Current risk factors and outcomes for the arterial switch operation.
The arterial switch operation is the preferred treatment for dextrotransposition of the great arteries and some forms of double-outlet right ventricle. All patients undergoing an arterial switch operation at a single institution from January 1, 1999, to September 1, 2005, were reviewed. Of the 168 patients, median age was 2 days (range, 0 to 358) and weight was 3.5 kg (range, 1.9 to 11.8 kg). Eleven patients were less than 36 weeks gestational age. Forty percent had coronary patterns other than usual. Mean cardiopulmonary bypass (CPB) time was 147 +/- 45 minutes, and mean cross-clamp time was 77 +/- 27 minutes. At a mean follow-up of 19 +/- 21 months, there were 10 (6%) hospital and 4 (3%) late deaths. Actuarial 1-month, 1-year, and 3-year survivals were 94%, 90%, and 89%, respectively. Bivariate analysis revealed weight less than 2.5 kg (p = 0.032), gestational age less than 36 weeks (p = 0.002), and CPB time greater than 150 minutes (p = 0.0075) decreased hospital survival. Intermediate-term survival was negatively impacted by weight less than 2.5 kg (p = 0.017), gestational age less than 36 weeks (p = 0.0096), CPB time greater than 150 minutes (p = 0.0050), and age at presentation greater than 4 weeks (p = 0.034). By multivariate analysis, gestational age less than 36 weeks (p = 0.0051) and CPB time greater than 150 minutes (p = 0.016) were independent risk factors for hospital mortality. Gestational age less than 36 weeks (p = 0.0096) and CPB time greater than 150 minutes (p = 0.005) were also independent predictors of intermediate-term mortality. Coronary anatomy could not be shown to affect survival, including no deaths among the 12 patients with intramural coronaries. The arterial switch operation can be performed with low mortality regardless of diagnosis or coronary pattern. The premature patient and minimizing CPB time remain as challenges to optimize outcomes for the arterial switch operation.


Automatized and desktop AC-susceptometer for the in situ and real time monitoring of magnetic nanoparticles' synthesis by coprecipitation.
The main purpose of this work was to design, develop, and construct a simple desktop AC susceptometer to monitor in situ and in real time the coprecipitation synthesis of magnetic nanoparticles. The design incorporates one pair of identical pick-up sensing coils and one pair of Helmholtz coils. The picked up signal is detected by a lock-in SR850 amplifier that measures the in- and out-of-phase signals. The apparatus also includes a stirrer with 45°-angle blades to promote the fast homogenization of the reaction mixture. Our susceptometer has been successfully used to monitor the coprecipitation reaction for the synthesis of iron oxide nanoparticles.


Zinc deficiency in the 11 day rat embryo: a scanning and transmission electron microscope study.
Zinc deficient rat embryos were obtained on the 11th day of pregnancy and examined by scanning and transmission electron microscopy. Scanning electron microscopy revealed an increase in the number of deformed embryos, as well as embryonic growth retardation. In addition, the epithelium of zinc deficient embryos displayed a marked increase in surface microvilli, as well as the presence of blebbing. Transmission electron microscopy indicated extensive cell death in the neural epithelium which was apparently more severely damaged by zinc deficiency than were mesenchymal cells. Mitochondrial cristae were affected to a greater degree than any other membrane of the cell and cristael disintigration appeared to represent the principal cellular lesion preceding necrosis of neural cells and neural tube teratology.


Are β3 -adrenoceptor gene polymorphisms relevant for urology?
β3 -adrenoceptors (ARs) are an important drug target for the treatment of overactive bladder syndrome (OAB) and are under investigation for other indications. The human β3 -AR gene is polymorphic; an exchange of amino acid tryptophan (Trp) for arginine (Arg) in position 64 of the receptor protein is the most frequent and best-studied polymorphism. A narrative review on the impact of β3 -AR polymorphisms on urological disease and its treatment is presented. Two out of four studies have reported that the 64Arg allele was found more frequently in subjects with OAB than in healthy controls. A large study in a highly selective population (men undergoing prostatectomy for cancer treatment) did not confirm this. On the other hand, studies examining symptom severity typically found little difference between 64Arg and 64Trp carriers. In vitro studies with endogenously expressed β3 -AR reported a decreased lipolytic response in human adipose tissue. Studies with heterologously expressed receptors sometimes found a decreased responsiveness to agonists including β3 -AR agonists, but others did not confirm that. The overall evidence points to carriers of the 64Arg genotype expressing fewer and/or hypofunctional β3 -ARs and being associated with the presence of OAB but such findings were only detected inconsistently. If this hypofunctionality exists, the consequences may be of insufficient magnitude to allow a robust detection. Only adequately powered studies comparing responses with a β3 -AR agonist in 64Arg carriers versus wild-type patients can address this.


A general nickel-catalyzed highly regioselective hydroarylation of unactivated alkenes enabled by the picolinamide auxiliary.
A practical method for regioselective hydroarylation of unactivated γ- or δ-vinyl alkylamines has been reported, enabling facile preparation of highly value-added ε- or ζ-aryl alkylamines. The protocol employs nickel catalysis, shows high functional group tolerance and can be used for modifying bio-related molecules.


Gene extraction for cancer diagnosis by support vector machines--an improvement.
To improve the performance of gene extraction for cancer diagnosis by recursive feature elimination with support vector machines (RFE-SVMs): A cancer diagnosis by using the DNA microarray data faces many challenges the most serious one being the presence of thousands of genes and only several dozens (at the best) of patient's samples. Thus, making any kind of classification in high-dimensional spaces from a limited number of data is both an extremely difficult and a prone to an error procedure. The improved RFE-SVMs is introduced and used here for an elimination of less relevant genes and just for a reduction of the overall number of genes used in a medical diagnostic. The paper shows why and how the, usually neglected, penalty parameter C and some standard data preprocessing techniques (normalizing and scaling) influence classification results and the gene selection of RFE-SVMs. The gene selected by RFE-SVMs is compared with eight other gene selection algorithms implemented in the Rankgene software to investigate whether there is any consensus among the algorithms, so the scope of finding the right set of genes can be reduced. The improved RFE-SVMs is applied on the two benchmarking colon and lymphoma cancer data sets with various C parameters and different standard preprocessing techniques. Here, decreasing C leads to the smaller diagnosis error in comparisons to other known methods applied to the benchmarking data sets. With an appropriate parameter C and with a proper preprocessing procedure, the reduction in a diagnosis error is as high as 36%. The results suggest that with a properly chosen parameter C, the extracted genes and the constructed classifier will ensure less overfitting of the training data leading to an increased accuracy in selecting relevant genes. Finally, comparison in gene ranking obtained by different algorithms shows that there is a significant consensus among the various algorithms as to which set of genes is relevant.


Initial fractal exponent of heart rate variability is associated with success of early resuscitation in patients with severe sepsis or septic shock: a prospective cohort study.
Heart rate variability (HRV) reflects autonomic nervous system tone as well as the overall health of the baroreflex system. We hypothesized that loss of complexity in HRV upon intensive care unit (ICU) admission would be associated with unsuccessful early resuscitation of sepsis. We prospectively enrolled patients admitted to ICUs with severe sepsis or septic shock from 2009 to 2011. We studied 30 minutes of electrocardiogram, sampled at 500 Hz, at ICU admission and calculated heart rate complexity via detrended fluctuation analysis. Primary outcome was vasopressor independence at 24 hours after ICU admission. Secondary outcome was 28-day mortality. We studied 48 patients, of whom 60% were vasopressor independent at 24 hours. Five (10%) died within 28 days. The ratio of fractal alpha parameters was associated with both vasopressor independence and 28-day mortality (P = .04) after controlling for mean heart rate. In the optimal model, Sequential Organ Failure Assessment score and the long-term fractal α parameter were associated with vasopressor independence. Loss of complexity in HRV is associated with worse outcome early in severe sepsis and septic shock. Further work should evaluate whether complexity of HRV could guide treatment in sepsis.


Effective extra-cellular recording from vertebrate neurons in culture using a new type of micro-electrode array.
We describe the fabrication and use of a new type of extracellular micro-electrode array mounted on a flexible transparent polyimide substrate that can be rapidly moved from one part of a culture of vertebrate neurons (rat nodose) to another, which permits co-culture of glia under the neurons and is easily and rapidly replaceable in the event of damage. The array can be mounted on a micromanipulator and moved into place whenever and wherever recordings with or without stimulation are needed. The basic electrode system consists of 20-30 microm diameter gold electrodes, with or without platinisation, exposed to the cells through openings in the polyimide and joined to the recording or stimulating circuitry through gold tracks embedded in the polyimide. If rigid control over neuron placement has been achieved the patterns of electrodes can be matched to the neuron positions.


Virus entry: open sesame.
Detailed information about the replication cycle of viruses and their interactions with host organisms is required to develop strategies to stop them. Cell biology studies, live-cell imaging, and systems biology have started to illuminate the multiple and subtly different pathways that animal viruses use to enter host cells. These insights are revolutionizing our understanding of endocytosis and the movement of vesicles within cells. In addition, such insights reveal new targets for attacking viruses before they can usurp the host-cell machinery for replication.


The changing face of informed consent.
All healthcare professionals are required to gain a patient's consent before proceeding with examination, investigation or treatment. Gone are the days when consent was about protecting the professional. Following a recent landmark Supreme Court case, 'informed' consent is now embedded in UK law. Patients have the right to high-quality information that allows them to be involved in making decisions about their care. Dentists have a duty of care to provide this information and guide their patients through the process. This paper reviews key ethical, legal, and professional guidance available to dentists about informed consent and concludes by discussing how shared decision-making is a model of healthcare delivery with much to offer dentist and patient alike.


Steps to a patient satisfaction survey.
Patient satisfaction must be assessed carefully in the current competitive climate. Seven steps are outlined for developing and testing a Patient Satisfaction Survey in a specialty unit of a general hospital.


[Immunologic reaction in parasitic invasion (author's transl)].
Among the manifold immunologic events which take place during parasitic invasions, production of autoantibodies and immune complexes can play a serious role during infections with African and American trypanosomes. The importance of complement deserves new attention. The increasing level of IgE induced by helminthic infections on a humoral basis seems to be caused by separate worm allergens; its involvement in self-cure phenomena together with processes on cellular levels is discussed. Destructive processes on various development stages of schistosoma and immunological events during cutaneous leishmaniasis are also cell mediated. Variations in the antigenic behaviour of parasites and their immunological mimicry by uptake of substances from the host and their immunosuppressive action are discussed. As reasons for absence of immunity against animal parasites this action can disturb either humoral or cellular immunological procedures or both.


