Cloning and functional expression of a human neuropeptide Y/peptide YY receptor of the Y1 type.
Neuropeptide Y (NPY) and peptide YY (PYY) are structurally related peptides that primarily function as neurotransmitter and gastrointestinal hormone, respectively. Previous functional and binding data have indicated the existence of at least three distinct receptor types, Y1, Y2, and Y3, for NPY and/or PYY in mammals. We describe here a human Y1 cDNA clone, hY1-5, isolated from a fetal brain library. The human Y1 receptor consists of 384 amino acids and has seven putative transmembrane domains like other members of the G-protein-coupled superfamily of receptors. In the region spanning the transmembrane domains, the Y1 receptor displays 29% sequence identity to human tachykinin receptors, but it only shows 21% and 23% homology with proposed bovine (LCR1) and Drosophila (PR4) NPY receptor clones, respectively. Northern blot analysis of a human neuroblastoma cell line, SK-N-MC, previously used by many investigators as a model system for studies on the Y1 receptor, revealed a single 3.5-kilobase mRNA species. Reverse transcriptase-polymerase chain reaction analysis indicated expression also in human cultured vascular smooth muscle cells, supporting the view that the Y1 receptor is associated with NPY/PYY-evoked vasoconstriction. When expressed in COS1 cells, hY1-5 conferred specific 125I-PYY binding sites with displacement patterns characteristic of the Y1 receptor, i.e. PYY greater than or equal to NPY greater than or equal to [Leu31,Pro34]NPY much greater than NPY2-36 greater than C2NPY greater than pancreatic polypeptide greater than NPY13-36 greater than NPY18-36. Moreover, in the Y1 receptor-transfected COS1 cells, but not in type 1 angiotensin II receptor-transfected control cells, NPY and PYY accelerated 45Ca2+ influx and inhibited forskolin-stimulated cAMP accumulation, both phenomena being characteristic of the mammalian Y1 receptor.


Potentially inappropriate medication use and mortality in patients with cognitive impairment.
Potentially inappropriate medications (PIMs) are associated with falls, hospitalization, and cognitive decline. Few studies have investigated the association between PIMs related to cognitive impairment (PIMCog) and mortality in dementia or mild cognitive impairment (MCI). This was a retrospective observational study. Patients diagnosed with MCI or dementia (DSM-IV criteria) presenting to a tertiary-referral memory clinic from 2013 to 2019 were eligible. The primary outcome was all-cause death. Secondary outcomes were vascular death and non-vascular death. The primary exposure variable of interest was PIMCog, defined as any medication in the Beers 2015 or STOPP criteria, classified as potentially inappropriate for patients with cognitive impairment. Anticholinergic burden was measured using the anticholinergic cognitive burden (ACB) scale. Polypharmacy was defined as ≥ 5 medications. Cox proportional hazard models were used to calculate hazard ratios (HRs) and 95% confidence intervals (95% CIs). Four hundred eighteen patients were included (n = 261 dementia, n = 157 MCI). The median age was 79 (interquartile range [IQR] 74-82) and median follow-up was 809 days (IQR 552-1571). One or more PIMCog was prescribed in 141 patients (33.4%). PIMCog use was associated with all-cause mortality after adjustment for age, sex, dementia severity, Charlson's Co-morbidity Index, chronic obstructive pulmonary disease, congestive cardiac failure, and peripheral vascular disease (HR 1.96, 95% CI 1.24-3.09). PIMCog use was associated with vascular death (HR 3.28, 95% CI 1.51-7.11) but not with non-vascular death (HR 1.40 95% CI 0.78-2.52). PIMCog use in patients with cognitive impairment is high. It is independently associated with all-cause mortality and vascular death. This is a potential modifiable risk factor for death in this patient cohort. Further research is required to independently validate this finding.


Contralateral noise has possible asymmetric frequency-sensitive effect on the 2F1-F2 otoacoustic emission in humans.
Contralateral acoustic stimulation has been shown to produce a suppressive effect on the 2F1-F2 distortion-product otoacoustic emission (DPOAE). According to reports by other studies, there is a frequency-specific suppressive region between 1 and 2kHz when the contralateral broadband or narrowband noise stimulation contains energy close to or near the F2 frequencies in that region. In general the broader the bandwidth of the noise stimulation, the greater is the suppressive effect. A frequency-sensitive region between 1 and 2kHz has also been suggested as narrowband noises with center frequencies remote from the F2 frequency seems to produce suppression in the same defined region. In order to test this hypothesis, narrowband noises wide enough to elicit suppression with center frequencies unmatched to the F2 frequencies were employed in the present study. For the right ear, a strong frequency-sensitive suppressive region was noted for the F2 frequency at 2kHz, but not at 1 and 4kHz. The left ear had some suppression at 1kHz, but this effect was not as robust as seen in the right ear. The apparent suppressive differences between ears suggest a possible MOC reflex asymmetry that has not previously been described for DPOAEs. These results suggest that physiological differences between ears and possibly handedness must be taken into account when examining contralateral suppression to narrowband noises. Furthermore, the frequency contributions of the contralateral stimulus may have a greater complex interaction with the cochlear physiology than previously considered.


Ultrastructure of serotonin-containing nerve fibres in the middle cerebral artery of the rat and evidence for its localization within catecholamine-containing nerve fibres by immunoelectron microscopy.
This study has examined the ultrastructural characteristics of serotonin (5-HT)-like immunoreactive (5-HT-I) nerves in the perivascular plexus of a major cerebral blood vessel and their relationship with catecholaminergic (CA) nerves. This was achieved by immunohistochemistry for 5-HT used alone and in combination with simultaneous 5-hydroxydopamine (5-OHDA) false transmitter preloading to demonstrate catecholaminergic nerves examined by electron microscopy. About 15% of perivascular nerves showed some 5-HT-like immunoreactivity. These nerves were characterised by a predominance of small pleomorphic vesicles with a diameter of 40-50 nm and a few large granular vesicles about 100-150 nm diameter. 5-HT-I nerves were found at all levels of the perivascular plexus, from the outer adventitia where they were formed together with non-reactive nerves into bundles by Schwann cells and their processes, to more singly at the adventitia-media border, sometimes in close apposition to smooth muscle cells, separated by some 125 nm. Examination of animals treated by 5-OHDA preloading revealed that 5-HT-I nerves were almost invariably identified as catecholaminergic. However, CA nerves did not always show 5-HT-like immunoreactivity, some 25% only being doubly labelled in this study. Treatment of animals with the catecholaminergic nerve uptake blocker desmethylimipramine resulted in abolition of 5-HT-like immunoreactivity. These results provide direct ultrastructural evidence for the localisation of 5-HT within the perivascular catecholaminergic nerves of a major cerebral vessel and suggest that the contained 5-HT is mainly derived by uptake rather than synthesis within them.


Correlation between decreased motor activity and dopaminergic degeneration in the ventrolateral putamen in monkeys receiving repeated MPTP administrations: a positron emission tomography study.
Parkinson's disease (PD) patients have remarkably reduced levels of dopaminergic biomarkers in the caudal putamen. However, the relationship between motor impairments and the localization of intrastriatal dopaminergic degeneration in monkey PD models remains unclear. To identify the striatal areas with dopaminergic dysfunction responsible for motor impairments, we measured changes in both general motor activity and in vivo dopaminergic biomarkers in three cynomolgus monkeys that repeatedly received 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), starting in the normal state and continuing until after tremor appearance. Binding of dopamine transporters (DAT) and D(2) receptors were measured by positron emission tomography (PET) using [(11)C]PE2I and [(11)C]raclopride, respectively. Region-of-interest-based regression analysis demonstrated the degree of general motor activity reduction to be explained by striatal DAT binding but not by D(2) receptor binding. Furthermore, voxel-based analysis revealed a significant correlation between reduced general motor activity and decreased DAT binding, specifically in the ventrolateral putamen, which corresponds to the area receiving upper body motor inputs from the primary motor cortex. These results suggest that specific functional deficits in PD models are closely related to the degeneration of dopaminergic terminals in the striatal subregion responsible for these functions and that the level of deficit is dependent on the degree of degeneration.


Differential effects of donor-specific alloantibody.
Alloantigen exposure typically provokes an adaptive immune response that can foster rejection of transplanted organs, and these responses present the most formidable biological barrier to kidney transplantation. Although most cellular alloimmune responses can be therapeutically controlled with T-cell-specific immunosuppressants, humoral alloimmune responses remain relatively untamed. Importantly, humoral immunity, typically manifesting as allospecific antibody production, is increasingly recognized for its variable appearance after kidney transplantation. Indeed, the appearance of alloantibody can herald the onset of rapid and destructive antibody-mediated rejection or have no demonstrable acute effects. The factors determining the end result of alloantibody formation remain poorly understood. This review will discuss the breadth of alloantibody responses seen in clinical kidney transplantation and provide an overview of potential factors explaining the phenotypic variability associated with humoral alloimmunity. We propose several avenues ripe for future investigation including the influence of innate immune components and the potential influence of heterologous immune responses in determining the ultimate clinical import of an alloantibody response.


Herpes virus infections in patients with neoplastic disease. Diagnosis and therapy.
Herpes viruses are among the most common and troublesome opportunistic pathogens infecting patients with neoplastic diseases. The recent development of partially effective and relatively nontoxic antiviral agents offers promise for the prophylaxis or therapy of these infections in high-risk groups. Vidarabine and acyclovir have shown efficacy in several herpes virus infections and are now licensed in the United States. Alpha interferon may also be useful in the prophylaxis or early therapy of certain herpes virus infections. Newer antiviral agents and combination therapies are under study. Early and rapid diagnosis of such infections is critical to the development of effective therapy.


LINE retrotransposition and host DNA repair machinery.
Long interspersed elements (LINEs), or non-long-terminal repeat (LTR) retrotransposons, are mobile genetic elements that exist in the genomic DNA of most eukaryotes, comprising a considerable portion of the host chromosomes. LINEs constitute endogenous mutagens that cause insertional mutations in host chromosomes and have a large impact on host genome evolution. Despite their importance, however, the molecular mechanism of LINE retrotransposition is not fully understood. Several studies suggest that host proteins that participate in the repair of DNA breaks modulate LINE retrotransposition. Recently, we provided evidence that there are 2 distinct pathways-annealing and direct-that join the 5'-end of LINEs to host chromosomal DNA. These pathways appear to be used distinctively by zebrafish LINEs and the human L1 in DT40 cells. In HeLa cells, only the annealing pathway appears to be used. This implies that different characteristics of the 2 LINEs and also host factors dictate which pathway is selected. Here, we discuss the 5'-end-joining pathways of LINE retrotransposition and propose that the pathways of LINE integration adopt certain host repair factors.


