mice. Diurnal behaviors, recorded with minimal experimental interference and analyzed using phenotypic array and temporal distribution analysis showed bimodal and unimodal profiles in the C57 and C3H mice, respectively. Phenotypic array analysis revealed distinct behavioral rhythms in Activity-Like Behaviors (i.e. walk, hang, jump, come down) (ALB), Exploration-Like Behaviors (i.e. dig, groom, rear up, sniff, stretch) (ELB), Ingestion-Like Behaviors (i.e. drink, eat) (ILB) and Resting-Like Behaviors (i.e. awake, remain low, rest, twitch) (RLB) of C3H and C57 mice. Temporal distribution analysis demonstrated that strain and time of day affects the magnitude and distribution of the spontaneous homecage behaviors. Wheel running activity, water and food measurements correlated with timing of homecage behaviors. Subcutaneous (3 mg/kg, sc) or oral (0.02 mg/ml, oral) melatonin treatments in C57 mice did not modify either the total 24 h magnitude or temporal distribution of homecage behaviors when compared with vehicle treatments. We conclude that C3H and C57 mice show different spontaneous activity and behavioral rhythms specifically during the night period which are not modulated by melatonin.


Circumscribed choroidal hemangioma: Clinical features and outcomes by age category in 458 cases.
To investigate features and outcomes of circumscribed choroidal hemangioma by patient age. Retrospective review of circumscribed choroidal hemangioma from 3/29/1967-6/4/2018 based on age at presentation (≤20 vs. >20-50 vs. >50 years). There were 458 circumscribed choroidal hemangiomas diagnosed at mean age (13 vs. 41 vs. 64 years, p < 0.001). The youngest age group had worse presenting visual acuity (20/400 vs. 20/150 vs. 20/100, p < 0.001), larger tumor basal diameter (13.5 vs. 6.6 vs. 6.2, p < 0.001), greater tumor thickness (5.8 vs. 3.1 vs. 2.9, p < 0.001), closer distance to foveola (0.5 vs. 1.4 vs. 1.2, p = 0.03), and greater extent of subretinal fluid (4 quadrants, 26% vs. 8% vs. 2%, p < 0.001). The youngest patients were less likely to be treated with primary observation (39% vs. 39% vs. 56%) or photodynamic therapy (10% vs. 27% vs. 22%) and more likely to be treated with plaque radiotherapy (26% vs. 6% vs. 3%) or external beam radiotherapy (13% vs. 1% vs. 0%) (p < 0.001). The youngest patients required greater total number of treatments (mean 4 vs. 2 vs. 1, p < 0.001). At mean follow-up (44 vs. 68 vs. 60 months, p = 0.37), the youngest patients had worse visual acuity (20/400 vs. 20/200 vs. 20/100, p = 0.03), but no difference in visual acuity loss of 3 or more Snellen lines (27% vs. 13% vs. 16%, p = 0.55). Younger patients (≤20 years) with circumscribed choroidal hemangioma present with worse visual acuity and larger, more posterior tumors. Future studies are needed to improve early detection and treatment for this subgroup of patients.


Influence of pocket gopher mounds on a Texas coastal prairie.
Effects of pocket gopher (Geomys attwateri) mound-building activity on plant community composition and soil nutrient concentrations were investigated in south Texas on both burned and unburned coastal prairie sites. Pocket gophers deposited large amounts of soil which were lower in nutrient content than randomly-collected samples. Above-ground plant biomass was greater around mounds than in random samples mainly because of increased dicots around mounds on the burned area when compared with random samples on the same area. Pocket gophers may have concentrated their activities (and therefore, mounds) in areas with higher dicot biomass on the burned area since they prefer perennial dicots as food, or the presence of mounds may have ameliorated the apparent negative effect of fire on dicots.


Environmental co-regulation of substance P, somatostatin and neurotransmitter synthesizing enzymes in cultured sympathetic neurons.
Mechanisms regulating peptide neurotransmitter metabolism were examined in dissociated cell cultures of the neonatal rat superior cervical ganglion (SCG). The pineal gland, a target of the SCG, produced a soluble factor (PCM) which increased substance P (SP) levels more than 15-fold in sympathetic neurons cultured in the presence of ganglion non-neuronal cells. Elimination of the non-neuronal cells decreased SP to negligible levels and abolished the stimulatory effects of PCM on SP expression. These observations suggest that ganglion non-neuronal cells stimulate sympathetic expression of SP, and that the pineal influences neuronal SP by acting on, or in concert with, ganglion support cells. PCM also influenced other neurotransmitter systems. In the presence of ganglion non-neuronal cells, PCM treatment increased choline acetyltransferase (CHAC) and decreased tyrosine hydroxylase (TOH) and somatostatin (SO). By contrast, PCM treatment of pure neuronal cultures resulted in negligible CHAC and SP levels and a doubling of SO with a small increase in TOH. In sympathetic neurons, SP expression may be associated with cholinergic development, whereas SO may be associated with noradrenergic phenotypic expression. Moreover, there is a reciprocal relationship between SP and SS expression by sympathetic neurons analogous to the previously described relationship between noradrenergic and cholinergic expression.


Determinants of late antenatal care follow up among pregnant women in Easter zone Tigray, Northern Ethiopia, 2018: unmatched case-control study.
The purpose of the study was to identify determinants of late antennal care at first visit in health facilities of eastern zone of Tigray, Northern Ethiopia 2018. Women with unplanned pregnancy (AOR = 4.03, 95%, CI 1.56-5.67), Participants whose previous first antenatal care was after 16 weeks (AOR = 3.9, 95% CI 1.98-7.68), Participants did not accompanied by their partner for antenatal visit (AOR = 1.29, 95%, CI 1.05-4.67), women recognized their current pregnancy at 3 months or late (AOR = 4.7, 95%, CI 2.49-9.04) and participants provided adequate time for their previous antenatal care by health professionals (AOR = 0.461, 95% CI 0.342-0.875) were found the determinant factors of late antenatal care at first Visit. Hence family planning utilization, times of first visit antenatal, information flow and supporting by partners have a great role in improving antenatal care at first visit. There for responsible bodies should give focuses on utilization of family planning, increasing awareness of pregnancy symptoms and health provisional provide adequate time during visits.


Alcohol consumption among recreational boaters: factors for intervention.
Recreational boating is a popular leisure time activity in many countries. It is estimated that, in Australia, boating incidents cause more harm than rail and air crashes combined and, in terms of transport, are second only to motor vehicle crashes as a cause of serious injury. The consumption of alcohol among recreational boaters is considered an important risk factor for fatalities and injuries among both operators and passengers. Using a database of all recreational vessels registered in Western Australia (WA), a sample of 500 adult boaters was recruited to participate in a telephone survey. The effects of demographic variables and boating characteristics upon the use of alcohol among recreational boaters on their last trip were explored using logistic regression. The odds of not having a drink were associated, after adjusting for age, with having completed a boating education course and with carrying children less than 12 years on board. The use of alcohol was not found to be prevalent among WA recreational boat owners. Based on these findings, it is recommended that efforts to decrease boating-related incidents, such as through education and legislation measures, be monitored over time to determine the effects of these strategies upon safety behaviours.


Enhanced removal of basic dye using carbon nitride/graphene oxide nanocomposites as adsorbents: high performance, recycling, and mechanism.
The presence of dyes in wastewater streams poses a great challenge for sustainability and brings the need to develop technologies to treat effluent streams. Here, we propose a mixture of high superficial area carbon-based nanomaterial strategy to improve the removal of basic blue 26 (BB26) by blending porous carbon nitride (CN) and graphene oxide (GO). We prepared CN and GO pristine materials, as well the nanocomposites with mass/ratio 30/70, 50/50, and 70/30, and applied them into BB26 uptake. Nanocomposite 50/50 CN/GO was found to be the better adsorbent, and the optimization of the adsorption revealed a fast equilibrium time of 30 min, after sonication for 2 min, nanocomposite 50/50, and BB26 dye loading of 0.1 g/L and 100 mg/L, respectively. The pH variation had great influence on BB26 uptake, and at ultrapure water pH, the dye removal capacity by the composite reached 917.78 mg/g. At pH 2, a remarkable removal efficiency of 3510.10 mg/g was obtained, probably due to electrostatic interactions among protonated amine groups of the dye and negatively charged CN/GO nanocomposite. The results obtained were best fitted to the pseudo-second-order kinetic model and the Dubinin-Radushkevich isotherm. The adsorption process was thermodynamically spontaneous, and physisorption was the main mechanism, which is based on weak electrostatic and π-π interactions. The dye attached to the CN/GO nanocomposite could be removed by washing with ethyl alcohol, and the adsorbent was reused for five consecutive cycles with high BB26 uptake efficiency. The CN/GO nanocomposite ability to remove the BB26 dye was 21 times higher than those reported in the literature, indicating CN/GO composites as potential filtering materials to basic dyes.


Survival of other fetuses after a fetal death in twin or triplet pregnancies.
To estimate the frequency of fetal death in multifetal pregnancies and the probability of survival to age 1 year for twins or triplets in which at least one fetal death occurred at 20 weeks' gestation or more. We used the Matched Multiple Birth File from the US National Center for Health Statistics, which included 152,233 sets of twins and 5356 sets of triplets registered from 1995 to 1997. The Cox proportional hazards model was used to estimate the adjusted relative risk of death before age 1 year for remaining twins and triplets. Fetal death at 20 weeks' gestation or later was uncommon, occurring in 2.6% of twin and 4.3% of triplet gestations. After adjustment for confounders, the survival of the remaining fetuses was inversely related to the time of the first fetal demise. Same-sex twins were two times more likely than opposite-sex twins to die after an intrauterine demise at 25-32 weeks' gestation and were more than three times more likely to die after a death at 33 weeks' gestation or more. After a fetal death in a multifetal pregnancy at 20 weeks' gestation or later, the survival of the remaining fetuses is inversely related to the time the death occurred. Among twins, survival also depends on sex concordance, with opposite-sex twins more likely than same-sex twins to survive.


Resource Allocation for Epidemic Control Across Multiple Sub-populations.
The number of pathogenic threats to plant, animal and human health is increasing. Controlling the spread of such threats is costly and often resources are limited. A key challenge facing decision makers is how to allocate resources to control the different threats in order to achieve the least amount of damage from the collective impact. In this paper we consider the allocation of limited resources across n independent target populations to treat pathogens whose spread is modelled using the susceptible-infected-susceptible model. Using mathematical analysis of the systems dynamics, we show that for effective disease control, with a limited budget, treatment should be focused on a subset of populations, rather than attempting to treat all populations less intensively. The choice of populations to treat can be approximated by a knapsack-type problem. We show that the knapsack closely approximates the exact optimum and greatly outperforms a number of simpler strategies. A key advantage of the knapsack approximation is that it provides insight into the way in which the economic and epidemiological dynamics affect the optimal allocation of resources. In particular using the knapsack approximation to apportion control takes into account two important aspects of the dynamics: the indirect interaction between the populations due to the shared pool of limited resources and the dependence on the initial conditions.