Simultaneous radioimmunoassay of serum testosterone and 5 alpha-dihydrotestosterone without chromatography.
When characterization of the specificity of an antiserum for radioimmunoassay (RIA) is performed by the conventional method, the conditions under which interference occurs are not respected because of the lack of specific antigen. We have studied the behavior of antisera reproducing the real environment existing in unknown samples, in which antigen, interferent and tracer complete simultaneously. A testosterone (T) antiserum and a 5 alpha-dihydrotestosterone (D) anti serum were characterized by setting up two distinct hapten recovery tests in the presence of both the hapten and the crossreactant added to steroid-free serum in various concentrations in order to reproduce multiple concentration ratios. These samples, together with the standard curves samples (prepared by 'spiking' steroid-free serum with known concentrations of T or D) were extracted and subjected to T-RIA and D-RIA without purification. The results have shown that the interferent-induced incremental ratio is a linear function of the ratio of the levels of cross-reactant and hapten via a proportionality factor inversely correlated to the antiserum specificity. By means of this function, the overestimated T and D levels found in samples after 'extraction only' have been corrected and the resulting values have shown acceptable correlation with the corresponding levels determined after column chromatography.


[Measuring three-dimensional displacement of tooth-supported rigid fixed bridge under oblique concentrated loading using digital laser speckle photography].
To measure three-dimensional displacements of rigid fixed bridge under oblique concentrated loading and to analyze the effect of displacement on the stability of abutment teeth. Right mandible of Beagle dog was used to establish the tooth-supported rigid fixed bridge. Digital laser speckle photography was employed to measure the three-dimensional displacements of the prosthesis under oblique concentrated loading ranging from 200 g to 3000 g. The displacements of prosthesis increased as the load increased. When the load was on the abutment, the buccolingual displacement of the abutment under loading was the biggest but no more than 100 microm. This was followed by mesiodistal and occlusallingual displacements. With slighter loadings, the buccolingual displacements of non-loaded abutment were greater than the mesiodistal displacements. However, as the loading increased greater mesiodistal displacements on non-loaded abutment were found compared to buccolingual displacements. When the bridge was loaded, the biggest displacements of the prosthesis were always shown on the buccolingual direction, followed by the mesiodistal direction and the occlusallingual direction. The displacements with loaded bridge were smaller than those when the load was applied on abutment. The displacements of rigid fixed bridge change with different loading positions. This has implications on the design of prosthesis. Lateral force should be minimized and early occlusion contact should be avoided, especially on the feeble abutment. This study has developed a new method for measuring bit shift of teeth.


An electroactive biofilm-based biosensor for water safety: Pollutants detection and early-warning.
Besides serving in wastewater treatment and energy generation fields, electroactive biofilm (EAB) has been employed as a sensitive bio-elements in a biosensor to monitor water quality by delivering electrical signals without additional mediators. Increasing studies have applied EAB-based biosensor in specific pollutant detection, typically biochemical oxygen demand (BOD) detection, as well as in early-warning of composite pollutants. Based on a comprehensive review of literatures, this study reveals how EAB outputs electrical signal, how we can evaluate and improve this performance, and what information we can expect from EAB-based biosensor. Since BOD detection and early-warning are normally confusing, this study manages to differentiate these two applications through distinguished purposes and metrics. Based on the introductions of progresses and applications of EAB-based biosensors so far, several novel strategies toward the future development of EAB-based biosensors are proposed.


Human hTM alpha gene: expression in muscle and nonmuscle tissue.
We have isolated a cDNA clone from a human skeletal muscle library which contains the complete protein-coding sequence of a skeletal muscle alpha-tropomyosin. This cDNA sequence defines a fourth human tropomyosin gene, the hTM alpha gene, which is distinct from the hTMnm gene encoding a closely related isoform of skeletal muscle alpha-tropomyosin. In cultured human fibroblasts, the hTM alpha gene encodes both skeletal-muscle- and smooth-muscle-type alpha-tropomyosins by using an alternative mRNA-splicing mechanism.


Contact lenses for visual rehabilitation after corneal laceration repair.
A contact lens fitting was tried after corneal laceration repair in 26 eyes. The mean patient age was 25 years; 22 eyes were aphakic and there were 23 males. Many patients had been referred for penetrating keratoplasty. Intact sutures in 13 eyes did not hinder contact lens fitting which was performed within 6 months in 12 cases. A contact lens was fit and successfully worn in 21 eyes (81%) and was fit in the remaining five patients, but not worn because of suboptimal vision (4 eyes) or an unsatisfactory fit (1 eye). Success rates were higher for small, peripheral lesions and in younger patients. Visual acuity was at least 20/30 in 12 patients and follow-up averaged 10.1 months. A contact lens can usually be fit after corneal lacerations, despite aphakia, with good visual results, thereby avoiding penetrating keratoplasty and intraocular lens implantation.


5-Year Follow-Up From the PARTNER 2 Aortic Valve-in-Valve Registry for Degenerated Aortic Surgical Bioprostheses.
The aim of this study was to report the outcomes of valve-in-valve (ViV) transcatheter aortic valve replacement (TAVR) at 5 years. TAVR for degenerated surgical bioprostheses in patients at high risk for reoperative surgery is an important treatment option that may delay or obviate the need for surgical intervention; however, long-term outcomes of this procedure are unknown. The PARTNER (Placement of Aortic Transcatheter Valves) 2 ViV and continued access registries prospectively enrolled patients with failed surgical bioprostheses at high risk for reoperation. Five-year clinical and echocardiographic follow-up data were obtained in 95.9% of patients. In 365 (96 registry and 269 continued access) patients, the mean age was 78.9 ± 10.2 years, the mean Society of Thoracic Surgeons predicted risk of surgical mortality score was 9.1 ± 4.7%, and New York Heart Association functional class was III or IV in 90.4%. At 5 years, the Kaplan-Meier rates of all-cause mortality and any stroke were 50.6% and 10.5%, respectively. Using Valve Academic Research Consortium 3 definitions, the incidence of structural valve deterioration, related hemodynamic valve deterioration, or bioprosthetic valve failure at 5 years was 6.6%. Aortic valve re-replacement was performed in 6.3% (n = 14), the majority of which was due to stenosis (n = 6) and combined aortic insufficiency/paravalvular regurgitation (n = 3). The mean gradient, Doppler velocity index, paravalvular regurgitation, and quality of life measured by Kansas City Cardiomyopathy Questionnaire scores in survivors remained stable from 30 days postprocedure through 5 years. At the 5-year follow-up, TAVR for bioprosthetic aortic valve failure in high surgical risk patients was associated with sustained improvement in clinical and echocardiographic outcomes.


Hazard Detection With Monocular Bioptic Telescopes in a Driving Simulator.
In most states, people with reduced visual acuity may legally drive with the aid of a bioptic telescope. However, concerns have been raised that the ring scotoma may impair detection of peripheral hazards. Using a driving simulator, we tested the hypothesis that the fellow eye would be able to compensate for the ring scotoma when using a monocular telescope. Sixteen bioptic users completed three drives with binocular viewing interleaved between three drives with monocular viewing. Forty pedestrians appeared and ran on the road for 1 second, including 26 within the ring scotoma, while participants were reading road signs through their own monocular telescopes. Head movements were analyzed to determine whether the pedestrian appeared before or only while using the telescope. For pedestrians that appeared only during bioptic use and were likely in the area of the ring scotoma, detection rates were significantly higher in binocular (fellow eye can compensate) than monocular (fellow eye patched) viewing (69% vs. 32%; P < 0.001); this was true for both current and noncurrent drivers. For pedestrians appearing before or after bioptic use, detection rates did not differ in binocular and monocular viewing. However, detection rates were even higher and reaction times shorter when the telescope was not being used. Both current and noncurrent drivers' fellow eyes were able to compensate, at least in part, for the ring scotoma. When using monocular telescopes, the fellow eye reduces the impact of the ring scotoma on hazard detection in binocular viewing.


The induction of cystitis and the implantation of tumours in rat and rabbit bladders.
A method of inducing tumour growth in rat and rabbit bladders is described. Tumour induction is dependent on a pre-induced cystitis. In rats, tumour also grows in the kidneys because of species characteristic vesicoureteric reflux. The possible significance is discussed in relation to the management of human bladder tumours.


DNA-binding and cleavage studies of novel copper(II) complex with L-phenylalaninate and 1,4,8,9-tetra-aza-triphenylene ligands.
DNA-binding properties of novel copper(II) complex [Cu(l-Phe)(TATP)(H(2)O)](+), where L-Phe=L-phenylalaninate and TATP=1,4,8,9-tetra-aza-triphenylene are investigated using electronic absorption spectroscopy, fluorescence spectroscopy, voltammetry and viscosity measurement. It is found that the presence of calf thymus DNA results in a hypochromism and red shift in the electronic absorption, a quenching effect on fluorescence nature of ethidium bromide-DNA system, an enhanced response on voltammograms of [Co(phen)(3)](3+/2+)-DNA system, and an obvious change in viscosity of DNA. From absorption titration, fluorescence analysis and voltammetric measurement, the binding constant of the complex with DNA is calculated. The latter two methods reveal the stronger binding of [Cu(l-Phe)(TATP)(H(2)O)](+) complex to double strand DNA by the moderate intercalation than [Co(phen)(3)](3+). Such a binding induces the cleavage of plasmid pBR322 DNA in the presence of H(2)O(2).


Association between age at onset and clinical features of schizophrenia: the Northern Finland 1966 birth cohort study.
To study the association between age at onset and the clinical picture of schizophrenia in an unselected young birth cohort. The study sample consists of 98 (64 males and 34 females) individuals with DSM-III-R schizophrenia collected from the Northern Finland 1966 birth cohort. Firstly, subjects were divided into very early- and young-onset subgroups by using the median age at onset (22 years in males and 20 in females), as a cut-off point. Secondly, we used age at onset as a continuous variable. Clinical features of schizophrenia were assessed using the Operational Criteria Checklist for Psychotic Illnesses (OCCPI). Inappropriate affect, positive thought disorder and deterioration from premorbid level of function associate with very early-onset schizophrenia, while slowed activity and dysphoria relate to young-onset. These symptoms correlate significantly with the age at onset. Differences in the clinical picture associating to the age at onset of schizophrenia are seen early. These findings indicate that certain symptoms of schizophrenia are dependent on the age at onset, and schizophrenia occurring initially in early life has some typical features. Using the age at onset as a continuous variable is independent of arbitrary cut-off points and produces more explicable results.