Correction to: Axial loading during MRI reveals insufficient effect of percutaneous interspinous implants (Aperius™ PerCLID™) on spinal canal area.
Unfortunately, the ninth reference was incorrectly published in the original publication. The complete correct reference is given below.


Vancomycin dosing in morbidly obese patients.
Vancomycin hydrochloride dosing requirements in morbidly obese patients with normal renal function were computed to determine the dose of vancomycin necessary to achieve target steady-state peak and trough concentrations and compared with a normal weight population. Morbidly obese patients [total body weight (TBW) 165 kg, ideal body weight (IBW) 63 kg] required 31.2 mg x kg(-1) x d(-1) TBW or 81.9 mg x kg(-1) x d(-1) IBW to achieve the target concentrations. Normal weight patients (TBW 68.6 kg) required 27.8 mg x kg(-1) x d(-1) to achieve the same concentrations. Because of altered kinetic parameters in the morbidly obese patients (obese: t1/2 = 3.3 h, V = 52 L, CL = 197 ml x min(-1); normal: t1/2=7.2 h, V=46 L, CL=77 ml x min(-1), 20 of 24 patients required q8h dosing (1938 mg q8h) compared with q12h dosing (954 mg q12h) in all normal weight patients in order to avoid trough concentrations that were too low for prolonged periods. There was a good correlation between TBW and CL, but only fair correlation between TBW and V. Doses required to achieve desired vancomycin concentrations are similar in morbidly obese and normal weight patients when TBW is used as a dosing weight for the obese (approximately 30 mg x kg(-1) x d(-1)). Shorter dosage intervals may be needed when dosing morbidly obese patients so that steady-state trough concentrations remain above 5 microg x ml(-1) in this population. Because of the large amount of variation in required doses, vancomycin serum concentrations should be obtained in morbidly obese patients to ensure that adequate doses are being administered. Dosage requirements for morbidly obese patients with renal dysfunction require further study.


Otosclerosis and otospongiosis: clinical and histological comparisons.
The simultaneous occurrence of otosclerosis and otospongiosis in the same lesion is well documented. The logical sequence of events would seem to be softening and destruction of the bone by the otospongiotic lesion, which is subsequently converted to otosclerosis. Examination of 46 temporal bones has led us to further classify the lesion as: 1. an active lesion, both otospongiotic and otosclerotic, 2. an inactive lesion, also both otosclerotic and otospongiotic, or 3. a fibrous lesion. Hyalinization of the spiral ligament only occurs adjacent to active otospongiotic or very large otosclerotic lesions. If the lesion adjacent to the spiral ligament is inactive, there is no hyalinization. It seems illogical that a hyalinized spiral ligament will return to normal when the lesion changes from active otospongiosis to inactive otosclerosis. Otospongiotic and otosclerotic lesions are found side by side at the periphery of lesions, and both are adjacent to normal bone. These findings have implications concerning the mode of action of sodium fluoride and other medications that are under evaluation for the control of the sensorineural hearing loss due to otosclerosis.


Meningococcal disease epidemiology in Brazil (2005-2018) and impact of MenC vaccination.
Meningococcal disease (MD) presents a substantial public health problem in Brazil. Meningococcal C conjugate (MenC) vaccination was introduced into the routine infant immunization program in 2010, followed by adolescent vaccination in 2017. We evaluated changes in national and regional MD incidence and mortality between 2005 and 2018, serogroup distribution and vaccine coverage. Data were obtained from national surveillance systems from 2005 to 2018. Age-stratified incidence and mortality rates were calculated and a descriptive time-series analysis was performed comparing rates in the pre-(2005-2009) and post-vaccination (2011-2018) periods; MD due to specific meningococcal serogroups were analyzed in the pre-(2007-2009) and post-vaccination (2011-2018) periods. From 2005 to 2018, 31,108 MD cases were reported with 6496 deaths; 35% of cases and deaths occurred in children < 5 years. Incidence and mortality rates declined steadily since 2012 in all age-strata, with significantly lower incidence and mortality in the post-vaccine introduction period in children aged < 1-year, 1-4 years, 5-9 years and 10-14 years. A significant decline in MenC disease in children < 5 years was observed following MenC vaccine introduction; infants < 1 year, from 3.30/100,000 (2007-2009) to 1.08/100,000 (2011-2018) and from 1.44/100,000 to 0.42/100,000 in 1-4-year-olds for these periods. Reductions in MenB disease was also observed. MenW remains an important cause of MD with 748 cases reported across 2005-2018. While initial infant vaccination coverage was high (>95% nationwide), this has since declined (to 83% in 2018); adolescent uptake was < 20% in 2017/18). Regional variations in outcomes and vaccine coverage were observed. A substantial decline in incidence and mortality rates due to MD was seen following MenC vaccine introduction in Brazil, especially among children < 5 years chiefly driven by reductions in MenC serogroup. While these benefits are considerable, the prevalence of MD due to other serogroups such as MenW and MenB remains a concern. A video summary linked to this article can be found on Figshare: https://doi.org/10.6084/m9.figshare.13379612.v1.


Cardiovascular event rates in patients with ST-elevation myocardial infarction were lower with early increases in mobilization of Oct4(high)Nanog(high) stem cells into the peripheral circulation during a 4-year follow-up.
Long-term clinical implications of embryonic stem cell markers such as Oct4 and Nanog have not been investigated in ST-elevation myocardial infarction (STEMI) patients. The aim of this study was to investigate the effects of early peripheral mobilization of stem cells with Oct4 and Nanog gene expression on major adverse cardiovascular events (MACEs) in patients with STEMI during a 4-year follow-up. Peripheral blood mononuclear cells (PBMCs) were isolated on days 0, 1 and 7 from patients with STEMI (n = 40) and healthy controls (n=20). The numbers of CD34+, CD117+, CD133+ and c-met+ stem cells were measured by flow-cytometry. Oct4 and Nanog gene expressions were analyzed by real-time PCR. MACEs such as non-fatal MI, death, stroke, target lesion and revascularization were observed. MACEs were significantly lower in patients with Oct4 gene expression ≥ 1.13 and Nanog gene expression ≥ 1.20 at admission. The numbers of CD34+, CD117+, CD133+ and c-met+ cells within 7 days after STEMI did not show significant differences in patients with or without MACE. Level of anti-inflammatory marker such as IL-10 was significantly higher within 7 days following STEMI in patients without MACE. Inflammatory and angiogenic markers such as CRP, IL-6, SCF, SDF-1α, and VEGF did not show significant differences in patients with or without MACE. mRNA levels of pluripotent embryonic stem cell markers such as Oct4 and Nanog were significantly higher in STEMI patients without MACEs during a 4-year follow-up. Baseline Oct4 and Nanog gene expression levels could be used as predictors of MACE in STEMI patients.


Novel applications of plant polyketide synthases.
The structurally and mechanistically simple type III polyketide synthases (PKSs) catalyze iterative condensations of CoA thioesters to produce a variety of polyketide scaffolds with remarkably diverse structures and biological activities. By exploiting the enzymes, we combined precursor-directed biosynthesis with nitrogen-containing substrates and structure-based enzyme engineering and generated unnatural, novel polyketide-alkaloid scaffolds with promising biological activities. The nucleophilic nitrogen atom and the engineered enzymes thus facilitated the formation of additional CC and CN bonds during the enzymatic transformations. The methodology will contribute to the further production of chemically and structurally divergent, unnatural natural products, as well as the rational design of novel biocatalysts with unprecedented catalytic functions.


Transformation mechanism of polycyclic aromatic hydrocarbons and hydrogen production during the gasification of coking sludge in supercritical water.
In this study, the characteristic of hydrogen production and polycyclic aromatic hydrocarbons (PAHs) transformation during supercritical water gasification (SCWG) of coking sludge (400 °C - 450 °C, 1 min-30 min) were explored. The total gas yield was between 0.62 mol/kg OM and 1.05 mol/kg OM (on dry basis), and the proportion of CH4 and H2 was only between 5.41% and 6.44%. PAH content were increased from 194.92 mg/kg to 326.04 mg/kg, and mainly high molecular weight PAHs, which were formed from the Diels-Adler reaction of single aromatic hydrocarbon and the addition reaction of low molecular weight PAHs. High reaction temperature favored more active PAH formation than reaction time. The possible control methods for PAH formation during SCWG of coking sludge was proposed. H2O2 and KOH addition effectively reduce PAHs amount in solid residues by 46.67% and 38.33%, and KOH performed positive effect on hydrogen production. The work revealed that the inhibition of PAHs and hydrogen production were achieved from SCWG of coking sludge with KOH addition.


[Effects of immunotherapy with infantile recurrent respiratory tract infection by Chinese materia medica].
To explore the effect of Chinese materia medica on immune intervention of infantile recurrent respiratory tract infection. Thirty-one children of recurrent respiratory tract infection were randomly divided into two groups: therapy group was treated with oral Chinese materia medica (b. i. d), control group was only treated with oral carboxymethyl liquor (< 4 years, 3 mL; 4-7 years, 5 mL; > 7 years, 7 mL, t. i. d). The change of IL-12,TNF-alpha, IL-6, IL-13, IL-6 and IL-4 in different time were observed and analyzes. Compared with the control group, the level of IL-12 and IL-2 was significantly increased after treatment of oral Chinese materia medica (P < 0.01), however, the level of TNF-alpha, IL-13, IL-4, and IL-6 was decreased after treatment (P < 0.01). During one years follow-up study, the frequency of respiratory infection every year of therapy group was significantly decreased than that of control group. Chinese materia medica could prevent infantile recurrent respiratory tract infection effectively, increase humoral immunity function and ensure normal growth in children.


[Social medical/public health assessment of bladder cancer after curative and adjuvant therapy].
Radical cystectomy and urinary diversion are a challenge for patients. Requirements for the successful participation of the patient are sufficient urinary diversion management and recuperation/recovery as the result of urological rehabilitation. A social medical assessment reviews the individual oncological prognosis and the rehabilitation results to determine the return to work.


Levels of choroidal body carbonic anhydrase activity and glycogen in farmed halibut.
We postulate that ocular abnormalities in farmed Atlantic halibut occur following aberrant gas production in the choroidal body associated with the enzyme carbonic anhydrase. We investigated choroidal body carbonic anhydrase activity in affected and normal halibut. Immunohistochemical detection of carbonic anhydrase allowed neither quantification of enzyme levels nor determination of enzyme activity. Histochemical detection of carbonic anhydrase, however, allowed microdensitometric quantification of enzyme activity. Mean enzyme activity in affected individuals was over twice that in normal fish. Glycogen, which we suggest is used to fuel oxygen production in the choroidal body, was evaluated by Best's carmine staining. Microdensitometry demonstrated glycogen levels of be significantly higher in affected fish. These findings show that posterior segment gas bubble and cyst formation is associated with aberrantly high choroidal enzyme activity and high choroidal glycogen concentrations.


