Early Mitochondrial Fragmentation and Dysfunction in a Drosophila Model for Alzheimer's Disease.
Many different cellular systems and molecular processes become compromised in Alzheimer's disease (AD) including proteostasis, autophagy, inflammatory responses, synapse and neuronal circuitry, and mitochondrial function. We focused in this study on mitochondrial dysfunction owing to the toxic neuronal environment produced by expression of Aβ42, and its relationship to other pathologies found in AD including increased neuronal apoptosis, plaque deposition, and memory impairment. Using super-resolution microscopy, we have assayed mitochondrial status in the three distinct neuronal compartments (somatic, dendritic, axonal) of mushroom body neurons of Drosophila expressing Aβ42. The mushroom body neurons comprise a major center for olfactory memory formation in insects. We employed calcium imaging to measure mitochondrial function, immunohistochemical and staining techniques to measure apoptosis and plaque formation, and olfactory classical conditioning to measure learning. We found that mitochondria become fragmented at a very early age along with decreased function measured by mitochondrial calcium entry. Increased apoptosis and plaque deposition also occur early, yet interestingly, a learning impairment was found only after a much longer period of time-10 days, which is a large fraction of the fly's lifespan. This is similar to the pronounced delay between cellular pathologies and the emergence of a memory dysfunction in humans. Our studies are consistent with the model that mitochondrial dysfunction and/or other cellular pathologies emerge at an early age and lead to much later learning impairments. The results obtained further develop this Drosophila model as a useful in vivo system for probing the mechanisms by which Aβ42 produces mitochondrial and other cellular toxicities that produce memory dysfunction.


Nutritional and management strategies for the prevention of fatty liver in dairy cattle.
Fatty liver occurs in dairy cattle during periods of elevated blood non-esterified fatty acids (NEFAs). Elevated blood NEFAs are associated with hormonal changes at parturition and negative energy balance. Approaches for preventing fatty liver include inhibition of fatty acid mobilization from adipose tissues and altering hepatic metabolism to enhance fatty acid oxidation or export as a constituent of very low-density lipoproteins (VLDL). Nutritional and management strategies to implement these approaches have been examined. Increasing energy density of diet, either by increasing non-fiber carbohydrate or fat, has failed to prevent fatty liver. Two nutritional supplements, ruminally-protected choline and propylene glycol, have proven effective at preventing fatty liver. Choline probably enhances hepatic VLDL secretion. Propylene glycol most likely reduces fatty acid mobilization from adipose tissue. Shortening or eliminating the dry period is a management strategy that reduces the magnitude of negative energy balance after calving and triglyceride accumulation in the liver.


The effect of the properties of cell nucleus and underlying substrate on the response of finite element models of astrocytes undergoing mechanical stimulations.
Astrocyte cells play a critical role in the mechanical behaviour of the brain tissue; hence understanding the properties of Astrocytes is a big step toward understanding brain diseases and abnormalities. Conventionally, atomic force microscopy (AFM) has been used as one of the most powerful tools to characterize the mechanical properties of cells. However, due to the complexities of experimental work and the complex behaviour of living cells, the finite element method (FEM) is commonly used to estimate the cells' response to mechanical stimulations. In this study, we developed a finite element model of the Astrocyte cells to investigate the effect of two key parameters that could affect the response of the cell to mechanical loading; the properties of the underlying substrate and the nucleus. In this regard, the cells were placed on two different substrates in terms of thickness and stiffness (gel and glass) with varying properties of the nucleus. The main achievement of this study was to develop an insight to investigate the response of the Astrocytes to mechanical loading for future studies, both experimentally and computationally.


Selective permeation and organic extraction of recombinant green fluorescent protein (gfpuv) from Escherichia coli.
Transformed cells of Escherichia coli DH5-alpha with pGFPuv, induced by IPTG (isopropyl-beta-d-thiogalactopyranoside), express the green fluorescent protein (gfpuv) during growth phases. E. coli subjected to the combination of selective permeation by freezing/thawing/sonication cycles followed by the three-phase partitioning extraction (TPP) method were compared to the direct application of TPP to the same culture of E. coli on releasing gfpuv from the over-expressing cells. Cultures (37 degrees C/100 rpm/ 24 h; mu = 0.99 h(-1)-1.10 h(-1)) of transformed (pGFP) Escherichia coli DH5-alpha, expressing the green fluorescent protein (gfpuv, absorbance at 394 nm and emission at 509 nm) were sonicated in successive intervals of sonication (25 vibrations/pulse) to determine the maximum amount of gfpuv released from the cells. For selective permeation, the transformed previously frozen (-75 degrees C) cells were subjected to three freeze/thaw (-20 degrees C/ 0.83 degrees C/min) cycles interlaid by sonication (3 pulses/6 seconds/25 vibrations). The intracellular permeate with gfpuv in extraction buffer (TE) solution (25 mM Tris-HCl, pH 8.0, 1 mM beta-mercaptoethanol beta-ME, 0.1 mM PMSF) was subjected to the three-phase partitioning (TPP) method with t-butanol and 1.6 M ammonium sulfate. Sonication efficiency was verified on the application to the cells previously treated by the TPP method. The intra-cell releases were mixed and eluted through methyl HIC column with a buffer solution (10 mM Tris-HCl, 10 mM EDTA, pH 8.0). The sonication maximum released amount obtained from the cells was 327.67 microg gfpuv/mL (20.73 microg gfpuv/mg total proteins-BSA), after 9 min of treatment. Through the selective permeation by three repeated freezing/thawing/sonication cycles applied to the cells, a close content of 241.19 microg gfpuv/mL (29.74 microg gfpuv/mg BSA) was obtained. The specific mass range of gfpuv released from the same cultures, by the three-phase partitioning (TPP) method, in relation to total proteins, was higher, between 107.28 microg/mg and 135.10 microg/mg. The selective permeation of gfpuv by freezing/thawing/sonication followed by TPP separation method was equivalent to the amount of gfpuv extracted from the cells directly by TPP; although selective permeation extracts showed better elution through the HIC column.


Severe localised periodontitis in an adult: initiation by orthodontic rubber bands.
An adult patient presented with advanced periodontitis around teeth 11 and 21. This was found to have been caused by the patient placing orthodontic bands 1 year previously around these teeth to close a mid-line diastema.


Diagnostic and predictive value of voiding diary data versus prostate volume, maximal free urinary flow rate, and Abrams-Griffiths number in men with lower urinary tract symptoms suggestive of benign prostatic hyperplasia.
To investigate the information of voiding data in relation to symptoms and well-being in men with lower urinary tract symptoms (LUTS) suggestive of benign prostatic hyperplasia (BPH) and to compare this information with that of prostate volume (Vprostate), maximal free urinary flow rate (Q(max,free)), and obstruction grade (OG). We performed mandatory tests, recommended tests, and pressure-flow studies in 384 consecutive men with LUTS suggestive of BPH. We estimated nocturia, diuria, and mean voided volume (Vmean) from their voiding diaries. Symptoms and well-being were quantified by American Urological Association symptom index (SI), quality-of-life score (QoL), symptom problem index (SPI), and BPH impact index (BII). We investigated the influence of Vprostate, Q(max,free), OG, Vmean, nocturia, and diuria on SI, QoL, SPI, and BII. We re-evaluated 48 men 6 months after transurethral resection of the prostate (TURP). We analyzed the predictive value of preoperative Q(max,free), Vprostate, OG, Vmean, nocturia, and diuria for the improvements of SI, QoL, SPI, and BII after TURP. We studied the improvements of Q(max,free), OG, Vmean, nocturia, and diuria after TURP and the improvements of SI, QoL, SPI, and BII. Prostate volume, Q(max,free), and OG were only slightly associated with SI, QoL, SPI, and BII, in contrast to Vmean, nocturia, and diuria. The predictive value of all parameters on the outcome of TURP was poor. Improvements of all parameters were strongly associated with improvements of SI, QoL, SPI, and BII after TURP. Voiding data should have a prominent role in the initial evaluation of men with LUTS suggestive of BPH.


miRNA-216 and miRNA-499 target cyb561d2 in zebrafish in response to fipronil exposure.
MicroRNA (miRNA) can regulate the expression of its target gene by mediating mRNA cleavage or by translational repression at a post-transcriptional level. Usually, one miRNA may regulate many genes as its targets, while one gene may also be targeted by many miRNAs. We previously demonstrated that cyb561d2, whose protein product is involved in cell defense, and chemical stress, is targeted by miR-155 in adult zebrafish (Danio rerio) when exposed to fipronil (5-amino-1-[2,6-dichloro-4-(trifluoromethyl) phenyl]-4-[(trifluoromethyl) sulphinyl]-1H-pyrazole-3-carbonitrile). Microcosm Targets prediction showed that the cyb561d2 gene is also highly possibly targeted by miR-194a, miR-216b, miR-429, and miR-499. These interactions need to be further validated experimentally. In this study, we evaluated the effects of fipronil on miR-194a, miR-216b, miR-429, miR-499 and cyb561d2 in zebrafish and investigated whether these four miRNAs could regulate the expression of cyb561d2 in both mRNA and protein levels. The expression of cyb561d2 was upregulated in both mRNA and protein level in a dose-dependent manner upon stimulation of fipronil, and miR-216b and miR-499 were downregulated concurrently, whereas there was no significant changes were observed in the expression level of miR-194a and miR-429. The dual luciferase report assay demonstrated that miR-216b and miR-499 interacted with cyb561d2 3'-untranslated regions (3'-UTR), miR-194a and miR-429 did not stimulate degradation of cyb561d2 mRNA. The expression of cyb561d2 was reduced in both mRNA and protein level when ZF4 cells were transfected with miR-499 mimic, whereas expression level of both mRNA and protein was increased when endogenous miR-499 was inhibited by transfection with miR-499 inhibitor. Likewise, the mRNA and protein level of cyb561d2 was affected by treatment with the mimics and the inhibitor of miR-216b. In contrast, when ZF4 cells were transfected with a mimic of miR-194a or miR-429, the expression of cyb561d2 mRNA was not significantly changed. As a result, cyb561d2 is targeted by miR-155, miR-216b and miR-499 upon fipronil exposure, and miR-194a and miR-429 can not target cyb561d2. The expression pattern of these 3 miRNAs presents novel fipronil responses that could be used as a toxicological biomarker.


Characteristics of nonparticipants differed based on reason for nonparticipation: a study involving the chronically ill.
Individuals who do and do not participate in studies often have different characteristics. The profile of nonparticipants may vary by type of nonparticipation. Few cross-sectional or longitudinal studies of chronically ill individuals identify these differences at baseline and of those that do; many do not have information on type of nonparticipation. We compared characteristics of chronically ill individuals to determine if they differ by type of nonparticipation. Data are from an evaluation of a chronic disease management program conducted in New South Wales, Australia during 2001 and 2002. Characteristics were compared using routinely collected hospital data. Reasons for nonparticipation were categorized as refusal, death, or untraceable. Individuals who refused to participate were older, female, discharged at risk, and have heart failure. Those untraceable were younger, not married, Indigenous, and receiving care during the intervention phase but not recruited. Individuals not participating due to death were older, male, not cohabiting, discharged at risk, and have a diagnosis of cancer or heart failure. The significant differences between untraceable, refused, and deceased individuals should be considered when designing studies and when adjusting for nonparticipation bias.