[Deletion mutation of beta-amyloid precursor protein decreases the secretion of beta-amyloid in SH-SY5Y neuroblastoma cells].
To explore the effect of A beta secretion lacking exons 2 and 3 of APP695 (delta APP695) on SH-SY5Y neuroblastoma cells. APP695 cDNA and delta APP695 cDNA eukaryotic expression vectors were constructed by gene recombination. Immunoprecipitation Western blotting was used to analyse the A beta secretion and MTT assay to observe the cell viability's change after being stably transfected with the plasmids by lipofectamine method. Stable transfectants were selected in 0.6 g.L-1 G418. delta APP695 cDNA construct transfection decreased significantly the production of beta amyloid in the SH-SY5Y compared with APP695 cDNA construct. It did not affect the cell viability by MTT method. This result suggests that the alternative splicing of APP exons 2 and 3 modulates the secretion of A beta by influencing the processing of APP.


NcoI TNF-β gene polymorphism and TNF expression are associated with an increased risk of developing Barrett's esophagus and esophageal adenocarcinoma.
Esophageal cancer development is a sequence that starts with reflux esophagitis (RE), followed by Barrett's esophagitis (BE), dysplasia, and finally esophageal adenocarcinoma (EAC). Tumor necrosis factor (TNF) is a potent anti-neoplastic agent, hence DNA polymorphisms that reduce TNF levels potentially enhance the development of BE and EAC. The aim of the study was to determine the impact of TNF gene variation on the RE-BE-EAC cascade. DNA from 887 Caucasian participants (197 controls, 305 RE, 257 BE, 128 EAC) was tested for the gene polymorphism TNF-β NcoI, and TNF production was determined by TNF-α specific immunohistochemistry on esophageal biopsies from these BE (n = 31) and EAC (n = 4) patients. As compared with healthy controls, the TNF-β NcoI A/A genotype was significantly more prevalent in BE (p = 0.04) and EAC patients (p = 0.02), but not in RE patients (p = 0.1). While TNF-α protein levels were invariably high in esophageal biopsies from EAC patients, most esophageal BE samples showed low to moderate TNF levels. Chronic inflammation, like in BE, markedly increase the risk of malignant transformation. In this study, the significantly higher frequency of the TNF-β NcoI A/A genotype and the local TNF expression indicate that the pro-inflammatory cytokine TNF plays a role in the development of BE and EAC.


Regulatory role of miR-125a/b in the suppression by selenium of cadmium-induced apoptosis via the mitochondrial pathway in LLC-PK1 cells.
Cadmium (Cd), a toxic heavy metal, is known to induce kidney damage and renal tubular dysfunction. Our previous studies have indicated that selenium (Se), as an essential trace element, protects against Cd-induced nephrotoxicity via the mitochondrial apoptotic pathway in vivo and in vitro. The available evidence suggests that the expression of miRNAs is altered after Cd exposure. However, the regulatory effects of miRNAs on Cd-induced apoptosis and on the suppression by Se of Cd-induced apoptosis remain unclear. The present study aimed to explore the effects of miRNAs on the suppression by Se of Cd-induced apoptosis in LLC-PK1 cells. miR-125a and miR-125b were downregulated in response to Cd exposure but were upregulated in the Se pretreatment group. Over-expression and low expression of miR-125a/b were simulated by their mimics and inhibitors, respectively. Both Se and over-expression of miR-125a/b attenuated Cd-induced apoptosis through significant enhancement of the anti-apoptotic protein Bcl-2, reduction of the pro-apoptotic proteins Bax and Bak, the release of cytochrome c and the inactivation of caspase-9 and caspase-3. The effects of downregulation of miR-125a/b on apoptosis were similar to those of Cd treatment, and both effects were inhibited by Se. Moreover, miR-125a/b were observed to target Bak and caspase-3 directly. In summary, miR-125a/b play an important role in the suppression of Cd-induced apoptosis by Se via the mitochondrial pathway in LLC-PK1 cells.


DT-Diaphorase Prevents Aminochrome-Induced Lysosome Dysfunction in SH-SY5Y Cells.
Aminochrome has been reported to induce lysosomal dysfunction by inhibiting the vacuolar H-type ATPase localized in lysosome membrane. DT-diaphorase has been proposed to prevent aminochrome neurotoxicity but it is unknown whether this enzyme prevents aminochrome-induced lysosomal dysfunction. In the present study, we tested the protective role of DT-diaphorase in lysosomal dysfunction by generating a cell line (SH-SY5YsiNQ7) with a stable expression of a siRNA against DT-diaphorase with only 10% expression of mRNA enzyme. The cells differentiated with retinoic acid and 12-o-tetradecanoylphorbol-13-acetate show a significant increase in the expression of tyrosine hydroxylase, vesicular monoamine transporter-2, and dopamine transporter. The incubation of SH-SY5YsiNQ7 cells with 10 μM aminochrome resulted in a significant decrease of lysosome pH determined by using acridine orange, while aminochrome has no effect on SH-SY5Y cells. These results support the proposed protective role of DT-diaphorase against aminochrome-induced lysosomal dysfunction.


Diagnosis of homozygosity and heterozygosity in congenital adrenal hyperplasia (CAH) and control of treatment.
In congenital adrenal hyperplasia the incidence of 21-hydroxylase deficiency is very high (approximately 1:7,000), whereas other enzyme defects such as 11-hydroxylase deficiency, 17-hydroxylase deficiency and 3 beta-hydroxysteroid dehydrogenase deficiency are much less frequent. The various forms of enzyme defects can be diagnosed by determining specific plasma steroids or specific urinary steroid metabolites. A new semi-automatic capillary gas liquid chromatography method has been introduced for the diagnosis of CAH and assessment of therapy. Heterozygous carriers of 21-hydroxylase deficiency can be detected in the general population by measuring 17-hydroxyprogesterone plasma levels after ACTH stimulation; however, results overlap with the general population. In families with a CAH index case, heterozygosity and homozygosity for the 21-hydroxylase deficiency gene can be detected by HLA-typing. 21-hydroxylase deficiency can be diagnosed prenatally by HLA-typing or by determining 17 OH-progesterone levels in the amniotic fluid.


Green synthesis and characterization of silver nanoparticles from Acacia nilotica and their anticancer, antidiabetic and antioxidant efficacy.
Both cancer and diabetes mellitus are serious health issues, accounting more than 11 million deaths worldwide annually. Targeted use of plant-mediated nanoparticles (NPs) in treatment of ailments has outstanding results due to their salient properties. The current study was designed to investigate the safe production of silver nanoparticles (AgNPs) from Acacia nilotica. Different concentrations of AgNO3 were tested to optimize the protocol for the synthesis of AgNPs from the bark extract. It was demonstrated that 0.1 M and 3 mM were found to be the optimum concentrations for the synthesis of AgNPs. Standard characterization techniques such as UV-vis spectrophotometry, SEM, SEM-EDX micrograph, spot analysis, elemental mapping and XRD were used for the conformation of biosynthesis of AgNPs. Absorption spectrum of plant-mediated AgNPs under UV-vis spectrophotometer showed a strong peak at 380 nm and 420 nm for AgNPs synthesized at 0.1 M and 3 mM concentration of salt. The SEM results showed that AgNPs were present in variable shapes within average particle size ranging from (20-50 nm). Anticancer, antidiabetic and antioxidant potential of green AgNPs was investigated and they showed promising results as compared to the positive and negative controls. Hence, AgNPs were found potent therapeutic agent against the human liver cancer cell lines (HepG2), strong inhibitor for α-glucosidase enzyme activity and scavenging agent against free radicals that cause oxidative stress. Further studies are however needed to confirm the molecular mechanism and biochemical reactions responsible for the anticancer and antidiabetic activities of the particles.


Ageing augments β-adrenergic cutaneous vasodilatation differently in men and women, with no effect on β-adrenergic sweating.
What is the central question of this study? β-Adrenergic receptor activation modulates cutaneous vasodilatation and sweating in young adults. In this study, we assessed whether age-related differences in β-adrenergic regulation of these responses exist and whether they differ between men and women. What is the main finding and its importance? We showed that ageing augmented β-adrenergic cutaneous vasodilatation, although the pattern of response differed between men and women. Ageing had no effect on β-adrenergic sweating in men or women. Our findings advance our understanding of age-related changes in the regulation of cutaneous vasodilatation and sweating and provide new directions for research on the significance of enhanced β-adrenergic cutaneous vasodilatation in older adults. β-Adrenergic receptor agonists, such as isoprenaline, can induce cutaneous vasodilatation and sweating in young adults. Given that cutaneous vasodilatation and sweating responses to whole-body heating and to pharmacological agonists, such as acetylcholine, ATP and nicotine, can differ in older adults, we assessed whether ageing also modulates β-adrenergic cutaneous vasodilatation and sweating and whether responses differ between men and women. In the context of the latter, prior reports showed that the effects of ageing on cutaneous vasodilatation (evoked with ATP and nicotine) and sweating (stimulated by acetylcholine) were sex dependent. Thus, in the present study, we assessed the role of β-adrenergic receptor activation on forearm cutaneous vasodilatation and sweating in 11 young men (24 ± 4 years of age), 11 young women (23 ± 5 years of age), 11 older men (61 ± 8 years of age) and 11 older women (60 ± 8 years of age). Initially, a high dose (100 µm) of isoprenaline was administered via intradermal microdialysis for 5 min to induce maximal β-adrenergic sweating. Approximately 60 min after the washout period, three incremental doses of isoprenaline were administered (1, 10 and 100 µm, each for 25 min) to assess dose-dependent cutaneous vasodilatation. Isoprenaline-mediated cutaneous vasodilatation was greater in both older men and older women relative to their young counterparts. Augmented cutaneous vasodilatory responses were observed at 1 and 10 µm in women and at 100 µm in men. Isoprenaline-mediated sweating was unaffected by ageing, regardless of sex. We show that ageing augments β-adrenergic cutaneous vasodilatation differently in men and women, without influencing β-adrenergic sweating.