Identification of neural pathways activated in dehydrated rats by means of Fos-immunohistochemistry and neural tracing.
The distribution of Fos-immunoreactivity (Fos-IR) was studied in the brains of rats deprived of water for 24 or 48 h and compared with that in brains of water-replete rats. Intense Fos-IR was observed in many neurons of the median preoptic nucleus (MnPO), organum vasculosum of the lamina terminalis (OVLT), supraoptic nucleus and hypothalamic paraventricular nucleus. There was less intense and sparse Fos-IR in the subfornical organ. In water-replete rats, Fos-IR was absent or very low in these regions. In other rats, cholera toxin B-gold conjugate was microinjected bilaterally into the supraoptic nucleus to identify retrogradely labelled neurons in the lamina terminalis projecting to the supraoptic nucleus. Approximately 30% of these retrogradely labelled neurons in the OVLT and MnPO also exhibited Fos-IR after 48 h of water deprivation. These data show that neurons in the MnPO, OVLT and, to a lesser extent, the subfornical organ probably play an important role in homeostatic responses to dehydration, such as vasopressin secretion.


Structure and mechanism of assembly line polyketide synthases.
Assembly line polyketide synthases (PKSs) are remarkable biosynthetic machines with considerable potential for structure-based engineering. Several types of protein-protein interactions, both within and between PKS modules, play important roles in the catalytic cycle of a multimodular PKS. Additionally, vectorial biosynthesis is enabled by the energetic coupling of polyketide chain elongation to the channeling of intermediates between successive modules. A combination of high-resolution analysis of smaller PKS components and lower resolution characterization of intact modules and bimodules has yielded insights into the structure and organization of a prototypical assembly line PKS. This review discusses our understanding of key structure-function relationships in this family of megasynthases, along with a recap of key unanswered questions in the field.


Optimization of microwave-assisted solvent extraction for volatile organic acids in tobacco and its comparison with conventional extraction methods.
In the present study, a new method using microwave-assisted solvent extraction (MASE) technique followed directly GC analysis was developed for the extraction of volatile organic acids (VOAs) in tobacco. The MASE conditions (heating time, volume of extracting solvent and extraction temperature) were optimized by means of an orthogonal array design (OAD) procedure. The results suggested that extractant, temperature and heating time were statistically the most significant factors. The extracts were directly analyzed with capillary GC operating in splitless-injection mode on an Agilent HP-FFAP capillary column. Under optimum operating conditions, MASE showed significantly better recoveries than those obtained by the conventional extraction method (ultrasonic and reflux extraction), ranging from 90.6% to 103.2%. In addition, a drastic reduction of the extraction time (20 min versus 4h) and solvent consumption (20 mL versus 100 mL) was achieved with an outstanding reproducibility (CV < or =5%).


[Histochemical study of the pilotropic action of 1-(chloromethyl)silatrane on guinea pig skin].
Histochemical methods have disclosed that external application of 5% ointment of 1-(chlormethyl)silatran leads to an increase of the glycogen and RNA content in guinea-pig skin, reduction of the heparin content in the basal membrane and external epithelial vagina. Factors that give rise to pilotropic activity of 1-(chlormethyl)silatran are discussed.


Effect of anticholinergic drug on long-term potentiation in rat hippocampal slices.
Long-term potentiation (LTP) was studied in CA1 neurons by tetanization of the Schaffer-commissural pathway in rat hippocampal slices. A brief tetanus (200 Hz for 1 s) caused an increase in amplitude of the population spike recorded from the CA1 area (typically about 200% of control), which lasted for more than 2 h. LTP was suppressed by perfusion of the muscarinic antagonist scopolamine (10(-5) M) from 5 min before to 15 min after the tetanus. If perfusion of the drug was begun after the tetanus, there was no affect on LTP. Scopolamine perfused without tetanus did not change the amplitude of the population spike. These results suggest that cholinergic system may affect the generation mechanisms of LTP.


Maternal type 1 diabetes reduces the risk of islet autoantibodies: relationships with birthweight and maternal HbA(1c).
The risk of type 1 diabetes is reduced in the children of mothers with type 1 diabetes compared with children of fathers with type 1 diabetes. We asked whether children of mothers with type 1 diabetes also have a decreased risk of developing islet autoantibodies, and which factors associated with maternal diabetes contribute to a reduced islet autoantibody risk in offspring. Singleton offspring of a mother (n = 1,008) or father with type 1 diabetes (n = 578) from the BABYDIAB study were included. Children were followed from birth for the development of islet autoantibodies defined as two or more autoantibodies to insulin, glutamic acid decarboxylase or insulinoma antigen 2 in two or more blood samples. Islet autoantibody risk was lower in children of mothers with type 1 diabetes (5 year risk, 3.2% vs 5.7% in children of fathers with type 1 diabetes; p = 0.04). Among factors that differed between pregnancies from mothers with and without type 1 diabetes, birthweight was associated with islet autoantibody risk. Risk was reduced in children with birthweights in the lower (adjusted HR 0.33; 95% CI 0.14-0.75; p = 0.009) and upper (HR 0.45; 95% CI 0.21-0.97; p = 0.04) tertiles compared with the middle tertile. A sub-analysis of maternal HbA(1c) suggested that moderately elevated third trimester maternal HbA(1c) was also associated with a reduced islet autoantibody risk in children of mothers with type 1 diabetes (5.7-7%; HR 0.38; 95% CI 0.15-0.96; p = 0.04 vs children of mothers with HbA(1c) < 5.7%). The risk of islet autoimmunity is modified by maternally influenced events such as birthweight.


[Prolonged release of chlorambucil and etoposide from poly-3-oxybutyrate-based microspheres].
Microspheres were obtained on the basis of poly(3-oxibutyrate) (POB) with the inclusion of the Chlorambucil and Etoposide cytostatic drugs in a polymer matrix, and the morphology, kinetics of drug release from microspheres, and the interaction between microspheres and tumor cells in vitro were studied. Data on the kinetics of drug release suggests that a prolonged release occurs by drug diffusion from the polymer matrix at the initial stage and at the expense of hydrolytic degradation of the polymer at a later stage. A study of the biocompatibility and biological activity of biopolymeric microspheres showed that chlorambucil operates actively and strongly inhibits the growth of cultured cells for a short time (24 h). Etoposide acts weaker (the percentage of cell growth suppression during 48 h does not exceed 50%), but subsequently it has a basis for the creation of new dosage forms with prolonged action of Etoposide and chlorambucil for cancer therapy.


[Plastic external ear reduction in bilateral macrotia].
We present the rare case of a bilateral reduction of macrotic ears by a helical advancement according to Davis. It is a quite simple and reliable technique, that permits to hide the scars in the relief of the pinna.


TRPM7 channel regulates PDGF-BB-induced proliferation of hepatic stellate cells via PI3K and ERK pathways.
TRPM7, a non-selective cation channel of the TRP channel superfamily, is implicated in diverse physiological and pathological processes including cell proliferation. Recently, TRPM7 has been reported in hepatic stellate cells (HSCs). Here, we investigated the contribution role of TRPM7 in activated HSC-T6 cell (a rat hepatic stellate cell line) proliferation. TRPM7 mRNA and protein were measured by RT-PCR and Western blot in rat model of liver fibrosis in vivo and PDGF-BB-activated HSC-T6 cells in vitro. Both mRNA and protein of TRPM7 were dramatically increased in CCl4-treated rat livers. Stimulation of HSC-T6 cells with PDGF-BB resulted in a time-dependent increase of TRPM7 mRNA and protein. However, PDGF-BB-induced HSC-T6 cell proliferation was inhibited by non-specific TRPM7 blocker 2-aminoethoxydiphenyl borate (2-APB) or synthetic siRNA targeting TRPM7, and this was accompanied by downregulation of cell cycle proteins, cyclin D1, PCNA and CDK4. Blockade of TRPM7 channels also attenuated PDGF-BB induced expression of myofibroblast markers as measured by the induction of α-SMA and Col1α1. Furthermore, the phosphorylation of ERK and AKT, associated with cell proliferation, decreased in TRPM7 deficient HSC-T6 cells. These observations suggested that TRPM7 channels contribute to perpetuated fibroblast activation and proliferation of PDGF-BB induced HSC-T6 cells via the activation of ERK and PI3K pathways. Therefore, TRPM7 may constitute a useful target for the treatment of liver fibrosis.


Durable sequence stability and bone marrow tropism in a macaque model of human pegivirus infection.
Human pegivirus (HPgV)-formerly known as GB virus C and hepatitis G virus-is a poorly characterized RNA virus that infects about one-sixth of the global human population and is transmitted frequently in the blood supply. We create an animal model of HPgV infection by infecting macaque monkeys with a new simian pegivirus (SPgV) discovered in wild baboons. Using this model, we provide a high-resolution, longitudinal picture of SPgV viremia where the dose, route, and timing of infection are known. We detail the highly variable acute phase of SPgV infection, showing that the viral load trajectory early in infection is dependent on the infecting dose, whereas the chronic-phase viremic set point is not. We also show that SPgV has an extremely low propensity for accumulating sequence variation, with no consensus-level variants detected during the acute phase of infection and an average of only 1.5 variants generated per 100 infection-days. Finally, we show that SPgV RNA is highly concentrated in only two tissues: spleen and bone marrow, with bone marrow likely producing most of the virus detected in plasma. Together, these results reconcile several paradoxical observations from cross-sectional analyses of HPgV in humans and provide an animal model for studying pegivirus biology.