Motivational concordance: an important mechanism in self-help therapeutic rituals involving inert (placebo) substances.
We tested the contribution of two mechanisms, response expectancy and motivational concordance, to reported psychological benefit from a popular, biologically inactive, self-help, complementary therapy (a placebo). Flower essences were taken by 251 people for self-selected symptoms and were randomized to receive three different kinds of information. When the flower essence was presented as a spiritual therapy, then baseline spirituality (beta=.35, P=.01) and expectancy (beta=.25, P=.03) independently predicted outcome. When flower essences were presented as an affirmation (i.e., nonspiritual) therapy, then spirituality negatively (beta=-.27, P=.03) and expectancy (beta=.33, P=.01) predicted outcome. For both groups, expectancy predicted outcome after controlling for spirituality and compliance, but did not after controlling for ease of task completion. Expectancy failed to predict outcome in the nonenhanced ritual group. The results suggest that motivational concordance is an important therapeutic mechanism for real-life placebos.


Nanoparticle-complexed antimiRs for inhibiting tumor growth and metastasis in prostate carcinoma and melanoma.
MiRNAs act as negative regulators of gene expression through target mRNA degradation or inhibition of its translation. In cancer, several miRNAs are upregulated and play crucial roles in tumorigenesis, making the inhibition of these oncomiRs an interesting therapeutic approach. This can be achieved by directly complementary single-stranded anti-miRNA oligonucleotides (antimiRs). A major bottleneck in antimiR therapy, however, is their efficient delivery. The nanoparticle formation with polyethylenimine (PEI) may be particularly promising, based on the PEI's ability to electrostatically interact with oligonucleotides. This leads to their protection and supports delivery. In the present study, we explore for the first time PEI for antimiR formulation and delivery. We use the branched low molecular weight PEI F25-LMW for the complexation of different antimiRs, and analyse tumor- and metastasis-inhibitory effects of PEI/antimiR complexes in different tumor models. In prostate carcinoma, transfection of antimiRs against miR-375 and miR-141 leads to tumor cell inhibition in 2D- and 3D-models. More importantly, an in vivo tumor therapy study in prostate carcinoma xenografts reveals anti-tumor effects of the PEI/antimiR complexes. In advanced melanoma and metastasis, we identify by a microRNA screen miR-150 as a particularly relevant oncomiR candidate, and validate this result in vitro and in vivo. Again, the systemic application of PEI/antimiR complexes inhibiting this miRNA, or the previously described antimiR-638, leads to profound tumor growth inhibition. These effects are associated with the upregulation of direct miRNA target genes. In a melanoma metastasis mouse model, anti-metastatic effects of PEI/antimiR treatment are observed as well. We thus describe PEI-based complexes as efficient platform for antimiR therapy, as determined in two different tumor entities using in vivo models of tumor growth or metastasis. Our study also highlights the therapeutic relevance of miR-375, miR-141, miR-150 and miR-638 as target miRNAs for antimiR-mediated inhibition.


Overview of Fusion Tags for Recombinant Proteins.
Virtually all recombinant proteins are now prepared using fusion domains also known as "tags". The use of tags helps to solve some serious problems: to simplify procedures of protein isolation, to increase expression and solubility of the desired protein, to simplify protein refolding and increase its efficiency, and to prevent proteolysis. In this review, advantages and disadvantages of such fusion tags are analyzed and data on both well-known and new tags are generalized. The authors own data are also presented.


Susceptibility of a polycaprolactone-based root canal filling material to degradation. II. Gravimetric evaluation of enzymatic hydrolysis.
Polycaprolactone is susceptible to enzymatic biodegradation via ester bond cleavage. This study examined the susceptibility of Resilon, a polycaprolactone-based root filling material to enzymatic hydrolysis. Resilon, gutta-percha, and polycaprolactone disks, prepared by compression molding, were incubated in phosphate-buffered saline, lipase PS or cholesterol esterase at 37 degrees C for 96 h. They were retrieved at different time intervals for gravimetric analysis and scanning electron microscopy. The materials exhibited slight weight gains when incubated in phosphate-buffered saline that can be attributed to water sorption. Gutta-percha showed similar weight gains in the two enzymes. Conversely, Resilon and polycaprolactone exhibited extensive surface thinning and weight losses after incubation in lipase PS and cholesterol esterase. Glass filler particles in Resilon were exposed following surface dissolution of the polymer matrix, creating rough surface topography. Biodegradation of Resilon by bacterial and salivary enzymes warrants further investigation of their activities using cultures of endodontically relevant microbes and human saliva extracts.


Clear Cell Carcinoma of Palatal Minor Salivary Gland Harboring a Novel EWSR1-ATF1 Fusion Gene: Report of a Case and Review of the Literature.
Clear cell carcinoma (CCC) is a rare low-grade malignant salivary gland carcinoma. EWSR1-ATF1 fusion has been characterized as a consistent finding in CCC, with breakpoints described between EWSR1 exon 11 and ATF1 exon 3. So far, over 100 cases of CCC harboring EWSR1 rearrangement arising from salivary gland of the oral cavity have been reported. Although EWSR1 involvement in these cases was confirmed by EWSR1 break-apart FISH indicating the translocation, sequence analysis for EWSR1-ATF1 fusion type has been reported only in three cases of CCC so far. Herein, we report a CCC case with novel EWSR1-ATF1 fusion (EWSR1 exon 15 and ATF1 exon 5) arising in minor salivary gland and review the role of the chimeric variants in some malignancies with EWSR1-ATF1 rearrangement. Current tumor was composed of the small nests of clear tumor cells and hyalized fibrous stroma. Immunohistochemically, the tumor was positive for AE1/AE3, CK5/6 and p63, negative for S100, Melan-A, SMA and CD10. After 8 months of follow-up, there are no evidence of recurrence.


Liquid-phase microextraction combined with high-performance liquid chromatography for the determination of local anaesthetics in human urine.
A simple liquid-phase microextraction (LPME) device combined with high-performance liquid chromatography (HPLC) is presented for the simultaneous analysis of local anaesthetics, lidocaine, bupivacaine, and tetracaine, from human urine sample. An organic solvent showed good compatibility with the mobile phase of the HPLC, o-dibutyl phthalate, was selected. Local anaesthetics are extracted from 6 ml of the feed aqueous solution and human urine sample into a water-immiscible organic solvent suspended at the needle tip of the microsyringe, then the organic solvent was directly introduced to a reversed-phase HPLC system. The kind of the organic extraction solvent, the stirring rate, the pH value of the aqueous feed solution, and the extraction time have been discussed. Under the optimized extraction conditions, high enrichment factors (more than 86.0-fold) and significant sample clean-up for all of studied local anaesthetics were achieved within 30 min. The detection limits (lower than 0.05 microg/ml) were comparable with previously reported gas chromatography methods. This method was applied to specimen of patient who was treated with extradural anaesthesia of lidocaine, bupivacaine, and tetracaine, and revealed that simultaneous determination of above three local anaesthetics in human urine was possible.


Confusion From the FDA on Compounding: Agency Tries to Expand List of Useable "Bulk Drug Substances".
Confusion, ire follow FDA interim policy on compounding.


Mechanism of the defect formation in supported graphene by energetic heavy ion irradiation: the substrate effect.
Although ion beam technology has frequently been used for introducing defects in graphene, the associated key mechanism of the defect formation under ion irradiation is still largely unclear. We report a systematic study of the ion irradiation experiments on SiO2-supported graphene, and quantitatively compare the experimental results with molecular dynamic simulations. We find that the substrate is, in fact, of great importance in the defect formation process, as the defects in graphene are mostly generated through an indirect process by the sputtered atoms from the substrate.


Essential thrombocythemia associated with fibrosis involving bilateral renal sinuses: A case report.
Essential thrombocythemia (ET) is associated with an increased risk of thrombosis and autoimmune renal involvement. We report an extremely rare case of an acute kidney injury (AKI) in the presence of bilateral renal pelvises fibrosis in a patient with a proven diagnosis of ET. A 48-year-old male patient with a past medical history of mild chronic kidney disease and ET was admitted to our hospital with AKI. The patient discontinued his hydroxyurea treatment for the past 2 months and laboratory data showed increasing serum creatinine levels and platelet counts with increased renal sizes, severe hydrocalyx, and bilateral renal sinuses' fibrosis in imaging. The patient started again on hydroxyurea therapy and showed improvement in all laboratory scales. ET and increased levels of platelet-derived growth factors could cause renal sinuses fibrosis and glomerulopathy. In ET patients with renal sinuses' fibrosis and glomerulopathy, initiating cytoreductive therapy could improve the outcome.


The impact of prior fragility fractures on long-term periprosthetic fracture risk following total knee arthroplasty.
The study found that patients undergoing total knee arthroplasty with prior fragility fracture had increased risk of subsequent fragility fracture and periprosthetic fracture within 8 years postoperatively when compared to those without a prior history. However, these patients were not at increased risk for all-cause revision within this period. The aim of this study was to characterize the association of prior FFs on long-term risk of secondary fragility fracture (FF), periprosthetic fracture (PPF), and revision TKA. Patients at least 50 years of age who underwent elective TKA were identified in the PearlDiver Database. Patients were stratified based on whether they sustained a FF within 3 years prior to TKA (7410 patients) or not (712,954 patients). Demographics and comorbidities were collected. Kaplan Meier analysis was used to observe the cumulative incidence of all-cause revision, PPF, and secondary FF within 8 years of TKA. Cox Proportional hazard ratio analysis was used to statistically compare the risk. In total, 1.0% of patients had a FF within three years of TKA. Of these patients, only 22.6% and 10.9% had a coded diagnosis of osteoporosis and osteopenia, respectively, at time of TKA. The 8-year cumulative incidence of secondary FF and periprosthetic fracture was significantly higher in those with a prior FF (27.5% secondary FF and 1.9% PPF) when compared to those without (9.1% secondary FF and 0.7% PPF). After adjusting for covariates, patients with a recent FF had significantly higher risks of secondary FF (HR 2.73; p < 0.001) and periprosthetic fracture (HR 1.86; p < 0.001) than those without a recent FF. Recent FF before TKA is associated with increased risk for additional FF and PPF within 8 years following TKA. Surgeons should ensure appropriate management of fragility fracture is undertaken prior to TKA to minimize fracture risk, and if not, be vigilant to identify patients with prior FF or other bone health risk factors who may have undocumented osteoporosis.


