Free physeal transplantation in the rabbit. An experimental approach to focal lesions.
We have developed a procedure to transplant free autogenous iliac-crest physeal grafts into defects created in the lateral aspect of the distal femoral physis of rabbits. Excision of a portion of the lateral part of the physis led consistently to formation of a bone bridge, growth arrest, and valgus deformity. There was no evidence of increased activity by the persisting physis to fill the defect or of differentiation of mesenchymal cells into a functioning physeal layer. We carried out preliminary studies to define the optimum conditions for obtaining and transplanting a free autogenous iliac-crest physeal graft into the femoral physeal defect. The iliac apophysis is composed of a fibrocartilaginous layer, cartilage similar to epiphyseal cartilage, and a physis. The graft included the physis and some overlying epiphyseal cartilage but the outermost fibrocartilaginous layer and the surrounding perichondrium were removed, as these two tissues were believed to inhibit diffusion. The graft was separated gently from the metaphysis. Histological studies showed that the separation occurred at the lower hypertrophic-chondrocyte zone. After optimum techniques had been developed for transplantation and the feasibility of the procedure had been established in twenty-one rabbits, the capacity of the physeal transplants to prevent bone-bridge formation, growth arrest, and valgus deformity was evaluated in thirty-nine rabbits, three to four months old, using the standardized transplantation procedure. The results were assessed in terms of the gross and radiographic appearance of the femur, measurements of length, and histological characteristics. The transplanted physis retained its normal morphology, united with the residual part of the original femoral physis, and participated in endochondral bone formation. Bone-bridge formation between the femoral epiphysis and metaphysis, growth arrest, and valgus deformity were prevented or minimized in most animals, and the physeal transplants yielded good or excellent results in 60 per cent of the procedures using stringent criteria. The results of this study show that free physeal transplants into focal defects can remain viable and function satisfactorily if they are fashioned properly and inserted gently. This study demonstrated that transplantation of an iliac-crest physis into a defect of the lateral aspect of the distal femoral physis can prevent bone-bridge formation, growth arrest, and the development of valgus deformity.(ABSTRACT TRUNCATED AT 400 WORDS)


A bioengineered arginine-depleting enzyme as a long-lasting therapeutic agent against cancer.
L-Arginine (L-Arg) depletion has attracted great attention in cancer therapy. Although two types of arginine-depleting enzymes, arginine deiminase (ADI) and human arginase I, are undergoing clinical trials, random site of PEGylation, low efficacy of heavy metal as co-factor, and immunogenicity limit the performance of these drugs and cause difficulty in a homogeneous production. Here we screened ten catalytic metal ions and have successfully produced a site-specific mono-PEGylated human arginase I mutant by conjugating the Cys45 residue to PEG-maleimide to minimize the decrease in activity and produce a homogeneous product. The catalytic efficiency trend of metal ion-enriched human arginase I mutant (HAI) was Co2+ > Ni2+ ≫ Mn2+. The overall kcat/KM values of Co-HAI and Ni-HAI were higher than Mn-HAI by ~ 8.7- and ~ 5.2-folds, respectively. Moreover, the results of enzyme kinetics and circular dichroism spectrometry demonstrated that the 20 or 40 kDa linear and branched PEG attached on the HAI surface did not affect the enzyme activity and the protein secondary structures. In vitro studies showed that both Co-HAI-PEG20L and Ni-HAI-PEG20L inhibited the growth of eight types of cancer cell lines. The pharmacodynamic study in mice demonstrated that the i.p. administration of Co-HAI-PEG20L at 13 mg/kg and Ni-HAI-PEG20L at 15 mg/kg was able to maintain a L-Arg level below its detection limit for over 120 h after one injection. The body weights of mice could return to normal levels within 5 days after injection, showing that the doses were well-tolerated. Therefore, both the Ni-HAI-PEG20L and Co-HAI-PEG20L are promising candidates for cancer therapy. KEY POINTS: • Mono-PEGylation applied on human arginase I mutant (HAI) successfully. • The catalytic efficiency of Co- and Ni-enriched HAI was higher than the wild type. • At least eight types of cancer cell lines were inhibited by Co- and Ni-HAI-PEG20L. • Co- and Ni-HAI-PEG20L were able to achieve weekly depletion of L-Arg. Graphical abstract.


Variations of structural components: specific intercultural differences in facial morphology, skin type, and structures.
Analysis of the differences in facial morphology and skin structure and tone among ethnic groups within the realm of plastic surgery is relevant due to the increasing number of ethnic individuals seeking cosmetic surgery. Previous classifications of ideal facial morphologic characteristics have been revised and challenged over the years to accurately reflect the differences in facial structure that are aesthetically pleasing to individuals of differing ethnic groups. The traditional neoclassic canons reflected the European Caucasian facial morphology and cannot be used to classify facial characteristics in ethnic groups due to drastic differences in measurement and proportion. In addition, differences in biophysiologic properties of ethnic skin types influence the progression of aging and the ability of skin to withstand environmental insults. Thickness of the stratum corneum, water content, and melanin composition are important factors that were analyzed in varying ethnic groups. Although it appears that Caucasian Americans are subject to earlier onset of skin wrinkling and sagging than are African Americans due to thinner stratum corneum layers and decreased water content, more research needs to be conducted to be inclusive of other ethnic groups. These data will enable plastic surgeons to treat these groups more effectively while preserving their unique characteristics.


Plasma membrane Ca2+-ATPase extrudes Ca2+ from hair cell stereocilia.
Mechanically sensitive hair cells of the auditory and vestibular systems use Ca2+ to control adaptation of mechanical transduction, to effect frequency tuning, to trigger neurotransmitter release, and to mediate efferent synaptic signaling. To determine the role that pumps play in regulation of Ca2+ in the hair bundle, the organelle responsible for mechanoelectrical transduction, we localized and quantified the plasma membrane Ca2+-ATPase (PMCA) of the bundle. We found that each hair bundle contains approximately 10(6) PMCA molecules or approximately 2000 per square micrometer of bundle membrane and that PMCA is the principal calmodulin binding protein of the bundle. Consistent with biochemical estimates of PMCA density, we measured with extracellular Ca2+-selective electrodes a substantial Ca2+ efflux from bundles. The number of bundle Ca2+ pumps and magnitude of resting Ca2+ efflux suggested that PMCA should generate a substantial membrane current as bundles expel Ca2+. Measurement of whole-cell currents revealed a transduction-dependent outward current that was consistent with the activity of PMCA. Finally, dialysis of hair cells with PMCA inhibitors led to a large increase in the concentration of Ca2+ in bundles, which suggests that PMCA plays a major role in regulating bundle Ca2+ concentration. Our data further indicate that PMCA could elevate the extracellular Ca2+ concentration close to hair bundles above the low level found in bulk endolymph.


[Radiation induced lipid peroxidation: factors which determine the oxidizability of lipids].
Lipids are the essential components of cell membranes and lipoproteins. Their peroxidation plays an important role in numerous pathologies in which oxidative stress is involved. Lipid peroxidation occurs through a chain reaction that contributes to membrane damage in cells. It results in the conversion of fatty acids to polar hydroperoxides and leads to the breakdown or malfunction of the membrane. Lipids are amphiphilic molecules that aggregate in aqueous solutions into micelles and liposomes. The effect of this structural organization is significant in studies of radiation-induced peroxidation damage in highly ordered biological systems such as biological membranes. In this paper, a synthesis of the data concerning radioinduced lipid peroxidation is completed by an original review of the different parameters that determine lipid oxidizability. In addition, the influence of lipid aggregation and the effect of molecular packing are discussed.


Disentangling the effects of geographic and ecological isolation on genetic differentiation.
Populations can be genetically isolated both by geographic distance and by differences in their ecology or environment that decrease the rate of successful migration. Empirical studies often seek to investigate the relationship between genetic differentiation and some ecological variable(s) while accounting for geographic distance, but common approaches to this problem (such as the partial Mantel test) have a number of drawbacks. In this article, we present a Bayesian method that enables users to quantify the relative contributions of geographic distance and ecological distance to genetic differentiation between sampled populations or individuals. We model the allele frequencies in a set of populations at a set of unlinked loci as spatially correlated Gaussian processes, in which the covariance structure is a decreasing function of both geographic and ecological distance. Parameters of the model are estimated using a Markov chain Monte Carlo algorithm. We call this method Bayesian Estimation of Differentiation in Alleles by Spatial Structure and Local Ecology (BEDASSLE), and have implemented it in a user-friendly format in the statistical platform R. We demonstrate its utility with a simulation study and empirical applications to human and teosinte data sets.


SIRT1 positively regulates breast cancer associated human aromatase (CYP19A1) expression.
Breast cancer remains one of the leading causes of death in women diagnosed with cancer. In breast cancer, aberrant expression of the CYP19A1 gene, which encodes the aromatase enzyme, contributes to increased intratumoral levels of estradiol. Regardless of whether this estrogen is produced by peripheral tissues or within specific subpopulations of cells within the breast tumor, it is clear that the aromatase enzymatic activity is critical for the growth of estrogen-dependent tumors. Currently, aromatase inhibitors have proven to be highly effective in blocking the growth of estrogen-dependent forms of breast cancer. CYP19A1 transcription is tightly controlled by 10 tissue-specific promoters. In breast cancer, however, aromatase transcription is driven by multiple promoters that somehow override the tissue-specific regulation of normal tissue. Here, we explore the role that the deacetylase, sirtuin-1 (SIRT1), plays in positively regulating aromatase in breast cancer. We demonstrate that the use of cambinol and the SIRT1/2 inhibitor VII, 2 small molecule inhibitors of SIRT1 and SIRT2, as well as small molecule inhibitors and small interfering RNA specific to SIRT1, all reduce the levels of aromatase mRNA. We further demonstrate that pharmacologic inhibition causes a marked reduction in aromatase protein levels. Additionally, by chromatin immunoprecipitation, we demonstrate that SIRT1 occupies the promoter regions PI.3/PII and PI.4, and its inhibition leads to increased acetylation of estrogen-related receptorα, a transcription factor that positively regulates CYP19A1 transcription in epithelial cells. Finally, we demonstrate by immunohistochemistry that SIRT1 is significantly up-regulated in invasive ductal carcinoma relative to normal tissue adjacent to tumor, further suggesting a role of SIRT1 in breast cancer. This work uncovers a new mechanism for the regulation of aromatase and provides rationale for further investigation of how the inhibition of specific sirtuins may provide a unique strategy for inhibiting aromatase that may complement or synergize with existing therapies.