Training and Delivery of Critical Care Medicine in India: Concerns Revealed by COVID-19 Pandemic.
How to cite this article: Ozair A, Agrawal A, Siddiqui SS. Training and Delivery of Critical Care Medicine in India: Concerns Revealed by COVID-19 Pandemic. Indian J Crit Care Med 2020;24(4):285-286.


From there to here: A journey through substance use disorder, prison, and recovery.
This narrative describes my experience with substance use disorder. It includes a trip through the criminal legal system, recovery, higher education, and a career in research. I convey key points from a history of using substances and committing crimes and how these created barriers to collegiate and professional aspirations, despite long-term recovery. The substance use services research field has come to value lived experience such as mine, lowering barriers for people with substance use and criminal histories. I hope that my work, built upon this experience, not only helps improve services, but also clears the way for more people in recovery to pursue a career in research.


The preparation and characterization of mono-iodinated photoreactive analogs of insulin.
Photoreactive derivatives of insulin (B29-(p-azidobenzoyl-insulin) iodinated primarily in either the B26 or A14 tyrosine of insulin were prepared by lactoperoxidase catalyzed iodination followed by separation on reverse-phase high-performance liquid chromatography. The binding affinities and photoaffinity labeling characteristics of these derivatives were studied in isolated rat adipocytes. Under nonreducing conditions, three forms of the insulin receptor were labeled equally by the B26-derivative, the A14-derivative, and the mixture of the iodinated derivatives. When dithiothreitol was used to reduce the radiolabeled receptors, the radioactivity associated with the binding subunit was much less than that in the intact receptor and the magnitude of the decrease was proportional to the amount of iodine in the A chain of the photoderivatives. Use of the photoreactive derivative iodinated primarily in the B26 position resulted in greater labeling of insulin receptor subunits since most of the radioactivity (80%) remained associated with the receptor upon reduction.


Antibodies to rheumatoid arthritis nuclear antigen (RANA) in Japanese patients with rheumatoid arthritis.
We studied antibodies to rheumatoid arthritis nuclear antigen (RANA) by the Ouchterlony method in 0.5% agarose plates, using soluble antigen extracted with 0.25 M sucrose solution from cultured Raji cells. Anti-RANA antibody was found in sera from 24 to 40 (60%) patients with rheumatoid arthritis (RA), from 4 of 20 (20%) patients with systemic lupus erythematosus (SLE), and from 2 of 30 (7%) healthy controls. When sucrose extracts from BJAB, Ramos, and JM cells were used as the cellular antigens, no anti-RANA precipitin lines were detected. Indirect immunofluorescence study, using Raji cells or human B lymphocytes transformed by EB virus as substrate tissues, demonstrated anti-RANA antibody as fine speckled nuclear staining. Although RA patients with positive anti-RANA antibody usually had high titers of anti-Epstein-Barr nuclear antigen (EBNA) and anti-viral capsid antigen (VCA) IgG antibodies, the Wilcoxon ranks sum test showed no close statistical correlation between the presence of anti-RANA antibodies and the titers of anti-EBNA or anti-VCA (IgG) antibodies. These data showed that the incidence of positivity of anti-RANA antibodies in Japanese RA patients is almost the same as that in American and European RA patients.


Analysis on the attenuation characteristics of PPCPs in surface water and their influencing factors based on a compilation of literature data.
The attenuation characteristics of PPCPs play an important part in predicting their environmental concentrations. However, considerable uncertainty remains in reported laboratory data on the attenuation characteristics of PPCPs. In this analysis, we compile information on laboratory-observed photodegradation half-lives (t1/2), biodegradation t1/2, the organic carbon normalized adsorption constant (KOC) and field-observed overall attenuation t1/2 for PPCPs in water bodies from more than 200 peer-reviewed studies. To mitigate the effects of such uncertainty, we derive representative values (RV) for PPCP degradability from these records to better compare the characteristics of different PPCPs. We further examine the influence of experimental conditions and environmental drivers on the determination of t1/2 using difference analysis and correlation analysis. The results indicate that for laboratory photodegradation tests, different light sources, initial concentration and volume significantly affect t1/2, whereas there is no significant difference between values obtained from tests conducted in pure water and natural water. For biodegradation, laboratory-measured t1/2 values in batch, flume and column studies gradually decrease, marking the controlling role of experimental setup. Redox condition, initial concentration and volume are also recognized as important influencing factors. For adsorption, water-sediment ratio is the primary reaction parameter. As two frequently investigated factors, however, pH and temperature are not significant factors in almost all cases. In field observations, the persistence of carbamazepine, typically used as a tracer, is in doubt. Water depth and latitude are the most correlated drivers of t1/2, indicating the predominant status of photodegradation in the overall attenuation rates. These findings call for caution when selecting experimental parameters and environmental drivers in determining PPCP's attenuation rates and establishing PPCP fate models in the field.


Risk Factors Related to COVID-19 Reinfection and Fatality During the Omicron (BA.1/BA.2) Period in Korea.
This study aimed to investigate the deaths due to coronavirus disease 2019 (COVID-19) reinfection and related risk factors. National cohort data were collected for a six-month period when omicron BA.1/BA.2 variant was dominant in South Korea. The long-term care facility residents (adjusted odds ratio, 3.11; 95% confidence interval [CI], 2.98-3.25) had significantly higher risk of reinfection than the general population. The risk of reinfection was significantly lower for persons with 2 or more vaccine doses compared to the unvaccinated. The risk of death was significantly higher in the reinfection group than in the primary infection group for persons in the 60-74 years age group (adjusted relative risk [aRR], 1.62; 95% CI, 1.19-2.20), and immunocompromised group (aRR, 4.56; 95% CI, 2.34-8.90). In these data, vaccination history was significantly related to reduced COVID-19 reinfection and severe progression, and scheduled vaccinations were important even with a history of infection.


The genome of anaplasma: DNA base composition and DNA/DNA hybridization.
The Tm value of DNA from Anaplasma centrale and Anaplasma marginale was found to be 87.1 degrees C and 89.3 degrees C, respectively. The G + C content, calculated from the Tm, was 45.1% for A. centrale and 48.5% for A. marginale. Identical hybridization patterns were obtained when the DNA from one species was hybridized to restriction endonuclease-digested DNA from the other species.


Predictive Drug Release Modeling Across Dissolution Apparatuses I and II using Computational Fluid Dynamics.
A modeling process is developed and validated with which active pharmaceutical ingredient (API) release is predicted across the United States Pharmacopeia (USP) dissolution apparatuses I and II based on limited experimental dissolution data (at minimum two dissolution profiles at different apparatus settings). The process accounts for formulation-specific drug release behavior and hydrodynamics in the apparatuses over the range of typical agitation rates and medium volumes. This modeling process involves measurement of experimental mass transfer coefficients via a conventional mass balance and the relationship of said mass transfer coefficients to hydrodynamics and apparatus setting via computational fluid dynamics (CFD). A novel 1-D model is hence established, which provided calibration data for a particular formulation, can model mass transfer coefficients and their corresponding drug release at apparatus configurations of interest. Based on validation against experimental data produced from five erosion-based formulations over a range of apparatus configurations, accuracy within 8 %LA (labelled amount of API) and an average root mean square deviation of 3 %LA is achieved. With this predictive capability, minimizing the number of dissolution experiments and the amount of chemical materials needed during method development appears feasible.


DNT cell inhibits the growth of pancreatic carcinoma via abnormal expressions of NKG2D and MICA in vivo.
This research aimed to investigate the effects of natural killer group 2 member D (NKG2D) and its ligands major histocompatibility complex class I chain-related molecules A(MICA) in DNT cell killing pancreatic carcinoma. Antibodies adsorption was used to separate DNT cell from human peripheral blood. Human pancreatic tumor models were established via implanting BXPC-3 cells into nude mice. Then randomly divided mice into blank group, gemcitabine group and DNT group. Mice weights and mice tumor volumes were measured every 5 days. 50 days later mice were euthanized at cervical dislocation method. Tumor weights were measured. Relative tumor volume and tumor inhibition rate were calculated. Western blot and qPCR were used to detect the expressions of NKG2D and MICA in the transplanted tumors of the three groups. DNT cell significantly increased over time. The blank group tumor volume and weight were significantly larger than the other groups (p < 0.001, p < 0.001), but there were no significantly difference between DNT group and gemcitabine group (p > 0.05). Gemcitabine and DNT cell tumor inhibition rate were 40.4% and 35.5%. Western blot and qPCR showed that MICA mRNA and protein levels in blank group were significantly higher than DNT group (p = 0.001, p = 0.003). NKG2D mRNA and protein levels in blank group were significantly lower than DNT cells group (p < 0.001, p = 0.001). In conclusion DNT cell can significantly inhibit the growth of pancreatic carcinoma in vivo, and the mechanism may be involved in abnormal expressions of MICA and NKG2D.


The Program for the Control of Visceral Leishmaniasis in Brazil: The Effect of the Systematic Euthanasia of Seropositive Dogs as a Single Control Action in Porteirinha, a Brazilian City with an Intense Transmission of Visceral Leishmaniasis.
Porteirinha is endemic for visceral leishmaniasis (VL), with intense disease transmission of the disease. We evaluated the impact of canine euthanasia as a single control measure on the incidence of VL in humans and canines. A prospective observational cohort study was carried out over four years (1998-2002) in 8 of the 12 neighborhoods of the city. The dynamics of canine visceral leishmaniasis (CVL) transmission were evaluated for 2 years, before beginning the screening-culling intervention. The comparative morbidity index (CMI) was used to stratify areas with the greatest risk of CVL, and the spatial distribution of human and canine VL cases was compared using univariate and bivariate K-functions. Human cases conglomerated in three neighborhoods. Spatial clusters were detected for CVL in 1998, 2000, and 2001, but not in 1999, when greater spatial dispersion occurred. The screening and culling intervention reduced the number of human VL cases and decreased the incidence of CVL, mainly in neighborhoods with a high CMI. The systematic euthanasia of seropositive dogs was shown to be an effective control action of the Program for Control of Visceral Leishmaniasis (PCLV) in Brazil. The fundamental role of domestic dogs in the epidemiological chain of VL was reaffirmed.