Estrogen Sulfotransferase Is an Oxidative Stress-responsive Gene That Gender-specifically Affects Liver Ischemia/Reperfusion Injury.
Estrogen sulfotransferase (EST) regulates estrogen homeostasis by sulfonating and deactivating estrogens. Liver ischemia and reperfusion (I/R) involves both hypoxia during the ischemic phase and oxidative damage during the reperfusion phase. In this report, we showed that the expression of EST was markedly induced by I/R. Mechanistically, oxidative stress-induced activation of Nrf2 was responsible for the EST induction, which was abolished in Nrf2(-/-) mice. EST is a direct transcriptional target of Nrf2. In female mice, the I/R-responsive induction of EST compromised estrogen activity. EST ablation attenuated I/R injury as a result of decreased estrogen deprivation, whereas this benefit was abolished upon ovariectomy. The effect of EST ablation was sex-specific because the EST(-/-) males showed heightened I/R injury. Reciprocally, both estrogens and EST regulate the expression and activity of Nrf2. Estrogen deprivation by ovariectomy abolished the I/R-responsive Nrf2 accumulation, whereas the compromised estrogen deprivation in EST(-/-) mice was associated with increased Nrf2 accumulation. Our results suggested a novel I/R-responsive feedback mechanism to limit the activity of Nrf2 in which Nrf2 induces the expression of EST, which subsequently increases estrogen deactivation and limits the estrogen-responsive activation of Nrf2. Inhibition of EST, at least in females, may represent an effective approach to manage hepatic I/R injury.


Dialysate-side urea kinetics. Neural network predicts dialysis dose during dialysis.
Determination of the adequacy of dialysis is a routine but crucial procedure in patient evaluation. The total dialysis dose, expressed as Kt/V, has been widely recognised to be a major determinant of morbidity and mortality in haemodialysed patients. Many different factors influence the correct determination of Kt/V, such as urea sequestration in different body compartments, access and cardiopulmonary recirculation. These factors are responsible for urea rebound after the end of the haemodialysis session, causing poor Kt/V estimation. There are many techniques that try to overcome this problem. Some of them use analysis of blood-side urea samples, and, in recent years, on-line urea monitors have become available to calculate haemodialysis dose from dialysate-side urea kinetics. All these methods require waiting until the end of the session to calculate the Kt/V dose. In this work, a neural network (NN) method is presented for early prediction of the Kt/V dose. Two different portions of the dialysate urea concentration-time profile (provided by an on-line urea monitor) were analysed: the entire curve A and the first half B, using an NN to predict the Kt/V and compare this with that provided by the monitor. The NN was able to predict Kt/V is the middle of the 4h session (B data) without a significant increase in the percentage error (B data: 6.69% +/- 2.46%; A data: 5.58% +/- 8.77%, mean +/- SD) compared with the monitor Kt/V.


[Meningiomas. Clinical case examples].
Using four case reports as examples the best treatment is discussed by considering the specific history and magnetic resonance imaging (MRI) findings. In many cases different therapeutic alternatives are possible for treatment of meningiomas.


The Figure-of-Eight Walk test: reliability and associations with stroke-specific impairments.
To investigate (1) the intra-rater, inter-rater and test-retest reliabilities of the Figure-of-Eight Walk (F8W) test times; (2) its correlation with other stroke-specific impairments; and (3) the cut-off scores best discriminating patients with stroke from the healthy elderly. Cross-sectional study. University-based rehabilitation centre. A convenience sample of 64 subjects: 35 subjects with chronic stroke and 29 healthy elderly. F8W test times, Fugl-Meyer Motor Assessment for the lower extremities (FMA-LE), hand-held dynamometer measurements of bilateral hip abductor and knee extensor isometric muscle strength, Five times Sit to Stand Test (FTSTST) times, 10-Meter Walk Test (10MWT), Timed Up and Go Test (TUGT) times, Berg Balance Scale (BBS) and Activities-specific Balance Confidence Scale (ABC) scores. Excellent intra-rater, inter-rater and test-retest reliabilities (intra-class correlation coefficient (ICC) range 0.944-0.999) of F8W test times were found. The F8W test times were also found to be significantly associated with FMA-LE, BBS, FTSTST, TUG scores and 10MWT. No significant correlation was found between F8W test times and either leg strength or ABC results. A F8W test time of 8.2 s was found to be the most representative for discriminating between healthy elderly and stroke subjects, with a sensitivity of 100% and a specificity of 89.7%. The F8W test time is a reliable measurement tool, which is able to differentiate the patients with stroke and healthy elderly subjects and correlated well with stroke-specific impairments and walking tests. The F8W is a reliable measurement tool for assessing the advanced walking performance of subjects with chronic stroke. Implication for Rehabilitation The F8W test times have excellent intra-rater, inter-rater and test-retest reliabilities in patients with chronic stroke. The F8W test times were also found to be significantly associated with FMA-LE, BBS, FTSTST, TUG scores and 10MWT. A F8W test time of 8.2 s was found to be the most representative for discriminating between healthy elderly and stroke subjects, with a sensitivity of 100% and a specificity of 89.7%. The F8W test time is a reliable and valid measure in assessing the advanced walking skill in patients with stroke.


Quantitative bone marrow magnetic resonance imaging through apparent diffusion coefficient and fat fraction in multiple myeloma patients.
Quantitative bone marrow (BM) MR sequences, as DWI and CSI, were used to evaluate BM water-fat composition. The aim of the study was to assess the potential usefulness of fat fraction (FF) and ADC, calculated by CSI or DWI, in diagnosing and classifying myeloma (MM) patients according to their different BM infiltration patterns. The study group included 43 MM patients (19F; 24M; mean age 64 years), 15 asymptomatic, 15 symptomatic with diffuse BM infiltration and 13 symptomatic with focal lesions (FLs). The control group was made up of 15 healthy subjects (7F; 8M; mean age 64 years). MRI examinations consisted of sagittal T1w TSE on the spinal column, axial DWI (b 50-400-800 mm2/s) and coronal T2 Dixon, on the whole body. Mean ADC and FF were calculated placing 1 ROI on 6 vertebras and 2 ROIs on either the pelvis or FL. ANOVA with Bonferroni's correction showed a significant difference in ADC values among the different groups of MM patients (P < 0.05), while FF was only significantly different between patients with diffuse infiltration and patients with FL (P = 0.002). ADC allowed distinguishing MM patients from normal BM patients with diffuse BM infiltration (cutoff value: 0.491 × 10-3 mm2/s; sensitivity 73%, specificity 80%). FF helped better discriminate healthy controls from normal BM patients (cutoff = 0.33, sensitivity 73%, specificity 92%) and patients with diffuse BM infiltration from those with FL (cutoff = 0.16, sensitivity 82%, specificity 92%). ADC and FF are potentially useful parameter for the quantitative evaluation of BM infiltration in MM patients.


Successful management of severe aortocaval compression in twin pregnancy.
In a patient with severe aortocaval compression, simultaneous brachial and femoral blood pressure measurements demonstrated the need for a 30 degrees left-down tilt to avoid significant obstruction of the vessels. When emergency cesarean section became necessary, proper positioning of the patient was readily accomplished.


Mitochondrial metabolism and DNA methylation: a review of the interaction between two genomes.
Mitochondria are controlled by the coordination of two genomes: the mitochondrial and the nuclear DNA. As such, variations in nuclear gene expression as a consequence of mutations and epigenetic modifications can affect mitochondrial functionality. Conversely, the opposite could also be true. However, the relationship between mitochondrial dysfunction and epigenetics, such as nuclear DNA methylation, remains largely unexplored. Mitochondria function as central metabolic hubs controlling some of the main substrates involved in nuclear DNA methylation, via the one carbon metabolism, the tricarboxylic acid cycle and the methionine pathway. Here, we review key findings and highlight new areas of focus, with the ultimate goal of getting one step closer to understanding the genomic effects of mitochondrial dysfunction on nuclear epigenetic landscapes.


Biomechanical model for appressorial design in Magnaporthe grisea.
The fungus Magnaporthe grisea, commonly referred to as the rice blast fungus, is responsible for destroying from 10% to 30% of the world's rice crop each year. The fungus attaches to the rice leaf and forms a dome-shaped structure, the appressorium, in which enormous pressures are generated that are used to blast a penetration peg through the rice cell walls and infect the plant. We develop a model of the appressorial design in terms of a bioelastic shell that can explain the shape of the appressorium, and its ability to maintain that shape under the enormous increases in turgor pressure that can occur during the penetration phase.


Expression and testing in plants of ArcLight, a genetically-encoded voltage indicator used in neuroscience research.
It is increasingly appreciated that electrical controls acting at the cellular and supra-cellular levels influence development and initiate rapid responses to environmental cues. An emerging method for non-invasive optical imaging of electrical activity at cell membranes uses genetically-encoded voltage indicators (GEVIs). Developed by neuroscientists to chart neuronal circuits in animals, GEVIs comprise a fluorescent protein that is fused to a voltage-sensing domain. One well-known GEVI, ArcLight, undergoes strong shifts in fluorescence intensity in response to voltage changes in mammalian cells. ArcLight consists of super-ecliptic (SE) pHluorin (pH-sensitive fluorescent protein) with an A227D substitution, which confers voltage sensitivity in neurons, fused to the voltage-sensing domain of the voltage-sensing phosphatase of C iona i ntestinalis (Ci-VSD). In an ongoing effort to adapt tools of optical electrophysiology for plants, we describe here the expression and testing of ArcLight and various derivatives in different membranes of root cells in Arabidopsis thaliana. Transgenic constructs were designed to express ArcLight and various derivatives targeted to the plasma membrane and nuclear membranes of Arabidopsis root cells. In transgenic seedlings, changes in fluorescence intensity of these reporter proteins following extracellular ATP (eATP) application were monitored using a fluorescence microscope equipped with a high speed camera. Coordinate reductions in fluorescence intensity of ArcLight and Ci-VSD-containing derivatives were observed at both the plasma membrane and nuclear membranes following eATP treatments. However, similar responses were observed for derivatives lacking the Ci-VSD. The dispensability of the Ci-VSD suggests that in plants, where H(+) ions contribute substantially to electrical activities, the voltage-sensing ability of ArcLight is subordinate to the pH sensitivity of its SEpHluorin base. The transient reduction of ArcLight fluorescence triggered by eATP most likely reflects changes in pH and not membrane voltage. The pH sensitivity of ArcLight precludes its use as a direct sensor of membrane voltage in plants. Nevertheless, ArcLight and derivatives situated in the plasma membrane and nuclear membranes may offer robust, fluorescence intensity-based pH indicators for monitoring concurrent changes in pH at these discrete membrane systems. Such tools will assist analyses of pH as a signal and/or messenger at the cell surface and the nuclear periphery in living plants.