Interaction of NMDA receptor and pacemaking mechanisms in the midbrain dopaminergic neuron.
Dopamine neurotransmission has been found to play a role in addictive behavior and is altered in psychiatric disorders. Dopaminergic (DA) neurons display two functionally distinct modes of electrophysiological activity: low- and high-frequency firing. A puzzling feature of the DA neuron is the following combination of its responses: N-methyl-D-aspartate receptor (NMDAR) activation evokes high-frequency firing, whereas other tonic excitatory stimuli (α-amino-3-hydroxyl-5-methyl-4-isoxazolepropionate receptor (AMPAR) activation or applied depolarization) block firing instead. We suggest a new computational model that reproduces this combination of responses and explains recent experimental data. Namely, somatic NMDAR stimulation evokes high-frequency firing and is more effective than distal dendritic stimulation. We further reduce the model to a single compartment and analyze the mechanism of the distinct high-frequency response to NMDAR activation vs. other stimuli. Standard nullcline analysis shows that the mechanism is based on a decrease in the amplitude of calcium oscillations. The analysis confirms that the nonlinear voltage dependence provided by the magnesium block of the NMDAR determine its capacity to elevate the firing frequency. We further predict that the moderate slope of the voltage dependence plays the central role in the frequency elevation. Additionally, we suggest a repolarizing current that sustains calcium-independent firing or firing in the absence of calcium-dependent repolarizing currents. We predict that the ether-a-go-go current (ERG), which has been observed in the DA neuron, is the best fit for this critical role. We show that a calcium-dependent and a calcium-independent oscillatory mechanisms form a structure of interlocked negative feedback loops in the DA neuron. The structure connects research of DA neuron firing with circadian biology and determines common minimal models for investigation of robustness of oscillations, which is critical for normal function of both systems.


The final piece of the Triangle of U: Evolution of the tetraploid Brassica carinata genome.
Ethiopian mustard (Brassica carinata) is an ancient crop with remarkable stress resilience and a desirable seed fatty acid profile for biofuel uses. Brassica carinata is one of six Brassica species that share three major genomes from three diploid species (AA, BB, and CC) that spontaneously hybridized in a pairwise manner to form three allotetraploid species (AABB, AACC, and BBCC). Of the genomes of these species, that of B. carinata is the least understood. Here, we report a chromosome scale 1.31-Gbp genome assembly with 156.9-fold sequencing coverage for B. carinata, completing the reference genomes comprising the classic Triangle of U, a classical theory of the evolutionary relationships among these six species. Our assembly provides insights into the hybridization event that led to the current B. carinata genome and the genomic features that gave rise to the superior agronomic traits of B. carinata. Notably, we identified an expansion of transcription factor networks and agronomically important gene families. Completion of the Triangle of U comparative genomics platform has allowed us to examine the dynamics of polyploid evolution and the role of subgenome dominance in the domestication and continuing agronomic improvement of B. carinata and other Brassica species.


Porcine parvovirus: virus purification and structural and antigenic properties of virion polypeptides.
Porcine parvovirus (PPV) was extensively purified from infected swine fetal homogenates by CaCl2 precipitation followed by CsCl density centrifugation. Two species of particles possessing PPV-specific hemagglutinating activity were observed banding at densities of 1.39 and 1.30 g/ml, representing full and empty 20-nm virion particles, respectively. Both classes of particles contained three major polypeptides. A, B, and C, with respective molecular weights of 83,000, 64,000, and 60,000. The amount of polypeptide A was similar in both species (approximately 10%); however, the B protein was most abundant in the 1.30-g/ml particles, whereas the C protein was the major polypeptide found in the 1.39-g/ml particles. Antisera generated to each sodium dodecyl sulfate-polyacrylamide gel-purified virion structural protein had reactivities qualitatively similar to those of conventional antisera raised against intact PPV in a variety of standard serological assays. The antisera generated against the individual sodium dodecyl sulfate-denatured PPV polypeptides were able to react with native, intact PPV virions and were capable of neutralizing virus infectivity.


Role of medical students in preventing patient harm and enhancing patient safety.
Substantial efforts are focused on the high prevalence of patient harm due to medical errors and the mechanisms to prevent them. The potential role of the medical student as a valuable member of the team in preventing patient harm has, however, often been overlooked. Four cases are presented from two US academic health centers in which medical students prevented or were in a position to prevent patient harm from occurring. The authors directly participated in each case. The types of harm prevented included averting non-sterile conditions, missing medications, mitigating exposure to highly contagious patients, and respecting patients' "do not resuscitate" requests. Medical students are often overlooked as valuable participants in ensuring patient safety. These cases show that medical students may be an untapped resource for medical error prevention. Medical students should be trained to recognize errors and to speak up when errors occur. Those supervising students should welcome and encourage students to actively communicate observed errors and near misses and should work to eliminate all intimidation by medical hierarchy that can prevent students from being safety advocates.


[Sarcomatoid carcinoma of the thymus].
Sarcomatoid carcinomas are rare malignancies composed of epithelial (carcinomatous) and sarcomatous elements. High-grade carcinoma is difficult to diagnose by light optic and immunohistochemical studies. The authors give the data available in the literature on sarcomatoid carcinoma of the thymus and also describe their own observation. Its histological pattern is represented by a combination of carcinomatous and sarcomatous elements with extensive necrotic foci. Immunohistologically, the tumor cells expressed total cytokeratin (AE1/AE3), CD99, and focally epithelial membrane antigen. Some tumor cells were vimentin- and SMA-positive. Markers, such as chromogranin A, bcl-2, bcl-6, S-100 protein and myogenin, were negative.


Maternal concern, social support, and health-related quality of life across childhood.
Mothers play a critical role in child and family health yet little is known about the concerns and health-related quality of life (HRQOL) of mothers beyond early childhood. The purpose of this study of 234 mothers of children up to 18 years of age was to examine relationships among maternal concern, social support, and HRQOL. Mothers reported a low level of concern, but concern was a significant predictor of HRQOL. The health burden for mothers was demonstrated primarily in the mental health components of HRQOL. The effect of social support was minimal and limited to the mental health domain of HRQOL. Results suggest regular assessment of maternal concerns and mental health coupled with education regarding coping strategies to support HRQOL for all mothers.


Stewart-Treves Syndrome: A Case Report and Review of the Literature.
The Stewart-Treves syndrome is a rare and deadly entity, which is defined as angiosarcoma arising in the setting of chronic lymphedema. It typically presents in women who develop lymphedema in the upper extremity secondary to axillary lymph node dissection for breast cancer surgery. It is extremely uncommon in the lower extremities as a result of idiopathic chronic lymphedema. Here, we present the case of a 63-year-old female patient with idiopathic chronic lymphedema of the lower extremities having morbid obesity (BMI 82.6) and multiple comorbidities. She developed multiple confluent, hemorrhagic and necrotic elevated purple-black papules in the lower extremities, for which the initial diagnosis was cellulitis. Because there was no improvement with antibiotics, a lower extremity ultrasound and biopsy was performed which showed multiple masses in the left inner upper calf with solid and cystic components. The pathology results of the punch biopsies were consistent with angiosarcoma. Immunohistochemical studies revealed positivity for CD31, FLI-1, and a high Ki-67 proliferation rate. Because of the patient's weight and medical comorbidities, no further extensive diagnostic tests were performed to detect metastatic disease, and because of contraindications, no further medical treatment was provided. The patient subsequently died 1 month after diagnosis.


Fluidity of brush border and basolateral membranes from human kidney cortex.
The physical state of lipids in brush border and basolateral membrane vesicles prepared from normal human kidney cortex was investigated by fluorescence polarization and electron spin resonance. At physiologic temperature, lipids were significantly less ordered, i.e., more fluid, in basolateral than in brush border membranes. This difference was also observed using corresponding liposomes made from total lipid extracts. For both brush border and basolateral membranes, temperature-dependent experiments revealed the existence of a broad thermotropic transition extending approximately from 20 to 42 degrees C. These data are interpreted to indicate that plasma membranes from human kidney cortex function physiologically at the upper critical temperature of a transition that probably corresponds to a liquid crystalline-to-gel lipid phase separation.


Molecular organization and comparative analysis of chromosome 5B of the wild wheat ancestor Triticum dicoccoides.
Wild emmer wheat, Triticum turgidum ssp. dicoccoides is the wild relative of Triticum turgidum, the progenitor of durum and bread wheat, and maintains a rich allelic diversity among its wild populations. The lack of adequate genetic and genomic resources, however, restricts its exploitation in wheat improvement. Here, we report next-generation sequencing of the flow-sorted chromosome 5B of T. dicoccoides to shed light into its genome structure, function and organization by exploring the repetitive elements, protein-encoding genes and putative microRNA and tRNA coding sequences. Comparative analyses with its counterparts in modern and wild wheats suggest clues into the B-genome evolution. Syntenic relationships of chromosome 5B with the model grasses can facilitate further efforts for fine-mapping of traits of interest. Mapping of 5B sequences onto the root transcriptomes of two additional T. dicoccoides genotypes, with contrasting drought tolerances, revealed several thousands of single nucleotide polymorphisms, of which 584 shared polymorphisms on 228 transcripts were specific to the drought-tolerant genotype. To our knowledge, this study presents the largest genomics resource currently available for T. dicoccoides, which, we believe, will encourage the exploitation of its genetic and genomic potential for wheat improvement to meet the increasing demand to feed the world.


Complete genomic sequence of Pasteurella multocida, Pm70.
We present here the complete genome sequence of a common avian clone of Pasteurella multocida, Pm70. The genome of Pm70 is a single circular chromosome 2,257,487 base pairs in length and contains 2,014 predicted coding regions, 6 ribosomal RNA operons, and 57 tRNAs. Genome-scale evolutionary analyses based on pairwise comparisons of 1,197 orthologous sequences between P. multocida, Haemophilus influenzae, and Escherichia coli suggest that P. multocida and H. influenzae diverged approximately 270 million years ago and the gamma subdivision of the proteobacteria radiated about 680 million years ago. Two previously undescribed open reading frames, accounting for approximately 1% of the genome, encode large proteins with homology to the virulence-associated filamentous hemagglutinin of Bordetella pertussis. Consistent with the critical role of iron in the survival of many microbial pathogens, in silico and whole-genome microarray analyses identified more than 50 Pm70 genes with a potential role in iron acquisition and metabolism. Overall, the complete genomic sequence and preliminary functional analyses provide a foundation for future research into the mechanisms of pathogenesis and host specificity of this important multispecies pathogen.