Liver Macrophages Stimulate the Expression of Hepatocyte Nuclear Factor-6 and Promote Hepatocyte Proliferation at the Early Stage of Liver Regeneration.
Hepatocyte nuclear factor (HNF-6) is a liver-specific protein and a key component in the differentiation process during the development of mature liver. The immunohistochemical staining and RT-PCR techniques were employed to examine the expression of HNF-6 and proliferation of Ki-67+ cells during the early regeneration of the liver on postsurgery in 3, 6, 12, and 24 h in original model of partial hepatectomy in rats. The earliest proliferating (Ki-67+) cells were observed in 3 h after surgery in liver sinusoids (liver macrophages) and then in liver parenchyma. Expression of HNF-6 in hepatocytes and epithelial cells of the bile ducts attained maximum in 6 h after surgery. At later terms, this parameter somewhat decreased, but still surpassed the control level.


Evolutionary history exposes radical diversification among classes of interaction partners of the MLLE domain of plant poly(A)-binding proteins.
Poly(A)-binding proteins (PABPs) are evolutionarily conserved proteins that have important functions in the regulation of translation and the control of mRNA stability in eukaryotes. Most PABPs encode a C-terminal domain known as the MLLE domain (previously PABC or CTC), which can mediate protein interactions. In earlier work we identified and predicted that four classes of MLLE-interacting proteins were present in Arabidopsis thaliana, which we named CID A, B, C, and D. These proteins encode transcription-activating domains (CID A), the Lsm and LsmAD domains of ataxin-2 (CID B), the CUE and small MutS-related domains (CID C), and two RNA recognition domains (CID D). We recently found that a novel class that lacks the LsmAD domain is present in CID B proteins. We extended our analysis to other classes of CIDs present in the viridiplantae. We found that novel variants also evolved in classes CID A and CID C. A specific transcription factor domain is present in a distinct lineage in class A, and a variant that lacks at least two distinct domains was also identified in a divergent lineage in class C. We did not detect any variants in Class D CIDs. This class often consists of four to six highly conserved RNA-binding proteins, which suggests that major redundancy is present in this class. CIDs are likely to operate as components of posttranscriptional regulatory assemblies. The evident diversification of CIDs may be neutral or may be important for plant adaptation to the environment and for acquisition of specific traits during evolution. The fact that CIDs subclasses are maintained in early lineages suggest that a presumed interference between duplicates was resolved, and a defined function for each subclass was achieved.


The medical history of adults with complex congenital heart disease affects their social development and professional activity.
In recent decades, advances in surgery and therapeutic catheterization have steadily increased the life expectancy and prevalence of adults with congenital heart disease (CHD). We assessed medical and psychosocial variables of adults with CHD, according to the disease complexity. We included, from a single-centre observational cohort study, 135 consecutive adults with CHD (median age of 40 years, interquartile range: 28.0-51.0) followed in our cardiology unit, who answered a questionnaire assessing daily activity and psychosocial functioning. Disease complexity was classified according to the Bethesda conference. Cardiac malformation complexity was simple in 61 (45.2%), moderate in 50 (37.0%) and complex in 24 (17.8%) patients. Cardiac surgery had been performed in 86.5% of moderate and complex patients. Complications (such as heart failure, arrhythmia and pulmonary hypertension) were mainly observed in the complex group (P=0.003). Physical activity was lower in the complex group (no activity in 58.8%, but sport previously contraindicated in 50% of these; P=0.03). Education level tended to be lower in the complex and moderate groups than in the simple group (respectively, 31.2% and 33.3% vs. 45.7% had passed the Baccalaureate; P=0.47). The pass rate was lower in patients with complications (P=0.037) or more than one cardiac surgery (P=0.03). In the complex group, 56.3% of patients were unemployed (P=0.048). Complexity of heart disease and medical history affect education level and employment of adults with CHD. Academic education of children with a complex defect and career counselling are important to prevent unemployment among adults with CHD.


The malpractice controversy and the quality of patient care.
The widespread doctor strikes of 1975 stimulated belated attention to a crisis in malpractice insurance. Most state legislatures responded only to a shadow crisis in insurance as they rallied to the defense of health care providers. The smouldering substantive crisis--the reality of malpractice--is now galvanizing institutions and professions into aggressive activities for quality assurance and renewal of trust between patient and doctor. New procedural experiments offer prospects for preserving economy and equity by containing the causes of malpractice suits within the health care system itself.


The role of probiotics in the inhibition of Campylobacter jejuni colonization and virulence attenuation.
Campylobacter jejuni is one of the most common bacterial causes of human gastroenterocolitis worldwide, leading to diarrhea and other serious post-infectious complications. Probiotics form an attractive alternative intervention strategy for most of the enteric infections. However, the role of probiotics in C. jejuni infections requires detailed investigations in order to delineate the probiotic strains that are effective against C. jejuni. Although there are several biological mechanisms involved in the inhibition of pathogenic bacterial growth, the strains of probiotics and their mechanisms of actions through which they combat C. jejuni invasion have not been studied in greater detail. This mini review details the factors that are involved in the colonization and establishment of C. jejuni infection, with special reference to chickens, the natural host of C. jejuni, and the studies that have investigated the effect of different probiotic strains against C. jejuni colonization and growth. This review has collated the studies conducted using probiotics to inhibit C. jejuni colonization and growth to date to provide a collective knowledge about the role of probiotics as an alternative intervention strategy for campylobacteriosis.


Acquired Madelung-like deformity in a gymnast.
High-level gymnastic training may cause clinical and radiographic changes in the wrist. A case is reported demonstrating bilateral closure of the ulnar side of the distal radius epiphyseal plate in a patient with a history of high-level gymnastic training. Cumulative microtrauma to the ulnar side of the distal radius epiphyseal plate may cause premature closure leading to a Madelung-like deformity.


Morphology and properties of thermoplastic starch blended with biodegradable polyester and filled with halloysite nanoclay.
The incorporation of halloysite clay nanotubes (HNTs) into thermoplastic starch/poly(butylene adipate-co-terephthalate) (TPS/PBAT) blends has been investigated with the aim of improving the compatibility and properties of the matrix. TPS/PBAT/HNTs nano-biocomposites with different TPS/PBAT weight fractions and HNTs contents were elaborated using a melt blending process, and their morphology and properties were investigated. The TPS80/PBAT20 and TPS20/PBAT80 blends exhibited dispersed phases of small droplets of PBAT or TPS, respectively, whereas the TPS50/PBAT50 blend presented a more homogeneous structure. Elongation at break of the TPS/PBAT/HNTs biocomposites with 5 wt% of HNTs significantly increased with increasing PBAT proportion, i.e., 6.5 %, 41.3 %, and 351.5 % for the composites based on TPS80/PBAT20, TPS50/PBAT50, and TPS20/PBAT80, respectively. The incorporation of 5 wt% of HNTs improved compatibility and increased Young's modulus of the TPS80/PBAT20, TPS50/PBAT50, and TPS20/PBAT80 blends approx. 350 %, 142 %, and 18 %, respectively. These results demonstrate that HNTs are promising nanofillers to improve properties of TPS-based blends.


Functional proteomics can define prognosis and predict pathologic complete response in patients with breast cancer.
To determine whether functional proteomics improves breast cancer classification and prognostication and can predict pathological complete response (pCR) in patients receiving neoadjuvant taxane and anthracycline-taxane-based systemic therapy (NST). Reverse phase protein array (RPPA) using 146 antibodies to proteins relevant to breast cancer was applied to three independent tumor sets. Supervised clustering to identify subgroups and prognosis in surgical excision specimens from a training set (n = 712) was validated on a test set (n = 168) in two cohorts of patients with primary breast cancer. A score was constructed using ordinal logistic regression to quantify the probability of recurrence in the training set and tested in the test set. The score was then evaluated on 132 FNA biopsies of patients treated with NST to determine ability to predict pCR. Six breast cancer subgroups were identified by a 10-protein biomarker panel in the 712 tumor training set. They were associated with different recurrence-free survival (RFS) (log-rank p = 8.8 E-10). The structure and ability of the six subgroups to predict RFS was confirmed in the test set (log-rank p = 0.0013). A prognosis score constructed using the 10 proteins in the training set was associated with RFS in both training and test sets (p = 3.2E-13, for test set). There was a significant association between the prognostic score and likelihood of pCR to NST in the FNA set (p = 0.0021). We developed a 10-protein biomarker panel that classifies breast cancer into prognostic groups that may have potential utility in the management of patients who receive anthracycline-taxane-based NST.


Radiological findings as favorable predictors of pain relief in patients with osteoporotic compression fractures after percutaneous vertebroplasty: a retrospective study of 156 cases.
We evaluated the relationships between pre-procedural radiological findings and short-term pain relief in patients with osteoporotic compression fractures after percutaneous vertebroplasty (PVP). A retrospective review of pre-procedural radiological images of 156 patients with painful osteoporotic compression fracture was performed. Pain was measured with a visual analogue scale (VAS). Complete pain relief was defined as a VAS pain score of 0 or 1 at 3 months after PVP. Statistical analyses were conducted to evaluate the relationship between the pre-procedural imaging factors and pain relief using Pearson's chi-squared test. Multivariate logistic regression analysis was also performed. Complete pain relief was obtained in 45.5% of patients. An intravertebral cleft larger than half the height of the fractured vertebral body (FVB) was a significant key factor in the complete pain relief group after 3 months. Further, ≥40% of the spinal canal occupied by bony fragments of the FVB was related to incomplete pain relief. A large intravertebral cleft was a favorable short-term outcome predictor in patients with osteoporotic compression fractures after PVP, while severe protrusion of the FVB causing lumbar spinal canal stenosis was not a favorable short-term outcome predictor of complete pain relief.


A coaxial nanocable textured by a cerium oxide shell and carbon core for sensing nitric oxide.
A corn-like CeO2/C coaxial cable textured by a cerium oxide shell and a carbon core was designed to sense NO. The carbon core possesses high electrical conductivity, and the CeO2 surface delivers excellent electrocatalytic activity. The sensor, typically operated at 0.8 V (vs. Ag/AgCl), exhibits a detection limit of 1.7 nM, which is 4-times lower than that of CeO2 nanotubes based one (at S/N = 3). It also displays wide linear response (up to 83 μM), a sensitivity of 0.81 μA μM-1 cm-2, and fast response (2 s). These values are highly competitive to that of a CeO2 tube (0.92 μA μM-1 cm-2 and 2 s). The sensor was used to quantify NO that is released by Aspergillus flavus. Graphical abstractSchematic representation of corn-like CeO2/C which can more sensitively and effectively detect NO released from A. flavus than when using CeO2 nanotubes, benefitting from its unique coaxial cable structure.


Pharmacokinetics of prednisolone in children with acute lymphoblastic leukaemia.
The pharmacokinetics of soluble oral prednisolone were studied during induction therapy in six children with acute lymphoblastic leukaemia. There was a three- to four-fold variation in the pharmacokinetics of total and free prednisolone. For total prednisolone, the mean elimination half-life was relatively short (1.37 h) and the total clearance, relatively high (15.1 ml min-1 kg-1). The mean free fraction was high (0.37).