[Results of the GECO series].
301 hip prostheses were implanted in patients under the age of 50 years. All patients were followed for a minimal period of 3 years or until revision. The young age is due to the etiologies: femoral head necrosis, congenital hip dislocation, post-traumatic arthrosis. The implanted prostheses were of different types: both components were usually cementless; most bearing surfaces were alumina-polyethylene, with a head diameter of 32 or 28 mm. Patients were followed for 3 to 18 years after implantation (mean: 7 years). At the last follow-up, 59 prostheses (20%) had been exchanged; 232 cases had a good functional result according to Merle d'Aubigné (77% of the whole series and 96% of the surviving prostheses). The non revised patients mostly had an excellent result with regard to pain, range of motion and gait, and 85% returned to work.The final X-ray examinations of surviving prostheses showed 22% acetabular and 25% femoral radiolucent lines without loosening, 7% severe acetabular polyethylene wear. 3% of acetabular and 9% of femoral prostheses had migrated, mostly during the first months after cementless implantation with subsequent stabilisation.Revisions were performed after 6 months to 16 years. Main reasons were primary instability of a cementless implant (14 cases), secondary instability of a cementless implant (18 cases) and loosening of a cemented implant (18 cases). The global survival rate is 67% after 10 years, and 47% after 14 years. The survival rate of cemented cups was higher than screwed uncemented cups after 12 years (73% vs 59% - p<0,05), and there was no difference with uncemented press-fit cups after 7 years (95% vs 90%). The survival rate of cemented cups was higher than uncoated uncemented cups after 13 years(67% vs 53% -p<0,05), and there was no difference with hydroxyapatite-coated uncemented cups after 5 years (97% vs 93%). There was no difference between survival rates of cemented or cementless femoral prosthesis after 12 years (63% vs 54%) with respect to the presence or absence of hydroxyapatite. There was no difference between the survival rates of all bearing surfaces after 5 and 11 years, but none of the zirconia-polyethylene bearings had been revised after 5 years. There was no difference between survival rates of 28 mm and 32 mm heads after 8 years (86% vs 78%).Cemented prostheses are clearly indicated for patients under 50. Screwed uncemented cups are no longer indicated. Uncemented prostheses showed no better survival than cemented ones. Zirconia-polyethylene bearings could lead to a better survival rate than alumina-polyethylene or steel-polyethylene bearings.


Validity of the Japanese version of functional assessment of cancer therapy-gastric (FACT-Ga) and its sensitivity to ascites volume change: a retrospective analysis of Japanese clinical trial participants.
The functional assessment of cancer therapy-gastric (FACT-Ga) questionnaire was designed to evaluate quality of life (QOL) in patients with gastric cancer. We aimed to explore the reliability and validity of FACT-Ga in Japanese patients, and assess the sensitivity of the gastric cancer subscale for detecting changes in cancer-related variables over time. The Japanese version of FACT-Ga was used, and data were obtained from Japanese patients who participated in either of two clinical trials: treatment for advanced or recurrent gastric cancer with ascites (advanced-GC group), or adjuvant chemotherapy after curative resection of gastric cancer (adjuvant group). Psychometric data including data used to determine reliability, internal consistency, and clinical validity were analyzed. Clinical validity was evaluated by comparing subscale scores for patients in the two groups, and by comparing subscale scores for patients with different performance status scores. Correlation between gastric cancer subscale scores and gastric cancer-related variables was also examined. In addition, sensitivity of the gastric cancer subscale to changes in ascites volume, abdominal girth, and Eastern Cooperative Oncology Group (ECOG) performance status (PS) was examined by evaluating their correlation in the advanced-GC group. We collected data on 156 patients (62 advanced-GC group patients and 94 adjuvant group patients). Response rates for the subscales were over 80 % at most time points for both the groups. Cronbach's coefficient alpha revealed good internal consistency for each subscale. At baseline, the adjuvant group had higher QOL scores than the advanced-GC group (P < 0.05), and QOL scores for patients with different performance status scores differed significantly. Changes in gastric cancer subscale scores showed statistically significant correlation with changes in ascites volume (Spearman's rank correlation coefficient, 0.5; P < 0.05). FACT-Ga is reliable and clinically valid for Japanese patients with gastric cancer. Detection of QOL changes that correlate with ascites volume changes suggests that it could be used more broadly; FACT-Ga scores could be used as an endpoint for patients with gastric cancer-related ascites.


Muscle blood flow after amputation with special reference to the influence of the amputation level. Assessed by 133 xenon and histamine. An animal experiment.
Amputation on rabbits was found to change the muscle blood flow (MBF) in the limb subjected to surgery as well as in the contralateral extremity, depending on the technique used for amputation and the level at which it was carried out. Amputation was immediately followed by a reduction in MBF in the stump. After amputation of the crus and knee disarticulation, the flow was normalized a few days later and exceeded the preoperative mean flow. After amputation of the femur such an increase in flow was not seen, but when osseous plugging of the medullary cavity was applied, the MBF in the stump was intensified even after amputation of the femur.


A Therapeutic Peptide Vaccine Against PCSK9.
Vaccination provides a promising approach for treatment of hypercholesterolemia and improvement in compliance. In this study, the appropriate virus-like particle (VLP)-peptide vaccines targeting proprotein convertase subtilisin/kexin type 9 (PCSK9) were screened. The screening criteria of target peptides were as follows: (1) located in catalytic domain of PCSK9, or regulating the binding of PCSK9 and LDL receptors (LDLR); (2) having low/no-similarity when matched with the host proteome; (3) possessing ideal antigenicity and hydrophilicity; (4) including the functional mutation site of PCSK9. It was found that mice vaccinated with VLP -PCSK9 peptide vaccines, especially PCSK9Qβ-003 vaccine, developed high titer IgG antibodies against PCSK9. PCSK9Qβ-003 vaccine obviously decreased plasma total cholesterol in both Balb/c mice and LDLR+/- mice. Also, PCSK9Qβ-003 vaccine decreased plasma PCSK9 level and up-regulated LDLR expression in liver. Additionally, PCSK9Qβ-003 vaccine injection was associated with significant up-regulation of sterol-regulatory element-binding protein-2 (SREBP-2), hepatocyte nuclear factor 1α (HNF-1α), and 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase in LDLR+/- mice. No obvious immune injury was detected in vaccinated animals. The PCSK9Qβ-003 vaccine, therefore, may be an attractive treatment approach for hypercholesterolemia through decreasing cholesterol and regulating lipid homeostasis.


Perceptions and concerns about sustainable healthcare of nursing students trained in sustainability and health: A cohort study.
To describe nursing students' perceptions of sustainable health education in the nursing curriculum and their concerns about sustainable healthcare and the impact of climate change on nursing. Sustainable health education involves education on the impact of climate change on health and the impact of healthcare on the environment. The effectiveness of sustainable health education in improving attitudes, knowledge and skills in sustainable healthcare has been demonstrated. However, there is a need to study students' perceptions of this and their concerns about achieving sustainable healthcare from the use and disposal of healthcare resources. A cohort study with an inductive content analysis of open-ended questions included in a survey. The study was carried out with undergraduate nursing students throughout their four-year undergraduate academic program using scenario-based learning and augmented reality related to sustainability, climate change and health. As students were exposed to three educational interventions, they completed a survey of open-ended questions about their perceptions of their environmental sustainability training in the nursing curriculum, their concerns about the resources' used in healthcare and their perceptions of the impact of climate change on the nursing profession. Students identified content in the nursing degree program on climate change and health and hospital waste segregation. They also demanded more content on 'low environmental impact nursing care' when their clinical practice training increased. Students were concerned about the excessive and unnecessary use of materials in healthcare, especially in the post-pandemic period, the lack of environmental awareness of healthcare professionals and the lack of power to change the situation. They recognised the lack of proper waste segregation in healthcare settings, no recycling bins and little reuse of materials. They were also concerned about the polluting disposal of material. They perceived important impacts of climate change on nursing, such as patient care due to increased pollution-related diseases, including foetal malformations and new health care needs arising from weather conditions. Finally, students were concerned about the impact this will have on nursing care work and require 'nursing leadership in environmental awareness'. Students demand more training in low environmental impact healthcare and innovative educational practices are effective in this regard. Appropriate Sustainable Healthcare Education can make future health professionals more environmentally aware and enable them to lead the shift towards climate-smart care. Students demand more training in low environmental impact healthcare and perceive significant impacts of climate change on nursing.


Effects of JO 1784, a selective sigma ligand, on the autoradiographic localization of M1 and M2 muscarinic receptor subtypes in trimethyltin treated rats.
The distribution patterns of M1 and M2 muscarinic receptor subtypes following TMT and JO 1784 administration in the male Sprague-Dawley rat were investigated. In the present study, JO 1784 was injected in doses of 1, 4 and 16 mg/kg i.p. for one week prior to the single injection of TMT (8 mg/kg i.p.) and subsequently for 33 days. The effects of JO 1784 on the density of muscarinic receptor sub-types (M1 and M2) in the control and trimethyltin (TMT) treated rats were then evaluated. The topographic distribution and changes in muscarinic (M1 and M2) receptor densities were determined by means of autoradiography using [3H]quinuclidinylbenzilate (QNB). Both sub-types of muscarinic receptors contributed to the observed decrease in total muscarinic receptor binding in TMT-treated rats. In control rats, JO 1784 alone decreased M1 receptor density in the amygdaloid nuclei, basal ganglia, cortex and hippocampus and decreased M2 receptor density in the amygdaloid nuclei, basal ganglia, cortex, hippocampus, hypothalamus and septal regions. In TMT treated rats, chronic JO 1784 administration has a "neuroprotective effect" on both M1 and M2 receptors subtypes. Thus, following chronic administration of JO 1784 to TMT treated rats, both increases and decreases in M1 receptor density were observed relative to TMT animals. A significant increase in M1 receptor density was found in the cortex, olfactory regions, septum, thalamus and basal forebrain nuclei. In the hippocampus (CA2 and CA3), a significant decrease in M1 receptor density was observed. In TMT-treated rats, JO 1784 produced a significant increase in M2, receptor density in several brain regions with the most marked effects occurring in the amygdaloid nuclei, basal ganglia, cortex, hippocampus and hypothalamus. The ability of the selective sigma ligand, JO 1784, to attenuate the loss of muscarinic receptors in TMT treated rats could be of importance in the development of novel neuroprotective drugs.