Measuring the rate of therapeutic adherence among outpatients with T2DM in Egypt.
The promotion of therapeutic adherence is considered as an integral component of pharmaceutical care practice and patient healthcare. It has been shown that despite effective methods of treatment, 50% of diabetic patients fail to achieve satisfactory glycemic control, which leads to accelerated development of complications and increased mortality. Clinical experience indicates that no improvement of metabolic control is possible without patients' adherence to medications. This study sought to examine the rate of medication adherence and different factors affecting it among Type 2 diabetic patients in Egypt. A total of 226 Type 2 diabetic patients who fulfilled the inclusion criteria were recruited in the current study. Adherence to the treatment was evaluated during patients' hospitalization in the Outpatient Clinics of Internal Medicine Department at University of Mansoura, Egypt. The medication adherence has been assessed during a personal interview with each patient using a multiple-choice graded questionnaire. In the study population, the adherence rates to medication, dietary/exercise and appointment were observed to be suboptimal. The most important social factors that were significantly affecting adherence rate to the prescribed oral hypoglycemic agent(s) included marital status (P < 0.01), family support (P < 0.01), and socio-economical level (P < 0.01). Other patient factors that were significantly affecting therapeutic adherence were patient knowledge about the disease (P < 0.01), patients' beliefs and motivation about prescribed drugs (P < 0.01), and regularity of patients' self monitoring of blood glucose level (P < 0.01). Among drug factors which found to affect significantly the rate of medication adherence are the number of drugs taken (P < 0.05), complexity of drug regimen (P < 0.01), and the presence of drug side effects (P < 0.01). Economical factor played an equally important role. Direct and indirect care costs in relation to patients' income were significantly affecting the rate of adherence to medication (P < 0.01). An improvement with the adherence to oral hypoglycemic agent(s) may be achieved through continuing patient education about diabetes, improvement of patients' economical levels as well as a reduction in the cost of medication. Pharmaceutical companies have to be involved and pharmacists have to be payed for helping chronically ill patients to take their medicines correctly for improving clinical outcomes.


Contaminants of emerging concern in drinking water: Quality assessment by combining chemical and biological analysis.
Drinking water quality is a priority issue of the environmental policy agenda, however regulation on Contaminants of Emerging Concern (CECs) is limited. A proposal to revise the Drinking Water Directive has recently been approved (EU Council 2020), which updates the quality standards and introduces the watch list mechanism, including for the first time endocrine disruptors and pharmaceuticals. The purpose of this study was to evaluate the occurrence of selected CECs in surface water at the entrance of drinking water treatment plants (DWTPs) and in treated water, ready for distribution in the network. Samples were collected at three different DWTPs (Italy) and CECs assessed by LC-MS/MS were the following: bisphenol A (BPA), nonylphenol (NP), octylphenol, perfluorooctanesulfonic and perfluorooctanoic acids (PFOS and PFOA), atenolol, caffeine (CFF), carbamazepine (CBZ), estrone, 17-β-estradiol, 17-α-ethinyl estradiol, diclofenac, and ibuprofen. In addition, biological analyses were performed to ascertain cumulative estrogenic and/or genotoxic potential of the samples. CFF, NP, PFOA, BPA, and CBZ were the most frequently detected contaminants, found in treated water in the following ranges: CFF 12.47-66.33 ng/L, NP 7.90-53.62 ng/L, PFOA <LOQ-12.66, ng/L, BPA <LOQ-6.27 ng/L, and CBZ <LOQ-1.20 ng/L. While treatments were generally efficacious in reducing BPA, CFF and CBZ, they were sometimes ineffective for NP and PFOA. According to the low concentrations and/or regulation limit for each single contaminant, the water analyzed met the criteria of good quality. No estrogenic or genotoxic activities were induced by the water assessed, with the exception of one sample. It has been suggested that drinking water may not represent a significant source of human exposure to CECs, however co-occurrence of different compounds may lead to additive/synergistic interactions causing unexpected effects on human health and the environment, and deserve implementation of strategies for detection and removal.


High-precision tumor resection down to few-cell level guided by NIR-IIb molecular fluorescence imaging.
In vivo fluorescence/luminescence imaging in the near-infrared-IIb (NIR-IIb, 1,500 to 1,700 nm) window under <1,000 nm excitation can afford subcentimeter imaging depth without any tissue autofluorescence, promising high-precision intraoperative navigation in the clinic. Here, we developed a compact imager for concurrent visible photographic and NIR-II (1,000 to 3,000 nm) fluorescence imaging for preclinical image-guided surgery. Biocompatible erbium-based rare-earth nanoparticles (ErNPs) with bright down-conversion luminescence in the NIR-IIb window were conjugated to TRC105 antibody for molecular imaging of CD105 angiogenesis markers in 4T1 murine breast tumors. Under a ∼940 ± 38 nm light-emitting diode (LED) excitation, NIR-IIb imaging of 1,500- to 1,700-nm emission afforded noninvasive tumor–to–normal tissue (T/NT) signal ratios of ∼40 before surgery and an ultrahigh intraoperative tumor-to-muscle (T/M) ratio of ∼300, resolving tumor margin unambiguously without interfering background signal from surrounding healthy tissues. High-resolution imaging resolved small numbers of residual cancer cells during surgery, allowing thorough and nonexcessive tumor removal at the few-cell level. NIR-IIb molecular imaging afforded 10-times-higher and 100-times-higher T/NT and T/M ratios, respectively, than imaging with IRDye800CW-TRC105 in the ∼900- to 1,300-nm range. The vastly improved resolution of tumor margin and diminished background open a paradigm of molecular imaging-guided surgery.


[Immune, inflammatory and hormonal changes in connection with laparoscopic surgery].
Analysis of the issue of laparoscopy on the intra and post-operative immune changes. Review article. Department of Obstetrics and Gynecology, Regional Hospital Kladno. Review of published facts with regard to impact laparoscopic surgery on immune function. The impact of surgery on the intra and post-operative immune, hormonal and inflammatory response is significantly less after laparoscopy than with an open approach.


Assessment and follow-up of the proportion of T315I mutant BCR-ABL transcripts can guide appropriate therapeutic decision making in CML patients.
Quantitative monitoring of imatinib mesylate (IM)-resistant, mutated BCR-ABL(+) cells during the follow-up of CML could be useful for optimizing therapeutic management. We retrospectively analyzed T315I mutated BCR-ABL clones throughout the CML history of two patients by nested-PCR-RFLP. At the time of progression, the T315I mutation represented 100% of the BCR-ABL transcripts. During follow-up, we showed that (i) despite a molecular response to IM, a high proportion of T315I transcripts were present (>85%) and predictive of relapse, (ii) interruption of IM and switching to other therapies resulted in a significant reduction in mutant transcript level while total BCR-ABL(+) transcripts remained stable.


Sexually inappropriate behaviors in seriously mentally ill children and adolescents.
This study examined the prevalence and clinical correlates of sexually inappropriate behaviors in all youth treated at a tertiary care public sector psychiatric hospital over a 5-year period. A retrospective chart review was completed on 499 subjects. Subjects were grouped in four mutually exclusive categories: no inappropriate sexual behaviors (n = 296), hypersexual (n = 82), exposing (n = 39) and victimizing (n = 82) behaviors. Those with histories of sexually inappropriate behaviors had much higher rates of being sexually abused (82 vs. 36%), and also had higher rates of physical abuse and neglect, behavior disorders, developmental problems, and family histories of antisocial behavior. They were less likely to have affective disorders. The hypersexual group had a higher proportion of females, and was associated in part with variables relating to sexual abuse and posttraumatic stress disorder. The more severe offending groups (exposing and victimizing) were associated with variables related to sexual abuse, developmental delays, lower IQ's, peer problems, and other acting-out behavior problems. These findings underscore the importance of evaluating for sexually inappropriate behaviors in seriously mentally ill youth, especially in those with histories of sexual abuse.


A simplified method for the measurement of gamma-butyrobetaine hydroxylase activity.
A method for the assay of gamma-butyrobetaine hydroxylase activity is described. The procedure is based on the measurement of 3H2O formed from [2,3-3H]gamma-butyrobetaine. The formation of 3H2O was essentially linear with time of incubation and enzyme concentration. Despite a significant isotope effect that causes the extent of hydroxylation to be underestimated, an appropriately determined correction factor permits one to relate quantitatively the degree of detritiation to the amount of carnitine formed. The assay is simple, rapid, specific, accurate, highly reproducible, and relatively sensitive. Its reliability and convenience represent an improvement over existing methods based on the tedious and time-consuming enzymatic radioisotopic determination of the carnitine formed or on the coupled decarboxylation of [1-14C]alpha-ketoglutarate, a method that cannot be used in crude extracts.


Regulation of opioid receptor trafficking and morphine tolerance by receptor oligomerization.
The utility of morphine for the treatment of chronic pain is hindered by the development of tolerance to the analgesic effects of the drug. Morphine is unique among opiates in its ability to activate the mu opioid receptor (MOR) without promoting its desensitization and endocytosis. Here we demonstrate that [D-Ala(2)-MePhe(4)-Gly(5)-ol] enkephalin (DAMGO) can facilitate the ability of morphine to stimulate MOR endocytosis. As a consequence, rats treated chronically with both drugs show reduced analgesic tolerance compared to rats treated with morphine alone. These results demonstrate that endocytosis of the MOR can reduce the development of tolerance, and hence suggest an approach for the development of opiate analogs with enhanced efficacy for the treatment of chronic pain.


Effects of organic matter-goethite interactions on reactive transport of nalidixic acid: Column study and modeling.
The fractionation of natural organic matter (NOM) and its impact on the binding of quinolones to mineral surfaces and transport behavior under flow-through conditions have been scarcely investigated. In this study, the sorption and transport of a widely used quinolone antibiotic, Nalidixic acid (NA), were investigated in goethite-coated sand (GCS) columns over a wide concentration range (5-50 mg/L) of Leonardite humic acid (LHA), a representative NOM. Simultaneous injection of NA and LHA in GCS columns mutually alter transport of each other, i.e. NA mobility and LHA molecular fractionation. Preloading of GCS column with LHA dramatically facilitated the transport behavior of NA, where nonspecific interactions with LHA-covered goethite surfaces controlled NA mobility. Simulations using a two-site nonequilibrium model showed that a modified sorption rate constant was required to accurately describe the breakthrough curves of NA under these conditions. This altered rate constant suggests that nonspecific interactions of NA on bound LHA may take place as an additional binding mechanism affecting adsorption kinetics. NOM fractionation alters sorption mechanisms and kinetics of quinolone antibiotics, which in turn affect their fractionation. These results may have important implications for an accurate assessment of the fate of these types of antibiotics in aquatic environments.


Ethical Challenges in Early-Phase Pediatric Research for Life-Limiting Illness.
Research with children is necessary to assure more effective treatments and potential cures of childhood illnesses. Ethical conduct of research requires minimizing the inherent risks of research, especially when it involves vulnerable populations such as children. Duchenne muscular dystrophy (DMD) is a progressive and fatal disease with no Food and Drug Administration-approved treatment. Clinical trials investigating so-called gene therapies are viewed by many in the DMD community with great promise. The goal of research is to secure generalizable knowledge and not directly benefit patients, yet many parents of boys with DMD hope, and even expect, that their sons will derive medical benefit by participating in early-phase "gene therapy" trials, raising concern for the therapeutic misconception. Physician-investigators must assist patient-subjects to distinguish realistic from unrealistic hope while maintaining reasonable expectations. In this article, I examine the therapeutic misconception and related concepts as framed within the context of "gene therapy" for DMD.