Comparing precision of distortion-compensated and stereophotogrammetric Roentgen analysis when monitoring fusion in the cervical spine.
Two methods to measure sagittal plane segmental motion in the cervical spine are compared. Translational and rotational motion was measured in nine cervical motion segments of nine patients by distortion-compensated (DCRA) as well as by stereophotogrammetric Roentgen analysis (RSA). To compare measurement precision of the new DCRA protocol with the established RSA technique under realistic clinical conditions and to discuss advantages and disadvantages of both methods in clinical studies. RSA constitutes the most precise method available to assess segmental motion or to monitor fusion in the cervical spine. Due to the invasive nature of the procedure there is an interest in alternative, non-invasive protocols, based on conventional, lateral radiographic views. In nine patients, segmental motion of nine cervical segments with spinal surgery and fusion had previously been assessed from stereo views by RSA. From the archive radiographs, sagittal plane segmental motion was re-assessed by DCRA. Results for sagittal plane translational and rotational motion obtained by both methods are compared. With respect to RSA, sagittal plane rotation was determined by DCRA with an error of 2.4 degrees and a mean difference not significantly different from zero. Sagittal plane translation was determined by DCRA with an error of less than 0.78 mm and a mean difference not significantly different from zero. As two methods are compared, these errors represent the combined (propagated) errors of RSA and DCRA. Averaged over the cohort investigated, measurement of sagittal plane segmental motion exhibited no significant difference between DCRA and RSA.


Reduced fading of Fast Blue fluorescence in the brain of the guinea-pig by treatment with sodium-nitroprusside.
Fading of the emitted fluorescent light of the retrogradely transported neuronal tracer Fast Blue is reduced considerably by treating sections for 20 min with sodium-nitroprusside (0.4% in 0.1 M acetate buffer; pH 3.3) and mounting sections from acetate buffer. Mounting from phosphate buffer abolishes the antifading effect of sodium-nitroprusside.


Bioevaluation study of 32P-CP-PLLA particle brachytherapy in a rabbit VX2 lung tumor model.
The purpose of this study was to investigate the therapy effects of intratumoral administration of (32)P-CP-PLLA particles in a rabbit VX2 lung tumor model. 16 rabbits with tumors were randomly divided into 4 groups. 4 rabbits served as untreated controls, and others received intratumoral administration of (32)P-CP-PLLA particles with CT guidance. The total radioactivities in treated groups were as follows: a low activity was 93 MBq (n=4) (group 1), a medium activity was 185 MBq (n=4) (group 2) and a high activity was 370 MBq (n=4) (group 3). Brachytherapy treated VX2 tumors underwent (18)F-FDG PET/CT at 0 day, 3 day, 7 day and 14 day postinjection. In control group, (18)F-FDG PET/CT images were acquired at the same time points but without any treatment. Bremsstrahlung SPECT images were performed at 14 days after intratumoral brachytherapy in treated groups. After Bremsstrahlung SPECT and last (18)F-FDG PET/CT imagings, the rabbits were euthanized and the tumors were removed for histological examination. Bremsstrahlung SPECT images study indicated that there was no leakage of (32)P out of the injection site at 14 days after treatment. Compared with the control, the tumor volumes in treated groups significantly decreased, and (32)P-CP-PLLA particle produced a reduction in maximum or mean SUV of VX2 tumor (p<0.05). The percentage changes in maximum and mean SUV gradually decreased in group 1 and group 2 from day 3 to day 14 (p<0.05). A transient increase in (18)F-FDG accumulation at group 3 occurred due to the inflammatory reaction elements. Activity dependence was seen in HE and PCNA staining after 14 days treatment among three treated groups (p<0.05). Our data suggested that (32)P-CP-PLLA particle localized on the injecting sites. This novel brachytherapy device efficiently suppressed the growth of the VX2 tumors implanted in the rabbit.


Adaptive significance of nitrogen storage in Bistorta bistortoides, an alpine herb.
Studies were conducted to examine the importance of nitrogen storage to seasonal aboveground growth in the alpine herb Bistorta bistortoides. Stored reserves accounted for 60% of the total nitrogen allocated to the shoot during the growing season. The stored nitrogen was equally partitioned between preformed buds of the shoot and the roots/rhizome. Reliance on stored N was similar in populations of a 105-day growing season site and of a 75-day growing season site. Contrary to our initial hypothesis, stored nitrogen reserves were not used to extend the growing season of this species into the late-spring when soils are still cold, and saturated with snow-melt water. The time at which stored nitrogen was used to initiate shoot growth coincided with the time of root initiation, rapid soil warming, and near maximum soil concentrations of NO inf3sup- and NH inf4sup+ . Thus, nitrogen demand and soil nitrogen supply were both high at the same time. The importance of nitrogen storage in this species appeared to be in satisfying the high demand of simultaneous vegetative and reproductive growth during the early-growing season after soils thawed. The initiation of rapid leaf and inflorescence growth occurred in mid-June in both sites. The maximum pool size of shoot nitrogen (maximum nitrogen demand) occurred only 12 days later in the long season site, and 28 days later in the short season site. The early-season utilization of nitrogen stores allows plants of this species to initiate reproductive allocation at the same time vegetative tissues are exhibiting maximal growth rates. By releasing vegetative and reproductive growth from competition for nitrogen, seeds could mature early in the alpine growing season, before the frost probability sharply increases in mid-August.


Inducible expression and pharmacological characterization of the mouse metabotropic glutamate 5b receptor.
The metabotropic glutamate receptor subtype 5 (mGlu5) and glutamatergic neurotransmission are associated with the pathophysiology of disorders such as anxiety, depression or chronic pain. Human and rat mGlu5 receptors have been cloned and characterized previously. We now describe the cloning of the mouse mGlu5b receptor gene from adult mouse brain and its expression using an ecdysone-inducible system. This subtype has an extra 96 bp sequence which is inserted to the cytoplasmic tail and is identical to the insert present in human and rat mGlu5b. Mouse mGlu5b receptor expression was induced in HEK-293EcR cells by incubation with ponasterone A, an analogue of the insect hormone ecdysone. A fluorometric calcium transient assay system was used to characterize the basic pharmacologic profile of an isolated stable cell line. Quisqualic acid was the most potent receptor agonist (EC(50) approximately 7 nM) although the cells also responded to l-glutamic acid and the Group I-selective receptor agonist, 3,5-dihydroxyphenylglycine (3,5-DHPG). The calcium transients stimulated by these agonists were potently inhibited by reference allosteric mGlu5 antagonists - 2-methyl-6-(phenylethynyl)pyridine (MPEP), 3-[(2-methyl-1,3-thiazol-4-yl)ethynyl]pyridine (MTEP) and 3-methoxy-5-(pyridine-2-ylethynyl)pyridine (methoxy-PEPy) (IC(50) ranges: 0.8-66 nM). The availability of this mouse mGlu5b receptor-expressing cell line will facilitate in vitro characterization of mGlu5 receptor-selective agonists or antagonists prior to in vivo pharmacologic testing.


Hydrogen sulfide attenuates hyperhomocysteinemia-induced mitochondrial dysfunctions in brain.
Hyperhomocysteinemia (HHcy) has been implicated in the development of neurodegenerative conditions and mild cognitive disorders. Mitochondrial dysfunctions are the major mechanisms involved in homocysteine (Hcy)-induced neurotoxicity. Although, hydrogen sulfide has been reported as potent antioxidant, its effects on Hcy-induced mitochondrial dysfunctions have not been studied. Therefore, the present study has been designed to evaluate the protective effect of NaHS on Hcy-induced mitochondrial dysfunctions in brain. NaHS supplementation decreased reactive oxygen species and nitrite levels in the cortex and hippocampus of animals with HHcy. NaHS supplementation increased the activity of mitochondrial electron transport chain components; NADH dehydrogenase, cytochrome c oxidase and F0-F1 ATPase in the cortex and hippocampus of HHcy animals along with in-gel activity of complex I - complex V in the mitochondria isolated from the cortex and hippocampus of HHcy animals. Moreover, NaHS supplementation also increased the mitochondrial complex I, II and IV mediated oxygen consumption rate in Hcy treated mitochondria. NaHS administration prevented the Hcy-induced mitochondrial damage as suggested by the decreased mitochondrial swelling in the cortex and hippocampus of HHcy animals. NaHS supplementation decreased the activity of lactate dehydrogenase isozymes (1-5) in the brain regions of HHcy animals. The expression of protein kinase c δ was also decreased in the brain regions of HHcy animals following NaHS supplementation. This was accompanied by reduced activity of caspase-3 indicating anti-apoptotic effect of H2S. Taken together, the findings suggest that H2S supplementation ameliorates Hcy-induced oxidative stress and mitochondrial dysfunctions suggesting H2S releasing drugs may be a novel therapeutic approach to treat HHcy associated neurological disorders.


MFG-E8 inhibits neutrophil migration through αvβ₃-integrin-dependent MAP kinase activation.
We have previously demonstrated the involvement of milk fat globule-epidermal growth factor-factor 8 (MFG‑E8) in reducing neutrophil infiltration in a murine model of acute lung injury (ALI). In the present study, we aimed to delineate the mechanisms through which MFG‑E8 attenuates neutrophil migration. Recombinant human MFG‑E8 (rhMFG‑E8) was expressed and purified in our facility. The human differentiated neutrophil cell line, dHL‑60, was treated with rhMFG‑E8 and cell migration assay was performed in a Boyden chamber using recombinant interleukin‑8 (IL‑8) as the chemoattractant. Surface CXCR2 and intracellular G protein‑coupled receptor kinase 2 (GRK2) levels were evaluated by flow cytometry or western blot analysis. The levels of mitogen‑activated protein (MAP) kinases were determined by western blot analysis. Treatment with rhMFG‑E8 resulted in a significant inhibition of dHL‑60 cell migration in a dose‑dependent manner. There was a 46% decrease in CXCR2 expression in the rhMFG‑E8‑treated dHL‑60 cells, which was associated with a 32% increase in GRK2 expression. In the dHL‑60 cells, treatment with rhMFG‑E8 promoted the phosphorylation of p38 and extracellular signal-regulated kinase (ERK) within 10‑30 min. The use of SB203580, a p38 inhibitor, and PD98059, an ERK inhibitor, resulted in the restoration of dHL‑60 cell migration which was significantly inhibited treatment with rhMFG‑E8. Furthermore, blocking the MFG‑E8 receptors, αvβ3/αvβ5‑integrins, by anti‑αv‑integrin neutralizing antibody (Ab) inhibited the activation of p38 and ERK, and reversed the rhMFG‑E8‑induced inhibition of dHL‑60 cell migration. Finally, treatment of the dHL‑60 cells with SB203580 and PD98059 neutralized the rhMFG‑E8‑induced downregulation of CXCR2 expression and upregulation of GRK2 expression, as well as the inhibitory effects on cell migration. Our findings reveal a novel mechanism of action of MFG‑E8 through which it inhibits neutrophil migration through αvβ3-integrin-dependent MAP kinase activation.