Exploring the relevance of social justice within a relational nursing ethic.
In the last few decades, a growing number of commentators have questioned the appropriateness of the 'justice view' of ethics as a suitable approach in health care ethics, and most certainly in nursing. Essentially, in their ethical deliberations, it is argued that nurses do not readily adopt the high degree of impartiality and objectivity that is associated with a justice view; instead their moral practices are more accurately reflected through the use of alternative approaches such as relational or care-based ethics. Yet, it has also been argued that this viewpoint does not necessarily 'do justice' to the broader moral responsibilities of nurses towards humanity in general, i.e. to the wider socio-cultural and socio-political issues in society, and to the concept of social justice in particular. This criticism has triggered a much closer examination of relational and care-based ethics in nursing at levels beyond individual responsiveness within relationships and brought into the spotlight the need for a more ethically refined nursing response to an increasingly complex set of socio-cultural inequalities. This article explores a relational ethic within nursing practices with contemporary ideas regarding social justice. In particular, it is argued that the synergy between the two actually produces an ethic that is capable of not only challenging the continuing predominance of justice-based ethics within health care, but of replacing it. Subsequently, in the discussion that follows, it is suggested that a combined social justice and relational care-based approach, as a social ethic, should guide the moral deliberations and actions of nurses. It is maintained that such an approach is not only possible, but crucial if nurses are to realize their full potential as ethical agents for individual and social good.


[Estimation of gasometric values at different altitudes above sea level in Mexico].
We calculated reference values for arterial blood gases at different altitudes in Mexico assuming that sea level PaCO2 is 40 Torr, and in Mexico City (2.24 km. above the sea level) is 31.13 Torr, average of reported reference values. With the previous two points, it is possible to calculate a linear regression: PaCO2 = 40-3.96(altitude in km.). The equation is very similar to that calculated from reports in alveolar gas in North-Americans (Fitzgerald < 5 km): PACO2 = 39.3-3.11(altitude in km), and from subjects acclimatized to acute altitude exposure (< 5 km): PACO2 = 38.3-2.5 (altitude in km). It is also similar to a alinear equation that can be calculated assuming that hyperventilation in permanent habitants of moderate altitudes is inversely proportional to inspired molar concentration of O2: PaCO2 = PIO2/3.74. On the other hand, the equation is very different than that obtained from Andean natives (Hurtado): PaCO2 = 40.4-1.35(altitude in km). The proposed linear equation for Mexico gives very similar results (< 2 Torr difference) than a complex curvilinear equation by Morris et al. appropriate only up to 2.3 km. Evidence from acute exposure to altitude (acclimatized) and in North-Americans (alveolar gas) supports a reasonably accurate linear relationship up to 4 km. and also that the increase in ventilation in response to moderate altitudes in adult permanent residents is inversely proportional to molar concentration of O2. PAO2 was calculated with alveolar gas equation and resting the P(A-a)O2 we obtained PaO2. In conclusion, according to reference values in Mexico City, PaCO2 decreases about 4 Torr per km of altitude above the sea level. The decrease is similar to that reported in North-Americans and in acute exposure to altitude (acclimatized), but much less than that reported in native Peruvians. Ventilation is inversely proportional to the molar concentration of O2 at least up to an altitude where SaO2 is at or above 90%.


Assessing the antioxidative properties and chemical composition of Linaria vulgaris infusion.
The ability of Linaria vulgaris (Scrophulariaceae) infusion to act as a scavenger of 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical, reactive oxygen species (superoxide radical, hydroxyl radical, hypochlorous acid (HOCl)) and nitric oxide was investigated. The obtained data indicate that the infusion has a good scavenging activity against superoxide radical and is a very potent nitric oxide and DPPH scavenger. In hydroxyl radical assay a pro-oxidant capacity was noticed, especially for concentrations higher than 31.25 microg mL(-1). No effect was found against HOCl. A phytochemical study of this extract was also performed. The HPLC/UV analysis allowed the identification and quantification of eight organic acids (oxalic, aconitic, citric, ketoglutaric, ascorbic, malic, shikimic and fumaric acids). The phenolic composition of the lyophilised infusion was also determined by HPLC/DAD and four compounds were quantified, but, despite its high content, only linarin was managed to be identified.


Traction Injury of Recurrent Laryngeal Nerve During Thyroidectomy.
Loss of the neuromonitoring signal (LOS) during thyroidectomy signifies recurrent laryngeal nerve (RLN) injury, which is one of the common complications, especially by traction injury. Transient intraoperative LOS means spontaneous recovery of nerve function during surgery or within 6-month post-surgery. Few articles discuss intraoperative recovery time and transient LOS, and there is no consensus on the risk factors for RLN traction injury and its recovery course; thus, we wanted to determine the maximum intraoperative recovery time. This retrospective study included patients who had undergone thyroidectomies at Tainan National Cheng Kung University Hospital between January 2015 and August 2018. A total of 775 patients (with 1000 nerves at risk) who underwent intermittent intraoperative neuromonitoring during thyroidectomy were included in this study. The LOS nerves were divided into 4 groups based on the LOS subtype and the intraoperative status of the recovery. The postoperative vocal cord function was determined by thyroid ultrasound and/or laryngoscope. All the patients would be followed up postoperatively in 2-3 days, 1 week, 2 weeks, and 4-6 weeks. LOS occurred in 67 of 775 (8.6%) patients and in 70 of 1000 nerves at risk (7.0%). There were 2 in 70 nerves (2.9%) with LOS type 1 (segmental nerve traction injury) with intraoperative recovery (Group 1), 5 (7.1%) with LOS type 1 without intraoperative recovery (Group 2), 47 (67.1%) with LOS type 2 (global injury) with intraoperative recovery (Group 3), and 16 (22.8%) with LOS type 2 without intraoperative recovery (Group 4). All LOS type 1 (segmental nerve injury) nerves had pathologic lesions near the RLN or vagus nerve, but none had invaded the nerves (p < 0.05). The resolving time intraoperatively in the 2 patients in Group 1 was 5 min and 10 min, respectively. The resolving time intraoperatively in Group 3 was 1-20 min, and the average time was 4.8 min. In Group 2, 3 injured nerves recovered within 6 weeks postoperatively, and 2 nerves in 12 weeks. In Group 4, all the 16 injured nerves recovered within 6 weeks postoperatively. Applying intermittent intraoperative neuromonitoring during thyroidectomy, traction recurrent laryngeal nerve injury still happened in 7.0%. 70% of the injured nerves recovered the function intraoperatively after releasing the traction, and the longest duration of recovery is 20 min.


Edward F. Adolph Distinguished Lecture: Skin-deep insights into vascular aging.
The skin is an accessible model circulation for studying vascular function and dysfunction across the lifespan. Age-related changes, as well as those associated with disease progression, often appear first in the cutaneous circulation. Furthermore, impaired vascular signaling and attendant endothelial dysfunction, the earliest indicators of cardiovascular pathogenesis, occur in a similar fashion across multiple tissue beds throughout the body, including the skin. Because microvascular dysfunction is a better predictor of long-term outcomes and adverse cardiovascular events than is large vessel disease, an understanding of age-associated changes in the control of the human cutaneous microcirculation is important. This review focuses on 1) the merits of using skin-specific methods and techniques to study vascular function, 2) microvascular changes in aged skin (in particular, the role of the endothelial-derived dilator nitric oxide), and 3) the impact of aging on heat-induced changes in skin vasodilation. While skin blood flow is controlled by multiple, often redundant, mechanisms, our laboratory has used a variety of distinct thermal provocations of this model circulation to isolate specific age-associated changes in vascular function. Skin-specific approaches and techniques, such as intradermal microdialysis coupled with laser-Doppler flowmetry (in vivo) and biochemical analyses of skin biopsy samples (in vitro), have allowed for the targeted pharmacodissection of the mechanistic pathways controlling skin vasoreactivity and study of the impact of aging and disease states. Aged skin has an attenuated ability to vasodilate in response to warm stimuli and to vasoconstrict in response to cold stimuli.


Long-Term Outcomes after Early Neonatal Hyperglycemia in VLBW Infants: A Systematic Review.
Long-term effects of early hyperglycemia in VLBW infants are poorly characterized. The objective of this study was to systematically review the effect of early hyperglycemia on growth, metabolic health, and neurodevelopment after neonatal intensive care unit discharge in VLBW infants. The systematic review was conducted in accordance with the PRISMA guidelines. A study protocol was registered in PROSPERO (CRD42019123335). Data sources included Ovid MEDLINE, Embase, Cochrane Library, CINAHL, and Scopus. Selected studies included infants with a blood glucose concentration >150 mg/dL (8.3 mmol/L) during the first 28 days of life, a gestational age (GA) <32 weeks, and/or a birth weight <1,500 g and longitudinal data on growth, metabolic health, or neurodevelopment outcomes. The GRADE system was used to assess quality of evidence. Eight studies (n = 987 infants) reported long-term outcomes from 4-month corrected GA to 7 years old. Most studies compared long-term outcomes of preterm infants with and without hyperglycemia. Two studies addressed outcomes related to interventions following early hyperglycemia. Some studies found differences in growth, metabolic health, and neurodevelopment outcomes between VLBW preterm infants with hyperglycemia and without hyperglycemia, while other studies found no differences between groups. The overall graded quality of evidence was low. Well-designed randomized controlled and prospective studies are necessary to determine the effect of early hyperglycemia and its treatment on later metabolic and neurodevelopmental outcomes in VLBW infants. Authors propose a potential study design for standardizing the assessment of long-term metabolic and neurodevelopmental outcomes following early hyperglycemia in preterm infants.


Atp7a determines a hierarchy of copper metabolism essential for notochord development.
The critical developmental and genetic requirements of copper metabolism during embryogenesis are unknown. Utilizing a chemical genetic screen in zebrafish, we identified small molecules that perturb copper homeostasis. Our findings reveal a role for copper in notochord formation and demonstrate a hierarchy of copper metabolism within the embryo. To elucidate these observations, we interrogated a genetic screen for embryos phenocopied by copper deficiency, identifying calamity, a mutant defective in the zebrafish ortholog of the Menkes disease gene (atp7a). Copper metabolism in calamity is restored by human ATP7A, and transplantation experiments reveal that atp7a functions cell autonomously, findings with important therapeutic implications. The gene dosage of atp7a determines the sensitivity to copper deprivation, revealing that the observed developmental hierarchy of copper metabolism is informed by specific genetic factors. Our data provide insight into the developmental pathophysiology of copper metabolism and suggest that suboptimal copper metabolism may contribute to birth defects.