Embryonal carcinoma with falsely positive elevation of serum alpha-fetoprotein after curative therapy: a case report.
An elevated serum alpha-fetoprotein is an important serum and cell marker in nonseminomatous testicular cancer. While the sensitivity of an elevated alpha-fetoprotein is 20 to 70 per cent, depending upon clinical stage subgrouping, the specificity is 100 per cent, since no falsely positive elevation of alpha-fetoprotein has been reported. We report a case of persistent falsely positive elevation of alpha-fetoprotein after curative therapy of embryonal carcinoma.


A polymorphism affecting MYB binding within the promoter of the PDCD4 gene is associated with severe asthma in children.
A previous genome-wide association study in asthma revealed putative associations that merit further investigation. In this study, the genome-wide significant associations of SNPs at the 5% false discovery rate were examined in independent groups of severe asthmatics. The panel consisted of 397 severe asthmatic adults, 116 severe asthmatic children, and a collection of 207 family-trios with an asthmatic proband. Three SNPs in the PDCD4 gene (rs6585018:G>A, rs1322997:C>A, and rs34104444:G>A) were significantly associated with severe childhood asthma (P values: 0.003, 0.002, 0.004) and total immunoglobulin E (IgE) levels (P values: 0.034, 0.041, 0.052). In an independent group of 234 asthmatic children and 652 controls, PDCD4 SNPs rs1407696:T>G and rs11195360:T>C were associated with total IgE levels (P values: 0.006, 0.014). In silico analysis of PDCD4 locus showed that rs6585018:G>A had the potential to affect MYB transcription factor binding, shown to act as a PDCD4-transcription inducer. Electromobility shift assays and reporter assays revealed that rs6585018:G>A alters MYB binding thereby influencing the expression of PDCD4. SNPs within MYB itself confer susceptibility to eosinophilia and asthma. Our association between a variant MYB binding site in PDCD4 and the severest form of childhood asthma therefore suggests that PDCD4 is a novel molecule of importance to asthmatic inflammatory responses.


Effectiveness of Lumbar Cerebrospinal Fluid Drain Among Patients With Aneurysmal Subarachnoid Hemorrhage: A Randomized Clinical Trial.
After aneurysmal subarachnoid hemorrhage, the use of lumbar drains has been suggested to decrease the incidence of delayed cerebral ischemia and improve long-term outcome. To determine the effectiveness of early lumbar cerebrospinal fluid drainage added to standard of care in patients after aneurysmal subarachnoid hemorrhage. The EARLYDRAIN trial was a pragmatic, multicenter, parallel-group, open-label randomized clinical trial with blinded end point evaluation conducted at 19 centers in Germany, Switzerland, and Canada. The first patient entered January 31, 2011, and the last on January 24, 2016, after 307 randomizations. Follow-up was completed July 2016. Query and retrieval of data on missing items in the case report forms was completed in September 2020. A total of 20 randomizations were invalid, the main reason being lack of informed consent. No participants meeting all inclusion and exclusion criteria were excluded from the intention-to-treat analysis. Exclusion of patients was only performed in per-protocol sensitivity analysis. A total of 287 adult patients with acute aneurysmal subarachnoid hemorrhage of all clinical grades were analyzable. Aneurysm treatment with clipping or coiling was performed within 48 hours. A total of 144 patients were randomized to receive an additional lumbar drain after aneurysm treatment and 143 patients to standard of care only. Early lumbar drainage with 5 mL per hour was started within 72 hours of the subarachnoid hemorrhage. Primary outcome was the rate of unfavorable outcome, defined as modified Rankin Scale score of 3 to 6 (range, 0 to 6), obtained by masked assessors 6 months after hemorrhage. Of 287 included patients, 197 (68.6%) were female, and the median (IQR) age was 55 (48-63) years. Lumbar drainage started at a median (IQR) of day 2 (1-2) after aneurysmal subarachnoid hemorrhage. At 6 months, 47 patients (32.6%) in the lumbar drain group and 64 patients (44.8%) in the standard of care group had an unfavorable neurological outcome (risk ratio, 0.73; 95% CI, 0.52 to 0.98; absolute risk difference, -0.12; 95% CI, -0.23 to -0.01; P = .04). Patients treated with a lumbar drain had fewer secondary infarctions at discharge (41 patients [28.5%] vs 57 patients [39.9%]; risk ratio, 0.71; 95% CI, 0.49 to 0.99; absolute risk difference, -0.11; 95% CI, -0.22 to 0; P = .04). In this trial, prophylactic lumbar drainage after aneurysmal subarachnoid hemorrhage lessened the burden of secondary infarction and decreased the rate of unfavorable outcome at 6 months. These findings support the use of lumbar drains after aneurysmal subarachnoid hemorrhage. ClinicalTrials.gov Identifier: NCT01258257.


The presence of tumor necrosis factor-alpha and its antibody in the sera of cachexic patients with gastrointestinal cancer.
Although cancer cachexia has been shown to involve several cytokines, the tumor necrosis factor-alpha (TNF) has rarely been detected in such patients. In this study, sera from 21 patients with cancer cachexia were examined for the presence of TNF and the anti-TNF antibody using an enzyme-linked immunosorbent assay (ELISA) and Western blotting, respectively. All of the patients had recurrent cancer and manifested the characteristics of progressive body weight loss. TNF was found in the sera of four patients (20%) at levels ranging from 10.4 to 53.1 pg/ml, while a positive reaction for the anti-TNF antibody was detected in the sera from six patients (30%), two of whom showed both TNF and its antibody. Thus, either TNF or the anti-TNF antibody was present in the sera from 8 of 21 patients (40%). The results of this study indicate that TNF may be present in the circulation of at least 40% of cachexic patients, and suggest that it may be one of the main mediators of cancer-associated cachexia.


Identification of late-onset hypogonadism in middle-aged and elderly men from a community of China.
In this study, we investigated the essential criteria for late-onset hypogonadism (LOH) syndrome based on the presence of symptoms associated with low testosterone levels in Han Chinese men. Blood tests for total testosterone (TT) and sex hormone-binding globulin (SHBG) were performed, and the aging male symptoms (AMS) questionnaire was conducted in a randomly selected cohort composed of 944 Chinese men aged 40 to 79 years from nine urban communities. Three sexual symptoms (decreased ability/frequency of sexual activity, decreased number of morning erections, and decreased libido) were confirmed to be related to the total and free testosterone levels. The thresholds for TT were approximately 12.55 nmol l-1 for a decreased ability/frequency to perform sex, 12.55 nmol l-1 for decreased frequency of morning erections, and 14.35 nmol l-1 for decreased sexual desire. The calculated free testosterone (CFT) thresholds for these three sexual symptoms were 281.14, 264.90, and 287.21 pmol l-1 , respectively. TT <13.21 nmol l-1 (OR = 1.4, 95%CI: 1.0-1.9, P = 0.037) or CFT <268.89 pmol l-1 (OR = 1.5, 95%CI: 1.1-20, P = 0.020) was associated with an increase in the aforementioned three sexual symptoms. The prevalence of LOH was 9.1% under the criteria, including all three sexual symptoms with TT levels <13.21 nmol l-1 and CFT levels <268.89 pmol l-1 . Our results may improve the diagnostic accuracy of LOH in older men.


Autoimmunity/inflammation in a monogenic primary immunodeficiency cohort.
Primary immunodeficiencies (PIDs) are rare inborn errors of immunity that have a heterogeneous phenotype that can include severe susceptibility to life-threatening infections from multiple pathogens, unique sensitivity to a single pathogen, autoimmune/inflammatory (AI/I) disease, allergies and/or malignancy. We present a diverse cohort of monogenic PID patients with and without AI/I diseases who underwent clinical, genetic and immunological phenotyping. Novel pathogenic variants were identified in IKBKG, CTLA4, NFKB1, GATA2, CD40LG and TAZ as well as previously reported pathogenic variants in STAT3, PIK3CD, STAT1, NFKB2 and STXBP2. AI/I manifestations were frequently encountered in PIDs, including at presentation. Autoimmunity/inflammation was multisystem in those effected, and regulatory T cell (Treg) percentages were significantly decreased compared with those without AI/I manifestations. Prednisolone was used as the first-line immunosuppressive agent in all cases, however steroid monotherapy failed long-term control of autoimmunity/inflammation in the majority of cases and additional immunosuppression was required. Patients with multisystem autoimmunity/inflammation should be investigated for an underlying PID, and in those with PID early assessment of Tregs may help to assess the risk of autoimmunity/inflammation.


miR169 and PmRGL2 synergistically regulate the NF-Y complex to activate dormancy release in Japanese apricot (Prunus mume Sieb. et Zucc.).
This study is the first to demonstrate that GA4-induced dormancy release is associated with the NF-Y complex, which interacts with gibberellin inhibitor RGL2 in Japanese apricot. Seasonal dormancy is not only vital for the survival in cold winter but also affects flowering of temperate fruit trees and the dormancy release depends on the accumulation of the cold temperatures (Chilling requirement-CR). To understand the mechanism of dormancy release in deciduous fruit crops, we compared miRNA sequencing data during the transition stage from paradormancy to dormancy release in the Japanese apricot and found that the miR169 family showed significant differentially up-regulated expression during dormancy induction and was down-regulated during the dormancy release periods. The 5' RACE assay and RT-qPCR validated its target gene NUCLEAR FACTOR-Y subunit A (NF-YA), which exhibited the opposite expression pattern. Further study showed that exogenous GA4 could inhibit the expression of the gibberellic acid (GA) signal transduction suppressor PmRGL2 (RGA-LIKE 2) and promote the expression of NF-Y. Moreover, the interaction between the NF-Y family and GA inhibitor PmRGL2 was verified by the yeast-two-hybrid (Y2H) system and a bimolecular fluorescence complementarity (BiFC) experiment. These results suggest that synergistic regulation of the NF-Y and PmRGL2 complex leads to the activation of dormancy release induced by GA4. These findings will help to elucidate the functional and regulatory roles of miR169 and NF-Y complex in seasonal bud dormancy induced by GA in Japanese apricot and provide new insights for the discovery of dormancy release mechanisms in woody plants.