Total protein patterns in human amniotic fluid.
Total protein patterns of amniotic fluid samples of different age groups, following polyacrylamide disc gel electrophoresis, showed a wide range of variation in early pregnancy, but no two cases had identical banding, indenticating their genetic variability. An assessment has been made here of the total protein, albumin and transferrin patterns from normal fluids of different age groups by gel electrophoresis, in order to observe the genetic variations and relationship with foetal age, if any.


Epidemiology of adverse reactions in anaesthesia in Germany and the Netherlands.
We consider 292 anaphylactoid reactions from the own material (1964-1980) out of two different Departments of anesthesiology to attempt to review the continental epidemiology of this life-threatening adverse drug reaction. We conclude the following: The specialty of this type of adverse reaction is likely to prove a special separating name i.e. "anaphylactoid reaction". The overall frequency can be estimated for 1 : 400-600 in anesthesiology, a preponderance of females may be possible. There is no predictability of the reaction, not even by sophisticated history. But it is possible to avoid the development of severe shock by early diagnosis and immediate full causative therapy. With growing experience in diagnosis and therapy the mortality of anaphylactoid reaction should be irrelevant in the near future.


Differentiation, regulation and function of regulatory T cells in non-lymphoid tissues and tumors.
Regulatory T cells (Tregs) play a substantial role in inhibiting excessive immune response. A large number of studies have focused on the tissue homeostasis maintenance and remodeling characteristics of Tregs in non-lymphoid tissues, such as the skin, colon, lung, brain, muscle, and adipose tissues. Herein, we overview the kinetics of Treg migration to non-lymphoid tissues and adaptation to the specific tissue microenvironment through the development of tissue-specific chemokine receptors, transcription factors, and phenotypes. Additionally, tumor-infiltrating Tregs (Ti-Tregs) play an important role in tumor generation and immunotherapy resistance. The phenotypes of Ti-Tregs are related to the histological location of the tumor and there is a large overlap between the transcripts of Ti-Tregs and those of tissue-specific Tregs. We recapitulate the molecular underpinnings of tissue-specific Tregs, which might shed new light on Treg-based therapeutic targets and biomarkers for inflammatory diseases and cancer.


Scientists Still Behaving Badly? A Survey Within Industry and Universities.
Little is known about research misconduct within industry and how it compares to universities, even though a lot of biomedical research is performed by-or in collaboration with-commercial entities. Therefore, we sent an e-mail invitation to participate in an anonymous computer-based survey to all university researchers having received a biomedical research grant or scholarship from one of the two national academic research funders of Belgium between 2010 and 2014, and to researchers working in large biomedical companies or spin-offs in Belgium. The validated survey included questions about various types of research misconduct committed by respondents themselves and observed among their colleagues in the last three years. Prevalences of misconduct were compared between university and industry respondents using binary logistic regression models, with adjustments for relevant personal characteristics, and with significance being accepted for p < 0.01. The survey was sent to 1766 people within universities and an estimated 255 people from industry. Response rates were 43 (767/1766) and 48% (123/255), and usable information was available for 617 and 100 respondents, respectively. In general, research misconduct was less likely to be reported by industry respondents compared to university respondents. Significant differences were apparent for one admitted action (gift authorship) and three observed actions (plagiarism, gift authorship, and circumventing animal-subjects research requirements), always with lower prevalences for industry compared to universities, except for plagiarism. This survey, based on anonymous self-report, shows that research misconduct occurs to a substantial degree among biomedical researchers from both industry and universities.


Long-term results using scleral-fixated posterior chamber intraocular lenses.
Transscleral ciliary sulcus fixation of a posterior chamber intraocular lens (IOL) is a technique for managing complicated cataract surgery, secondary IOL implantation, and IOL exchange. We performed a retrospective review of 32 patients who had a minimum of six months and a mean of 13.3 months follow-up after transscleral ciliary sulcus fixation of a posterior chamber IOL. In 30 patients, vision was maintained or improved following surgery. Complications were not uncommon; they were visually significant in three patients and necessitated a reoperation for late subluxation of the IOL in one other patient. Although the long-term visual results are encouraging, the postoperative complication rate of scleral-fixated posterior chamber IOLs is between that of posterior chamber IOLs inserted when the posterior capsule is intact and that of closed-loop anterior chamber IOLs.


The role of polyols in the pathophysiology of hypergalactosemia.
Cellular accumulation of galactitol has been suggested to cause the apparent dietary-independent, long-term complications in classic galactosemia. Experimental animals rendered hypergalactosemic by galactose feeding accumulate tissue galactitol, as well as millimolar quantities of galactose, and manifest biochemical, physiological and pathological abnormalities which are generally eliminated or curtailed by the concomitant administration of an aldose reductase inhibitor. This includes reduced cellular content of the cyclic polyol, myo-inositol, which like galactitol may function as an alternate intracellular osmolyte. However, the abnormalities detected in experimental galactosemic animals are more compatible with findings in experimental diabetes mellitus than in human galactosemia. Because patients with galactokinase deficiency fail to manifest the CNS and ovarian complications which characterize classic galactosemia, yet during long-term lactose restriction excrete comparable urinary quantities of galactitol, this polyol alone is not likely to play an important role during postnatal life in the pathogenesis of long-term complications. Notwithstanding, a role for either galactitol or myo-inositol in an intrauterine toxicity cannot be dismissed.


Extracellular matrix organization modulates fibroblast growth and growth factor responsiveness.
To learn more about the relationship between extracellular matrix organization, cell shape, and cell growth control, we studied DNA synthesis by fibroblasts in collagen gels that were either attached to culture dishes or floating in culture medium during gel contraction. After 4 days of contraction, the collagen density (initially 1.5 mg/ml) reached 22 mg/ml in attached gels and 55 mg/ml in floating gels. After contraction, attached collagen gels were well organized; collagen fibrils were aligned in the plane of cell spreading; and fibroblasts had an elongated, bipolar morphology. Floating collagen gels, however, were unorganized; collagen fibrils were arranged randomly; and fibroblasts had a stellate morphology. DNA synthesis by fibroblasts in contracted collagen gels was suppressed if the gels were floating in medium but not if the gels were attached, and inhibition was independent of the extent of gel contraction. Therefore, growth of fibroblasts in contracted collagen gels could be regulated by differences in extracellular matrix organization and cell shape independently of extracellular matrix density. We also compared the responses of fibroblasts in contracted collagen gels and monolayer culture to peptide growth factors including fibroblast growth factor, platelet-derived growth factor, transforming growth factor-beta, and interleukin 1. Cells in floating collagen gels were generally unresponsive to any of the growth factors. Cells in attached collagen gels and monolayer culture were affected similarly by fibroblast growth factor but not by the others. Our results indicate that extracellular matrix organization influenced not only cell growth, but also fibroblast responsiveness to peptide growth factors.


Lawyers' delights and geneticists' nightmares: at forty, the double helix shows some wrinkles.
The National Institutes of Health (NIH) request to patent the base sequences of incomplete and uncharacterized fragments of DNA copied on messenger RNAs (cDNAs) extracted from human tissues, the refusal by the patent office, and the appeal placed by NIH, have incited a violent controversy, fueled by rational, as well as emotional elements. In a compromising mode between liberalism and protectionism, I propose that legal protection be considered only for those RNA/DNA sequences, either natural or artificial, which can generate practical applications per se, and not through their expression products. Another controversy is developing around a popular tool for genomic research: the fidelity of yeast artificial chromosome (YAC) libraries being distributed worldwide for physical mapping is being questioned. Some of these libraries have been shown to be affected by surprisingly high levels of co-cloning, in addition to more common gene reshuffling instances. Also in this case, scientific as well as non-scientific components have to be considered. Possible remedies for the underlying problems may be found in the proper use of kinetic, enzymatic and microbiological variables in the production of YACs. Here too, a sharper distinction between the secular and scientific gratifications of research could help.


Predictors of Mortality Among HIV-exposed Infants Through 18 Months of Age in Kenya: A Retrospective Review of Programmatic Data.
We identified mortality predictors among HIV-exposed uninfected infants and infants living with HIV in Kenyan early infant diagnosis services between 2012 and 2017. Younger maternal age and absence of antenatal antiretroviral therapy among HIV-exposed uninfected infants (n = 2366) and travel time to hospital and delayed infant testing among infants living with HIV (n = 130) predicted mortality, highlighting the importance of supporting engagement in maternal/pediatric HIV services.


Differential modulation of nerve growth factor receptor (p75) and cholinergic gene expression in purified p75-expressing and non-expressing basal forebrain neurons by BMP9.
The synthesis of acetylcholine and its release from basal forebrain cholinergic neurons (BFCN) that innervate the cerebral cortex and hippocampus are considered essential processes for normal learning, memory and attention. We have developed a purification and cell culture method of BFCN in order to examine the regulation of their cholinergic phenotype. Cells isolated from the septal region of late embryonic mice were purified by fluorescence-activated cell sorting based on their expression of the nerve growth factor receptor (p75), a surface marker for mature BFCN. Consistent with previous reports, p75-positive (p75+) cells were enriched in choline acetyltransferase (ChAT) and the high-affinity choline transporter (ChT), as measured by reverse transcriptase PCR. In culture, these cells maintained their gene expression of p75, ChAT and ChT, while p75-negative (p75-) cells had a low expression of these genes. Incubation of the cells with BMP9 not only increased p75 and ChAT gene expression in p75- cells, but also augmented the expression of these genes in p75+ cells. Conversely, BMP9 decreased ChT gene expression in p75+ cells and had no such effect in p75- cells. Immunostaining confirmed that p75 protein expression was modulated by BMP9 in a similar way as p75 mRNA, and also revealed that only a subset of p75- cells respond to BMP9 in this manner. These data suggest that mature BFCN in culture may express their cholinergic phenotype in the absence of exogenous trophic input, but that BMP9 can further modulate this phenotype. Moreover, BMP9 induces the cholinergic phenotype in a set of basal forebrain non-cholinergic neurons or precursor cells.