Superselective injection of BCNU through a latex calibrated-leak balloon.
Carmustine (BCNU) has been used extensively to treat glioblastomas by injection through a catheter placed in the cervical internal carotid artery. The technique causes severe pain to the eye and has resulted in ipsilateral blindness. The use of a latex calibrated-leak balloon positioned above the ophthalmic artery in the internal carotid artery or in one of its branches appears to circumvent the ocular complications mentioned. At an infusion rate at 125 ml/hr the balloon does not inflate and does not occlude the artery. This new technique has been used to treat 10 patients without complications.


Immunity and other defenses in pea aphids, Acyrthosiphon pisum.
Recent genomic analyses of arthropod defense mechanisms suggest conservation of key elements underlying responses to pathogens, parasites and stresses. At the center of pathogen-induced immune responses are signaling pathways triggered by the recognition of fungal, bacterial and viral signatures. These pathways result in the production of response molecules, such as antimicrobial peptides and lysozymes, which degrade or destroy invaders. Using the recently sequenced genome of the pea aphid (Acyrthosiphon pisum), we conducted the first extensive annotation of the immune and stress gene repertoire of a hemipterous insect, which is phylogenetically distantly related to previously characterized insects models. Strikingly, pea aphids appear to be missing genes present in insect genomes characterized to date and thought critical for recognition, signaling and killing of microbes. In line with results of gene annotation, experimental analyses designed to characterize immune response through the isolation of RNA transcripts and proteins from immune-challenged pea aphids uncovered few immune-related products. Gene expression studies, however, indicated some expression of immune and stress-related genes. The absence of genes suspected to be essential for the insect immune response suggests that the traditional view of insect immunity may not be as broadly applicable as once thought. The limitations of the aphid immune system may be representative of a broad range of insects, or may be aphid specific. We suggest that several aspects of the aphid life style, such as their association with microbial symbionts, could facilitate survival without strong immune protection.


A Comprehensive Review on Drug Therapies and Nanomaterials Used in Orthodontic Treatment.
Orthodontic treatment typically requires an extended duration of 1-2 years to complete the treatment. Accelerating the rate of tooth movement during orthodontic treatment is essential for shortening the overall treatment duration. After the completion of orthodontic treatment, a prominent concern arises in the form of orthodontic relapse, where the teeth tend to revert to their original positions. This issue affects approximately 60% of the global population, underscoring the importance of implementing effective measures to address orthodontic relapse. An approach in this regard involves the targeted administration of herbal and synthetic drugs applied directly to the specific area of interest to facilitate tooth movement and prevent orthodontic relapse. Apart from this, researchers are investigating the feasibility of utilizing different types of nanoparticles to improve the process of orthodontic tooth movement. In recent years, there has been a noticeable increase in the number of studies examining the effects of various drugs on orthodontics. However, the currently available literature does not provide significant evidence relating to orthodontic tooth movement. In this review, the authors provide valuable information about the drugs and nanomaterials that are capable of further enhancing the rate of orthodontic tooth movement and reducing the risk of orthodontic relapse. However, a notable hurdle remains, i.e., there is no marketed formulation available that can enhance orthodontic tooth movement and reduce treatment time. Therefore, researchers should try herbal-synthetic approaches to achieve a synergistic effect that can enhance orthodontic tooth movement. In this nutshell, there is an urgent need to develop a non-invasive, patient-compliant, and cost-effective formulation that will provide quality treatment and ultimately reduce the treatment time. Another critical issue is orthodontic relapse, which can be addressed by employing drugs that slow down osteoclastogenesis, thereby preventing tooth movement after treatment. Nevertheless, extensive research is still required to overcome this challenge in the future.


MDCT in the preoperative planning of abdominal perforator surgery for postmastectomy breast reconstruction.
This study aimed to evaluate the utility of MDCT in planning abdominal perforator surgery for breast reconstruction in patients who have undergone mastectomy. One hundred twenty-six consecutive patients scheduled for postmastectomy breast reconstruction using deep inferior epigastric perforator flaps underwent MDCT. The images were evaluated to identify, characterize, and map the dominant musculocutaneous perforator vessels of the deep inferior epigastric artery. In the first 36 patients, we compared the intraoperative findings with the preoperative MDCT findings. In the latter 90 patients, the dominant perforator vessels were directly selected on the basis of MDCT findings. We found an exact correlation between the intraoperative and radiologic findings in the first 36 cases. In the following 90 cases, the average operating time saved per patient was 1 hour 40 minutes and there was a significant reduction in postsurgical complications. The preoperative evaluation by MDCT confirmed the wide range of variability in the vascular anatomy of the abdominal wall previously described in anatomic studies. MDCT provides valuable information before surgery about the arterial anatomy of the inferior abdominal wall. It enables accurate identification of the most suitable dominant perforator vessel and makes surgical perforator flap procedures for breast reconstruction faster and safer.


Potential Medication Treatment According to Pathological Mechanisms in Abdominal Aortic Aneurysm.
Abdominal aortic aneurysm (AAA) is a vascular disease with high mortality. Because of the lack of effective medications to stop or reverse the progression of AAA, surgical operation has become the most predominant recommendation of treatment for patients. There are many potential mechanisms, including inflammation, smooth muscle cell apoptosis, extracellular matrix degradation, oxidative stress, and so on, involving in AAA pathogenesis. According to those mechanisms, some potential therapeutic drugs have been proposed and tested in animal models and even in clinical trials. This review focuses on recent advances in both pathogenic mechanisms and potential pharmacologic therapies of AAA.


Inhibition of cancer cell growth in cisplatin-resistant human oral cancer cells by withaferin-A is mediated via both apoptosis and autophagic cell death, endogenous ROS production, G2/M phase cell cycle arrest and by targeting MAPK/RAS/RAF signalling pathway.
Oral cancer is one of the common oral cavity and pharynx cancers and its treatment is limited by early metastasis, late diagnosis and emergence of drug resistance. Herein we investigated the anticancer effects of Withaferin-A against the cisplatin-resistant SCC-4 human oral cancer cells. The proliferation rate of the human oral cancer cells was evaluated by CCK-8 assay. Apoptotic cell death was studied by acridine orange (AO) / ethidium bromide (EB) staining as well as by flow cytometry using annexin V/propidium iodide (PI) staining. Cell cycle analysis was performed by flow cytometry. Reactive oxygen species (ROS) examination was carried out by flow cytometry. Protein expressions were determined by immunoblotting. The results of the MTT assay showed that Withaferin-A caused decrease in the proliferation of SCC-4 cells and exhibited an IC50 of 14 µM. The anticancer effects of Withaferin-A were mainly due to the induction of apoptosis which was linked with upsurge of Bax and depletion of BCl-2. Annexin V/PI assay showed that the apoptotic cells were 0.75, 5.8, 12.4 and 22.66% at 0,7,14 and 28 µM concentrations of Withaferin-A. Withaferin-A also caused increase in the ROS and decrease in the MMP levels of the SCC-4 cells. Western blot analysis showed that the expression of LC3B II increased while of p62 decreased remarkably upon treatment with Withaferin-A, suggestive of autophagic cell death. Cell cycle analysis by flow cytometry showed that Withaferin-A caused increase in the G2/M phase cells triggering arrest of cancer cells at the G2/M checkpoint of the cell cycle. Finally, western blot analysis showed that Withaferin-A blocked the MAPK/RAS/RAF signalling pathway in the SCC-4 cells. These results suggest that Withaferin-A may prove a potent lead molecule in oral cancer treatment and warrants further investigations.


Core-shell ZnO@CoO nitrogen doped nano-composites as highly sensitive electrochemical sensor for organophosphate pesticides detection.
Core-shell ZIF-8@ZIF-67 was synthesized by growing a cobalt-based ZIF-67 on a ZIF-8 seed particle. Herein, through selective etching of the ZIF-8@ZIF-67 core and subsequent direct carbonization, core-shell hollow ZnO@CoO nitrogen-doped nanoporous carbon (HZnO@CoO-NPC) nanocomposites were prepared. HZnO@CoO-NPCs possessed a high nitrogen content, large surface area, high degree of graphitization and excellent electrical conductivity, all of which were attributed to successfully integrating the unique advantages of ZIF-8 and ZIF-67. HZnO@CoO-NPCs were used to assemble acetylcholinesterase (AChE) biosensors for organophosphorus pesticides (OPs) detection. The low detection limit of 2.74 × 10-13 M for chlorpyrifos and 7.6 × 10-15 M for parathion-methyl demonstrated the superior sensing performance. The results showed that the electrochemical biosensor constructed by HZnO@CoO-NPC provided a sensitive and efficient electrochemical strategy for OPs detection.


Genetic epidemiology and pathology of raccoon-derived Sarcoptes mites from urban areas of Germany.
The raccoon, Procyon lotor (Carnivora: Procyonidae), is an invasive species that is spreading throughout Europe, in which Germany represents its core area. Here, raccoons mostly live in rural regions, but some urban populations are already established, such as in the city of Kassel, or are starting to build up, such as in Berlin. The objective of this study was to investigate Sarcoptes (Sarcoptiformes: Sarcoptidae) infections in racoons in these two urban areas and to identify the putative origin of the parasite. Parasite morphology, and gross and histopathological examinations of diseased skin tissue were consistent with Sarcoptes scabiei infection. Using nine microsatellite markers, we genotyped individual mites from five raccoons and compared them with Sarcoptes mites derived from fox, wild boar and Northern chamois, originating from Italy and Switzerland. The raccoon-derived mites clustered together with the fox samples and were clearly differentiated from those of the wild boar and chamois samples, which suggests a fox origin for the raccoon mange infection. These results are evidence of the cross-transmission of S. scabiei among wild carnivores. Although our results cannot elucidate whether raccoons became infected by frequent interaction with endemically or epidemically infected foxes or whether these cases resulted from occasional contacts among these animal species, they do nevertheless show that pathogens can be shared among urban populations of native and invasive carnivores.