Impaired spatial coding within objects but not between objects in prosopagnosia.
Patients with prosopagnosia from occipitotemporal lesions have impaired perception of the configuration of facial features. This may be an example of impaired "within-object" spatial coding, which others propose to be distinct from "between-object" spatial coding. To determine whether the prosopagnosic deficit in perceiving spatial configuration was specific to within-face and not between-face spatial coding and whether the deficit was face-selective or extended to objects other than faces. Six prosopagnosic patients were tested using an oddity paradigm in which they detected which of three simultaneously seen stimuli was an altered target. In the "within-face" task, the target face had altered interocular distance or mouth position. In the "between-face" task, the target face was located farther away from the other two. In the "within-object" task, the stimulus was a two-dot pattern, and the target pattern had altered interdot distance. Spatial judgments were impaired within faces for all six patients and within the two-dot pattern for five of six patients. However, all six had normal between-face spatial perception. Impaired perception of spatial relations in prosopagnosia is selective to the spatial structure within individual objects and spares the perception of spatial location of objects. It is not specific to faces. It reveals a process involved in analyzing object structure, consistent with the patients' deficits in recognizing facial identity, and illustrates a different type of "visuospatial" defect.


Changes in Corneal Morphology with Age in Asian Population: A Multicenter Study of 30,618 Cases.
To evaluate normal reference ranges for corneal morphological parameters and investigate age-related changes in these parameters in Asian subjects with healthy eyes in order to provide reference data for preoperative evaluation of corneal refractive surgery and the early differential diagnosis of subclinical and asymptomatic keratoconus. This cross-sectional, multicenter, observational study was conducted in five provinces of China, from January 2014 through October 2019. It is a retrospective analysis. Examiner-blinded clinical measurements were performed after stratification of the subjects into the following age groups: < 18, 18-30, 31-40, 41-50. We evaluated 30,618 healthy eyes of Chinese subjects who exhibited a normal corneal morphology, had no history of eye surgery or trauma, stopped wearing soft contact lenses for at least 2 weeks (rigid contact lenses for at least 4 weeks), and underwent topographic studies for both eyes on the same day. While the anterior and posterior corneal curvatures (K1 and K2) increased with age, corneal astigmatism of the anterior and posterior surfaces (ΔK) and central, minimum, and overall corneal thicknesses decreased with age. Age-related decrease of the overall corneal thickness was more obvious toward the periphery. The anterior and posterior corneal surface heights exhibited a decrease and an increase, respectively. Both index of height asymmetry (IHA) and index of vertical asymmetry (IVA) exhibited an increase with age. The cornea exhibits overall thinning with age and gradually changes from a flat ellipse to an elongated ellipse in Asian individuals with healthy eyes. However, the anterior and posterior surfaces become smoother with age. Owing to the very large number of cases, these small differences are statistically significant. The results obtained are consistent with the hypothesis that a normal cornea seems to withstand quite well the effect of IOP, external pressures, and the natural cross-linking.


FloodIMG: Flood image DataBase system.
A breakthrough in building models for image processing came with the discovery that a convolutional neural network (CNN) can progressively extract higher-level representations of the image content. Having high-resolution images to train CNN models is a key for optimizing the performance of image segmentation models. This paper presents a new dataset-called Flood Image (FloodIMG) database system-that was developed for flood related image processing and segmentation. We developed various Internet of Things Application Programming Interfaces (IoT API) to gather flood-related images from Twitter, and US federal agencies' web servers, such as the US Geological Survey (USGS) and the Department of Transportation (DOT). Overall, >9200 images of flooding events were collected, preprocessed, and formatted to make the dataset applicable for CNN training. Bounding boxes and polygon primitives were also labeled on each image to localize and classify an object in the image. Two use cases of FloodIMG are presented in this paper, where the Fast Region-based CNN (R-CNN) algorithm was used to estimate flood severity and depth during recent flooding events in the US. As of >9200 images, 7,400 were categorized as training sets, whereas >1,800 images were used for the R-CNN testing. Users can access the FloodIMG database freely through Kaggle platform to create more accessible, accurate, and optimized image segmentation models. The FloodIMG workflow concludes with a visualization of colors and labels per image that can serve as a benchmark for flood image processing and segmentation.


[Pathogenesis and treatment of presbyacusis. Current status and future perspectives].
Factors responsible for presbyacusis include physiological ageing processes as well as endogenous or exogenous causes. In the industrial countries, two main exogenous causes are exposure to loud noise and obesity. Pathomechanisms contributing to presbyacusis are hypoxia/ischemia, reactive species formation and oxidative stress, apoptotic and necrotic death of hair cells and spiral ganglion cells as well as inherited and acquired mutations in the mitochondrial DNA. Important for the successful treatment of presbyacusis is a timely fitting of hearing aids on both ears to improve communication and provide the auditory system with acoustic information. Using the hearing aids will also elevate the detection threshold of an existing tinnitus signal. At present, several therapeutic strategies based on pharmacological intervention are under discussion. The application of antioxidants or caloric restriction are considered to prevent or reduce oxidative stress-induced damage. Animal experiments evidenced that superoxide dismutase 2 (SOD2) strongly decreases in age; thus, a further approach may be the overexpression or modulation of the SOD2 within the cochlea. Adenoviral-mediated gene transfer technology would be a tempting approach to address this type of therapy. Finally, hair cell regeneration could be a possible treatment of presbyacusis in the future.


Stability of influenza sub-unit vaccine. Does a couple of days outside the refrigerator matter?
In this study 27 full scale production batches of influenza sub-unit vaccine were evaluated on their stability. The batches varied with respect to the strains they contained and regarding the presence of the preservative thiomersal in the solution. The stability study showed that haemagglutinin content was the most sensitive parameter. An increase in the storage temperature strongly increased the degradation rate of haemagglutinin. The degradation rate of the haemagglutinin differed for the different strains tested. However, statistical evaluation of the data obtained for the most sensitive strain tested showed that even exposure during a 2 week period of the vaccine to room temperature would not adversely affect the shelf life claim of the influenza subunit vaccine of 1 year in the refrigerator. Moreover, this study showed that the presence of thiomersal in the solution has no effect on the stability of the vaccine.


Novel applications of histopathological markers to distinguish prognostic subgroups in colorectal adenocarcinoma.
To explore the novel applications of histological factors by stratifying the prognostic markers of the overall CRC patients in subgroups. A total of 17 histopathological and molecular factors were retrospectively collected and systematically analyzed for the prediction of CRC prognosis in the overall and stratified subgroups by using the Kaplan-Meier curve analysis as well as the Cox regression test. The χ2 test was used to analyze the correlation of the prognostic markers with other factors. The histopathological markers including the lymph node metastasis (LNM), perineural/venous invasion (PVI), TNM stage, the local recurrence or distant metastasis after surgery (R/M) and the molecular markers Ki-67 expression as well as KRAS mutation were identified to be the independent prognostic biomarkers in the overall CRC. The differential prognosis of LNM was found to be significant in age, tumor site, histological classification (histo_classification), cell differentiation, and KRAS/NRAS/BRAF (KNB) mutation stratified subgroups. The PVI was discovered to differently predict survival for patients in age, histo_classification, differentiation, and R/M stratified subgroups. Same as LNM and PVI, TNM was also found to demonstrate differential prognosis in age, tumor site, histo_classification, differentiation, R/M status and KRAS/KNB mutation stratified subgroups. More importantly, R/M was firstly identified not to be terrible for patients in age, histo_classification, LNM, TNM, Ki-67, and KRAS/KNB stratified subgroups. Besides, KRAS mutation was innovatively found to show differential prognosis in age, differentiation, and LNM stratified subgroups. The stratification analyses of prognostic markers in CRC patients indicate novel applications of the above histopathological and molecular markers in clinic and the findings provide new insights into future investigations of precision pathology.


Elevated blood eosinophil count is a valuable biomarker for predicting late acute cellular rejection after liver transplantation.
Recent studies have indicated the value of increased blood eosinophil counts for the diagnosis of acute cellular rejection (ACR) after orthotopic liver transplantation (OLT). However, the relationship between eosinophil count and late ACR at more than 6 months after OLT is still unclear. We sought to retrospectively analyzed the ACR predictive value of eosinophil counts. In the day before or the day of biopsy among 40 biopsies performed on 37 patients beyond 6 months after OLT. Relative eosinophil count was significantly higher in the ACR (n = 24) than the non-ACR cohort, albeit with no significant difference in absolute eosinophil count. Receiver operating characteristic (ROC) analysis showed an absolute eosinophil count of 0.145 × 10(9)/L and a relative eosinophil count of 2.3% to show the highest Youden index with area under the ROC curves of 0.746 and 0.813, respectively. When absolute eosinophil count ≥ 0.145 × 10(9)/L or relative eosinophil count ≥ 2.3% was defined to be elevated, the sensitivity and specificity to predict ACR were 45.8% and 87.5%, and 75% and 87.5%, respectively. When the absolute eosinophil count ≥ 0.285 × 10(9)/L or relative eosinophil count ≥ 3% was defined as elevated, the sensitivity and specificity were 25% and 100%, and 50% and 100%, respectively. All patients with an absolute eosinophil count ≥ 0.285 × 10(9)/L showed a relative eosinophil count ≥ 3%. Elevated blood eosinophil count was a valuable biomarker to predict late ACR after OLT.


Metal mobilization from metallurgical wastes by soil organic acids.
Three types of Cu-slags differing in chemical and mineralogical composition (historical, shaft furnace, and granulated slags) and a matte from a lead recovery process were studied with respect to their susceptibility to release Cu, Zn and Pb upon exposure to organic acids commonly encountered in soil environments. Leaching experiments (24-960 h) were conducted with: i) humic acid (20 mg/L) at pH t0 = 4.4, ii) fulvic acid (20 mg/L) at pH t0 = 4.4, iii) an artificial root exudates (ARE) (17.4 g/L) solution at pH t0 = 4.4, iv) ARE solution at pH t0 = 2.9 and v) ultrapure water (pH t0 = 5.6). The results demonstrated that the ARE contribute the most to the mobilization of metals from all the wastes analyzed, regardless of the initial pH of the solution. For example, up to 14%, 30%, 24% and 5% of Cu is released within 960 h from historical, shaft furnace, granulated slags and lead matte, respectively, when exposed to the artificial root exudates solution (pH 2.9). Humic and fulvic acids were found to have a higher impact on granulated and shaft furnace slags as compared to the ultrapure water control and increased the release of metals by a factor up to 37.5 (Pb) and 20.5 (Cu) for granulated and shaft furnace slags, respectively. Humic and fulvic acids amplified the mobilization of metals by a maximal factor of 13.6 (Pb) and 12.1 (Pb) for historical slag and lead matte, respectively. The studied organic compounds contributed to different release rates of metallic contaminants from individual metallurgical wastes under the conditions tested.