Mechanism of non-steroidal anti-androgen-induced liver injury: Reactive metabolites of flutamide and bicalutamide activate inflammasomes.
Flutamide is a non-steroidal anti-androgen agent, which is mainly used for the treatment of prostate cancer. Flutamide is known to cause severe adverse events, which includes idiosyncratic liver injury. However, details of the mechanism of these adverse reactions have not been elucidated. We investigated whether flutamide induces the release of damage-associated molecular patterns (DAMPs) that activate inflammasomes. We also tested bicalutamide, enzalutamide, apalutamide, and darolutamide for their ability to activate inflammasomes in differentiated THP-1 cells. The supernatant from the incubation of flutamide and bicalutamide with human hepatocarcinoma functional liver cell-4 (FLC-4) cells increased caspase-1 activity and production of IL-1ß by differentiated THP-1 cells. In the supernatant of FLC-4 cells with flutamide and bicalutamide, the heat shock protein (HSP) 40 or 60 was significantly increased. Addition of a carboxylesterase or a CYP inhibitor to the FLC-4 cells prevented release of HSPs from the FLC-4 cells. These results suggested that the reactive metabolites of flutamide and bicalutamide can cause the release of DAMPs from hepatocytes and activate inflammasomes. Inflammasome activation may be an important step in the activation of the immune system by flutamide or bicalutamide, which in some patients, can cause immune-related adverse events.


Locomotory and physiological responses induced by clove and cinnamon essential oils in the maize weevil Sitophilus zeamais.
Plant essential oils have been suggested as a suitable alternative for controlling stored pests worldwide. However, very little is known about the physiological or behavioral responses induced by these compounds in insect populations that are resistant to traditional insecticides. Thus, this investigation evaluated the toxicity (including the impacts on population growth) as well as the locomotory and respiratory responses induced by clove, Syzygium aromaticum L., and cinnamon, Cinnamomum zeylanicum L., essential oils in Brazilian populations of the maize weevil Sitophilus zeamais. We used populations that are resistant to phosphine and pyrethroids (PyPhR), only resistant to pyrethroids (PyR1 and PyR2) or susceptible to both insecticide types (SUS). The PyPhR population was more tolerant to cinnamon essential oil, and its population growth rate was less affected by both oil types. Insects from this population reduced their respiratory rates (i.e., CO2 production) after being exposed to both oil types and avoided (in free choice-experiments) or reduced their mobility on essential oil-treated surfaces. The PyR1 and PyR2 populations reduced their respiratory rates, avoided (without changing their locomotory behavior in no-choice experiments) essential oil-treated surfaces and their population growth rates were severely affected by both oil types. Individuals from SUS population increased their mobility on surfaces that were treated with both oil types and showed the highest levels of susceptibility to these oils. Our findings indicate that S. zeamais populations that are resistant to traditional insecticides might have distinct but possibly overlapping mechanisms to mitigate the actions of essential oils and traditional insecticides.


Enzyme stabilization by glutaraldehyde crosslinking of adsorbed proteins on aminated supports.
The stabilization achieved by different immobilization protocols have been compared using three different enzymes (glutaryl acylase (GAC), D-aminoacid oxidase (DAAO), and glucose oxidase (GOX)): adsorption on aminated supports, treatment of this adsorbed enzymes with glutaraldehyde, and immobilization on glutaraldehyde pre-activated supports. In all cases, the treatment of adsorbed enzymes on amino-supports with glutaraldehyde yielded the higher stabilizations: in the case of GOX, a stabilization over 400-fold was achieved. After this treatment, the enzymes could no longer be desorbed from the supports using high ionic strength (suggesting the support-protein reaction). Modification of the enzymes immobilized on supports that did not offer the possibility of react with glutaraldehyde showed the same stability that the non modified preparations demonstrating that the mere chemical modification did not have effect on the enzyme stability. This simple strategy seems to permit very good results in terms of immobilization rate and stability, offering some advantages when compared to the immobilization on glutaraldehyde pre-activated supports.


Development and proposal of a scoring system for giant cell tumour of the bone around the knee.
The purpose of this study was to provide the surgeons with effective and reliable guidelines for surgical decision-making by establishing a scoring system for giant cell tumour (GCTSS) based on evidence and expert opinion. The modified Delphi technique and analytic hierarchy process were used to establish the GCTSS. The GCTSS was defined and classified based on different surgical methods using data from 207 patients collected retrospectively between October 2003 and December 2014. Finally, prospective data of 40 patients between December 2014 and October 2015 were used to analyze concordance between score categorization and experts' consensus on surgical procedure. A novel GCTSS included pathological fracture, cortical bone destruction, tumour size, and articular surface involved. The total scores ranged from 1 to 12 points. The strategy for each patient was decided: a total score of 1-4 suggested intralesional curettage alone for excellent post-operative function; 5-9 points indicated intralesional curettage with internal fixation for less surgery-related complications; and 10-12 points indicated prosthesis replacement for long-term local control. The κ-statistic for the predictive validity of total score was 0.611. The κ coefficient of each group represented moderate or substantial agreement, which was acceptable. The intraclass correlation coefficient for inter- and intra-observer reliability of total score was 0.831 and 0.740, respectively. The novel GCTSS is a comprehensive scoring system with content validity that can aid surgeons in assessing the aggressiveness or severity of giant cell tumour and might become a prognostic tool for surgical decision-making.


Statins decrease vascular epithelial growth factor expression via down-regulation of receptor for advanced glycation end-products.
Statins, inhibitors of 3-hydroxy-3-methylglutaryl coenzyme A reductase, possess pleiotropic effects that have been extended to modulation of various cellular behaviors. This study aimed to examine whether statins modulate vascular endothelial growth factor A (VEGF-A) expression in human retinal pigment epithelium (RPE) cells. Human RPE cells (h1RPE7), damaged by hydroquinone (HQ) + advanced glycation endproducts (AGE) in an in vitro AMD model, were treated with atorvastatin or lovastatin for 24 h. The expression of VEGF-A and receptor for AGE (RAGE) was evaluated by real-time RT-PCR. VEGF-A secretion was measured by ELISA. To investigate the impact of RAGE on VEGF-A expression, small interfering RNA (siRNA) for RAGE (siRAGE) was introduced into h1RPE7 cells and VEGF-A expression was measured by real-time RT-PCR. Deletions of VEGF-A and RAGE promoters were performed and transcriptional activities were measured after the addition of statins to HQ + AGE-damaged RPE cells. The mRNA levels of VEGF-A and RAGE and the levels of VEGF-A in the culture medium were increased by HQ + AGE. Both atorvastatin and lovastatin attenuated HQ + AGE-induced VEGF-A and RAGE expression. These statins also decreased VEGF-A levels in the culture medium. RNA interference of RAGE attenuated the up-regulation of VEGF-A in the HQ + AGE treated cells. The deletion analysis demonstrated that these statins attenuated RAGE promoter activation in HQ + AGE-damaged RPE cells. Statins attenuated HQ + AGE-induced VEGF expression by decreasing RAGE expression. As VEGF is an important factor in developing wet AMD, statins could decrease the risk of wet-type AMD and be used as preventive medicines.


Bending stiffness of a crystalline actin bundle.
The acrosomal process of the sperm of the horseshoe crab (Limulus polyphemus) is a unique crystalline actin bundle, consisting of multiple actin filaments cross-linked by the actin-bundling protein, scruin. For successful fertilization, the acrosomal bundle must penetrate through a 30 microm thick jelly coat surrounding the egg and thus it must be sufficiently stiff. Here, we present two measurements of the bending stiffness of a single crystalline bundle of actin. Results from these measurements indicate that the actin:scruin composite bundle has an average elastic modulus of 2 GPa, which is similar to that of a single actin filament, and a bending stiffness that is more than two orders of magnitude larger than that of a bundle of uncross-linked actin filaments due to stiffening by the scruin matrix.


CD28-mediated signalling co-stimulates murine T cells and prevents induction of anergy in T-cell clones.
Occupancy of the T-cell antigen receptor is insufficient to induce T-cell activation optimally; a second co-stimulatory signal is required. Exposure of T-cell clones to complexes of antigen with major histocompatibility complex molecules in the absence of the co-stimulatory signal induces a state of clonal anergy. This requirement for two stimuli for T-cell activation could have an important role in vivo in establishing peripheral tolerance to antigens not encountered in the thymus. The receptor on T cells required for the co-stimulatory stimulus involved in the prevention of anergy has not been identified. The human T-cell antigen CD28 provides a signal that can synergize with T-cell antigen receptor stimulation in activating T cells to proliferate and secrete lymphokines. Here we report that a monoclonal antibody against the murine homologue of CD28 (ref. 7; J.A.G. et al., manuscript in preparation) can provide a co-stimulatory signal to naive CD4+ T cells and to T-cell clones. Moreover, we demonstrate that this co-stimulatory signal can block the induction of anergy in T-cell clones.


The development of delusion revisited: a transdiagnostic framework.
This study proposes a transdiagnostic framework for delusion development, analysing psychiatric (schizophrenia, bipolar disorder, major depressive disorder) and neurological disorders (stroke, and neurodegenerative diseases) in which delusions are predominant. Our aim is to identify a transdiagnostic core of neural and cognitive alterations associated with delusions across distinct clinical disorders. Reviewed empirical evidence suggests delusions are associated: on the neural level with changes in the ventromedial prefrontal cortex (vmPFC) networks, and on the neuropsychological level with dysfunction in the processes (generation of affective value, the construction of internal models of the world, and the reflection about Self and/or Other's mental states) that these network mediate. The concurrent aberration of all these processes could be critical for the clinical transition to a psychotic delusional state. In particular, delusions could become clinically manifest when (1) stimuli are attributed an aberrant affective salience, that (2) is explained by the patient within distorted explanatory internal models that (3) are poorly inhibited by cognitive control systems. This framework extends the two-factor account of delusion model and suggests that common neural mechanisms for the delusions in psychiatric and in neurological disorders.


Predictive value of the single-breath nitrogen test for hospitalization due to respiratory disease.
The relationship between indices of the single-breath nitrogen test (SBNT) measured in 1974 and hospitalization in the 9 year period 1977-1986 was examined in a random population sample of 876 men aged 46-69 years. Men who could not perform acceptable SBNT tracings had an increased risk of hospitalization due to respiratory disease in general. When age and smoking habits were controlled for, slope of phase III was significantly related to hospitalization due to respiratory disease in general and chronic obstructive pulmonary disease (COPD), whereas closing volume and closing capacity were marginally related to hospitalization due to respiratory disease in general but not to hospitalization due to COPD. The relationship between slope of phase III and hospitalization due to COPD remained significant after the forced expiratory volume in 1 sec (FEV1) was controlled for: odds ratio 1.4 per % N2/L (95% confidence interval 1.1-1.7). The effect of the slope of phase III was considered to be clinically insignificant, and we conclude that in a random population sample indices from only 1 SBNT do not provide prognostic information concerning hospitalization in addition to that provided by FEV1.