Determination of Femoral Neck Angle and Torsion Angle Utilizing a Novel Three-Dimensional Modeling and Analytical Technology Based on CT Datasets.
Exact knowledge of femoral neck inclination and torsion angles is important in recognizing, understanding and treating pathologic conditions in the hip joint. However, published results vary widely between different studies, which indicates that there are persistent difficulties in carrying out exact measurements. A three dimensional modeling and analytical technology was used for the analysis of 1070 CT datasets of skeletally mature femurs. Individual femoral neck angles and torsion angles were precisely computed, in order to establish whether gender, age, body mass index and ethnicity influence femoral neck angles and torsion angles. The median femoral neck angle was 122.2° (range 100.1-146.2°, IQR 117.9-125.6°). There are significant gender (female 123.0° vs. male 121.5°; p = 0.007) and ethnic (Asian 123.2° vs. Caucasian 121.9°; p = 0.0009) differences. The median femoral torsion angle was 14.2° (-23.6-48.7°, IQR 7.4-20.4°). There are significant gender differences (female 16.4° vs. male 12.1°; p = 0.0001). Femoral retroversion was found in 7.8% of the subjects. Precise femoral neck and torsion angles were obtained in over one thousand cases. Systematic deviations in measurement due to human error were eliminated by using automated high accuracy morphometric analysis. Small but significant gender and ethnic differences were found in femoral neck and torsion angles.


Strain differences in activity and emotionality do not account for differences in learning and memory performance between C57BL/6 and DBA/2 mice.
This study examined emotionality, activity, learning and memory, as well as the influence of emotionality and activity on learning and memory performance in C57BL/6 and DBA/2 mice using a mouse-test battery. DBA/2 mice performed more poorly than C57BL/6 mice in complex learning tasks such as the water maze and object recognition tasks. In contrast, C57BL/ 6 mice showed attenuated habituation tonovelty in the open field apparatus and poorer performance in the step-down passive avoidance task. The C57BL/6 mice were less exploratory and more anxious than the DBA/ 2 mice. The anxiety score (open arm entries in the elevated plus maze) was significantly correlated with all measures of learning and memory in the object recognition task, and some measures in the passive avoidance and water maze tasks. Analysis of covariance (with open arm entries as a covariate) revealed that some measures on trial 1 of the object recognition task, but not the memory scores on trial 2,were confounded by anxiety. No confounding factors of anxiety were found in the water maze or passive avoidance tasks. Similar results were obtained with the activity scores (line crossing and rearing in the open field). In conclusion, strain differences in activity and anxiety did not account for strain differences in learning and memory performance of C57BL/6 and DBA/2 mice. Nonetheless, the importance of using complete behavioural test batteries should be stressed to ensure that strain differences in learning and memory tasks are not confounded by non-cognitive factors.


UV but not X rays stimulate homologous recombination between sister chromatids and homologs in a Saccharomyces cerevisiae mec1 (ATR) hypomorphic mutant.
MEC1, the essential yeast ATM/ATR homolog, prevents replication fork collapse and is required for the cellular response to DNA damage. We had previously observed higher rates of spontaneous SCE, heteroallelic recombination and translocations in mec1-21 mutants, which still retain some G2 checkpoint function, compared to mec1 null mutants, which are completely defective in checkpoint function, and wild type. However, the types of DNA lesions that are more recombinogenic in mec1-21, compared to wild type, are unknown. Here, we measured DNA damage-associated SCE, homolog (heteroallelic) recombination, and homology-directed translocations in mec1-21, and characterized types of DNA damage-associated chromosomal rearrangements that occur in mec1-21. Although frequencies of UV-associated recombination were higher in mec1-21, the mutant was defective in double-strand break-associated SCE and heteroallelic recombination. Over-expression of Rad53 in mec1-21 reduced UV-associated recombination but did not suppress the defect in X-ray-associated recombination. Both X ray and UV exposure increased translocation frequencies in mec1-21, but the majority of the UV-associated products were non-reciprocal translocations. We suggest that although recombinational repair of double-stand breaks is less efficient in mec1 mutants, recombinants may be generated by other mechanisms, such as break-induced replication.


MiR-15a-3p regulates ferroptosis via targeting glutathione peroxidase GPX4 in colorectal cancer.
Colorectal cancer (CRC) is the second most common cancer-related deaths throughout the world. Ferroptosis is a recently regulated form of cell death, lately gains attention. MicroRNA-15a-3p (miR-15a-3p) plays a regulatory role in various kinds of cancers. However, the role of miR-15a-3p in cellular ferroptosis is still unclear. Here, we aimed to clarify whether miR-15a-3p could regulate the ferroptosis of CRC. Here we identified miR-15a-3p positively regulates ferroptosis via directly targeting glutathione peroxidase glutathione peroxidase 4 (GPX4) in CRC. Overexpression of miR-15a-3p repressed GPX4 through binding to the 3'-untranslated region of GPX4, resulting in increased reactive oxygen species level, intracellular Fe2+ level, and malondialdehyde accumulation in vitro and in vivo. Correspondingly, suppression of miR-15a-3p reduced the sensitivity of CRC cells to erastin and GPX4. Taken together, these data demonstrate that miR-15a-3p regulates ferroptosis through targeting GPX4 in CRC cells, illustrating the novel role of microRNA in ferroptosis.


Translocation (6;9)(p23;q34) in smoldering leukemia and acute nonlymphocytic leukemia.
Two further cases with myeloproliferative disorders (a child with smoldering leukemia and a young male with acute nonlymphocytic leukemia of FAB type M2) and a translocation t(6;9)(p23;q34) are described. Special attention is paid to environmental factors and the early age of onset of patients within the group of leukemias with this specific translocation. In one of our cases a secondary chromosomal anomaly has arisen, which is comparable with another case from the literature.


Calcium oxalate stone disease: effects and side effects of cellulose phosphate and succinate in long-term treatment of absorptive hypercalciuria or hyperoxaluria.
Absorptive hypercalciuria was treated in 27 patients with cellulose phosphate. In all patients urinary calcium decreased and stone formation virtually ceased. The most striking side effect was an excessive hyperoxaluria, necessitating withdrawal of the drug in 8 patients. Succinate decreased the hyperoxaluria in 14 of 19 patients. All patients had mild hypercalciuria and hypermagnesiuria. This study was done to determine the therapeutic value and the side effects in the treatment of absorptive hypercalciuria with sodium cellulose phosphate and of hyperoxaluria with succinate.


Assembly and disassembly of a Zn10 high-nuclearity circular helicate.
A nano-scale decanuclear Zn(II) circular helicate is synthesized without the aid of counteranions during the assembly process, and can be totally disassembled into its reactants by specific anions.


Posttraumatic stress disorder following critical illness.
Posttraumatic stress disorder (PTSD) is a common psychiatric condition that can occur after a traumatic event. Individuals exposed to traumatic events are at risk for PTSD and other psychologic morbidity, such as depression, panic disorder, generalized anxiety disorder, and substance abuse. The burden of PTSD can be high, with inability to work or return to prior levels of functioning. Emerging literature suggests a significantly increased risk for symptoms of PTSD among survivors of critical illness and the families of patients who survive or die after critical illness. This review summarizes these studies and provides suggestions for current clinical implications and for future research.


Directional takeoff, aerial righting, and adhesion landing of semiaquatic springtails.
Springtails (Collembola) have been traditionally portrayed as explosive jumpers with incipient directional takeoff and uncontrolled landing. However, for these collembolans that live near the water, such skills are crucial for evading a host of voracious aquatic and terrestrial predators. We discover that semiaquatic springtails, Isotomurus retardatus, can perform directional jumps, rapid aerial righting, and near-perfect landing on the water surface. They achieve these locomotive controls by adjusting their body attitude and impulse during takeoff, deforming their body in midair, and exploiting the hydrophilicity of their ventral tube, known as the collophore. Experiments and mathematical modeling indicate that directional-impulse control during takeoff is driven by the collophore's adhesion force, the body angle, and the stroke duration produced by their jumping organ, the furcula. In midair, springtails curve their bodies to form a U-shape pose, which leverages aerodynamic forces to right themselves in less than ~20 ms, the fastest ever measured in animals. A stable equilibrium is facilitated by the water adhered to the collophore. Aerial righting was confirmed by placing springtails in a vertical wind tunnel and through physical models. Due to these aerial responses, springtails land on their ventral side ~85% of the time while anchoring via the collophore on the water surface to avoid bouncing. We validated the springtail biophysical principles in a bioinspired jumping robot that reduces in-flight rotation and lands upright ~75% of the time. Thus, contrary to common belief, these wingless hexapods can jump, skydive, and land with outstanding control that can be fundamental for survival.


Highly Multiplexed, Semiautomated Nextera Next-Generation Sequencing (NGS) Library Preparation.
High-throughput, inexpensive DNA sequencing is an essential component of large-scale DNA assembly operations. Using traditional and acoustic liquid-handling robotics, Illumina's Nextera Tagmentation reactions can be miniaturized and paired with custom PCR index primers to produce highly multiplexed NGS libraries for pooled sequencing. This chapter describes a high-throughput protocol that enables the simultaneous sequencing of thousands of DNA constructs in a single sequencing run at a dramatically reduced cost compared to bench-top methods.


Role of intramolecular interaction in connexin50: mediating the Ca2+-dependent binding of calmodulin to gap junction.
Gap junction channels formed by connexin50 (Cx50) are critical for maintenance of eye lens transparency. Cleavage of the carboxyl terminus (CT) of Cx50 to produce truncated Cx50 (Cx50trunc) occurred naturally during maturation of lens fiber cells. The mechanism of its altered properties is under confirmation. It has been suggested that calmodulin (CaM) participates in gating some kinds of gap junction. Here, we performed confocal colocalization and co-immunoprecipitation experiments to study the relationships between Cx50 and CaM. Results exhibited that the CaM could colocalize Ca2+ dependently with CT in the linear area of cell-to-cell contact formed by Cx50trunc, while it could not localize in the linear area without expression of CT. Further study indicated that the CT could interact Ca2+ independently with the cytoplasmic loop (CL) of Cx50. These data put forward the importance of Ca2+-independent intramolecular interaction between CT and CL of Cx50, which mediate the Ca2+-dependent binding of CaM to Cx50. These intra- and intermolecular interactions may further improve our understanding of biological significance of the Cx50 in the eye lens.