Executive dysfunction in school-age children with ADHD.
The study examined executive function deficits (EFD) in school-age children (7 to 14 years) with ADHD. A clinical sample of children diagnosed with ADHD (n = 49) was compared to a population sample (n = 196) on eight executive function (EF) measures. Then, the prevalence of EFD in clinical and non-clinical children was examined at the individual level according to three methods previously applied to define EFD, and a fourth method was included to control for the effect of age on performance. Children with ADHD were significantly more impaired on measures of EF than children without ADHD at the group level. However, only about 50% of children with ADHD were found to have EFD at the individual level, and results appeared relatively robust across methods applied to define EFD. As a group, children with ADHD displayed more problems on neuropsychological measures of EF than non-clinical children; at the individual level, there appeared to be heterogeneity in EF impairment.


Simultaneous Modeling of Biomarker and Toxicity Response Predicted Optimal Regimen of Guadecitabine (SGI-110) in Myeloid Malignancies.
Guadecitabine (SGI-110) is a novel next-generation hypomethylating agent (HMA) administered as s.c. injection with extended decitabine exposure. Dose/exposure-response analyses of longitudinal measures of long interspersed nucleotide element-1 (LINE-1) methylation and absolute neutrophil counts (ANC) pooled from 79 and 369 patients in 2 phase I/II trials, respectively, were performed to assist, through modeling and simulation, the selection of dosing regimens for phase III. Simulation of ANC predicted a decrease after a 5-day regimen of 60 mg/m2 with partial recovery before the next cycle, whereas the nadir of 90 mg/m2 on the same schedule was below 100/µl. ANC following a 60 mg/m2 10-day regimen was predicted to be suppressed below 100/µl as long as treatment continued without recovery. The developed models provided useful tools to assist simultaneous evaluation of the relative dynamics of the two effects (DNA demethylation and the effect on ANC).


Refractive outcomes of combined phacovitrectomy and delayed cataract surgery in retinal detachment.
To compare the accuracy of refractive outcomes between combined pars plana vitrectomy (PPV) and cataract surgery and delayed cataract surgery after PPV in cases with rhegmatogenous retinal detachment (RD). Retrospective case series. Thirty-eight eyes underwent combined phacovitrectomy (combined group) and 25 eyes underwent delayed cataract surgery after PPV (delayed group). RD height was measured using optical coherence tomography. Refractive outcomes were evaluated using mean absolute error (MAE; the difference between final refractive error and target refractive error). Combined group showed significant myopic shift (mean error; -0.40 ± 1.07 vs 0.07 ± 0.56 D, p = 0.028) and large MAE (0.81 ± 0.81 vs 0.48 ± 0.29 D, p = 0.028) compared with delayed group. Multiple logistic regression analysis revealed that only RD height was significantly associated with MAE greater than 2 D after combined surgery (in 100-µm unit, odds ratio 3.23, 95% CI 1.04-10.02, p = 0.042). RD height was also significantly correlated with the difference in axial length (AL) between 2 eyes of the patients (p = 0.006, r = 0.406) and the difference in AL measured at pre- versus post-RD repair in the delayed group (p < 0.001, r = 0.774). Combined phacovitrectomy in patients with rhegmatogenous RD induced significant myopic shift because of underestimation of AL, especially in patients with high RD height. Thus, in cases with high temporal RD or large AL differences between eyes, either delayed cataract surgery or combined cataract surgery using the contralateral AL is recommended.


Inhibition of c-Src/p38 MAPK pathway ameliorates renal tubular epithelial cells apoptosis in db/db mice.
Renal tubular epithelial cells (RTEC) apoptosis, which plays a key role in the pathogenesis and progression of diabetic nephropathy (DN), is believed to be contributive to the hyperglycemia-induced kidney failure, though the exact mechanisms remain elusive. In this study, we investigated how inhibition of c-Src/p38 MAPK pathway would affect RTEC apoptosis. The c-Src inhibitor PP2 i.p. administered every other day for 8 weeks to diabetic db/db mice significantly reduced their kidney weights, daily urinary volumes, blood glucose, blood urea nitrogen, serum creatinine, triglyceride and urine albumin excretion, whereas deactivation of c-Src and p38 MAPK were also observed, along with decreases in both Bax/Bcl-2 ratio and cleaved caspase-3 level in the kidneys. In vitro, exposure of HK-2 cells (a human RTEC line), to high glucose (HG) promoted phosphorylation of c-Src and p38 MAPK, and subsequently, as revealed by western blotting, TUNEL assay and flow cytometry, increased cell death, which can be inhibited by PP2. Especially, a specific p38 MAPK inhibitor, SB203580, that both attenuated HG-induced c-Src activation and abrogated the expression of PPARγ and CHOP, also reduced apoptosis. Taken together, PP2 inhibits c-Src and therefore reduces apoptosis in RTEC, which at least in part, is due to suppressed p38 MAPK activation in diabetic kidney.


Transient Electrical Currents Mediated by the Na+/K+-ATPase: A Tour from Basic Biophysics to Human Diseases.
The Na+/K+-ATPase is a chemical molecular machine responsible for the movement of Na+ and K+ ions across the cell membrane. These ions are moved against their electrochemical gradients, so the protein uses the free energy of ATP hydrolysis to transport them. In fact, the Na+/K+-ATPase is the single largest consumer of energy in most cells. In each pump cycle, the protein sequentially exports 3Na+ out of the cell, then imports 2K+ into the cell at an approximate rate of 200 cycles/s. In each half cycle of the transport process, there is a state in which ions are stably trapped within the permeation pathway of the protein by internal and external gates in their closed states. These gates are required to open alternately; otherwise, passive ion diffusion would be a wasteful end of the cell's energy. Once one of these gates open, ions diffuse from their binding sites to the accessible milieu, which involves moving through part of the electrical field across the membrane. Consequently, ions generate transient electrical currents first discovered more than 30 years ago. They have been studied in a variety of preparations, including native and heterologous expression systems. Here, we review three decades' worth of work using these transient electrical signals to understand the kinetic transitions of the movement of Na+ and K+ ions through the Na+/K+-ATPase and propose the significance that this work might have to the understanding of the dysfunction of human pump orthologs responsible for some newly discovered neurological pathologies.


Nurse practitioner led pain management the day after caesarean section: A randomised controlled trial and follow-up study.
Pain on the day after caesarean section is often treated with controlled-release oxycodone to supplement the decline in analgesia from intrathecal opioids. Evidence suggests that caesarean birth is a biopsychosocial experience where a comprehensive approach is needed that promotes control and participation in pain management. This study compared immediate-release oxycodone integrated with supportive educational strategies to controlled-release oxycodone. A follow-up phase aimed to explore pain over three months. This study was a two-group parallel randomised controlled trial. A metropolitan hospital in Australia with a birthing suite, operating rooms, and a postnatal unit. English-speaking women scheduled for elective caesarean section were mailed trial information. Exclusion criteria included contraindications to intrathecal analgesia, herpes simplex infection, a history of chronic pain, opioid tolerance, or substance abuse. A total of 131 participants were recruited and randomised out of 298 eligible participants. Group allocation was undertaken using sequentially numbered opaque sealed envelopes. The nurse practitioner intervention commenced on the day after surgery with immediate-release oxycodone alongside supportive strategies. The control group received scheduled doses of controlled-release oxycodone. All participants could request additional oxycodone or tramadol. Primary outcomes were pain intensity and secondary outcomes included patient global impression of change, pain interference, opioid consumption, and maternal perception of control. A follow-up phase evaluated pain outcomes over three months. The final sample size was 122, with 61 participants in each group. Pain intensity scores were analysed by linear mixed regression models. There were no statistical differences over 24h between the control and intervention groups at rest (p=0.40, 95% CI - 4.8mm, 11.9mm) or on sitting or moving (p=0.561, 95% CI -15.2mm, 8.3mm). Patient global impression of change was significant over three hours (p=0.014, OR=2.5, 95% CI 1.2, 5.3). The intervention group reported less pain interference while consuming less oxycodone (p<0.05). There was no difference between groups in terms of perceived control over pain management (p=0.273, 95% CI -16.2mm, 4.6mm). The follow-up analysis graded 5.9% of participants as experiencing severe pain interference. Chronic pain following caesarean was associated with postnatal depression (p<0.001). The research showed that a nurse practitioner intervention can improve pain management following caesarean section. The results underscore the influence of biological, psychological, and social factors on acute pain. Hence, this study reinforces the need for a biopsychosocial approach to acute pain management following caesarean delivery.


Copper toxicity in the marine copepod Tigropus japonicus: low variability and high reproducibility of repeated acute and life-cycle tests.
The intertidal copeopod Tigriopus japonicus, which is abundant and widely distributed along the coasts of Western Pacific, has been suggested to be a good marine ecotoxicity testing organism. In this study, a series of experiments were conducted to investigate the reproducibility and variability of copper (Cu) sensitivity of T. japonicus so as to evaluate its potential to serve as an appropriate test species. To understand the seasonal variation of Cu sensitivity, individuals of T. japonicus were collected from the field in summer and winter, and subjected to standard 96 h acute (static renewal) toxicity tests. 96 h-LC50 values of T. japonicus collected from the two seasons were marginally different (p = 0.05), with an overall coefficient of variation (CV) of 33%. Most importantly, our results indicated that chronic Cu sensitivity of T. japonicus was highly reproducible. The CVs of intrinsic rates of increase in the population of the control and Cu treatment (10 microg Cu l(-1)) groups were only 10-11% between 10 runs of a standardised complete life-cycle test. Moreover, different Cu(II) salts generally resulted in a similar 96 h-LC50 value while Cu(I) chloride was consistently slightly less toxic than Cu(II) salts. Given such a high reproducibility of toxic responses, it is advocated to use T. japonicus as a routine testing organism.


Comparison of quasi-Rayleigh waves and Rayleigh waves, and clarifying the cut-off frequency of quasi-Rayleigh waves.
The Partial Wave Method is unique in that it establishes a foundation on which various elastodynamic guided waves can be compared. In this paper, the method is used to compare quasi-Rayleigh waves and Rayleigh waves, and investigate the eccentricities of the Partial Wave Method at phase velocities equal to the Rayleigh wave speed. The comparison results in the definition of two types of quasi-Rayleigh waves and an explanation for quasi-Rayleigh wave behavior reported in the literature at frequencies that do not satisfy Viktorov's quasi-Rayleigh wave condition. These conclusions are also verified by the superposition of A0 and S0 mode wave-structures calculated using the semi-analytical finite element method.