Phase II study of weekly paclitaxel in patients with non-small cell lung cancer who have failed previous treatments.
New effective therapy is desirable for patients with non-small cell lung cancer (NSCLC) who have failed previous treatments. Fractionated administration of paclitaxel may be less toxic and more active against NSCLC. The aim of this study was to evaluate the activity and toxicity of weekly paclitaxel therapy for NSCLC in a second-line setting. Patients with pathological or cytological diagnosis of NSCLC, measurable lesions, and one or more prior therapies were enrolled. We administered weekly infusions of 80 mg/m2 paclitaxel 3 times in a 4-week cycle. In the absence of progressive disease or intolerable toxicity, each patient was treated for a minimum of 4 cycles. Of 39 patients enrolled, 1 patient achieved complete response and 11 patients achieved partial response (response rate, 31%: 95% confidence interval, 17-48%). The median survival time was 43 weeks (range, 7-128 weeks). Grade 3 or 4 leukopenia occurred in only 7 patients (18%). Neurotoxicity was the most frequent adverse effect (grades 1 and 2.26 and 5%, respectively). Although all patients recovered rapidly with corticosteroid treatment, drug-induced pneumonitis was observed in 3 patients (8%). Low-dose weekly paclitaxel is a promising therapy with high effectiveness for advanced NSCLC in patients with NSCLC who have failed previous treatments.


The smoothened agonist SAG reduces mitochondrial dysfunction and neurotoxicity of frataxin-deficient astrocytes.
Friedreich's ataxia is a rare hereditary neurodegenerative disease caused by decreased levels of the mitochondrial protein frataxin. Similar to other neurodegenerative pathologies, previous studies suggested that astrocytes might contribute to the progression of the disease. To fully understand the mechanisms underlying neurodegeneration in Friedreich's ataxia, we investigated the reactivity status and functioning of cultured human astrocytes after frataxin depletion using an RNA interference-based approach and tested the effect of pharmacologically modulating the SHH pathway as a novel neuroprotective strategy. We observed loss of cell viability, mitochondrial alterations, increased autophagy and lipid accumulation in cultured astrocytes upon frataxin depletion. Besides, frataxin-deficient cells show higher expression of several A1-reactivity markers and release of pro-inflammatory cytokines. Interestingly, most of these defects were prevented by chronically treating the cells with the smoothened agonist SAG. Furthermore, in vitro culture of neurons with conditioned medium from frataxin-deficient astrocytes results in a reduction of neuronal survival, neurite length and synapse formation. However, when frataxin-deficient astrocytes were chronically treated with SAG, we did not observe these alterations in neurons. Our results demonstrate that the pharmacological activation of the SHH pathway could be used as a target to modulate astrocyte reactivity and neuron-glia interactions to prevent neurodegeneration in Friedreich's ataxia.


Oral charcoal adsorbent (AST-120) prevents progression of cardiac damage in chronic kidney disease through suppression of oxidative stress.
Chronic kidney disease (CKD) is an important risk factor for cardiovascular disease (CVD). Increased oxidative stress plays a role in the pathogenesis of CVD in CKD patients. The oral charcoal adsorbent AST-120 attenuates the progression of CKD possibly by removing uraemic toxins such as indoxyl sulfate (IS), and reduces oxidative stress. We investigated the relationship between oxidative stress and cardiac damage in CKD and its prevention by AST-120. Male Lewis rats were administered adriamycin at 8 weeks of age, and the right kidney was removed at 12 weeks of age. From 14 weeks of age, the rats were treated daily with AST-120 (n = 8) or were untreated (control group, n = 8). At 34 weeks of age, the rats were killed and urinary and blood biochemical tests as well as cardiac histological analyses were performed. At 14 weeks of age, there were no significant differences in blood pressure, renal function (creatinine clearance: 1.54 +/- 0.28 mL/min versus 1.60 +/- 0.22 mL/min), oxidative stress markers or other biochemical data between the control and AST-120 groups. At 34 weeks, despite similar blood pressure and renal function (creatinine clearance: 0.78 +/- 0.46 mL/min versus 0.75 +/- 0.54 mL/min), serum concentrations of IS and urinary excretion of 8-hydroxydeoxyguanosine (8-OHdG), acrolein and IS were significantly lower in the AST-120 group than in the control group. Heart volume, left ventricular volume and cardiac fibrosis were significantly smaller in the experimental AST-120 group than in the control group. Immunohistological analysis revealed that the numbers of 8-OHdG- and acrolein-positive cardiomyocytes and the degrees of myocardial and perivascular fibrosis were ameliorated by AST-120 administration. The myocardial fibrosis score was significantly associated with the 8-OHdG- (r = 0.848, P < 0.001) and acrolein-positive (r = 0.812, P < 0.001) cell scores. The perivascular fibrosis score was also significantly associated with the 8-OHdG- (r = 0.906, P < 0.0001) and acrolein-positive (r = 0.789, P < 0.001) cell scores. Oxidative stress is suggested to play a key role in the development of cardiac hypertrophy and fibrosis in CKD. AST-120 may suppress oxidative stress and reduce cardiac damage in CKD.


Cerebral venous thrombosis in the mediterranean area in children.
Cerebral Venous Sinus (sinovenous) Thrombosis (CSVT) is a serious and rare disorder, increasingly recognized and diagnosed in pediatric patients. The etiology and pathophisiology has not yet been completely clarified, and unlike adults with CSVT, management in children and neonates remains controversial. However, morbidity and mortality are significant, highlighting the continued need for high-quality studies within this field. The following review will highlight aspects of CSVT in the mediteranian area in children.


Biofouling of reverse-osmosis membranes under different shear rates during tertiary wastewater desalination: microbial community composition.
We investigated the influence of feed-water shear rate during reverse-osmosis (RO) desalination on biofouling with respect to microbial community composition developed on the membrane surface. The RO membrane biofilm's microbial community profile was elucidated during desalination of tertiary wastewater effluent in a flat-sheet lab-scale system operated under high (555.6 s(-1)), medium (370.4 s(-1)), or low (185.2 s(-1)) shear rates, corresponding to average velocities of 27.8, 18.5, and 9.3 cm s(-1), respectively. Bacterial diversity was highest when medium shear was applied (Shannon-Weaver diversity index H' = 4.30 ± 0.04) compared to RO-membrane biofilm developed under lower and higher shear rates (H' = 3.80 ± 0.26 and H' = 3.42 ± 0.38, respectively). At the medium shear rate, RO-membrane biofilms were dominated by Betaproteobacteria, whereas under lower and higher shear rates, the biofilms were dominated by Alpha- and Gamma- Proteobacteria, and the latter biofilms also contained Deltaproteobacteria. Bacterial abundance on the RO membrane was higher at low and medium shear rates compared to the high shear rate: 8.97 × 10(8) ± 1.03 × 10(3), 4.70 × 10(8) ± 1.70 × 10(3) and 5.72 × 10(6) ± 2.09 × 10(3) copy number per cm(2), respectively. Interestingly, at the high shear rate, the RO-membrane biofilm's bacterial community consisted mainly of populations known to excrete high amounts of extracellular polymeric substances. Our results suggest that the RO-membrane biofilm's community composition, structure and abundance differ in accordance with applied shear rate. These results shed new light on the biofouling phenomenon and are important for further development of antibiofouling strategies for RO membranes.


A soluble C1b protein and its regulation of soluble type 7 adenylyl cyclase.
Adenylyl cyclase (AC) is a prototypical cell-signaling molecule expressed in virtually all organisms from bacteria to man. While C1b, a poorly conserved region within mammalian AC, has been implicated in numerous isoform-specific regulatory properties, no one has purified the C1b region as a functional protein to homogeneity in order to study its role in enzyme function. We hypothesize that C1b is an internal regulatory subunit. To pursue this hypothesis, we constructed several soluble C1b proteins from type VII AC, arriving at one, 7C1b-S, which can be expressed and purified from Escherichia coli. 7C1b-S is relatively stable, as demonstrated by limited proteolytic analysis, circular dichroism, and UV Raman spectroscopy. Using size-exclusion chromatography and co-immunoprecipitation we demonstrate that 7C1b-S interacts with a cardinal activator of AC (Gsalpha) and with the conserved first catalytic domain (C1a) of type VII AC. We show that 7C1b-S inhibits Gsalpha-stimulated and Gsalpha-forskolin stimulated activity in our soluble ACVII model system. On the basis of these results, we suggest that 7C1b-S meets basic criteria to serve as a model protein for the C1b region and may be used as a prototype to develop other isoform C1b soluble model proteins to further investigate the role of this domain in isoform-specific regulation of adenylyl cyclase.


Extensive Independent Amplification of Platy-1 Retroposons in Tamarins, Genus Saguinus.
Platy-1 retroposons are short interspersed elements (SINEs) unique to platyrrhine primates. Discovered in the common marmoset (Callithrix jacchus) genome in 2016, these 100 bp mobile element insertions (MEIs) appeared to be novel drivers of platyrrhine evolution, with over 2200 full-length members across 62 different subfamilies, and strong evidence of ongoing proliferation in C. jacchus. Subsequent characterization of Platy-1 elements in Aotus, Saimiri and Cebus genera, suggested that the widespread mobilization detected in marmoset (family Callithrichidae) was perhaps an anomaly. Two additional Callithrichidae genomes are now available, a scaffold level genome assembly for Saguinus imperator (tamarin; SagImp_v1) and a chromosome-level assembly for Saguinus midas (Midas tamarin; ASM2_v1). Here, we report that each tamarin genome contains over 11,000 full-length Platy-1 insertions, about 1150 are shared by both Saguinus tamarins, 7511 are unique to S. imperator, and another 8187 are unique to S. midas. Roughly 325 are shared among the three callithrichids. We identified six new Platy-1 subfamilies derived from Platy-1-8, with the youngest new subfamily, Platy-1-8c_Saguinus, being the primary source of the Saguinus amplification burst. This constitutes the largest expansion of Platy-1 MEIs reported to date and the most extensive independent SINE amplification between two closely related species.


The development of human exposure standards for radio-frequency fields.
Biological effects and health implications of radio frequency (RF) electromagnetic fields have been a subject of scientific investigation for more than 50 years. It has become a focus of attention because of the expanded use of RF radiation in the frequency range between 300 MHz and 6 GHz for wireless communication over the past decade. Another cause for the attention is the uncertainty of some observed responses and lack of understanding of the mechanism of interaction of RF electromagnetic fields with biological systems. At present, considerable efforts are devoted to developing and revising RF exposure standards. Each of these efforts should aim to make explicit the philosophy and process by which they reason and decide guidelines for deeming exposure as safe. Furthermore, the reconciliation of philosophies of protection will definitely be an asset, in practice, to those interested in international harmonization of RF exposure standards.