HEI-OC1 cells as a model for investigating drug cytotoxicity.
The House Ear Institute-Organ of Corti 1 (HEI-OC1) is one of the few, and arguable the most used, mouse auditory cell line available for research purposes. Originally proposed as an in vitro system for screening of ototoxic drugs, it has been used to investigate, among other topics, apoptotic pathways, autophagy and senescence, mechanism of cell protection, inflammatory responses, cell differentiation, effects of hypoxia, oxidative and endoplasmic reticulum stress, and expression of molecular channels and receptors. However, the use of different techniques with different goals resulted in apparent contradictions on the actual response of these cells to some specific treatments. We have now performed studies to characterize the actual response of HEI-OC1 cells to a battery of commonly used pharmacological drugs. We evaluated cell toxicity, apoptosis, viability, proliferation, senescence and autophagy in response to APAP (acetaminophen), cisplatin, dexamethasone, gentamicin, penicillin, neomycin, streptomycin, and tobramycin, at five different doses and two time-points (24 and 48 h), by flow cytometry techniques and caspase 3/7, MTT, Cytotoxicity, BrdU, Beclin1, LC3 and SA-β-galactosidase assays. We also used HEK-293 and HeLa cells to compare some of the responses of these cells to those of HEI-OC1. Our results indicate that every cell line responds to the each drug in a different way, with HEI-OC1 cells showing a distinctive sensitivity to at least one of the mechanisms under study. Altogether, our results suggest that the HEI-OC1 might be a useful model to investigate biological responses associated with auditory cells, including auditory sensory cells, but a careful approach would be necessary at the time of evaluating drug effects.


Complete Genome Sequence of a Virulent Duck Enteritis Virus (DEV/India/IVRI-2016) Isolated from Southern India.
Molecular characterization of Indian isolates of duck enteritis virus (DEV) so far has been limited to a few selected genomic regions. Here, we report the complete genome sequence of an isolate, DEV/India/IVRI-2016, from southern India that is 158,091 bp in length.


The impact of presumed consent laws and institutions on deceased organ donation.
This article purports to advance the literature on the impact of presumed consent laws on deceased donation rates by examining the interactions between a presumed consent legal regime and other customs and institutions, using data on health expenditure, death rates caused by cerebro vascular diseases, motor vehicle accidents and homicides, legislation, legal systems, family consent, civil rights and liberties and donor registry systems, for 24 countries over a 14-year period. Countries in which presumed consent is enacted exhibit significantly higher donation rates only if family consent is routinely sought and a combined registry is maintained or neither practice is administered. Otherwise, presumed consent legislation does not have a sizeable impact on deceased donation rates.


After urgent drainage of an obstructed kidney by internal ureteric stenting; is ureteroscopic stone extraction always needed?
To assess the probability of spontaneous stone passage and its predictors after drainage of obstructed kidney by JJ stent, as insertion of an internal ureteric stent is often used for renal drainage in cases of calcular ureteric obstruction. Between January 2011 and June 2013, patients for whom emergent drainage by ureteric stents were identified. The patients' demographics, presentation, and stone characteristics were reviewed. The primary endpoint for this study was stone-free status at the time of stent removal, where all patients underwent non-contrast spiral computed tomography (NCCT) before stent removal. Ureteroscopic stone extraction was performed for CT detectable ureteric stones at the time of stent removal. Potential factors affecting the need for ureteroscopic stone extraction at the time of stent removal were assessed using univariate and multivariate statistical analyses. Emergent ureteric stents were undertaken in 196 patients (112 males, 84 females) with a mean (SD) age of 53.7 (16.2) years, for renal obstruction drainage. At the time of stent removal, 83 patients (42.3%) were stone free; with the remaining 113 patients (57.7%) undergoing ureteroscopic stone extraction. On multivariate analysis, stone width [odds ratio (OR) 15.849, 95% confidence interval (CI) 2.83; P = 0.002) and radio-opaque stones (OR 12.035, 95% CI 4.65; P < 0.001) were independent predictors of the need for ureteroscopic stone extraction at the time of stent removal. Spontaneous ureteric stone passage is possible after emergent drainage of an obstructed kidney by ureteric stenting. Stone opacity, larger stone width, and positive preoperative urine culture are associated with a greater probability of requiring ureteroscopic stone extraction after emergent drainage by ureteric stenting.


Pseudotumor cerebri syndrome with different types of hormonal contraceptives in women of child-bearing age.
There is a lack of comparative safety data on the risk of pseudotumor cerebri syndrome (PTCS) associated with different hormonal contraceptives. We sought to quantify the risk of PTCS associated with eight different types of hormonal contraceptives compared with oral levonorgestrel. We conducted a retrospective cohort study, with a case-control analysis of 4 871 504 women aged 15-45 years in the period 2008-2015, using IQVIA Ambulatory Electronic Medical Records data in the USA. Patients who used nine different contraceptive agents including intrauterine levonorgestrel, medroxyprogesterone injection, etonogestrel/ethinyl estradiol vaginal ring and combination oral contraceptives (COCs) that contained ethinyl estradiol and the progestins levonorgestrel, norgestimate, desogestrel, norethindrone and drospirenone, were included. Diagnosis of PTCS was defined using the first International Classification of Diseases, 9th or 10th revision, code for intracranial hypertension in patients who had also received an imaging code in the 30 days prior to the index date. A total of 3323 PTCS cases and 13 292 matched controls were identified. No increase in risk was found when analysing intrauterine levonorgestrel or COCs containing desogestrel, norethindrone, drospirenone, norgestimate or norgestrel versus COC levonorgestrel. The adjusted incidence rate ratio for etonogestrel/etonogestrel/ethinyl estradiol vaginal ring and medroxyprogesterone suspension compared with levonorgestrel COC was 4.45 [95% confidence interval (CI) 1.98-9.96] and 2.20 (95% CI 1.33-3.64), respectively. This study found an elevated risk for PTCS among users of etonogestrel vaginal ring and medroxyprogesterone suspension when compared with oral levonorgestrel. Future studies are needed to confirm these findings.


Peritoneal carcinomatosis in patients with ovarian cancer: enhanced CT versus 18F-FDG PET/CT.
The study aimed to evaluate the efficacy of PET/CT in detecting peritoneal carcinomatosis in patients with ovarian cancer and to compare the diagnostic accuracy of FDG PET/CT with that of enhanced abdominal CT. The medical records of 46 consecutive patients with ovarian cancer were reviewed, and the presence of peritoneal tumor on (18)F-FDG PET/CT and enhanced abdominal CT scans was also assessed. Imaging results were compared with the pathologic findings obtained by surgery. Pathologic results were positive for peritoneal carcinomatosis in 26 patients and negative in 20 patients. PET/CT correctly detected 25 of 26 patients with peritoneal carcinomatosis and enhanced abdominal CT correctly detected 23 of 26 patients. Sensitivity and specificity for the diagnosis of peritoneal carcinomatosis were 96.2% and 90%, respectively, for PET/CT and 88.5% and 65%, respectively, for enhanced abdominal CT. The accuracy of PET/CT was statistically higher than that of enhanced abdominal CT (93.5% vs 78.3%, P = 0.039). Four distinctly abnormal PET/CT findings of single nodular, multiple nodular, diffuse, and mixed FDG uptakes were identified and corresponded to pathologic findings. (18)F-FDG PET/CT imaging is efficient in the diagnosis of peritoneal carcinomatosis and its performance is superior to that of enhanced abdominal CT.


Hemolytic transfusion reactions after administration of intravenous immune (gamma) globulin: a case series analysis.
This case series summarizes our observations of hemolytic reactions after the administration of large amounts of intravenous immune (gamma) globulin (IVIG). Cases of hemolysis were identified by a decrease in hemoglobin not otherwise explained following IVIG administration. Sixteen cases were identified over a 2 1/2-year period at the Ottawa Hospital of approximately 1000 patients receiving IVIG (1.6%). Characteristics of these patients include a large dose of IVIG, female sex, non-O blood group, and underlying inflammatory state. Significant hemolysis may occur after the administration of large doses of IVIG. A two-step mechanism of hemolysis is proposed, sensitization by ABO isohemagglutinins followed by phagocytosis by activated macrophages. A simple protocol to facilitate the early detection of such cases is presented.


Sodium alginate/graphene oxide aerogel with enhanced strength-toughness and its heavy metal adsorption study.
Ordered porous sodium alginate/graphene oxide (SAGO) aerogel was fabricated by in situ crosslinking and freeze-drying method. GO, as reinforcing filler, can be easily incorporated with SA matrix by self-assembly via hydrogen bonding interaction. Compared with pure SA aerogel, the as-prepared SAGO exhibited excellent mechanical strength and elasticity, and the compression strength of SAGO can reach up to 324 kPa and remain 249 kPa after five compression cycles when 4 wt% GO was added, which were considered significant improvements. SEM result presents that the addition of GO obviously improves the porous structures of aerogel, which is beneficial for the enhancement of strength-toughness and adsorbability. As a consequence, the adsorption process of SAGO is better described by pseudo-second-order kinetic model and Langmuir isotherm, with maximum monolayer adsorption capacities of 98.0 mg/g for Cu2+ and 267.4 mg/g for Pb2+, which are extremely high adsorption capacities for metal ions and show far more promise for application in sewage treatment.


An extremely uncommon complication of acute pancreatitis: spontaneous pancreatic cutaneous fistula.
We report the case of a 67-year-old male who was referred to our hospital due to severe abdominal pain and fever. His past medical history included a total gastrectomy surgery for stomach cancer. Abdominal examination showed intense pain on deep palpation in the epigastric region. Laboratory test revealed elevated C-reactive protein (235 mg/l) and amylase (1,789 U/l). Computerized tomography reported a pancreatic collection with the presence of air bubbles inside and areas of necrotic pancreatic parenchyma.


Retroperitoneal abscess cured by percutaneous drainage with ultrasonography.
This report presents an unusual case of an extensive retroperitoneal abscess caused by appendicitis in a child. A 3-year-old boy who presented with right lower abdominal pain and a high fever was found, by abdominal ultrasonography (US) and computed tomography (CT), to have a retroperitoneal abscess. Percutaneous drainage of the abscess under US guidance was successfully performed. There was no sign of recurrence of appendicitis or the abscess 12 months after the drainage.


Acute reversible renal failure secondary to renal candidiasis.
A diabetic patient presenting with bilateral enlarged kidneys and acute renal failure was found to have renal parenchymal candidiasis without obstruction. Treatment with amphotericin B resulted in return of renal function and decrease in renal size.