Effects of interferons and cytokines on melanoma cells.
This review summarizes recent information on the effects of immunomodulatory cytokines on human melanoma cells. The action of interferon (IFN)-alpha, -beta, and -gamma has been extensively examined in melanoma and melanocyte cultures in vitro, and increasing information on the action of other cytokines is now available. All IFNs revealed a dose-dependent antiproliferative effect on melanoma cells with the highest growth inhibition caused by IFN-beta. Proliferation was also inhibited by interleukin (IL) 1-alpha and -beta, and tumor necrosis factor (TNF)-alpha. For IL-4, both growth-stimulatory and -inhibitory properties have been reported. Cellular differentiation in terms of melanin synthesis, formation of dendritelike structures, and antigenic changes was not affected by IFN-alpha or -beta. IFN-gamma, however, induced a more dedifferentiated and biologically more aggressive phenotype of melanoma cells. Histocompatibility antigen (HLA) class I molecules were found upregulated by all IFNs and by TNF-alpha, associated with a marked increase of melanoma cell lysis by tumor infiltrating lymphocytes in vitro. HLA class II molecules were de novo expressed or enhanced by IFN-gamma and TNF-alpha. The adhesion molecules ICAM-1, LFA-3, and VLA-2 were upregulated by IFN-gamma, TNF-alpha, and IL-1-beta, whereas melanoma-associated antigens were hardly affected by cytokines. It seems that both antiproliferative and immunomodulatory effects may contribute to the antitumoral activity of cytokines in vivo. In vivo application of cytokines as well as combinations with cytotoxic drugs, therefore, may be promising for future treatment strategies.


STEMI patients and nonculprit lesions: To treat or not to treat? and when? A review of most recent literature.
Patients with ST segment elevation myocardial infarction and multivessel disease represent a high percentage of ischemic patient with a worse outcome than patient with single coronary artery disease. Therefore, initial management of these patients is of high importance, but unfortunately this is not clarified yet. We analyze the available literature trying to afford current doubts to determine which way of revascularization is to be preferred. © 2015 Wiley Periodicals, Inc.


[Results and predictors of long-term outcomes of off-pump coronary artery bypass grafting: 2 831 cases from a single center].
To describe the long-term survival of off-pump coronary artery bypass grafting (CABG) and to analysis the risk factors of operative mortality and long-term survival. From January 2001 to December 2012, 2 831 patients undergoing off-pump CABG in Peking University People's Hospital, 2 099 cases (74.1%) of them were male, the average age was (63±9) years. The perioperative data was retrospectively collected. Binary Logistic regression was used to find the risk factors which affect the operative mortality. Follow-up evaluation was completed regularly. Kaplan-Meier survival curve, Log-rank test and Cox regression model were used to find out factors which affect the long-term result. Totally 2 831 patients underwent isolating off-pump CABG, in whom 45 patients died perioperative, 2 786 patients discharged successfully. Binary Logistic regression showed that sex (female) (χ2=4.4, OR=2.307, P=0.035), peripheral vascular disease (χ2=17.4, OR=6.616, P=0.000), New York Heart Association (NYHA) class grade≥3 (χ2=10.5, OR=3.491, P=0.001), ejection fraction≤40% (χ2=16.9, OR=5.230, P=0.000), emergency surgery (χ2=11.9, OR=5.127, P=0.001) are risk factors of operative mortality. The follow-up time was (74±44) months. Totally 107 patients were lost from follow-up, 109 patients died in follow-up. The survival rate at 1, 3, 5 , 8 and 10 years was 97.2%, 95.5%, 94.3%, 93.6%, 92.1%, respectively. Univariate analysis showed that age (>65 years), hypertension, renal insufficiency, peripheral vascular disease, history of myocardial infarction, NYHA class grade≥3 and emergency surgery were risk factors of the long-term survival (χ2=8.150 to 88.241, P<0.05). Cox regression analysis showed that age (>65 years) (χ2=12.1, RR=2.295, P=0.000), renal insufficiency (χ2=12.3, RR=3.160, P=0.000), peripheral vascular disease (χ2=42.5, RR=5.626, P=0.000), NYHA class grade≥3 (χ2=9.1, RR=1.994, P=0.002) and emergency surgery (χ2=5.5, RR=2.247, P=0.019) were independent risk factors that affect the long-term survival. Sex (female), peripheral vascular disease, NYHA class grade≥3, ejection fraction≤40%, emergency surgery are risk factors of operative mortality. Age (>65 years), renal insufficiency, peripheral vascular disease, NYHA class grade≥3 and emergency surgery are independent risk factors that affect the long-term survival. Off-pump CABG has favorable perioperative and long-term outcome, and it definitely is a very safe and effective technique for coronary artery revascularization.


A Novel Molecular Cytogenetic Finding of Leiomyomatosis Peritonealis Disseminata.
Leiomyomatosis peritonealis disseminata (LPD) is a rare disease characterised by the subperitoneal proliferation of smooth muscle cells that form benign nodules. A few studies have aimed to reveal the pathogenesis of LPD without reaching a clear explanation. Karyotype analysis and array-comparative genomic hybridization (aCGH) of a human LPD case were performed to evaluate the role of chromosomal abnormalities in LPD pathogenesis. The LPD nodules showed a 45, XX, del(7p), t(11; 17) (q23;q25),-22 de novo karyotype, and the aCGH analysis confirmed these deletions at 7p22.3-p12.1 (1,862,362-52,766,911 bp) and 22q11.23-q13.33 (21,973,915-49,265,116 bp) with lengths of 50.9 Mb and 27.3 Mb, respectively. In this study, we described two large novel aberrations - deletions in chromosome 7 and 22 - that might play an important role in LPD disease. These findings might contribute to new insights to unravel the pathogenesis of LPD and develop further clinical treatments. © 2015 S. Karger AG, Basel.


Necrotizing pneumonia in children: report of 41 cases between 2006 and 2011 in a French tertiary care center.
Forty-one children hospitalized for necrotizing pneumonia were retrospectively analyzed. Necrotizing pneumonia represented 0.8% of community-acquired pneumonia and 6% of hospitalized community-acquired pneumonia. The chest radiograph revealed necrosis on admission in onethird of cases. Twenty-one cases (51%) were documented, including 13 Staphylococcus aureus, all Panton-Valentine leukocidin positive, 7 Streptococcus pneumoniae and 1 Fusobacterium nucleatum.


Targeting CD19 for diffuse large B cell lymphoma in the era of CARs: Other modes of transportation.
CD19 is nearly ubiquitously expressed on B-lymphocytes and in B-cell malignancies. Although CD19-directed CAR T cells have greatly improved outcomes in B-cell malignancies, there are significant limitations with this therapy. CD19 can also be effectively targeted by other drug classes, such as monoclonal antibodies, antibody-drug conjugates, and bispecific T cell engagers or antibodies. However, the optimal patient selection and sequencing of these novel therapies has not yet been established. In this review, we discuss the utilization of CD19 as a target for the treatment of DLBCL, focusing on tafasitamab, loncastuximab tesirine, and blinatumomab. We provide a comprehensive review of the pivotal clinical trials, discussing the strength and limitations of the data for each agent. We explore the emerging evidence that CD19 expression is retained following exposure to these agents and that patients can be successfully re-challenged with anti-CD19 therapies of a different drug class upon disease relapse post-CAR T cells. Finally, we discuss how these drugs potentially fit into the most current treatment paradigm for DLBCL.


[Carcinoid of the appendix. Consideration on 5 incidentally diagnosed cases].
5 cases of carcinoid of the appendix discovered during 1094 appendectomies are described. Neither carcinoid syndrome or local metastasis was observed in any of the patients. The importance of the histological examination of all appendices removed is emphasised. Where there is no metastasis, radical surgery is appropriate. The prognosis is excellent but patients should be monitored by the measurement of 5HIIA in the urine.


Methylation changes of SIRT1, KLF4, DAPK1 and SPG20 in B-lymphocytes derived from follicular and diffuse large B-cell lymphoma.
Diffuse large-B cell lymphomas (DLBCL) and follicular lymphomas (FL) are the most represented subtypes among mature B-cell neoplasms and originate from malignant B lymphocytes. Methylation represents one of the major epigenetic mechanisms of gene regulation. Silent information regulator 1 (SIRT1) is a class III lysine-deacetylase playing several functions and considered to be a context-dependent tumor promoter. We present the quantitative methylation, gene expression and tissue distribution of SIRT1 and some key mediators related to lymphoma pathogenesis in B lymphocytes purified from biopsies of follicular hyperplasias, FL and DLBCL. SIRT1 mRNA levels are higher in FL than follicular hyperplasias and DLBCL. B cell lymphoma 6 (BCL6) positively correlates with SIRT1. SIRT1 promoter shows a methylation decrease in the order: follicular hyperplasia - FL - DLBCL. Kruppel-like factor 4 (KLF4), Death-associated protein kinase 1 (DAPK1) and Spastic Paraplegia 20 (SPG20) methylation increase significantly in FL and DLBCL compared to follicular hyperplasias. Gene expression of DAPK1 and SPG20 inversely correlates with their degree of methylation. Our findings evidence a positive correlation between SIRT1 and BCL6 expression increase in FL. SIRT1 methylation decreases in FL and DLBCL accordingly and this parallels the increase of KLF4, DAPK1 and SPG20 methylation.


Intra-abdominal packing does not increase infection risk or mandate longer presumptive antibiotic therapy.
Damage control laparotomy allows for resuscitation and reversal of coagulopathy with improved mortality. In-tra-abdominal packing is often used to limit hemorrhage. Temporary abdominal closure is associated with increased rates of subse-quent intra-abdominal infection. The effect of increased duration of antibiotics is unknown on these infection rates. We sought to determine the role of antibiotics in damage control surgery. A retrospective analysis of all trauma patients requiring damage control laparotomy on admission to an ACS verified level one trauma center from 2011 to 2016 was performed. Demographic and clinical data including ability and time to attain primary fascial closure, as well as complication rates, were recorded. The primary outcome measure was intra-abdominal abscess formation following damage control laparotomy. Two-hundred and thirty-nine patients underwent DCS during the study period. A majority were packed (141/239, 59.0%). No differences existed in demographics or injury severity between groups, and infection rates were similar (30.5% vs. 38.8%, P=0.18). Patients with infection were more likely to have suffered gastric injury (23.3% vs. 6.1%, P=0.003) than those without complication. There was no significant association between gram negative and anaerobic (Odds Radio [OR] 0.96, 95% confidence interval [CI] 0.87-1.05) or antifungal therapy (OR 0.98, 95% CI 0.74-1.31) and infection rate, regardless of duration on multivariate regression CONCLUSION: Our study offers the first review of the effect of antibiotic duration on intra-abdominal complications following DCS. Gastric injury was more commonly identified in patients who developed intra-abdominal infection. Duration of antimicrobial therapy does not affect infection rate in patients who are packed following DCS.