The communication of pharmacogenetic research results: participants weigh in on their informational needs in a pilot study.
In this brief investigation, the informational needs of research participants [n = 62; mothers who had breastfed, taken codeine, and participated in a pharmacogenetic study] were probed during a counselling session in which they received their CYP2D6 pharmacogenetic research results and overall study results. In addition to the standard information, developed by a multidisciplinary team and provided to the participants, 38% of individuals had further questions related to potential adverse effects in babies, future codeine or medication use, heredity, and consequences for policies and programmes. The diversity and complexity of the questions raised support the need to communicate the results in the context of personalized genetic counselling information sessions.


Impact of hepatitis C and liver fibrosis on antiretroviral plasma drug concentrations in HIV-HCV co-infected patients: the HEPADOSE study.
To compare plasma antiretroviral concentrations in HIV-HCV co-infected and in matched HIV mono-infected patients. This was a cross-sectional, observational study. Antiretroviral trough concentrations (C(min)) in plasma were measured in HIV-HCV co-infected patients with liver disease documented by liver biopsy, matched with HIV mono-infected patients according to gender and antiretroviral treatment. C(min) values in serum were measured using an HPLC method. Statistical analysis was performed using the Wilcoxon test. Seventy-three HIV-HCV co-infected patients and 66 HIV-infected patients were enrolled; 70% of patients were receiving a protease inhibitor (PI)- and 30% a non-nucleoside reverse transcriptase inhibitor (NNRTI)-based regimen. Among the 73 co-infected patients, 27 had a fibrosis score (Fibrotest(®)) of F4. Abacavir was the only nucleoside reverse transcriptase inhibitor whose trough concentrations differed between the co-infected and mono-infected groups. PI median plasma C(min) values were not different in the two groups, except for lopinavir, with a lower C(min) in the co-infected group than in the HIV-infected group (median 3673 versus 5990 ng/mL, P=0.04), and nelfinavir, with significantly higher concentrations in the co-infected group. Seventy-five percent of co-infected patients scoring F4, 33% of those scoring F0-F3 and 12% of HIV-infected patients were underdosed (P=0.02). Co-infected patients receiving an NNRTI had a higher plasma C(min) than HIV-infected patients; median C(min) was 3583 versus 1494 ng/mL (P=0.025) and 5331 versus 3954 ng/mL (P=0.10) for efavirenz and nevirapine, respectively. Overall, there was a greater proportion of co-infected patients with high concentrations of both NNRTIs (15/23) compared with HIV mono-infected patients (5/21) (P=0.008), especially in co-infected patients with an advanced liver fibrosis stage. Median plasma C(min) values differed significantly between HIV and HIV-HCV co-infected patients for abacavir, lopinavir and efavirenz. NNRTIs were strongly overdosed in HIV-HCV co-infected patients.


Antagonistic effects of hexose 1,6-bisphosphates and fructose 2,6-bisphosphate on the activity of 6-phosphofructokinase purified from honey-bee flight muscle.
6-Phosphofructokinase purified from honey-bee flight muscle is inhibited by ATP and, unusually, by glucose 1,6-bisphosphate and fructose 1,6-bisphosphate. The inhibition by either of the bisphosphates is not relieved by AMP, but is relieved by fructose 6-phosphate and especially by fructose 2,6-bisphosphate. Lack of effect by AMP is consistent with a low activity of adenylate kinase in this muscle.


Gastric fluid volume and pH in elective patients following unrestricted oral fluid until three hours before surgery.
This clinical study was designed to assess the results of new preoperative fasting guidelines in which patients are instructed that they must not eat any solid food after midnight, but that they may drink unrestricted amounts of clear fluid until three hours before their scheduled time of surgery. We studied 199 healthy, elective surgical inpatients aged 18-70 yr to determine whether there was any correlation between the ingestion interval or the volume of fluid ingested, with the volume and pH of residual gastric fluid at induction of anaesthesia. Pregnant patients, and those with gastric disorders or who were taking medications that affect gastric motility or secretion, were excluded. Either no premedication was given, or oral diazepam 5-15 mg was given 90 min preoperatively. Of the 199 patients, 105 ingested 50-1200 ml on the morning of surgery. The ingestion-induction interval was less than three hours in 12 patients whose actual surgery time was ahead of schedule. The remaining 94 patients did not drink because they were scheduled for surgery before 11:00 (n = 51), they did not want to drink (n = 24), or they were advised not to drink by their nurse or surgeon (n = 16). Following induction of anaesthesia, gastric fluid was aspirated through a #18 Salem sump orogastric tube, the volume was recorded and pH was measured with a calibrated pH meter. Patients were divided retrospectively into four groups (in three of which patients ingested fluid) according to the ingestion-induction interval (1.3-3.0 hr, 3.1-5.0 hr, 5.1-8.0 hr, and nothing by mouth after midnight).(ABSTRACT TRUNCATED AT 250 WORDS)


Dissecting regulatory networks of filopodia formation in a Drosophila growth cone model.
F-actin networks are important structural determinants of cell shape and morphogenesis. They are regulated through a number of actin-binding proteins. The function of many of these proteins is well understood, but very little is known about how they cooperate and integrate their activities in cellular contexts. Here, we have focussed on the cellular roles of actin regulators in controlling filopodial dynamics. Filopodia are needle-shaped, actin-driven cell protrusions with characteristic features that are well conserved amongst vertebrates and invertebrates. However, existing models of filopodia formation are still incomplete and controversial, pieced together from a wide range of different organisms and cell types. Therefore, we used embryonic Drosophila primary neurons as one consistent cellular model to study filopodia regulation. Our data for loss-of-function of capping proteins, enabled, different Arp2/3 complex components, the formin DAAM and profilin reveal characteristic changes in filopodia number and length, providing a promising starting point to study their functional relationships in the cellular context. Furthermore, the results are consistent with effects reported for the respective vertebrate homologues, demonstrating the conserved nature of our Drosophila model system. Using combinatorial genetics, we demonstrate that different classes of nucleators cooperate in filopodia formation. In the absence of Arp2/3 or DAAM filopodia numbers are reduced, in their combined absence filopodia are eliminated, and in genetic assays they display strong functional interactions with regard to filopodia formation. The two nucleators also genetically interact with enabled, but not with profilin. In contrast, enabled shows strong genetic interaction with profilin, although loss of profilin alone does not affect filopodia numbers. Our genetic data support a model in which Arp2/3 and DAAM cooperate in a common mechanism of filopodia formation that essentially depends on enabled, and is regulated through profilin activity at different steps.


Engaging School Mental Health Professionals to Deliver Evidence-Based Interventions to Hispanic Families.
Research has demonstrated that a number of evidence-based programs can be effectively implemented in different community settings, such as schools, to target Hispanic youth and their families; however, successful implementation of such programs represents a challenge for practitioners. This article describes experiences and strategies associated with recruiting, training, and supervising school mental health professionals in the school-based implementation of an evidence-based, family-centered prevention program for Hispanic families. School mental health professionals were recruited and given intensive training, weekly supervision for adherence monitoring, and ongoing technical assistance, in addition to intervention manuals and materials. We emphasize how strategies based on the prevention program itself were used to recruit, engage, and train school mental health professionals to deliver a family-based evidence-based program, blending research and practice in a large public school system. Implications of lessons learned are discussed, as well as the specific strategies to overcome challenges when engaging and training community partners in delivering a manualized intervention with rigorous adherence to the program.


Axial muscle weakness.
Axial myopathy is a rare neuromuscular disorder characterised by selective involvement of the paraspinal muscles, and presenting either as a bent spine and/or dropped head syndrome. The axial muscles can be involved in various conditions, including neuromuscular disease, movement disorders, spinal disease and metabolic disorders. There have been recent descriptions of disorders with selective axial muscle involvement, but overall axial myopathy remains under-recognised. Here, we review disorders of axial muscle function, provide guidance on interpreting axial muscles imaging and suggest a diagnostic algorithm to evaluate patients with axial muscles weakness.


Osteoinduction and long-term osseointegration promoted by combined effects of nitrogen and manganese elements in high nitrogen nickel-free stainless steel.
High nitrogen nickel-free stainless steel (HNS) has attracted particular attention in recent years, because of its potential application in orthopedic implants. However, the effects of HNS on osteoblast differentiation of bone marrow mesenchymal stem cells and long-term osseointegration are still unclear. In this study, a newly developed HNS with high manganese (Mn) content (Fe-21Cr-16Mn-2Mo-0.93N) was successfully manufactured. The HNS exhibited the improved bio-corrosion resistance and surface hydrophilicity compared to 316L stainless steel (316L SS) due to its high nitrogen (N) content. The hydrophilic surface of HNS could promote initial adhesion and spreading of rat bone marrow mesenchymal stem cells (rBMSCs) via the upregulation of the gene expression of integrin α1 and integrin β1. The high Mn and nickel-free content of HNS facilitated rBMSC proliferation. More importantly, it was revealed that ionic manganese from HNS could enhance the expression levels of osteoblast-related genes (RUNX-2, ALP, OCN and OPN) of rBMSCs, as well as promote ALP activity and OCN protein expression. Moreover, the activation of extracellular signal-related kinases (ERK) 1/2 pathway, via which HNS might induce osteogenic differentiation, was observed in rBMSCs cultured on HNS. Finally, based on HNS implantation in rabbit tibiae for 4, 12 and 26 weeks with 316L SS as a control, Micro-CT evaluation, sequential fluorescent labeling and histological analysis consistently demonstrated that HNS evidently promoted new bone formation and bone-implant contact around implants. Biomechanical tests also confirmed the increased bonding strength at the bone-HNS implant interface in both early and late stages. These results suggest that HNS can induce osteogenic differentiation of rBMSCs and promote rapid and long-term osseointegration of implants, which may be attributed to the combined effects of N and Mn elements.


Preliminary karyotype and chromosomal localization of ribosomal DNA sites in white spruce using fluorescence in situ hybridization.
We have localized the major ribosomal DNA (rDNA) loci on metaphase chromosomes and in interphase nuclei of white spruce (2n = 24) by fluorescence in situ hybridization. Hybridization sites of the biotin-labelled rDNA probe were detected using antibody-fluorochrome conjugates and a confocal laser scanning microscope. White spruce has at least 12, and possibly as many as 14, rDNA sites, 1 site present on each of seven separate chromosome pairs. This is one of the highest numbers of rDNA loci yet reported among plant species. The position of the rDNA loci together with secondary constriction patterns permit, for the first time, all homologous pairs of white spruce chromosomes to be distinguished. We discuss the application of molecular cytogenetics in studies relating to the organization and evolution of DNA sequences within conifer genomes.