Application of enteric viruses for fecal pollution source tracking in environmental waters.
Microbial source tracking (MST) tools are used to identify sources of fecal pollution for accurately assessing public health risk and implementing best management practices (BMPs). This review focuses on the potential of enteric viruses for MST applications. Following host infection, enteric viruses replicate and are excreted in high numbers in the hosts' feces and urine. Due to the specificity in host infection, enteric viruses have been considered one of the most accurate library-independent culture-independent MST tools. In an assessment of molecular viral assays based on sensitivity, specificity and the density of the target virus in fecal-impacted samples, human adenovirus and human polyomavirus were found to be the most promising human-specific viral markers. However, more research is needed to identify promising viral markers for livestock because of cross-reactions that were observed among livestock species or the limited number of samples tested for specificity. Other viral indicators of fecal origin, F+ RNA coliphage and pepper mild mottle virus, have also been proposed as potential targets for developing MST markers. Enhancing the utility of enteric viruses for MST applications through next generation sequencing (NGS) and virus concentration technology is discussed in the latter part of this review. The massive sequence databases generated by shotgun and gene-targeted metagenomics enable more efficient and reliable design of MST assays. Finally, recent studies revealed that alternative virus concentration methodologies may be more cost-effective than standard technologies such as 1MDS; however, improvements in the recovery efficiency and consistency are still needed. Overall, developments in metagenomic information combined with efficient concentration methodologies, as well as high host-specificity, make enteric viruses a promising tool in MST applications.


Parallel mixing of photolithographically defined nanoliter volumes using elastomeric microvalve arrays.
Portable microfluidic systems provide simple and effective solutions for low-cost point-of-care diagnostics and high-throughput biomedical assays. Robust flow control and precise fluidic volumes are two critical requirements for these applications. We have developed a monolithic polydimethylsiloxane (PDMS) microdevice that allows for storing and mixing subnanoliter volumes of aqueous solutions at various mixing ratios. Filling and mixing is controlled via two integrated PDMS microvalve arrays. The volumes of the microchambers are entirely defined by photolithography, hence volumes from picoliter to nanoliter can be fabricated with high precision. Because the microvalves do not require an energy input to stay closed, fluid can be stored in a highly portable fashion for several days. We have confirmed the mixing precision and predictability using fluorescence microscopy. We also demonstrate the application of the device for calibrating fluorescent calcium indicators. Due to the biocompatibility of PDMS, the device will have broad applications in miniaturized diagnostic assays as well as basic biological studies.


The reaction of amorphous iron sulfide with Mo(VI) under different pH conditions.
Iron sulfide (FeS) is an important scavenger for hexavalent molybdate (Mo(VI)) in an anoxic environment; it plays a crucial role in the mobilization and transformation of Mo(VI), although the underlying reaction mechanisms between Mo(VI) and FeS remain unclear. This study investigates the Mo(VI) reaction kinetics with the amorphous FeS over a pH range 5.0-9.0 and Mo's chemical form on the FeS surface. It is found that the Mo(VI) reaction kinetics with FeS follow a pseudo first-order model, and the reaction rate constant (kobs) increases with a decrease in the pH value. The kobs at pH 5.0 is 0.027 min-1, which is about 38 times higher than that at pH 9.0. The rapid Mo(VI) removal under acidic conditions is due to quick Mo(VI) transformation into stable MoS2 and thiomolybdate (MoVOxSy). The amount of MoS2 formed on the surface of FeS increases with a decrease in the pH value. Under neutral and alkaline conditions, Mo(VI) is not transformed into MoS2 by FeS because the precipitation of iron oxy-hydroxide passivates the active sites of FeS. The study also investigates the effect of the initial dosage of FeS (20-200 mg L-1) and Mo(VI) (10-50 mg L-1) on the reaction kinetics of Mo(VI) with FeS. The results provides important information on the environmental fate of Mo(VI) in the anoxic environment containing amorphous FeS.


Isolation of the insoluble straight fibrils of Pick's disease.
This paper describes the isolation and partial purification of the straight fibrils that occur in the neurons of Pick's disease. Pick fibrils are highly insoluble in a variety of solvents. These fibrils were shown to be sodium dodecyl sulfate insoluble even in the presence of a reducing agent at elevated temperatures. This allowed the selective isolation of the fibrils using the SDS boiling procedure and sucrose gradient centrifugation that have been described for isolation of paired helical filaments of Alzheimer's disease. The isolated fibrils retained the native morphology seen in tissue sections, but some appeared to become unraveled to yield a paired helical appearance. These results indicate that the Pick fibrils have many chemical and structural characteristics in common with Alzheimer paired helical filaments, and suggest that these two diseases may be closely related.


Changes with age in rat central monoaminergic system responses to cold stress.
Changes with age in responses to stress of certain central monoaminergic systems were investigated. Three groups of rats, 4, 18 and 29 months old, were exposed to cold and the effect of this stress on hypothalamic tyrosine hydroxylase, and on the metabolism of DA and 5HT in different brain regions was evaluated. Senescent rats were unable for several hours to compensate the loss of body heat. Corticosterone secretion however was equally stimulated. Hypothalamic tyrosine hydroxylase activity was enhanced in the young rats but not in the old ones. However, the two groups of senescent rats did not show the increase in HVA levels noted in striata of young rats 2 hours after cold exposure. In contrast, the 18 and 29-month-old rats presented enhanced serotonergic tonus, indicated by the greater increase in 5HIAA determined by stress.


Differentiating psychiatrically disturbed children on the basis of a structured interview.
A structured psychiatric interview designed for both clinical assessment and data collection was given to 50 pairs of well-matched pediatric and psychiatric clinic children and their mothers. The two groups were shown to be clearly distinguished on the basis of numbers of symptoms. Behavior and school symptoms gave the sharpest distinction between the two groups. Neurotic and somatic symptoms were the poorest discriminators. The psychiatric children showed significantly more antisocial symptoms than their matched pediatric peers. Age differences showed the expected trends with the widest divergence in the number of symptoms being in the oldest age group.


Omental infarction--a mimicker of acute appendicitis in children.
The aim of this study was to review the presenting features of pediatric patients found to have omental infarction with a normal appendix at surgery. A retrospective review was conducted of all patients with surgically proven omental infarction treated at KK Women's and Children's Hospital between May 1997 and January 2004. Twelve children (10 boys and 2 girls) were treated for primary omental infarction. The mean age was 9.0 +/- 1.42 years. The mean weight was 41.3 +/- 9.65 kg, with 6 (50%) above the 97th percentile and the remainder were between the 50th and 97th percentile. All 12 children presented with right-sided abdominal pain. At presentation, 9 had low-grade fever of 37.3 degrees C or higher, with the maximum temperature recorded at 37.9 degrees C. The mean total white blood cell count was 13.3 +/- 2.66. Significant neutrophilia (relative percentage, > 70%; absolute neutrophils, > 8000) was noted in 9 children. Eight children underwent radiological imaging, which included ultrasonography (n = 3) and/or computed tomography (CT) (n = 7) of the abdomen. Preoperative diagnosis was made on 4 CT scans. Surgical resection resulted in immediate resolution of symptoms. Omental infarction often mimics acute appendicitis preoperatively, although CT may be diagnostic. Surgical resection of the affected omentum expedites clinical recovery.


Comparisons of chewing rhythm, craniomandibular morphology, body mass and height between mothers and their biological daughters.
To study and compare the relationships between mean chewing cycle duration, selected cephalometric variables representing mandibular length, face height, etc., measured in women and in their teenage or young-adult biological daughters. Daughters were recruited from local high schools and the University of Michigan School of Dentistry. Selection criteria included healthy females with full dentition, 1st molar occlusion, no active orthodontics, no medical conditions nor medication use that could interfere with normal masticatory motor function. Mothers had to be biologically related to their daughters. All data were obtained in the School of Dentistry. Measurements obtained from lateral cephalograms included: two "jaw length" measures, condylion-gnathion and gonion-gnathion, and four measures of facial profile including lower anterior face height, and angles sella-nasion-A point (SNA), sella-nasion-B point (SNB) and A point-nasion-B point (ANB). Mean cycle duration was calculated from 60 continuous chewing cycles, where a cycle was defined as the time between two successive maximum jaw openings in the vertical dimension. Other variables included subject height and weight. Linear and logistic regression analyses were used to evaluate the mother-daughter relationships and to study the relationships between cephalometric variables and chewing cycle duration. Height, weight, Co-Gn and Go-Gn were significantly correlated between mother-daughter pairs; however, mean cycle duration was not (r(2)=0.015). Mean cycle duration was positively correlated with ANB and height in mothers, but negatively correlated with Co-Gn in daughters. Chewing rate is not correlated between mothers and daughters in humans.


A Study of the Influence of the HCl Concentration on the Composition and Structure of (Hydroxy)Arylsiloxanes from the Hydrolysis-Condensation Reaction of Aryltrichlorosilanes.
The hydrolysis-condensation reactions of m-tolyl, m-chlorophenyl, and α-naphtyl-trichlorsilanes, (1, 2, and 3, respectively) in water-acetone solutions were examined for how they were influenced by the change in the concentration of HCl (CHCl). The composition of the products was monitored by 29Si NMR spectroscopy and atmospheric pressure chemical ionization mass spectrometry (APCI-MS). The acidity of the medium was shown to affect the yields of the products, and so, what products were formed. For 3, e.g., APCI-MS showed peaks of α-naphtyl-T8 and α-naphtyl-T10 as the most abundant in the spectra taken after 48 and 240 h for the reaction conducted at CHCl = 0.037 mol L-1. Unlike this, at CHCl = 0.15 mol L-1, those peaks were of [α-naphtyl(HO)2SiO]2(α-naphtyl)(HO)Si and/or [α-naphtyl(HO)Si]3, [α-naphtyl(HO)Si]4,5, and α-naphtyl-T8 after 192 h. However, at both CHCl values, the main product (and an intermediate) after 24 h was trans-1,1,3,3-tetrahydroxy-1,3-di-α-naphtyldisiloxane. It was isolated and its structure established by 1H-, 29Si-NMR, and X-ray powder diffraction.


Nursemaid's Elbow Reduction.
Nursemaid's elbow, also known as radial head subluxation, is a common childhood orthopedic injury that can easily be diagnosed and reduced by the advanced practice nurse. It is most common in children 1-4 years of age and typically occurs as the result of a pulling mechanism on an outstretched arm. This leads to subluxation of the radial head at the annular ligament. The child subsequently refuses to use the affected arm, leading the caregiver to present for evaluation. This article explores epidemiology, pathophysiology, clinical presentation, reduction techniques, and parent education.