Placebo versus allopurinol for renal calculi.
Certain patients with recurrent calcium oxalate calculi will show a mild uricacidemia and/or uricosuria. A prospective blind study was begun in 1970, comparing placebo to allopurinol in the management of these patients. The 92 patients had been followed for a minimum of 6 months to 5 years before the code was broken. It is demonstrated clearly that there is a placebo effect in the management of renal calculous disease. It also has been shown that the only event that clearly separates the 2 groups is complete cessation of calculous formation. Allopurinol is effective in 61 per cent of the patients and these successes usually can be identified after 1 year of treatment.


Kidney function in male and female rats chronically exposed to potassium dichromate.
Male and female Wistar rats were given 25 mg l-1 chromium (as potassium dichromate) in drinking water for 6 months. Lactate dehydrogenase, lysozyme, total proteins, N-acetyl-beta-D-glucosaminidase, albumin and beta 2-microglobulin (beta 2-m) were measured in 24-h urine after 3 and 6 months of exposure. Body and kidney weight and chromium excretion were also examined. Except for the chromium excretion, no statistically significant changes were observed in the exposed male rats. In female rats there were significant increases in the urinary excretion of albumin after 3 and 6 months of exposure and the urinary excretion of beta 2-m after 3 months of exposure.


Carbon/PEEK composite materials as an alternative for stainless steel/titanium hip prosthesis: a finite element study.
Total hip replacement (THR) has been ranked within the most typical surgical processes in the world. The durability of the prosthesis and loosening of prosthesis are the main concerns that mostly reported after THR surgeries. In THR, the femoral prosthesis can be fixed by either cement or cementless methods in the patient's bones. In both procedures, the stability of the prosthesis in the hosted bone has a key asset in its long-term durability and performance. This study aimed to execute a comparative finite element simulation to assess the load transfer between the prosthesis, which is made of carbon/PEEK composite and stainless steel/titanium, and the femur bone. The mechanical behavior of the cortical bone was assumed as a linear transverse isotropic while the spongy bone was modeled like a linear isotropic material. The implants were made of stainless steel (316L) and titanium alloy as they are common materials for implants. The results showed that the carbon/PEEK composites provide a flatter load transfer from the upper body to the leg compared to the stainless steel/titanium prosthesis. Furthermore, the results showed that the von Mises stress, principal stress, and the strain in the carbon/PEEK composites prosthesis were significantly lower than that made of the stainless steel/titanium. The results also imply that the carbon/PEEK composites can be applied to introduce a new optimum design for femoral prosthesis with adjustable stiffness, which can decrease the stress shielding and interface stress. These findings will help clinicians and biomedical experts to increase their knowledge about the hip replacement.


Comparison of total cortisol, free cortisol, and surrogate markers of free cortisol in diagnosis of adrenal insufficiency in patients with stable cirrhosis.
Measurements of serum levels of total cortisol can overestimate the prevalence of adrenal dysfunction in patients with cirrhosis because they have low concentrations of corticosteroid-binding globulin and albumin. We used measurements of serum total cortisol and serum free cortisol after the low-dose short Synacthen test (LDSST) to assess adrenal dysfunction. We studied 79 patients with stable cirrhosis; adrenal dysfunction was defined by peak concentrations of total cortisol ≤494 mmol/L and/or peak concentrations of free cortisol ≤33 nmol/L after the LDSST. We determined free cortisol index (FCI) scores and calculated free cortisol levels by using Coolens' equation. The Cox regression model was used to assess the relationship between adrenal dysfunction and outcomes (death or liver transplant). On the basis of measurement of total cortisol, 34% of patients had adrenal dysfunction, and on the basis of measurement of free cortisol, 29% had adrenal dysfunction. There was agreement between total cortisol and free cortisol levels in 22% of patients; in 13%, adrenal dysfunction was diagnosed only on the basis of total cortisol and in 6% only on the basis of free cortisol (κ coefficient, 0.56; P < .01). Low concentrations of corticosteroid-binding globulin (21 vs 54 μg/mL, P < .01) led to an overestimation of adrenal dysfunction that was based on measurement of total cortisol. Measurements of calculated free cortisol constantly overestimated free cortisol concentrations, with variations as large as 87% for baseline values and up to 84% after stimulation. Adrenal insufficiency, defined by FCI scores <12, was detected in 30% of patients; among them, 23% also had subnormal peak levels of free cortisol (κ coefficient, 0.70; P < .001). Adrenal dysfunction was not significantly associated with patient outcomes, on the basis of Cox model analysis. Adrenal insufficiency, defined by LDSST, is frequent in patients with stable cirrhosis, on the basis of measurements of total and free cortisol. FCI scores are better than measurement of total cortisol in assessing adrenal function in patients with cirrhosis. We did not associate adrenal dysfunction with outcome, but further studies are needed.


Convergence of light and chloroplast signals for de-etiolation through ABI4-HY5 and COP1.
Seedling de-etiolation prepares plants to switch from heterotrophic to photoautotrophic growth, a transition essential for plant survival. This delicate de-etiolation process is precisely controlled by environmental and endogenous signals. Although intracellular plastid-derived retrograde signalling is essential for the de-etiolation process, the molecular nature of these retrograde signals remains elusive(1-3). Here we show that chloroplast and light signals antagonistically fine-tune a suite of developmental and physiological responses associated with de-etiolation through a transcriptional module of ABA INSENSITIVE 4 (ABI4) and ELONGATED HYPOCOTYL 5 (HY5). Moreover, ABI4 and HY5 antagonistically regulate the expression of CONSTITUTIVE PHOTOMORPHOGENIC1 (COP1) and the subsequent greening process. In turn, ABI4 and HY5 are targeted for degradation by COP1 in the light and dark, respectively, to ensure a proper interplay of ABI4 and HY5 actions during seedling de-etiolation. Our study provides a new molecular mechanism for understanding how chloroplast signals converge with light signals to optimize early plant development.


Road to a Genetic Model of Gelatinous Drop-Like Corneal Dystrophy.
Gelatinous drop-like corneal dystrophy (GDLD; OMIM: 204870) is an autosomal recessive Mendelian-inherited disease. Subepithelial amyloid deposition results in severe visual impairment. Its pathogenesis remains unknown. Identification of the responsible gene is a useful approach to investigating the pathogenesis of genetic diseases. Such an identification is also necessary for the creation of genetic models. For this purpose, our group performed positional cloning using homozygosity mapping. The results revealed that GDLD maps to the short arm of chromosome 1. Subsequent mutation analyses identified tumor-associated calcium signal transducer 2 (TACSTD2; former name, M1S1) as the gene responsible for GDLD. After identification of the gene, we examined the phylogenetical distribution of TACSTD2 in the genomes of vertebrates.


Clinical evaluation of a single-pass implantable electrode for all modes of pacing. The "Crown of Thorns" lead.
Physiological pacing was instituted in 17 patients (11 men and 6 women), mean age 67 years (range 33-77 years), using a variety of multiprogramable generators attached to a permanent single-pass dual chamber electrode. Eleven patients were paced in VAT mode (Cordis 208A or Siemens-Elema 625 generator), two patients in DVI mode (Intermedics Cyberlith IV generator) and four patients in DDD mode (Siemens-Elema 664/P33 or Telectronics Autima unit). Mean intracardiac P wave amplitude was 2.0 mV +/- 0.78/SD, range 0.7-3.6 mV. and mean atrial and ventricular pacing thresholds were 1.0 V and 0.5 V, respectively. Fourteen patients had completely successful A-V pacing during a follow-up period of 4-13 months (mean 7 +/- 2.7 months). Two failures were associated with malposition of the atrial crown and occurred exclusively with the Cordis 208A generator. In both patients generator replacement using a more sensitive unit (Siemens-Elema 625) resulted in successful VAT pacing for most of the time. Complete failure of A-V pacing occurred in only one patient who died from coronary artery disease after four months of follow-up. Thus, all of the remaining 16 patients achieved long-term (6 months) satisfactory physiological pacing using this new lead. We conclude that the "Crown of Thorns" electrode is a successful single-pass unipolar lead and can be used with all types of dual chamber generator for all modes of pacing.


Acquired iron overload associated with antitransferrin monoclonal immunoglobulin: a case report.
We describe a patient with an unusual combination of hypersideremia (700 microg/dL), hypertransferrinemia (570 mg/dL), hyperferritinemia (800 microg/L), and monoclonal gammopathy of undetermined significance (MGUS), in which the monoclonal immunoglobulin showed specific transferrin-binding activity. Liver histology revealed hepatic iron overload, prominent in periportal hepatocytes, suggesting intestinal iron hyperabsorption. We demonstrate that low urinary hepcidin, likely due to impaired iron delivery to erythroid cells via the transferrin cycle pathway over time, may be the mechanism for iron loading. We suggest that MGUS associated with monoclonal antibodies with antitransferrin activity should be added to the list of acquired causes of hemochromatosis.


Evidence for the different physiological significance of the 6- and 2-minute walk tests in multiple sclerosis.
Researchers have recently advocated for the 2-minute walk (2MW) as an alternative for the 6-minute walk (6MW) to assess long distance ambulation in persons with multiple sclerosis (MS). This recommendation has not been based on physiological considerations such as the rate of oxygen consumption (V·O2) over the 6MW range. This study examined the pattern of change in V·O2 over the range of the 6MW in a large sample of persons with MS who varied as a function of disability status. Ninety-five persons with clinically-definite MS underwent a neurological examination for generating an Expanded Disability Status Scale (EDSS) score, and then completion of the 6MW protocol while wearing a portable metabolic unit and an accelerometer. There was a time main effect on V·O2 during the 6MW (p=.0001) such that V·O2 increased significantly every 30 seconds over the first 3 minutes of the 6MW, and then remained stable over the second 3 minutes of the 6MW. This occurred despite no change in cadence across the 6MW (p=.84). The pattern of change in V·O2 indicates that there are different metabolic systems providing energy for ambulation during the 6MW in MS subjects and steady state aerobic metabolism is reached during the last 3 minutes of the 6MW. By extension, the first 3 minutes would represent a test of mixed aerobic and anaerobic work, whereas the second 3 minutes would represent a test of aerobic work during walking.