C1-based biomanufacturing: Advances, challenges and perspectives.
One-carbon (C1) compounds have emerged as a key research focus due to the growth of metabolic engineering and synthetic biology as affordable and sustainable nonfood sugar feedstocks for energy-efficient and environmentally friendly biomanufacturing. This paper summarizes and discusses current developments in C1 compounds for biomanufacturing. First, two primary groups of microbes that use C1 compounds (native and synthetic) are introduced, and the traits, categorization, and functions of C1 microbes are summarized. Second, engineering strategies for C1 utilization are compiled and reviewed, including reconstruction of C1-utilization pathway, enzyme engineering, cofactor engineering, genome-scale modeling, and adaptive laboratory evolution. Third, a review of C1 compounds' uses in the synthesis of biofuels and high-value compounds is presented. Finally, potential obstacles to C1-based biomanufacturing are highlighted along with future research initiatives.


Patients With Metabolic Syndrome May Experience More Tooth Loss and Poorer Functional Dentition Than Those Without Metabolic Syndrome.
Association between metabolic syndrome and tooth loss: A systematic review and meta-analysis. Souza ML, Massignan C, Glazer Peres K, Aurélio Peres M. J Am Dent Assoc 2019;150(12):1027-1039.e7. Government. Foundation for Research and Innovation Support of Santa Catarina State (Brazil), Coordination of Improvement of Higher Level Personnel (Brazil). Systematic review with meta-analysis of data.


The hydrodeoxygenation of bioderived furans into alkanes.
The conversion of biomass into fuels and chemical feedstocks is one part of a drive to reduce the world's dependence on crude oil. For transportation fuels in particular, wholesale replacement of a fuel is logistically problematic, not least because of the infrastructure that is already in place. Here, we describe the catalytic defunctionalization of a series of biomass-derived molecules to provide linear alkanes suitable for use as transportation fuels. These biomass-derived molecules contain a variety of functional groups, including olefins, furan rings and carbonyl groups. We describe the removal of these in either a stepwise process or a one-pot process using common reagents and catalysts under mild reaction conditions to provide n-alkanes in good yields and with high selectivities. Our general synthetic approach is applicable to a range of precursors with different carbon content (chain length). This allows the selective generation of linear alkanes with carbon chain lengths between eight and sixteen carbons.


Analysis of reproducibility and robustness of a renal proximal tubule microphysiological system OrganoPlate 3-lane 40 for in vitro studies of drug transport and toxicity.
Microphysiological systems are an emerging area of in vitro drug development, and their independent evaluation is important for wide adoption and use. The primary goal of this study was to test reproducibility and robustness of a renal proximal tubule microphysiological system, OrganoPlate 3-lane 40, as an in vitro model for drug transport and toxicity studies. This microfluidic model was compared with static multiwell cultures and tested using several human renal proximal tubule epithelial cell (RPTEC) types. The model was characterized in terms of the functional transport for various tubule-specific proteins, epithelial permeability of small molecules (cisplatin, tenofovir, and perfluorooctanoic acid) versus large molecules (fluorescent dextrans, 60-150 kDa), and gene expression response to a nephrotoxic xenobiotic. The advantages offered by OrganoPlate 3-lane 40 as compared with multiwell cultures are the presence of media flow, albeit intermittent, and increased throughput compared with other microfluidic models. However, OrganoPlate 3-lane 40 model appeared to offer only limited (eg, MRP-mediated transport) advantages in terms of either gene expression or functional transport when compared with the multiwell plate culture conditions. Although OrganoPlate 3-lane 40 can be used to study cellular uptake and direct toxic effects of small molecules, it may have limited utility for drug transport studies. Overall, this study offers refined experimental protocols and comprehensive comparative data on the function of RPETCs in traditional multiwell culture and microfluidic OrganoPlate 3-lane 40, information that will be invaluable for the prospective end-users of in vitro models of the human proximal tubule.


Management of upper urinary tract calculi with ureteroscopic techniques.
To evaluate our experience in treating 155 patients with upper urinary tract calculi ureteroscopically. The treatment of urinary calculi has remained the most frequent application of ureteroscopy. Miniaturization of semirigid and flexible ureteroscopes has permitted easier access to calculi throughout the urinary tract. Ureteroscopic stone treatment was attempted in 155 patients with upper urinary tract calculi between November 1995 and March 1997. Fifty-nine (38.1%) patients had renal calculi, 82 (52.9%) ureteral, and 14 (9%) had both renal and ureteral calculi. Both semirigid and flexible ureteroscopes were used for treatment (rigid alone in 21 [13.5%], flexible in 64 [41.3%], and both rigid and flexible in 70 [45.2%] patients). Lithotripsy was required in 122 (79%) of the patients. The holmium:yttrium-aluminum-garnet laser was used in 113 (92.6%) of these patients. All patients with ureteral calculi (29 proximal, 19 mid, and 34 distal) were successfully cleared after one endoscopic procedure except for 1 patient with a proximal ureteral calculus who had a 4-mm residual fragment in the kidney. Of the 59 patients with renal calculi, 47 (79.7%) were totally clear of stones 1 month after treatment. The remaining 12 (20.3%) patients had evidence of residual calculi 3 to 4 mm or less in diameter. In patients with combined renal and ureteral calculi, 1 1 of 14 (78.6%) were rendered stone free. The remaining 3 (21.4%) patients had evidence of residual calculi 4 mm in diameter. Overall, 95% of the patients were treated in an outpatient setting. Morbidity was low, with no evidence of stricture. Ureteroscopy and laser lithotripsy in experienced hands are a safe and reliable method for the treatment of ureteral and even intrarenal calculi.


Pancreatic insufficiency in Cystic Fibrosis.
Pancreatic insufficiency (PI) affects about 85% of the cystic fibrosis population. Although most are PI soon after birth, some will have pancreatic sufficiency (PS) for some or all of their life. Understanding the clinical presentation, diagnosis, and management of PI is crucial to the care of people with cystic fibrosis.


An overview of the internalization and effects of microplastics and nanoplastics as pollutants of emerging concern in bivalves.
Microplastic and nanoplastic pollution in aquatic environments is a topic of emerging concern due to the internalization, retention time and effects of these particles in aquatic biota. Bivalves are considered bioindicators due to their wide distribution, sessile behaviour, occupation of ecological niches and ability to filter a large water volume. The study of microplastics and nanoplastics in bivalves has revealed the uptake mechanisms, internalization, distribution and depuration of these particles as well as their effects on physiological parameters, morphological alterations, immunotoxicity and changes in gene expression and proteomic profiles. In this review, we examine the primary characteristics of microplastics and nanoplastics (type of material, size, coating, density, additives and shapes) involved in their possible toxicity in bivalves. Furthermore, secondary characteristics such as the suspension media, aggregation stage and adsorption of persistent pollutants were also recorded to assess the impact of these materials on bivalves. Here, we have highlighted the efforts exerted thus far and the remaining gaps in understanding the extent of microplastic and nanoplastic impacts on bivalves on the basis of laboratory experiments and mesocosm bioassays and in the field. Furthermore, further microplastic and nanoplastic toxicological studies are proposed to facilitate the realistic assessment of environmental risk.


Microscopic Lesions in Canine Eyes with Primary Glaucoma.
Although the clinical classification of primary glaucoma in dogs is quite simple, the phenotypes of glaucoma in most of the species are indeed multiple. Ophthalmologists can often evaluate the dynamic changes of clinical signs at different times in the course of the disease, whereas pathologists are often presented with globes that have undergone abundant therapies and are at the end stage. Therefore, an open collaboration between clinicians and pathologists can produce the most accurate interpretation in the pathology report and improve patient outcomes. This article focuses on the histomorphologic elements that characterize, and are important to, canine primary glaucomas.


The bridge beyond: strengthening nursing practice in attitudes towards death, dying, and the terminally ill, and helping the spouses of critically ill patients.
Death and dying are often surrounded by a conspiracy of silence. Usually it is an unconsciously organised silence, leaving those whose lives are threatened by terminal illness or an accelerated death with a physiological insult and without much-needed communication. The silence seems self-protecting for relatives, medical and nursing staff. Silence is a false reassurance and offers no emotional growth or understanding of the events which surround death in its finality. People who fear death of a close family member often begin the process of grieving their loss before actual loss occurs, and can remain locked in grief for years unless helped by the right communication. According to Hampe (1975) this phenomenon is known as 'anticipating grief' or grieving. If medical staff and nurses are to feel confident in anything to do with death and dying, or to feel comfortable in dealing with terminal illness, they should have some understanding of death itself. They should also feel confident in opening up and maintaining an effective dialogue. This requires training in the relevant interviewing, assessment and counselling skills (Maguire, 1985). A systematic approach is necessary to provide good care for the spouses of critically ill patients, who may suffer anticipatory grief. Excerpts from a very helpful paper by Breu (1982) and an adapted care plan are included (with permission) at the end of this paper.


Three-dimensional ultrastructure of the endolymphatic sac.
The subcellular structures of the epithelial cells of the guinea pig endolymphatic sac were studied. By using a newly developed scanning electron microscopy technique, the intracellular organelles could be studied three-dimensionally and the topographic relationships analyzed. The light epithelial cell has an extensive network of endoplasmic reticulum which is characteristically arranged in a baso-apical direction. The connections between the inner surface of the plasmalemma and the endoplasmic reticulum were observed, as were connections between the Golgi complex and the endoplasmic reticulum. Our findings support the hypothesis that the endoplasmic reticulum might form transcellular channels through which the cell can transport water and ions from the lumen of the endolymphatic sac out into the subepithelial tissue. The dark epithelial cells seen in particular contained the smooth type of endoplasmic reticulum. Lysosomes were also observed in the dark cells, indicating that these cells probably have more of a secretory function.


Lung solute permeability and lung liquid absorption in premature ventilated fetal goats.
In 21 premature fetal goats ventilated at low tidal volumes, measurements were made of alveolar surfactant activity by bubble stability tests on alveolar liquid, lung epithelium solute permeability using radiolabeled tracer molecules, lung liquid absorption from wet and dry lung weights, effectiveness of lung gas exchange, and lung mechanics. Arterial pO2 and residual gas volume were higher and arterial pCO2 and specific compliance were lower in animals whose bubble stability test was greater than 1:5 on alveolar liquid aspirated following one hr of ventilation. Animals without detectable surfactant in fetal lung liquid before ventilation had transfer of all size tracer solutes between alveolar liquid and blood, whereas those with detectable surfactant did not allow tracers 14 A or larger in radius to cross the lung epithelium. Lung liquid absorption rate in animals without lung liquid surfactant in fetal lung liquid before ventilation averaged 25% hr, whereas those with surfactant absorbed liquid at only 2%/hr in this protocol. All differences in solute permeability and lung liquid absorption were related to preventilation surfactant levels in fetal lung liquid, and not to postventilation surfactant levels or to lung function.