Late-onset islet autoimmunity in childhood: the Diabetes Autoimmunity Study in the Young (DAISY).
We sought to assess the frequency, determinants and prognosis for future diabetes in individuals with islet autoimmunity and whether these factors differ depending on the age of onset of islet autoimmunity. A prospective cohort (n = 2547) of children from the general population who had a high-risk HLA genotype and children who had a first-degree relative with type 1 diabetes were followed for up to 21 years. Those with the persistent presence of one or more islet autoantibodies were categorised as early-onset (<8 years of age, n = 143, median 3.3 years) or late-onset (≥8 years of age, n = 64, median 11.1 years), and were followed for a median of 7.4 and 4.7 years, respectively. Progression to diabetes was evaluated by Kaplan-Meier analysis with logrank test. Factors associated with progression to diabetes were analysed using the parametric accelerated failure time model. Children with late-onset islet autoimmunity were more likely to be Hispanic or African-American than non-Hispanic white (p = 0.004), and less likely to be siblings of individuals with type 1 diabetes (p = 0.04). The frequencies of the HLA-DR3/4 genotype and non-HLA gene variants associated with type 1 diabetes did not differ between the two groups. However, age and HLA-DR3/4 were important predictors of rate of progression to both the presence of additional autoantibodies and type 1 diabetes. Late-onset islet autoimmunity was more likely to present with a single islet autoantibody (p = 0.01) and revert to an antibody-negative state (p = 0.01). Progression to diabetes was significantly slower in children with late-onset islet autoimmunity (p < 0.001). A late onset of islet autoimmunity is more common in African-American and Hispanic individuals. About half of those with late-onset islet autoimmunity progress to show multiple islet autoantibodies and develop diabetes in adolescence or early adulthood. Further investigation of environmental determinants of late-onset autoimmunity may lead to an understanding of and ability to prevent adolescent and adult-onset type 1 diabetes.


Detoxification of coir pith through refined vermicomposting engaging Eudrilus eugeniae.
Hazardous coir industrial waste, coir pith has been subjected to 50 days vermicomposting with Eudrilus eugeniae by amending nitrogenous legume plant, Gliricidia sepium together with cattle dung in different combinations, after 21 days precomposting using Pleurotus sajor-caju spawn. An increase in electrical conductivity, total NPK and calcium, and a decrease in organic matter, total organic carbon, C/N ratio, C/P ratio and total phenolic content in the final vermicompost were observed. Dehydrogenase, urease and cellulase activity peaked up to 30 days of vermicomposting and then declined. The phytotoxicity studies with Brassica juncea, C/N ratio and enzyme activities confirmed the stability and maturity of vermicompost. The results also demonstrated that the 2:3:1 ratio (coir pith + Gliricidia sepium + cow dung) is a suitable effective combination for nutrient-rich (N: 2.43%; P: 0.92%; K: 2.09%) vermicompost production. The total phenolic contents declined during the vermicomposting with a lower final content of 21.26 mg/g GAE in 2:3:1 combination of substrates from the initial level (105.56 mg/g GAE). Besides, the concentration of total phenol contents inversely related to the germination index of Brassica juncea (r = -0.761), indicating that the phenolic content could also play an important role in phytotoxicity. Growth and fecundity of Eudrilus eugeniae in 2:3:1 combination revealed the acceptability and rapid decomposition of coir pith substrate into vermifertilizer.


Endoplasmic reticulum stress-responsive microRNAs are involved in the regulation of abiotic stresses in wheat.
ER stress-responsive miRNAs, tae-miR164, tae-miR2916, and tae-miR396e-5p, are essential in response to abiotic stress. Investigating ER stress-responsive miRNAs is necessary to improve plant tolerance to environmental stress. MicroRNAs (miRNAs) play vital regulatory roles in plant responses to environmental stress. Recently, the endoplasmic reticulum (ER) stress pathway, an essential signalling pathway in plants in response to adverse conditions, has been widely studied in model plants. However, miRNAs associated with ER stress response remain largely unknown. Using high-throughput sequencing, three ER stress-responsive miRNAs, tae-miR164, tae-miR2916, and tae-miR396e-5p were identified, and their target genes were confirmed. These three miRNAs and their target genes actively responded to dithiothreitol, polyethylene glycol, salt, heat, and cold stresses. Furthermore, in some instances, the expression patterns of the miRNAs and their corresponding target genes were contrasting. Knockdown of tae-miR164, tae-miR2916, or tae-miR396e-5p using a barley stripe mosaic virus-based miRNA silencing system substantially enhanced the tolerance of wheat plants to drought, salt, and heat stress. Under conditions involving these stresses, inhibiting the miR164 function by using the short tandem target mimic approach in Arabidopsis thaliana resulted in phenotypes consistent with those of miR164-silenced wheat plants. Correspondingly, overexpression of tae-miR164 in Arabidopsis resulted in a decreased tolerance to drought stress and, to some extent, a decrease in tolerance to salt and high temperature. These results revealed that tae-miR164 plays a negative regulatory role in wheat/Arabidopsis in response to drought, salt, and heat stress. Taken together, our study provides new insights into the regulatory role of ER stress-responsive miRNAs in abiotic stress responses.


Combination therapy with bosentan and sildenafil for refractory digital ulcers and Raynaud's phenomenon in a 30-year-old woman with systemic sclerosis: Case report and literature review.
Systemic sclerosis is a rare autoimmune disease characterized by skin and organ fibrosis, and vasculopathy. Raynaud's phenomenon is almost universally present in systemic sclerosis and can be the most debilitating symptom. Raynaud's phenomenon may lead to the development of digital ulcers, potentially complicated by infection, tissue necrosis, and auto-amputation. Recommended treatments have variable efficacy. We report the case of a 30-year-old woman with diffuse systemic sclerosis suffering from severe Raynaud's phenomenon and digital ulcers with digital tissue necrosis who was treated with combination therapy of an endothelin receptor antagonist and phosphodiesterase 5 inhibitor. In addition, we reviewed the literature on the topic. Previous therapy with calcium-channel blockers, intravenous iloprost, and bosentan had all failed to control symptoms. We added sildenafil in combination with bosentan and observed a rapid and sustained treatment effect. Raynaud's phenomenon severity, number of attacks, and attack duration decreased within 2 weeks of initiating treatment. Furthermore, this resulted in the healing of established digital ulcers. Our case report suggests that combination therapy may be a feasible treatment for the most severely affected and refractory patients. In our literature review, we found one retrospective study and three additional cases with similarly encouraging results.


Validation of the cone beam computed tomography-based stereolithographic surgical guide aiding autotransplantation of teeth: clinical case-control study.
To compare the outcome of cone beam computed tomography (CBCT)-based surgical planning and transfer technique for tooth autotransplantation versus conventional autotransplantation. The study material comprised 40 pediatric subjects in whom 48 teeth were transplanted following a case-control design. While the study group (mean age 11 years) underwent CBCT imaging for surgical planning and transfer via stereolithographic tooth replica fabrication, the historical control group (mean age 12 years) was subjected to conventional autotransplantation. The CBCT-based preoperative planning and the use of a tooth replica decreased the extra-alveolar time and reduced the number of positioning trials with the donor tooth. In the control group, 6 patients showed 1 or more complications, while this was noticed for only 2 study patients. CBCT-based surgical planning of tooth autotransplantation may benefit from a shorter surgical time, while being a less invasive technique, causing fewer failures than a conventional approach.


Metabolic acidosis after bladder replacement: comparison of severity and reversibility in ileal and colonic reservoirs.
Metabolic acidosis developed frequently after ureterosigmoidostomy and rectosigmoid bladder construction but has been reported rarely after the newer methods of continent urinary diversion which also employ intestinal reservoirs. We created an animal model in which to compare the metabolic effects of bladder replacement with segments of ileum or colon and the potential for reversing these derangements with nicotinic acid and chlorpromazine. One year after six dogs' bladders were replaced by colon (three) or ileum (three), all dogs appeared in excellent health and were free of urinary tract obstruction and clinical infection. Both groups of dogs were severely acidotic with diminished arterial pH and arterial and venous total CO2 concentrations although normal serum electrolytes and creatinine concentrations were maintained. Both groups of dogs absorbed approximately one half the urinary sodium, chloride and urea presented to their intestinal reservoirs. After treatment with nicotinic acid and chlorpromazine, the metabolic status of both groups of animals improved. Although nicotinic acid reduced urinary excretion of electrolytes more effectively than chlorpromazine, nicotinic acid was not more effective for reversing metabolic acidosis. When nicotinic acid was provided as an adjunct to sodium bicarbonate therapy in two animals acidosis was corrected at reduced doses of sodium bicarbonate. Based upon this work in an animal model, there does not appear to be a metabolic advantage to intestinal reservoirs which incorporate ileum versus colon. However, asymptomatic patients with normal serum electrolytes and creatinine concentrations may be acidotic. The effects of long term mild acidosis are unknown. However, if therapy is required to prevent diminution of whole body buffers or changes in bone density specific therapy with nicotinic acid or chlorpromazine may reduce the requirement for alkali for correction of metabolic acidosis.


Intra-articular injection of loaded sPL sustained-release microspheres inhibits osteoarthritis and promotes cartilaginous repairs.
Osteoarthritis is a chronic inflammatory disease of the joints associated with significant morbidity and lower quality of life. Current treatment strategies focus on reducing cartilage degeneration but fail to restore their proliferative ability. Super-activated platelet lysate (sPL) is an enhanced form of platelet-rich plasma that can be easily inactivated. The purpose of this study is to evaluate whether sPL-loaded PLGA/chitosan/gelatin microspheres can prevent and treat osteoarthritis. Features of biological microspheres were detected by SEM and ELISA. Osteoarthritis chondrocytes were co-cultured with hydrogel loaded with sPL. The effect of biological microspheres on chondrocyte proliferation was evaluated using a CCK-8 cell proliferation test. Cell morphology and cell necrosis were measured with a microscope. The gene expression levels of cartilage-related markers type 2 collagen, aggrecan (ACAN), and SRY type high mobility group box-9 (SOX9) were determined by real-time quantitative polymerase chain reaction (Rt-PCR). A rat osteoarthritis model was established. Micro-CT was used to characterize cartilaginous changes after the injection of biological microspheres. Histopathological HE staining, Safranin-O Fast Green staining and staining scores, type II collagen staining, and proteoglycan staining were used to evaluate the degree of cartilaginous repair. Biological microspheres were able to continuously release biological factors. Exposure to loading sPL microspheres significantly increased chondrocyte proliferation, reduced cell necrosis, and increased the expression of cartilage markers type 2 collagen, ACAN, and SOX9 in osteoarthritic chondrocytes. In vivo experiments found that biological microspheres also smoothen cartilage surfaces, promote the expression of proteoglycan and type 2 collagen while also increasing cartilaginous integrity as evaluated using Safranin-O Fast Green staining. PLGA/chitosan/gelatin hydrogel loaded with sPL is a promising tool for effective and non-invasive articular cartilage repair in osteoarthritis. Biological microspheres loaded with sPL release various biological factors to promote chondrocyte proliferation and upregulate chondrocyte functionalization genes (SOX9, CoX II, ACAN), leading to an overall enhanced cartilaginous matrix.


Correction to: Characterization of oogonia stem cells in mice by Fragilis.
In the original publication the labelling of Figure 1D, Y-axis is incorrectly published. The correct labeling should be read as Fragilis+/SSEA1+ and the correct figure is provided in this correction.