Potentiometric and voltammetric investigations of H2/H+ catalysis by periplasmic hydrogenase from Desulfovibrio gigas immobilized at the electrode surface in an amphiphilic bilayer assembly.
Interactions of an enzyme with an organized amphipilic bilayer are explored as a general means of enzyme immobilization in electroenzymatic systems. Immobilization of Desulfovibrio gigas hydrogenase at the electrode surface involves hydrophobic interactions of the enzyme with the bilayer assembly consisting of octadecyltrichlorosilane and octadecylviologen (C18MV2+) molecules. Due to a hydrophobic character of the enzyme, these interactions direct the enzyme to occupy a central position in the bilayer's hydrocarbon region and lead to immobilization of 3 pmol/cm2 of the enzyme in the plane of the bilayer. This corresponds to 50% surface coverage. The immobilized enzyme catalyzes H2 oxidation mediated by the C18MW2+/.+ couple. This electroenzymatic scheme functions under steady-state voltammetric as well as potentiometric conditions in the pH range 3.5-10. Coupling of enzymatic activity to the electrode surface is accomplished via lateral diffusion of the octadecylviologen molecules along the bilayer assembly.


Low-Temperature Charging and Aging Mechanisms of Si/C Composite Anodes in Li-Ion Batteries: An Operando Neutron Scattering Study.
The addition of Si compounds to graphite anodes has become an attractive way of increasing the practical specific energies in Li-ion cells. Previous studies involving Si/C anodes lacked direct insight into the processes occurring in full cells during low-temperature operation. In this study, a powerful combination of operando neutron diffraction, electrochemical tests, and post-mortem analysis is used for the investigation of Li-ion cells. 18650-type cylindrical cells in two different aging states are investigated by operando neutron diffraction. The experiments reveal deep insights and important trends in low-temperature charging mechanisms involving intercalation, alloying, Li metal deposition, and relaxation processes as a function of charging C-rates and temperatures. Additionally, the main aging mechanism caused by long-term cycling and interesting synergistic effects of Si and graphite are elucidated.


[The medicinal treatment of anxiety in alcoholism in the withdrawal stage (author's transl)].
The benzodiazepine preparation Camazepam was tested for its anxiolytic activity and found to be effective. It differs from preparations which have already been in use for a long time, especially by the lack of any hypnotic and muscle relaxant effects. In the clinical trial it was also free from side effects so far as they could be detected by the usual routine investigations of the technical laboratory.


Evaluating life history trade-offs through the presence of linear enamel hypoplasia at Pueblo Bonito and Hawikku: A biocultural study of early life stress and survival in the Ancestral Pueblo Southwest.
Due to the indelible nature of enamel, bioarchaeologists use linear enamel hypoplasia (LEH) to detect early investments in surviving stress and have identified an association between LEH presence and constraints in growth and maintenance as well as an increased susceptibility to future stress events. This study evaluates heterogenous frailty and susceptibility to death in relation to episodes of early life stress, as reflected by LEH presence, in the Ancestral Pueblo Southwest. This study hypothesizes that LEH presence will be associated with decreased survivorship and an increased likelihood of mortality in both samples. This study uses two samples, one from Pueblo Bonito (A.D. 800-1200; n = 28) and the second from Hawikku (A.D. 1300-1680; n = 103). Kaplan-Meier survival analysis with a log-rank test was used to evaluate the effect of LEH presence on survivorship for the two samples. Survival analysis reveals statistically significant differences in mortality risk between individuals with and without LEH for the Hawikku sample, but no significant differences for the Pueblo Bonito sample. The results demonstrate differences in the response to early life stress at the Hawikku and Pueblo Bonito sites, likely reflecting context. The Pueblo Bonito sample represents a high-status group, and survival following LEH may be the result of cultural buffering. Hawikku dates to a period associated with increased levels of disease and malnutrition as well as Spanish colonization. This environment may have exacerbated mortality risk for individuals in the region who survived early life stress and signifies the consequences of European colonialism in the New World.


Facilitatory role of central dopamine in the osmotic release of vasopressin.
In conscious rats, intracerebroventricular (i.c.v.) injection of the dopamine antagonist haloperidol (0.15 mumol) blocked increases in plasma vasopressin (AVP) and arterial pressure caused by i.c.v. administration of a hypertonic solution (990 mOsm/kg, 10 microliters), without affecting plasma osmolality, electrolytes or hematocrit. The application of the alpha-adrenergic antagonist phenoxybenzamine (0.15 mumol), however, did not significantly inhibit the responses of plasma AVP and arterial pressure to the hypertonic solution. We concluded that brain dopamine may play a facilitatory role in the osmotically stimulated AVP secretion.


A cross-sectional survey of the acceptability of data collection processes for validation of a European point prevalence survey of healthcare-associated infections and antimicrobial use.
Statistical measurements alone are insufficient to ensure robust data for point prevalence surveys (PPS) of healthcare-associated infections (HAI). Data quality is determined by the type of data, data collection methods and available resources. Data collectors' views regarding the acceptability of data collection process for validation studies are also important to consider. To explore data collectors' views on the acceptability of data collection processes used for a European validation PPS of HAI and antimicrobial use (AMU). An anonymous online survey was conducted with 67 data collectors from 10 European countries involved in the study. Twenty-five (64.1%) participants viewed AMU data collection as easy/quite easy whereas only five (12.8%) thought HAI data collection was easy/quite easy. Six (17%) participants indicated that incentives and 21 (56.8%) that disincentives were possibly/definitely present for reporting cases of HAI. Engagement of staff was not thought to have adversely affected data collection as only one (2.6%) and five (15.4%) participants thought involvement of hospital PPS teams and administration was low/very low, respectively. Participants believed the approaches used were appropriate but that more training was required prior to data collection, some case definitions should be reviewed and the number of variables reduced.


Mechanism of the CO-sensing heme protein CooA: new insights from the truncated heme domain and UVRR spectroscopy.
The bacterial CO-sensing heme protein CooA activates expression of genes whose products perform CO-metabolism by binding its target DNA in response to CO binding. The required conformational change has been proposed to result from CO-induced displacement of the heme and of the adjacent C-helix, which connects the sensory and DNA-binding domains. Support for this proposal comes from UV Resonance Raman (UVRR) spectroscopy, which reveals a more hydrophobic environment for the C-helix residue Trp110 when CO binds. In addition, we find a tyrosine UVRR response, which is attributable to weakening of a Tyr55-Glu83 H-bond that anchors the proximal side of the heme. Both Trp and Tyr responses are augmented in the heme domain when the DNA-binding domain has been removed, apparently reflecting loss of the inter-domain restraint. This augmentation is abolished by a Glu83Gln substitution, which weakens the anchoring H-bond. The CO recombination rate following photolysis of the CO adduct is similar for truncated and full-length protein, though truncation does increase the rate of CO association in the absence of photolysis; together these data indicate that truncation causes a faster dissociation of the endogenous Pro2 ligand. These findings are discussed in the light of structural evidence that the N-terminal tail, once released from the heme, selects the proper orientation of the DNA-binding domain, via docking interactions.


Loss of TAX1BP1-Directed Autophagy Results in Protein Aggregate Accumulation in the Brain.
Protein aggregates disrupt cellular homeostasis, causing toxicity linked to neurodegeneration. Selective autophagic elimination of aggregates is critical to protein quality control, but how aggregates are selectively targeted for degradation is unclear. We compared the requirements for autophagy receptor proteins: OPTN, NBR1, p62, NDP52, and TAX1BP1 in clearance of proteotoxic aggregates. Endogenous TAX1BP1 is recruited to and required for the clearance of stress-induced aggregates, whereas ectopic expression of TAX1BP1 increases clearance through autophagy, promoting viability of human induced pluripotent stem cell-derived neurons. In contrast, TAX1BP1 depletion sensitizes cells to several forms of aggregate-induced proteotoxicity. Furthermore, TAX1BP1 is more specifically expressed in the brain compared to other autophagy receptor proteins. In vivo, loss of TAX1BP1 results in accumulation of high molecular weight ubiquitin conjugates and premature lipofuscin accumulation in brains of young TAX1BP1 knockout mice. TAX1BP1 mediates clearance of a broad range of cytotoxic proteins indicating therapeutic potential in neurodegenerative diseases.


Pantomime in aphasia: effects of stimulus characteristics.
The effects of three types of stimuli (objects, line drawings, action pictures) on the comprehension and production of pantomimes in aphasic patients were assessed. A significantly higher number of correct pantomime responses was found for objects and action pictures than for line drawings. Significant positive correlations were found between aphasia subtest scores and pantomimic scores. The theoretical and clinical implications of these findings are discussed.


SnoRNP is essential for thermospermine-mediated development in Arabidopsis thaliana.
Polyamines have been discovered for hundreds of years and once considered as a class of phytohormones. Polyamines play critical roles in a range of developmental processes. However, the molecular mechanisms of polyamine signaling pathways remain poorly understood. Here, we measured the contents of main types of polyamines, and found that endogenous level of thermospermine (T-Spm) in Arabidopsis thaliana is comparable to those of classic phytohormones and is significantly lower than those of putrescine (Put), spermidine (Spd), and spermine (Spm). We further found a nodule-like structure around the junction area connecting the shoot and root of the T-Spm biosynthetic mutant acl5 and obtained more than 50 suppressors of acl5nodule structure (san) through suppressor screening. An in-depth study of two san suppressors revealed that NAP57 and NOP56, core components of box H/ACA and C/D snoRNPs, were essential for T-Spm-mediated nodule-like structure formation and plant height. Furthermore, analyses of rRNA modifications showed that the overall levels of pseudouridylation and 2'-O-methylation were compromised in san1 and san2 respectively. Taken together, these results establish a strong genetic relationship between rRNA modification and T-Spm-mediated growth and development, which was previously undiscovered in all organisms.


Dissecting the role of lactate metabolism LncRNAs in the progression and immune microenvironment of osteosarcoma.
The process of lactate metabolism has been proved to play a critical role in the progression of various cancers and to influence the immune microenvironment, but its potential role in osteosarcoma remains unclear. We have acquired transcriptomic and clinical data from 84 osteosarcoma samples and 70 normal bone samples from the TARGET and GTEx databases. We identified differentially expressed lactate metabolism-related LncRNAs (LRLs) in osteosarcoma and performed Cox regression and LASSO regression to establish LRLs prognostic signature (LRPS). The reliability of LRPS performance was examined by separate prognostic analysis, viability curves and receiver operating characteristic (ROC) curves. Furthermore, the effects of LRPS on the immune microenvironment of osteosarcoma were investigated, and the functions of the focal genes were experimentally validated. A total of 856 differentially expressed LRLs were identified and 5 of them were selected to construct LRPS, which was a better prognostic predictor for osteosarcoma compared with other published prognostic signatures (AUC up to 0.947 and 0.839 in the training and test groups, respectively, with adj-p<0.05 for KM curves). We found that LRPS significantly affected the immune infiltration of osteosarcoma, while RP11-472M19.2 significantly promoted the metastasis of osteosarcoma, which was well validated experimentally. Encouragingly, a number of sensitive drugs were identified for LRPS and RP11-472M19.2 high-risk groups. Our study shows that lactate metabolism plays a crucial role in the development of osteosarcoma and has been well validated experimentally, providing extremely important insights into the clinical treatment and in-depth research of osteosarcoma.