Neuropeptide Y inhibition of calcium channels in PC-12 pheochromocytoma cells.
We previously demonstrated, using rat PC-12 pheochromocytoma cells differentiated to a sympathetic neuronal phenotype with nerve growth factor (NGF), that neuropeptide Y (NPY) inhibits catecholamine synthesis as well as release. Inquiry into the mechanisms of these inhibitions implicated distinct pathways involving reduction of Ca2+ influx through voltage-activated Ca2+ channels. In the present investigation the effects of NPY on whole cell Ba2+ currents were examined to obtain direct evidence supporting the mechanisms suggested by those studies. NPY was found to inhibit the voltage-activated Ba2+ current in NGF-differentiated PC-12 cells in a reversible fashion with an EC50 of 13 nM. This inhibition was pertussis toxin sensitive and resulted from NPY modulation of L- and N-type Ca2+ channels. The inhibition of L-type channels was not seen with < 1 nM free intracellular Ca2+ or when protein kinase C (PKC) was inhibited by chelerythrine or PKC-(19-31). Furthermore, the effect of NPY on L-type channels was mimicked by the PKC activator phorbol 12-myristate 13-acetate. These studies demonstrate that, in addition to inhibition of N-type Ca2+ channels, in NGF-differentiated PC-12 cells NPY inhibits L-type Ca2+ channels via an intracellular Ca(2+)- and PKC-dependent pathway.


When care is left to roam: carers' experiences of grassroots nonprofit services in Ireland.
Increasingly countries are turning to nonprofit organisations to provide health and social care, particularly for people with disabilities. Alongside this change, debates continue about how states should manage the relationship with such organisations. Should features of the old-style "welfare" model be retained? Should aspects of the "new public management" model be chosen to measure the impact of the work? Yet others argue that grassroots organisations should form the basis of a service provision system. In the context of these debates, Ireland serves as an interesting case study of the system of care that can emerge when the state operates a "relaxed control" approach. This paper takes the perspectives of users themselves: family carers who are accessing services for a disabled adult child, to examine the effects of this approach on the ground. We show how geography played a central role in shaping these experiences, and discuss how we can learn from the Irish context. Rather than arguing for narrowly defined contractual measures, we conclude by proposing a renewed focus on relationship building with the aim of effective system operation, in the future of care services.


[B-cell lymphomas with concurrent myc and bcl-2/IgH or bcl-6 translocations].
To identify and investigate clinicopathological features of B cell lymphomas with concurrent myc and bcl-2/IgH or bcl-6 translocations ("double-hit" lymphoma). Tissue microarray was constructed from formalin-fixed and paraffin-embedded tissue samples of aggressive B cell lymphomas diagnosed between 2009 and 2012, including 129 cases of diffuse large B cell lymphoma (DLBCL), 5 cases of B-cell lymphoma, unclassifiable with features intermediate between diffuse large B-cell lymphoma and Burkitt lymphoma (BCLU), 7 cases of Burkitt lymphoma and 4 cases of high-grade follicular lymphoma with diffuse large B cell lymphoma component. Interphase fluorescence in-situ hybridization (FISH) was performed with a panel of probes including myc, bcl-2/IgH and bcl-6 to document related gene translocation and copy number changes. Medical record review was performed and follow-up data was recorded. Among 145 cases, 5 cases (3.4%) of B cell lymphomas with concurrent myc and bcl-2/IgH or bcl-6 rearrangements (double-hit lymphomas) were identified, including 2 cases involving myc and bcl-2 translocations (1 DLBCL and 1 BCLU), and 3 cases involving myc and bcl-6 translocations (all DLBCLs). Three cases with concurrent bcl-2/IgH and bcl-6 translocations were found. Single gene translocations or increase of copy numbers were found in 66 cases, representing 51.2% (66/129) of all de novo DLBCLs. Ki-67 index of the 5 "double-hit" lymphomas ranged from 60% to 100%. Clinical follow-up data were available in 4 of the 5 "double-hit" lymphoma patients, three of whom died within 2 years and 1 patient was alive after 36 months of follow-up. "Double-hit" B-cell lymphomas are rare and can only be identified by molecular detection. They should not be considered synonymous with BCLU morphologically, and may present entities within other morphological spectra. Most of the patients have a poor prognosis. Further in-depth studies of larger case numbers are required to determine the pathologic and genetic variables of the lesion.


[An indicator of needs for the regional allocation of public health resources in Quebec].
This paper proposes an indicator of needs for regional resource allocation in public health in Quebec. In the literature, the only reported measure of needs for regional resource allocation in public health has two parameters: size of population and Potential Years of Life Lost for preventable causes of death (APVP-EV). We verified whether APVP-EV were associated with various indices of socioeconomic conditions and lifestyle, preventive behaviours, satisfaction with social life, healthy life expectancy and some clienteles specific to public health. Our analysis indicates that the rate of Potential Years of Life Lost by avoidable causes is significantly related to all variables except satisfaction with social life. We therefore propose using size of population and its rate of APVP-EV to determine regional allocation of resources in public health.


Modification of the frequency- and voltage-dependent effects of quinidine when administered in combination with tocainide in canine Purkinje fibers.
Frequency- and voltage-dependent modification of drug-induced inhibition of maximal upstroke velocity of the action potential (Vmax) by the combined administration of two class I antiarrhythmic drugs was studied in canine Purkinje fibers, taking depression of upstroke velocity as an indicator of sodium channel blockade. The kinetics of onset of drug-induced Vmax depression and the time course of Vmax recovery were studied after exposure to therapeutic concentrations of tocainide (50 microM) and quinidine (5 microM) both singly and in combination. The rate constant for onset of block during a drive train at a cycle length of 600 msec was 0.95 +/- 0.32 pulses in the presence of tocainide and 5.61 +/- 0.50 pulses in the presence of quinidine. The magnitude of block was three times greater with quinidine than with tocainide. The magnitude of block produced by the combination was no greater than that produced by quinidine alone and may be partly due to abbreviation of action potential duration by tocainide. Onset of block in the presence of the combination was best fitted by a double exponential with rate constants of 0.88 +/- 0.19 and 6.47 +/- 1.36 pulses. Vmax recovery after termination of a rapid train of impulses was delayed by both drugs. Poststimulation recovery from either tocainide- or quinidine-induced block was characterized by a single time constant (1.04 +/- 0.49 and 4.81 +/- 0.76 sec, respectively), while that of the combination was characterized by two time constants (0.43 +/- 0.22 and 5.94 +/- 0.56 sec), presumably corresponding to dissociation of each drug from the sodium channel receptor. The mixture of the two drugs produced a large depression of Vmax of early diastolic premature responses without producing much further depression of Vmax than that produced by quinidine alone at longer coupling intervals. The time constant of recovery from tocainide-induced block was greatly dependent upon membrane potential. After steady-state changes in frequency, the combination produced a greater depression of Vmax at rapid heart rates compared with that produced by quinidine alone, but abbreviated action potential duration more at slower heart rates. Addition of tocainide to fibers equilibrated with quinidine shifted the Vmax-membrane potential relationship to more hyperpolarized potentials, resulting in greater depression of Vmax at more depolarized membrane potentials with little or no additional depression of Vmax at more negative membrane potentials. The results provide a rationale for a possible enhanced antiarrhythmic efficacy of a combination of two class I drugs that have different kinetics of interaction with the sodium channel.


Factor analysis with a priori knowledge--application in dynamic cardiac SPECT.
Two factor analysis of dynamic structures (FADS) methods for the extraction of time-activity curves (TACs) from cardiac dynamic SPECT data sequences were investigated. One method was based on a least squares (LS) approach which was subject to positivity constraints. The other method was the well known apex-seeking (AS) method. A post-processing step utilizing a priori information was employed to correct for the non-uniqueness of the FADS solution. These methods were used to extract 99mTc-teboroxime TACs from computer simulations and from experimental canine and patient studies. In computer simulations, the LS and AS methods, which are completely different algorithms, yielded very similar and accurate results after application of the correction for non-uniqueness. FADS-obtained blood curves correlated well with curves derived from region of interest (ROI) measurements in the experimental studies. The results indicate that the factor analysis techniques can be used for semi-automatic estimation of activity curves derived from cardiac dynamic SPECT images, and that they can be used for separation of physiologically different regions in dynamic cardiac SPECT studies.


Impact of piperaquine resistance in Plasmodium falciparum on malaria treatment effectiveness in French Guiana: a descriptive epidemiological study.
Plasmodium falciparum is an apicomplexan parasite responsible for lethal cases of malaria. According to WHO recommendations, P falciparum cases are treated with artemisinin-based combination therapy including dihydroartemisinin-piperaquine. However, the emergence of resistant parasites against dihydroartemisinin-piperaquine was reported in southeast Asia in 2008 and, a few years later, suspected in South America. To characterise resistance emergence, a treatment efficacy study was performed on the reported patients infected with P falciparum and treated with dihydroartemisinin-piperaquine in French Guiana (n=6, 2016-18). Contemporary isolates collected in French Guiana were genotyped for P falciparum chloroquine resistance transporter (pfCRT; n=845) and pfpm2 and pfpm3 copy number (n=231), phenotyped using the in vitro piperaquine survival assay (n=86), and analysed through genomic studies (n=50). Additional samples from five Amazonian countries and one outside the region were genotyped (n=1440). In field isolates, 40 (47%) of 86 (95% CI 35·9-57·1) were resistant to piperaquine in vitro; these phenotypes were more associated with pfCRTC350R (ie, Cys350Arg) and pfpm2 and pfpm3 amplifications (Dunn test, p<0·001). Those markers were also associated with dihydroartemisinin-piperaquine treatment failure (n=3 [50%] of 6). A high prevalence of piperaquine resistance markers was observed in Suriname in 19 (83%) of 35 isolates and in Guyana in 579 (73%) of 791 isolates. The pfCRTC350R mutation emerged before pfpm2 and pfpm3 amplification in a temporal sequence different from southeast Asia, and in the absence of artemisinin partial resistance, suggesting a geographically distinctive epistatic relationship between these genetic markers. The high prevalence of piperaquine resistance markers in parasite populations of the Guianas, and the risk of associated therapeutic failures calls for caution on dihydroartemisinin-piperaquine use in the region. Furthermore, greater attention should be given to potential differences in genotype to phenotype mapping across genetically distinct parasite populations from different continents. Pan American Health Organization and WHO, French Ministry for Research, European Commission, Santé publique France, Agence Nationale de la Recherche, Fundação de Amparo à Pesquisa do Estado do Amazonas, Ministry of Health of Brazil, Oswaldo Cruz Foundation, and National Institutes of Health. For the French and Portuguese translations of the abstract see Supplementary Materials section.