[Psoriasis SCID-mouse model].
Psoriasis is characterized by a complex phenotype and pathogenesis along with polygenic determination. Several psoriasis animal models have only been able to incompletely reproduce the disease. A xenogeneic transplantation approach, grafting skin from psoriatic patients onto mice with a severe combined immunodeficiency (SCID), was the first to meet the criteria for a psoriasis model. During the last 10 years, this psoriasis SCID-mouse model not only allowed telling experiments focusing on pathogenetic aspects, but also proved being a powerful tool for drug discovery with a good predictive value.


Neuroprotective metabolites via fungal biotransformation of a novel sapogenin, cyclocephagenol.
Cyclocephagenol (1), a novel cycloartane-type sapogenin with tetrahydropyran unit, is only encountered in Astragalus species. This rare sapogenin has never been a topic of biological activity or modification studies. The objectives of this study were; (i) to perform microbial transformation studies on cyclocephagenol (1) using Astragalus endophyte, Alternaria eureka 1E1BL1, followed by isolation and structural characterization of the metabolites; (ii) to investigate neuroprotective activities of the metabolites; (iii) to understand structure-activity relationships towards neuroprotection. The microbial transformation of cyclocephagenol (1) using Alternaria eureka resulted in the production of twenty-one (2-22) previously undescribed metabolites. Oxidation, monooxygenation, dehydration, methyl migration, epoxidation, and ring expansion reactions were observed on the triterpenoid skeleton. Structures of the compounds were established by 1D-, 2D-NMR, and HR-MS analyses. The neuroprotective activities of metabolites and parent compound (1) were evaluated against H2O2-induced cell injury. The structure-activity relationship (SAR) was established, and the results revealed that 1 and several other metabolites had potent neuroprotective activity. Further studies revealed that selected compounds reduced the amount of ROS and preserved the integrity of the mitochondrial membrane. This is the first report of microbial transformation of cyclocephagenol (1).


Manipulating the baby biome: What are the issues?
Establishing a baby biome through the controversial practice of 'vaginal seeding' has generated interest among the general public and healthcare providers alike. We discuss the potential risks of this practice and offer a harm minimisation approach to managing women requesting vaginal microbiome transfer after delivery by caesarean section.


Studies on the metabolism of 2,4'-isobutylphenylpropionic acid (ibuprofen) by gas chromatography and mass spectrometry. Dialysis fluid, a convenient medium for studies on drug metabolism.
2,4'-Isobutylphenylpropionic acid (ibuprofen) has previously been demonstrated to yield four urinary metabolites, formed by omega 1-, omega 2- and omega 3-hydroxylation and by a further oxidation of the primary alcohol of the omega 1-hydroxylated metabolite to a carboxyl group. By synthesis and gas chromatography--mass spectrometry the suggested structure of the omega 3-hydroxylated metabolite was verified in the present study. Moreover, a new metabolite, 2,4'-carboxyphenylpropionic acid, was demonstrated to be present in substantial amounts in dialysis fluid from a nephrectomized patient. In such patients ingested drugs cannot be excreted in the urine, but are metabolized to end products. Thus, dialysis fluid may be a convenient medium for studies on drug metabolism.


Skin movement artefact assessment and compensation in the estimation of knee-joint kinematics.
In three dimensional (3-D) human movement analysis using close-range photogrammetry, surface marker clusters deform and rigidly move relative to the underlying bone. This introduces an important artefact (skin movement artefact) which propagates to bone position and orientation and joint kinematics estimates. This occurs to the extent that those joint attitude components that undergo small variations result in totally unreliable values. This paper presents an experimental and analytical procedure, to be included in a subject-specific movement analysis protocol, which allows for the assessment of skin movement artefacts and, based on this knowledge, for their compensation. The effectiveness of this procedure was verified with reference to knee-joint kinematics and to the artefacts caused by the hip movements on markers located on the thigh surface. Quantitative validation was achieved through experimental paradigms whereby prior reliable information on the target joint kinematics was available. When position and orientation of bones were determined during the execution of a motor task, using a least-squares optimal estimator, but the rigid artefactual marker cluster movement was not dealt with, then knee joint translations and rotations were affected by root mean square errors (r.m.s.) up to 14 mm and 6 degrees, respectively. When the rigid artefactual movement was also compensated for, then r.m.s errors were reduced to less than 4 mm and 3 degrees, respectively. In addition, errors originally strongly correlated with hip rotations, after compensation, lost this correlation.


The adjuvanticity of gamma inulin.
Gamma-inulin (g-IN) is a polymorph identified as the active component of inulin preparations that specifically activates the alternative pathway of complement (APC). The APC is central to many leucocyte functions, including B cell activation. We show here that g-IN, when formulated as a pure, endotoxin-free, fine suspension insoluble at 37 degrees C and given at 50-100 micrograms per mouse, is a potent adjuvant for both humoral and cell-mediated responses to a variety of antigens. g-IN increased secondary IgG responses five- to 28-fold (P less than 0.001), using as antigen phosphorylcholine coupled to keyhole limpet haemocyanin; subclasses IgG 2a, 2b, and 3 were boosted several hundred-fold, IgG 1 10-fold. IgM and IgA were increased four- to six-fold. Delayed hypersensitivity, by footpad swelling after secondary challenge with sheep red blood cells (SRBC), was increased more than two-fold (P less than 0.001) if g-IN was included with the primary SRBC, equivalent to increasing primary doses 10-fold. g-IN was equally active if given 5 days before the primary SRBC. Thus it is an immune stimulant rather than a depot or vehicle for antigen. Mice primed subcutaneously with 30-300 HA units of H2N2 influenza virus (strain A/JAP) and challenged intranasally with a lethal dose of H1N1 virus (strain A/WSN) all died, but if g-IN was given with the primary antigen 50% of the mice survived (P less than 0.001), a deduced but not proven boost to cytotoxic T cell-mediated immunity. Unpublished work has shown that g-IN has no adverse effects at adjuvant-active doses.(ABSTRACT TRUNCATED AT 250 WORDS)


Working with complex care patients.
The following article provides the authors' definition of complex care patients. It is based on what they have learned about these special patients from the literature and from their personal clinical experiences. When I received this article for review, I decided to do a literature search on the term complex care patient and had a difficult time finding relevant articles. I asked a wonderful librarian from the Boston Children's Hospital, Alison Clapp, to help me with the search, and she had to maneuver through a maze of clinical subheadings. She described the search as "a difficult one." It is ironic that the search for information pertaining to these patients mirrors the complexity of care that they require from us. Are complex care patients the same as complex patients? The insertion of the word care may make a difference: You decide. There are complex patients who challenge us technically and intellectually, and then there are those who require even more from us--they challenge us technically, intellectually, physically, emotionally, and spiritually. The term complex care patient encompasses all these care dimensions. I think it is an excellent choice of words to make us consider what we are increasingly expected to do--and how we cannot do it alone.--Maura MacPhee, RN, PhD, Clinical Practice Column Editor.


CT Appearance Pattern After Stereotactic Body Radiation Therapy Predicts Outcomes in Early-Stage Non-Small-Cell Lung Cancer.
Computed tomography (CT) appearance pattern after lung tumor stereotactic body radiation therapy(SBRT) might predicts survival. This study aimed to investigate the correlation between CT appearance pattern after SBRT and outcomes in patients with early-stage non-small-cell lung cancer (NSCLC). Clinical data of inoperable patients with early-stage NSCLC undergoing SBRT were retrospectively analyzed from 2012 to 2015 at the Zhejiang Cancer Hospital. The relationship between CT appearance pattern after SBRT and patient's survival was analyzed. The data from 173 patients with early-stage lung cancer treated with SBRT were analyzed. One month after SBRT, diffuse consolidation was seen in 17 patients, patchy consolidation in 28 patients, diffuse ground-glass opacity (GGO) in 10 patients, and patchy GGO in 22 patients. The survival time was significantly longer in the "no evidence of increased density" group compared with the "consolidation or GGO" group [2-year overall survival (OS) rate, 96.1% vs 89.3%; hazard ratio (HR), 0.36; 95% confidence interval (CI), 0.16-0.85; P = 0.015]. A similar trend was found in the progression-free survival (PFS) analysis (2-year PFS rate, 91.3% vs 85.0%; HR, 0.35; 95% CI, 0.13-0.95; P = 0.015) and distant metastasis free survival(DMFS) (2-year DMFS rate, 93.3% vs 87.1%; HR, 0.41; 95% CI, 0.20-0.86; P = 0.031). However, no significant difference was found in recurrence-free survival between the two groups (P = 0.212). One month after SBRT, the radiological change "no evidence of increased density" was prevalent. The OS, PFS, and DMFS were significantly longer in the "no evidence of increased density" group compared with the "consolidation or GGO" group. Further studies are needed to validate these findings.


Proteolysis of extracellular matrix by invadopodia facilitates human breast cancer cell invasion and is mediated by matrix metalloproteinases.
Breast cancer cell lines vary in invasive behavior and one highly invasive cell line (MDA-MB-231) proteolytically degrades extracellular matrix with invadopodia (Thompson et al. 1992, J Cell Physiol, 150, 534-44; Chen et al 1994, Breast Cancer Res Treat, 31, 217-26). Invadopodial proteolysis of extracellular matrix is thought to be necessary for invasion; however, this has not been demonstrated directly. To obtain such evidence, normal (HBL-100) and malignant (MCF-7, MDA-MB-231) breast cells were evaluated for invadopodial proteolysis of extracellular matrix and invasive behavior. We report that invadopodial proteolysis of immobilized fibronectin is positively correlated with invasion of cells into type I collagen gels. Moreover, reducing the proteolytic activity of invadopodia with the metalloproteinase inhibitor, batimastat (BB-94), also decreases invasion indicating that breast cancer cell invasion is dependent upon proteolytically active invadopodia.


Use of dried poultry litter in the diet of pregnant and lactating Awassi ewes.
Two groups of 45 Awassi ewes, were fed a control diet (group C) or a diet containing 0.30 poultry litter from laying birds (group PL). The experiment began when the rams were put with the ewes and continued through pregnancy and lactation. Mean weight changes of ewes of both groups were small and not significantly different. The numbers of ewes that lambed and the numbers of lambs weaned, as proportions of the number of ewes mated, were 0.91 and 0.73 respectively for group C, and 0.93 and 0.70 for group PL. The mean weaning weight of lambs of group C (20.6 kg) was just significantly greater than the value for group PL (18.4 kg) but the estimated mean daily milk yields, 0.716 and 0.626 kg respectively, did not differ significantly. Differences in breeding and lactation performance, between ewes given the control and those given the poultry litter diet, were small for all the data obtained. Also, there was no disease problem related to the use of poultry litter; and the food products milk and cheese, from ewes given poultry litter, were just as acceptable as those from ewes given the control diet.