The influence of handling procedures during mouse oocyte and embryo recovery on viability and subsequent development in vitro.
The effect on mouse oocyte and embryo viability of prolonging the time that they spend in the oviduct prior to flushing has been investigated. Oocytes and embryos from oviducts that were flushed quickly and with as little temperature variation as possible were compared with oocytes and embryos flushed from oviducts that had been left for 30 min, either at 37 degrees C in saline or in the intact dead mouse before dissection. It was found that retention of the eggs and embryos in the oviducts leads to a lower viability at recovery and also affects the subsequent developmental potential of embryos in culture.


Ophthalmologic manifestations in primary antiphospholipid syndrome patients: A cross-sectional analysis of a primary antiphospholipid syndrome cohort (APS-Rio) and systematic review of the literature.
There is a broad spectrum of eye involvement in antiphospholipid syndrome (APS). The majority of descriptions are presented as case reports that include mostly APS patients secondary to systemic lupus erythematosus (SLE), with no compelling evidence in primary APS (PAPS). This study aimed to describe ocular manifestations in our well-defined PAPS cohort (APS-Rio) and then perform a systematic literature review (SLR) of ocular manifestations in patients with APS or positivity to aPL without SLE. We retrospectively analyzed PAPS patients followed at our outpatient clinics. All patients fulfilled Sydney APS classification criteria (2006). We evaluated them for ocular symptoms and previous ocular diagnoses. Antiphospholipid antibodies and clinical APS manifestations were compared between patients with and without ocular manifestations. For the SLR, electronic databases were searched up to November 2019. We studied 105 PAPS patients; 90.5% were female and 56.2% were Caucasian. We found ocular manifestations in 37.1% of our cohort. Thrombosis was the main criteria manifestation (95.2%) and lupus anticoagulant was the most prevalent antibody. Ophthalmologic diagnoses were present in 7 patients, with 5 having retinal vessels thromboses. Amaurosis fugax was the leading complaint, present in 30 patients. In the univariate analysis, amaurosis fugax was related to livedo (p = 0.005), Raynaud's phenomenon (p = 0.048) and the presence of anticardiolipin antibody (≥40 GPL/MPL) (p = 0.041). Hemianopia was associated with arterial hypertension (p = 0.049). In the multivariate analysis, the only association found was between livedo and amaurosis fugax (OR 4.09, 95%CI 1.5-11.11, p = 0.006). Our SLR incorporated 96 articles of ocular manifestations in patients with PAPS or positivity to aPL without SLE. Ocular findings varied from 5 to 88%, including anterior and posterior segments, orbital and neuro-ophthalmologic changes. There is little evidence on ocular manifestations in PAPS. We described an association between livedo and amaurosis fugax. Prospective studies are needed to promote the best treatment and avoid blindness in PAPS patients.


Injuries to the Biceps Pulley.
The biceps reflection pulley is a soft tissue sling that stabilizes the long head of the biceps tendon (LHB) before it enters the bicipital groove. Injuries to the biceps pulley and related instability of the LHB are common diagnoses in patients with anterior shoulder pain. This article summarizes the current concepts for treatment of injuries to the biceps pulley. Clinical and radiological presentation, arthroscopic assessment, and current treatment options are outlined.


Application of Bayesian genomic prediction methods to genome-wide association analyses.
Bayesian genomic prediction methods were developed to simultaneously fit all genotyped markers to a set of available phenotypes for prediction of breeding values for quantitative traits, allowing for differences in the genetic architecture (distribution of marker effects) of traits. These methods also provide a flexible and reliable framework for genome-wide association (GWA) studies. The objective here was to review developments in Bayesian hierarchical and variable selection models for GWA analyses. By fitting all genotyped markers simultaneously, Bayesian GWA methods implicitly account for population structure and the multiple-testing problem of classical single-marker GWA. Implemented using Markov chain Monte Carlo methods, Bayesian GWA methods allow for control of error rates using probabilities obtained from posterior distributions. Power of GWA studies using Bayesian methods can be enhanced by using informative priors based on previous association studies, gene expression analyses, or functional annotation information. Applied to multiple traits, Bayesian GWA analyses can give insight into pleiotropic effects by multi-trait, structural equation, or graphical models. Bayesian methods can also be used to combine genomic, transcriptomic, proteomic, and other -omics data to infer causal genotype to phenotype relationships and to suggest external interventions that can improve performance. Bayesian hierarchical and variable selection methods provide a unified and powerful framework for genomic prediction, GWA, integration of prior information, and integration of information from other -omics platforms to identify causal mutations for complex quantitative traits.


Human natural killer cells and cytotoxic T lymphocytes require cell surface carbohydrate determinants for lytic function.
Cloned and uncloned populations of natural killer (NK) cells and cytotoxic T lymphocytes (CTL) were treated with tunicamycin, an antibiotic that inhibits N-linked glycosylation, in order to study the potential role of cell surface carbohydrate determinants in lytic function. It is shown that tunicamycin-treated NK and CTL effector cells lose killer function in a dose-dependent manner. This effect is reversible; cells washed free of tunicamycin begin to recover their killer activity within 2 to 3 days after initial treatment. Conjugate experiments indicate that killer-target cell binding is not affected by tunicamycin treatment of the NK cells. It is also shown that tunicamycin treatment of target cells does not significantly affect their ability to be lysed by NK or CTL effector cells. These studies provide evidence that carbohydrate determinants are important in the lytic mechanism of both CTL and NK cells, rather than in specific effector-target cell binding.


A clinicopathologic case study of two patients with pediatric orbital IgG4-related disease.
The purpose of this report is to describe the clinical, radiographica, and histopathologic findings in two pediatric patients with orbital IgG4-related disease.


Effect of the rider position during rising trot on the horse׳s biomechanics (back and trunk kinematics and pressure under the saddle).
Knowledge about the horse-saddle-rider interaction remains limited. The aim of this study was to compare the effect of the rider׳s position at rising trot on the pressure distribution, spine movements, stirrups forces and locomotion of the horse. The horse׳s back movements were measured using IMUs fixed at the levels of thoracic (T6, T12, T16) and lumbar (L2, L5) vertebrae, the pressure distribution using a pressure mat and stirrups forces using force sensors. The horse׳s and rider׳s approximated centres of mass (COM) were calculated using 2D reflective markers. To compare both trot phases (rider seated/rider standing), three horses were trotted at the rising trot by the same rider. Means±SD of each parameter for sitting and standing were compared using a Student׳s t test (p=0.05). Stirrups forces showed two peaks of equal magnitude in every stride cycle for left and right stirrups but increased during the standing phase. Simultaneously, the pressure for the whole mat significantly increased by +3.1kPa during the sitting phase with respect to standing phase. The T12-T16 and T16-L2 angular ranges of motion (ROM) were significantly reduced (-3.2° -1.2°) and the T6-T12 and L2-L5 ROM were significantly increased (+1.7° +0.7°) during sitting phase compared to standing phase. During rising trot, the sitting phase does not only increase the pressure on the horse׳s back but also reduces the back motion under the saddle compared to the standing phase. These results give new insights into the understanding of horse-rider interactions and equine back pain management.


An in vitro embryotoxicity assay based on the disturbance of the differentiation of murine embryonic stem cells into endothelial cells. II. Testing of compounds.
The embryonic stem cell test (EST) developed by Spielmann et al. [Spielmann, H., Pohl, I., Doering, B., Liebsch, M., Moldenhauer, F., 1997. The embryonic stem cell test, an in vitro embryotoxicity test using two permanent mouse cell lines: 3T3 fibroblasts and embryonic stem cells. In Vitro. Toxicol. 10, 119-127] is currently the most promising in vitro assay to predict the embryotoxic potential of compounds. In this assay the disturbance of the differentiation of embryonic stem (ES) cells into contracting cardiomyocytes by test compounds as well as the direct cytotoxicity of the test compounds on ES cells and 3T3 fibroblasts is analyzed. On the basis of these results and by applying a biostatistical prediction model (PM) [Genschow, E., Scholz, G., Brown, N., Piersma, A., Brady, M., Clemann, N., Huuskonen, H., Paillard, F., Bremer, S., Becker, K., Spielmann, H., 2000. Development of prediction models for three in vitro embryotoxicity tests in an ECVAM validation study. In Vitr. Mol. Toxicol. 13, 51-66; Genschow, E., Spielmann, H., Scholz, G., Pohl, I., Seiler, A., Clemann, N., Bremer, S., Becker, K., 2004. Validation of the embryonic stem cell test in the international ECVAM validation study on three in vitro embryotoxicity tests. Altern. Lab. Anim. 32, 209-244; Genschow, E., Spielmann, H., Scholz, G., Seiler, A., Brown, N., Piersma, A., Brady, M., Clemann, N., Huuskonen, H., Paillard, F., Bremer, S., Becker, K., 2002. The ECVAM international validation study on in vitro embryotoxicity tests: results of the definitive phase and evaluation of prediction models. European Centre for the Validation of Alternative Methods. Altern. Lab. Anim. 30, 151-176] test compounds can be classified as non-embryotoxic, weakly or strongly embryotoxic. In order to introduce a further endpoint into the EST, the disturbance of vasculogenesis and/or angiogenesis, a protocol to differentiate ES cells into endothelial cells, was established in the accompanying paper. PECAM-1 and VE-Cadherin gene expressions, quantified by real-time TaqMan PCR, were shown to be appropriate molecular markers for the differentiation of ES cells into endothelial cells. In the present study, the disturbance of the differentiation of ES cells into endothelial cells (i.e. the reduction in the expression of PECAM-1 and VE-Cadherin) by six test compounds with known embryotoxic potential was investigated: all-trans-retinoic acid (RA) and 5-fluorouracil (5-FU) are strongly embryotoxic, diphenylhydantoin (DPH) and valproic acid (Val) are weakly embryotoxic and saccharin (Sacch) and penicillin G (Pen G) are non-embryotoxic. In a first step the concentration of the test compound resulting in a 50% inhibition of PECAM-1 and VE-Cadherin gene expression and the concentration leading to a 50% decrease in the viability of ES cells and 3T3 fibroblasts were determined. In a second step and in a first attempt to assess the predictive potential of the newly developed test system the concentration values obtained were applied in the PM of the established EST to classify the selected test compounds. All six test compounds were correctly classified (i.e. the data obtained in vitro correlated with their known embryotoxic potential in vivo). Taken together it can be concluded that the disturbance of the differentiation of murine ES cells into endothelial cells represents a very promising new endpoint in a broadened EST with PECAM-1 and VE-Cadherin as specific differentiation marker genes.