The impact of weekend hospital admission on the timing of intervention and outcomes after surgery for spinal metastases.
Many studies have found that patients admitted on the weekend have inferior outcomes compared with those admitted on a weekday, which may be due partially to decreased availability of procedures. To evaluate the impact of weekend admission on the timing of intervention and outcomes after surgery for metastatic spine disease. Data from the Nationwide Inpatient Sample (2005-2008) were retrospectively extracted. Patients were included if they had metastatic disease and underwent spine surgery; elective hospital admissions were excluded. Multivariate logistic regression analyses were conducted to calculate the odds of undergoing early surgery, in-hospital death, and the development of a complication for patients admitted on the weekend compared with those admitted on a weekday. All analyses were adjusted for differences in age, sex, comorbid disease, primary tumor histology, myelopathy, visceral metastases, and expected primary payer, as well as hospital volume, bed size, and teaching status. We evaluated 2714 admissions. Weekend admission was associated with a significantly lower adjusted odds of receiving surgery within 1 day (odds ratio, 0.66, 95% confidence interval, 0.54-0.81; P < .001) and within 2 days (odds ratio, 0.68; 95% confidence interval, 0.56-0.83; P < .001) of admission. The adjusted odds of in-hospital death and developing a postoperative complication were not significantly different for those admitted on the weekend. In this nationwide study examining patients with spinal metastases, those admitted on the weekend were significantly less likely to receive early intervention. Future studies are needed to delineate the reasons for differences in the timing of surgery.


Transfection of apoptosis related gene Fas ligand in human hepatocellular carcinoma cells and its significance in apoptosis.
To evaluate the expression of apoptosis related gene Fas ligand (FasL) in human hepatocellular carcinoma (HCC) cells HepG2 and its significance in apoptosis. Levels of soluble Fas ligand (sFasL) in a group of patients with hepatitis B virus (HBV)-induced chronic hepatitis, HBV-positive liver cirrhosis and HCC were evaluated. In a further study, the recombinant eukaryotic expression plasmid pcDNA3.1hisB-FasL was transfected into HCC cells HepG2 by lipofection, and then soluble FasL was examined in the supernatant of culture cells by EIA, FasL expression in HepG2 cells was detected by immunohistochemistry. After being stained by annexin V and propidium iodine, cells were passed through a flow cytometer and examined by a fluorescence microscope and a laser scanning microscope. The sFasL levels were significantly lower in patients with HCC when compared to the patients with hepatitis or liver cirrhosis. In comparison with untransfected cells, the soluble FasL could be detected in the supernatant of transfected cells. FasL was expressed on the membranes and cytoplasm of transfected cells. The apoptotic cell rate was 36.30% in transfected cells, and was 11.53% in untransfected cells. Moreover, the different stage of apoptotic cells could be distinguished by annexin V and propidium iodine staining. Fas ligand is an apoptotic pathway of HCC cells.


Longitudinal vaginal septum: a proposed classification and surgical management.
To demonstrate various types of longitudinal vaginal septa (LVS), their classification, and the surgical management of typical and unique morphologic conditions of LVS. Video presentation of clinical appearance and surgical techniques for treatment of LVS. University hospital and two private. Representative cases from 121 consecutive women treated from 2013 to 2018 with LVS as a part of complex uterovaginal malformations or in isolated forms with [1] typical morphologic configuration of LVS, [2] rarer variants, or [3] specific anatomic restrictions. Resection of LVS performed as a main surgical procedure in cases with didelphys and bicornuate uterus in symptomatic women and as a part of corrective surgery of complete septate uterus. The three main nonsuturing techniques used were speculoscopy and septum excision using three different electrosurgical modalities; speculoscopy with laparoscopic devices; and vaginoscopy with hysteroscopic instruments. Clinical appearance and suggested classification, feasibility of surgery, and perioperative and anatomic results in a short follow-up period (3 months). We identified distinct types of longitudinal vaginal septa. Considering clinical appearance, we suggest classification of LVS based on four main features: [2] completeness of vaginal division: partial and complete type; [2] the symmetricity: symmetric and asymmetric position (with dominant left and right side); [3] association with the cervix: merged and isolated forms; and [4] concomitant vaginal openings: normal, and narrow openings: vaginal stenosis and hymen persistent (Fig. 1). Vaginoscopic techniques by hysteroscope were successful in atraumatic treatment of women with substantial anatomic restrictions, and all of the presented techniques can be effectively used for typical LVS. However, vessel-sealing systems allow for bloodless surgery in contrast with other methods. This study was based on previously acquired data during large prospective study approved by the local ethics committee, and written informed consent to participate in the prospective study and permit publishing anonymous data regarding the medical images, videos of procedures, and results was obtained from all patients. A new classification of longitudinal vaginal septum allows better characterization compared with the currently available classification systems. Different surgical modalities are discussed with their respective advantages and disadvantages. Vaginoscopic incision using resectoscope is a reasonable alternative for women with an intact hymen and vaginal stenosis. The impact of vaginal septum resection on obstetric, reproductive, and sexual outcomes should be assessed in randomized controlled trials and large well-designed studies.


Neu2000, an NR2B-selective, moderate NMDA receptor antagonist and potent spin trapping molecule for stroke.
Excess activation of ionotropic glutamate receptors, primarily N-methyl-D-aspartate (NMDA) receptors and free radicals, evoke nerve cell death following hypoxic-ischemic brain injury in various animal models. However, clinical trials in stroke patients using NMDA receptor antagonists have failed to show efficacy primarily due to the limited therapeutic time window for neuroprotection and a narrow therapeutic index. In comparison, antioxidants prolonged the time window for neuroprotection in animal models of ischemic stroke and showed greater therapeutic potential in clinical trials for ischemic stroke. Excess activation of NMDA receptors and free radicals mediate the two separate pathways of nerve cell death in stroke and a safe and multifunctional drug that can block both routes in the brain will likely provide a better therapeutic outcome in patients with stroke. Derivatives of the lead structures of sulfasalazine and aspirin have led to the discovery of a new molecule, Neu2000, that has demonstrated excellent neuroprotection against NMDA- and free radical-induced cell death. Neu2000 is an NR2B-selective, moderate NMDA receptor antagonist with potent cell-permeable, spin trapping antioxidant action even at nanomolar concentrations. Nonclinical and human phase I studies demonstrated that Neu2000 can be translated to treat patients with stroke with better efficacy and therapeutic time window.


Oxygenation response to positive end-expiratory pressure predicts mortality in acute respiratory distress syndrome. A secondary analysis of the LOVS and ExPress trials.
Previous trials of higher positive end-expiratory pressure (PEEP) for acute respiratory distress syndrome (ARDS) failed to demonstrate mortality benefit, possibly because of differences in lung recruitability among patients with ARDS. To determine whether the physiological response to increased PEEP is associated with mortality. In a secondary analysis of the Lung Open Ventilation Study (LOVS, n = 983), we examined the relationship between the initial response to changes in PEEP after randomization and mortality. We sought to corroborate our findings using data from a different trial of higher PEEP (ExPress, n = 749). The oxygenation response (change in ratio of arterial partial pressure of oxygen to fraction of inspired oxygen: P/F) after the initial change in PEEP after randomization varied widely (median, 9.5 mm Hg; interquartile range, -16 to 47) and was only weakly related to baseline P/F or the magnitude of PEEP change. Among patients in whom PEEP was increased after randomization, an increase in P/F was associated with reduced mortality (multivariable logistic regression; adjusted odds ratio, 0.80 [95% confidence interval, 0.72-0.89] per 25-mm Hg increase in P/F), particularly in patients with severe disease (baseline P/F [less-than-or-equal-to] 150 mm Hg). Changes in compliance and dead space were not associated with mortality. These findings were confirmed by a similar analysis of data from the ExPress trial. Patients with ARDS who respond to increased PEEP by improved oxygenation have a lower risk of death. The oxygenation response to PEEP might be used to predict whether patients will benefit from higher versus lower PEEP.


[Etiology and pathogenesis of epiphyseal necrosis in childhood as exemplified with the hip].
In many cases the etiology of juvenile necrosis of the femoral head has not been unequivocally explained. Essentially, however, it is due to reduced arterial blood flow, although venous congestion also has to be considered. Epiphyseal necrosis is frequently a result of known underlying diseases. These include, for example, both specific and nonspecific forms of coxitis and rheumatic coxitis, hormonal dysfunctions, metabolic disorders such as mucopolysaccharidosis, blood diseases (e.g. hemophilia, sickle cell anemia), leukosis, and tumors and necrosis following cortisone and cytostatic therapy. The following four factors may directly or indirectly cause arterial vascular damage and thus necrosis: 1) primarily exogenous and 2) primarily endogenous factors, 3) genetic, and 4) skeletal maturation factors. These etiological factors are described in detail and discussed. Epiphyseal deformation occurs in a strict sequence of five phases, which are described. It should be emphasized that while necrotic bone tissue is being gradually resorbed, deposits of new bone matrix are already accumulating on dead trabecula and will subsequently form new bone fibre. In other word, a gradual substitution takes place. Two important clinical facts derive from this. First, epiphyseal crash is not due to the load of the body's weight. Since active resorption takes place, there is no justification for weight-relieving therapy in the basic therapeutic concept for Perthes' disease. Second, in light of our studies, surgical containment therapy appears to be the most suitable form of treatment if performed at the right time, i.e. in the golden period of remodelling, depending on the patient's age, the extent of necrosis and the degree of dislocation. Where stringent criteria are fulfilled we perform a three-dimensional intertrochanteric osteotomy, i.e. an extension-derotation reduction osteotomy.


Factors affecting stress among Indian dental students.
The aim of this study was to investigate the perceived sources of stress and the role of parents in its etiology among dental students in a private dental school in India. A modified Dental Environment Stress (DES) Questionnaire was administered to 256 dental students. The main sources of stress were found to be fear of facing parents after failure, full loaded day, and fear of failing course or year. Students whose first choice of admission was dentistry experienced less stress than students whose first choice was another field. Also the students who joined dentistry due to parental pressure showed greater stress than those who joined of their own accord. Male students experienced greater stress than females. The results of this study indicate that a congenial environment needs to be created for dental education and parents also need to be counseled against forcing their children to join an educational program that is not of their choice.