N-terminal sequence tagging using reliably determined b2 ions: a useful approach to deconvolute tandem mass spectra of co-fragmented peptides in proteomics.
With the recent introduction of higher-energy collisional dissociation (HCD) in Orbitrap mass spectrometry, the popularity of that technique has grown tremendously in the proteomics society. HCD spectra, however, are characterized by a limited distribution of bn-type ions, which permit the generation of reliable sequence tags based on complementary b,y pairs both for de novo sequencing and sequence tagging strategies. Instead, most peptide HCD spectra (~95%) are dominated with b2 ions. In this work, we analyzed positive predictive values of b2 ions in HCD, and found that b2 ions can be determined with >97% certainty in the presence of a2 and its complementary yn-2 ions. Analytically, b2 ions provide information on the composition of the first two N-terminal amino acids in peptides. Their utilization in N-terminal sequence tagging leads to a significant decrease in false discovery rate by filtering out false positives while retaining true positive identifications. As a consequence, the number of peptide spectrum matches (PSMs) increased by 4.8% at fixed FDR (1%). This approach allows for deconvolution of mixture spectra and increased the number of PSM to 9.2% in a complex human sample and to 24% in a complex sample of synthetic peptides at 1% FDR.


Nitrate-dependent control of root architecture and N nutrition are altered by a plant growth-promoting Phyllobacterium sp.
Both root architecture and plant N nutrition are altered by inoculation with the plant growth-promoting rhizobacteria (PGPR) Phyllobacterium strain STM196. It is known that NO3- and N metabolites can act as regulatory signals on root development and N transporters. In this study, we investigate the possible interrelated effects on root development and N transport. We show that the inhibition of Arabidopsis lateral root growth by high external NO3- is overridden by Phyllobacterium inoculation. However, the leaf NO3- pool remained unchanged in inoculated plants. By contrast, the Gln root pool was reduced in inoculated plants. Unexpectedly, NO3- influx and the expression levels of AtNRT1.1 and AtNRT2.1 genes coding for root NO3- transporters were also decreased after 8 days of Phyllobacterium inoculation. Although the mechanisms by which PGPR exert their positive effects remain unknown, our data show that they can optimize plant development independently from N supply, thus alleviating the regulatory mechanisms that operate in axenic conditions. In addition, we found that Phyllobacterium sp. elicited a very strong induction of AtNRT2.5 and AtNRT2.6, both genes preferentially expressed in the shoots whose functions are unknown.


Lung cancer and rehabilitation--what are the barriers? Results of a questionnaire survey and the development of regional lung cancer rehabilitation standards and guidelines.
Evidence supports the role of rehabilitation in the management of lung cancer symptoms. Previous research reports that rehabilitation needs are inadequately recognised and managed, which may adversely affect patients' quality-of-life and create burden for caregivers. This study aims to explore the perceptions of palliative care and respiratory multidisciplinary team (MDT) members about the role of rehabilitation for lung cancer patients, examine patterns of referral for lung cancer patients to rehabilitation services, and highlight the barriers which prevent the referral of lung cancer patients to rehabilitation services. Questionnaires were completed by MDT members within a regional cancer network during June 2010. Fifty-nine healthcare professionals participated. Ninety-four per cent of respondents perceived their patients had rehabilitation needs. Referral most commonly occurred during the palliative (29.6 %) and post-treatment (23.7 %) disease phases. Barriers to referral included "lack of knowledge of services or referral mechanisms" (28.8 %), "waiting lists" (28.8 %) and the perception that patients "do not desire rehabilitation" (22 %). Rehabilitation needs were most frequently discussed at palliative in-patient MDT meetings [reported as "often" by 37 (62.7 %) respondents] and least discussed at lung MDTs (half of respondents reporting that rehabilitation was "never" discussed). Rehabilitation services were considered adequate by 39 % of respondents. Long waiting times and lack of knowledge of services are among several factors that may prevent lung cancer patients being offered rehabilitation. In order to improve the quality of care delivered to cancer patients, it is important to remove barriers that affect delivery of rehabilitation services.


Haddad syndrome presenting with abdominal distension associated with long-segment aganglionosis.
Haddad syndrome is a form of neurocristopathy characterized by a combination of congenital central hypoventilation syndrome and Hirschsprung disease (HD). Although Haddad syndrome is extremely rare, awareness of the combination of long-segment HD in Haddad syndrome as well as radiographic manifestations may help to ensure a timely diagnosis as well as to facilitate optimal treatment of this unusual condition. We report a case of Haddad syndrome with long-segment, intestinal aganglionosis in a newborn infant. This report emphasizes the features of HD in children with Haddad syndrome and suggests that specific attention be given to its interpretation on plain radiographs.


The contribution of inflammatory astrocytes to BBB impairments in a brain-chip model of Parkinson's disease.
Astrocyte dysfunction has previously been linked to multiple neurodegenerative disorders including Parkinson's disease (PD). Among their many roles, astrocytes are mediators of the brain immune response, and astrocyte reactivity is a pathological feature of PD. They are also involved in the formation and maintenance of the blood-brain barrier (BBB), but barrier integrity is compromised in people with PD. This study focuses on an unexplored area of PD pathogenesis by characterizing the interplay between astrocytes, inflammation and BBB integrity, and by combining patient-derived induced pluripotent stem cells with microfluidic technologies to generate a 3D human BBB chip. Here we report that astrocytes derived from female donors harboring the PD-related LRRK2 G2019S mutation are pro-inflammatory and fail to support the formation of a functional capillary in vitro. We show that inhibition of MEK1/2 signaling attenuates the inflammatory profile of mutant astrocytes and rescues BBB formation, providing insights into mechanisms regulating barrier integrity in PD. Lastly, we confirm that vascular changes are also observed in the human postmortem substantia nigra of both males and females with PD.


Catalytic and Medical Potential of a Phyto-Functionalized Reduced Graphene Oxide-Gold Nanocomposite Using Willow-Leaved Knotgrass.
In the current study, a simple, environmentally friendly, and cost-effective reduced graphene oxide-gold nanoparticle (rGO-AuNP) nanocomposite was successfully phytosynthesized using the aqueous leaf extract of a common weed found on the Nile banks, Persicaria salicifolia, for the first time. The phytosynthesis of rGO-AuNPs was first confirmed via the color transformation from brown to black as well as throughvarious techniques such as transmission electron microscopy (TEM) and Raman spectroscopy. Two UV-vis peaks at 275 and 530 nm were observed for the nanocomposite with a typical particle size of mostly spherical AuNPs of 15-20 nm. However, other shapes were occasionally detected including rods, triangles, and rhomboids. Existing phytoconstituents such as flavonoids and glycosides in the plant extract were suggested to be responsible for the phytosynthesis of rGO-AuNPs. The excellent catalytic efficacy of rGO-AuNPs against MB degradation was confirmed, and a high antibacterial efficiency against Escherichia coli and Klebsiella pneumonia was also confirmed. Promising antioxidant performance of rGO-AuNPs was also proved. Furthermore, it was concluded that rGO-AuNPs acquired higher efficiency than AuNPs synthesized from the same plant extract in all of the studied applications.


Direct observation of single molecule conformational change of tight-turn paperclip DNA triplex in solution.
DNA triplex modulates gene expression by forming stable conformation in physiological condition. However, it is not feasible to observe this unique molecular structure of large molecule with 54 oligodeoxynucleotides directly by conventional nuclear magnetic approach. In this study, we observed directly single molecular images of paperclip DNA triplexes formation in a buffer solution of pH 6.0 by atomic force microscopy (AFM). Meanwhile, a diffuse "tail" of unwound DNA was observed in pH 8.0 solution. This designable approach in visualizing the overall structures and shapes of oligo-DNAs at the single molecular level, by AFM, is applicable to other biopolymers as well.


Risk stratification of CAD with SPECT-MPI in women with known estrogen status.
To compare exercise tolerance testing (ETT) with gated single photon emission computed tomography-myocardial perfusion imaging (SPECT-MPI) risk stratification in women with an intermediate to high CAD pretest risk and known estrogen status (ES). SPECT-MPI is an effective test for risk stratifying patients with stable angina. However in women, the current guidelines recommend the exercise tolerance testing (ETT) as first line test. Further, the relationship of stress imaging to ES, an independent risk indicator for CAD, is unknown. 2,194 women with an intermediate to high CAD pre-test risk were referred for a clinically indicated ETT with gated SPECT-MPI. Duke treadmill scores (DTS) and summed stress score (SSS) were calculated. SSS were classified as normal (SSS < 3), mildly abnormal (SSS 4-8), or moderate-severely abnormal (SSS > 8). The ES was assessed as premenopausal, postmenopausal on hormone replacement therapy (HRT) as ES+ while postmenopausal not on HRT were ES-. An annualized cardiac event rate of a composite of cardiac death, unstable angina (UA) leading to hospitalization, non-fatal myocardial infarction, or late coronary revascularization was calculated for all the groups. The annualized cardiac event rate was 1.3% PPY, 2.1% PPY, and 3.2% PPY for low, intermediate, and high risk DTS (P = .2). Patients with intermediate DTS and mildly abnormal or moderate-severely abnormal gated SPECT-MPI had a significantly higher cardiac event rates (5.3% PPY and 10.8% PPY, respectively) than those with a normal gated SPECT-MPI (1.2%, PPY, P = .01). This was also demonstrated on further Cox-regression analysis. Risk stratification of SPECT-MPI over DTS was independent of ES. Gated SPECT-MPI provides risk stratification beyond standard exercise stress testing for women with suspected coronary artery disease, especially in patients with intermediate DTS and is independent of ES.


Clinicopathologic and Survival After Cystectomy Outcomes in Squamous Cell Carcinoma of the Bladder.
Squamous cell carcinoma of the bladder (SqCC) is a rare disease with limited management data. Thus, we sought to characterize the clinicopathologic and survival outcomes amongst patients with SqCC and explore the association of squamous differentiation within urothelial carcinoma (UC w/Squam), as compared to muscle invasive pure UC. We conducted a single-center retrospective cohort study of patients, stratified by histology, who underwent cystectomy for MIBC. Baseline clinicopathologic characteristics were compared, and overall survival was assessed using Kaplan-Meier method. We identified 1,034 patients; 37 (3.58%) with SqCC histology, 908 (87.81%) with UC histology, and 89 (8.61%) with UC w/ Squam histology. Among SqCC patients, a higher proportion were Black and similarly a higher proportion were women; amongst patients with UC w/ Squam a higher proportion had lower BMI; and amongst patients with UC a higher proportion had lower clinical (c) T, cN, pathological (p) T, and pN stages. Patients presenting with UC were more likely to receive intravesical therapy; patients presenting with SqCC were less likely to receive neoadjuvant chemotherapy (NAC). Adjuvant chemotherapy rates were similar. With post-hoc Bonferroni analysis, overall survival, cancer-specific survival, and recurrence-free survival were significantly worse for the UC w/ Squam cohort. UC w/ Squam histology was associated with worse survival outcomes after cystectomy for muscle invasive bladder cancer compared to UC. Our results suggest that UC w/ Squam is associated with more advanced disease compared to UC, warranting further prospective work on consideration of combination therapies for patients with this disease state.