Integrated proteogenomic characterization of urothelial carcinoma of the bladder.
Urothelial carcinoma (UC) is the most common pathological type of bladder cancer, a malignant tumor. However, an integrated multi-omics analysis of the Chinese UC patient cohort is lacking. We performed an integrated multi-omics analysis, including whole-exome sequencing, RNA-seq, proteomic, and phosphoproteomic analysis of 116 Chinese UC patients, comprising 45 non-muscle-invasive bladder cancer patients (NMIBCs) and 71 muscle-invasive bladder cancer patients (MIBCs). Proteogenomic integration analysis indicated that SND1 and CDK5 amplifications on chromosome 7q were associated with the activation of STAT3, which was relevant to tumor proliferation. Chromosome 5p gain in NMIBC patients was a high-risk factor, through modulating actin cytoskeleton implicating in tumor cells invasion. Phosphoproteomic analysis of tumors and morphologically normal human urothelium produced UC-associated activated kinases, including CDK1 and PRKDC. Proteomic analysis identified three groups, U-I, U-II, and U-III, reflecting distinct clinical prognosis and molecular signatures. Immune subtypes of UC tumors revealed a complex immune landscape and suggested the amplification of TRAF2 related to the increased expression of PD-L1. Additionally, increased GARS, related to subtype U-II, was validated to promote pentose phosphate pathway by inhibiting activities of PGK1 and PKM2. This study provides a valuable resource for researchers and clinicians to further identify molecular pathogenesis and therapeutic opportunities in urothelial carcinoma of the bladder.


Synthetic biology approaches to access renewable carbon source utilization in Corynebacterium glutamicum.
Corynebacterium glutamicum (C. glutamicum), an important industrial workhorse, is capable of efficiently producing a variety of value-added chemicals and fuels beyond amino acids. C. glutamicum has a broad natural substrate spectrum and can simultaneously utilize various carbon sources in blends. The substrate spectrum of C. glutamicum has been further extended by detailed knowledge of carbon core metabolism and well-established genetic tools and engineering strategies. At present, many pathways have been successfully engineered in C. glutamicum for access to alternative renewable sources to produce natural or non-natural products, making C. glutamicum a promising and favorable microbial cell factory. In this review, we mainly focus on synthetic biology and metabolic engineering strategies for developing synthetic strains that grow on renewable sources to produce the target products. At the same time, we also explore the promotion and future challenges of existing synthetic biology platforms for industrial platform microorganism metabolic engineering efforts.


Synthesis of methyl 4'-O-methyl-13C12-beta-D-cellobioside from 13C6-D-glucose. Part 1: reaction optimization and synthesis.
A high yielding synthetic route for methyl 4'-O-methyl-beta-D-cellobioside starting from d-glucose was established. The reaction conditions optimized with nonlabeled materials were used for the synthesis of methyl 4'-O-methyl-13C12-beta-D-cellobioside, a compound having more than 99% 13C enrichment at each of the twelve pyranose carbon atoms. The labeled compound is required to study the hydrogen bond network of cellodextrins and cellulose by CPMAS NMR experiments.


Interdisciplinary approach for somatoprosthetic rehabilitation of a patient with clino-syndactyly and unusual dermatoglyphics.
Traumatic injuries are the most common cause of finger amputations. Therapeutic amputations are less common but may be essential in some patients with congenital malformations of digits. A 21-year-old female patient reported for esthetic management of malformed middle and ring finger of right hand. Examination revealed clino-syndactyly with unusual dermatoglyphics. She was successfully rehabilitated by collaborative efforts of plastic surgeon and prosthodontist, by therapeutic amputation with surgical recontouring and fabrication of somatoprosthesis. The patient was satisfied with esthetics of the prosthesis and somatoprosthesis improved function. At 6-month recall, no alterations or repair of prosthesis was required. Individualized treatment planning with integration of modifications involving a multidisciplinary approach is required to meet needs of patients with congenital digital malformations. The duplication of unusual dermatoglyphics made the prosthesis more realistic. An individualized treatment plan with modifications to conventional techniques is required to cater to rehabilitative needs of patients with congenital digital malformations.


The wandering Mirena: laparoscopic retrieval.
Levonorgestrel-containing intrauterine contraceptive devices, marketed as Mirena (Bayer HealthCare Pharmaceuticals, Inc. Australia) are widely used in contemporary gynecology, primarily as an effective method for contraception and for control of menstrual disorders like menorrhagia and dysmenorrhea. In this article, the authors report 2 cases of Mirena migration following intrauterine insertion by general practitioners (family physicians). In the first case, the contraceptive device had moved to the patient's right iliac fossa just anterior to the cecum and, in the second, within the peritoneal cavity close to the left leaf of the diaphragm. Both patients underwent uneventful laparoscopic retrieval of the devices.


Autoimmune hemolytic anemia caused by anti "e": A challenge: A case report with review of literature.
Autoimmune hemolytic anemia (AIHA) is featured by short red cell survival due to autoantibodies. AIHA caused by anti 'e' is a tough clinical situation as antigen 'e' is a highly prevalent antigen. The present case highlights the same and different issues related to it.


[Genetic mechanisms of cell transdifferentiation].
Studies have been considered, which concern identification of regulatory genes in adult newts and their expression during retinal and lens regeneration. B.L. Astaurov repeatedly urged to join efforts of geneticists and embryologists in studies of the mechanisms underlying biological phenomena. This was also true for studies of regeneration. Such studies became possible only after introduction of molecular biology methods. Studies of the mechanisms underlying regeneration have been recently carried out jointly by geneticists and developmental biologists. This review presented at the conference dedicated to the 100th anniversary of B.L. Astaurov deals with these aspects in studies of regeneration.


[Incidence of intravenous colistin nephrotoxicity in hospitalized patients].
The increase in infections with multidrug resistant bacteria has forced to return to the use of colistin, antibiotic with known nephrotoxicity. The aim of the study is to determine the incidence of colistin nephrotoxicity nowadays. Retrospective-observational-unicentric study was collected hospitalized patients in intravenous colistin treatment during the years 2018-2019. Nephrotoxicity was defined according to the RIFLE scale. The variables to determine it were serum creatinine (sCr) and glomerular filtration (GF). The variables analyzed were age, sex, treatment duration, loading and cumulative dose, empirical/targeted treatment, chronic kidney disease, concomitant use of intravenous contrast and nephrotoxic drugs. A total of 90 patients (60% men) were included, with an average age of 58.2±18.1 years. The mean duration of treatment was 9±8.3 days, with an average cumulative dose of 69.8±71MU. There were no differences between sCr and GF at the beginning and end of treatment. The incidence of nephrotoxicity was 1.73 cases/100 days of treatment (prevalence of 15.56%). Colistin nephrotoxicity has an important incidence, without developing severe illness.


A genotype-phenotype correlation study in a group of 54 patients with X-linked agammaglobulinemia.
X-linked (Bruton's) agammaglobulinemia (XLA) is a rare immunodeficiency caused by a block in B-cell development caused by mutations in the Bruton's tyrosine kinase (BTK) gene. Many aspects of XLA and BTK function remain unresolved; atypical presentations have been reported, and no clear genotype-phenotype correlation has been established. We sought to contribute to the understanding of XLA through the phenotypic and biochemical characterization of a large group of Spanish patients with agammaglobulinemia. We also sought to classify the mutations according to their severity to analyze a genotype-phenotype correlation. Clinical and analytic data were collected from the clinical records. We studied the BTK gene, protein expression, and function, and the findings were correlated with the phenotypic information. Fifty-four patients were given diagnoses of XLA. We identified 38 different mutations in BTK, 26 not described in other patients, and several uncommon clinical phenotypes or analytic characteristics were found. The statistical analysis shows that less severe mutations or minimal detection of protein by means of flow cytometry are associated with decreased severity in clinical and analytic data, demonstrating a clear relation between the type of mutation and the disease expressivity. However, some exemptions to this rule were noted. XLA is a variable disease. Globally, a genotype-phenotype correlation is observed, but individual discrepancies between the severity of the mutation and the clinical and analytic phenotype suggest that other loci or ambient factors significantly influence the disease presentation and evolution.


Toward a comprehensive view of the skills involved in word reading in Grades 4, 6, and 8.
Research to date has proposed four main variables involved in reading development: phonological awareness, naming speed, orthographic knowledge, and morphological awareness. Although each of these variables has been examined in the context of one or two of the other variables, this study examines all four factors together to assess their unique contribution to reading. A sample of children in Grades 4, 6, and 8 (ages 10, 12, and 14 years) completed a battery of tests that included at least one measure of each of the four variables and two measures of reading accuracy. Phonological awareness, orthographic knowledge, and morphological awareness each contributed uniquely to real word and pseudoword reading beyond the other variables, whereas naming speed did not survive these stringent controls. The results support the sustained importance of these three skills in reading by older readers.


Designing, structural determination and biological effects of rifaximin loaded chitosan- carboxymethyl chitosan nanogel.
Due to the poor solubility and permeability of rifaximin (RFX), it is not effective against intracellular pathogens although it shows strong activity against most bacteria. To develop an effective mucoadhesive drug delivery system with a targeted release in bacterial infection site, RFX-loaded chitosan (CS)/carboxymethyl-chitosan (CMCS) nanogel was designed and systematically evaluated. FTIR, DSC, and XRD demonstrated that the nanogel was formed by interactions between the positively charged NH3+ on CS and CMCS, and the negatively charged COO on CMCS. RFX was encapsulated into the optimized nanogel in amorphous form. The nanogel was a uniform spherical shape with a mean diameter of 171.07 nm. It had excellent sustained release, strong mucin binding ability, and pH-responsive properties of quicker swelling and release at acidic pH. It showed low hemolytic ratio and high antioxidant activity. The present investigation indicated that the CS-nanogel could be potentially used as a promising bacterial responsiveness drug delivery system.