Donation after circulatory death transplant outcomes using livers recovered by local surgeons.
Donation after circulatory death (DCD) liver transplantation (LT) outcomes have been attributed to multiple variables, including procurement surgeon recovery techniques. Outcomes of 196 DCD LTs at Mayo Clinic Arizona were analyzed based on graft recovery by a surgeon from our center (transplant procurement team [TPT]) versus a local procurement surgeon (non-TPT [NTPT]). A standard recovery technique was used for all TPT livers. The recovery technique used by the NTPT was left to the discretion of that surgeon. A total of 129 (65.8%) grafts were recovered by our TPT, 67 (34.2%) by the NTPT. Recipient age (p = 0.43), Model for End-Stage Liver Disease score (median 17 vs. 18; p = 0.22), and donor warm ischemia time (median 21.0 vs. 21.5; p = 0.86) were similar between the TPT and NTPT groups. NTPT livers had longer cold ischemia times (6.5 vs. 5.0 median hours; p < 0.001). Early allograft dysfunction (80.6% vs. 76.1%; p = 0.42) and primary nonfunction (0.8% vs. 0.0%; p = 0.47) were similar. Ischemic cholangiopathy (IC) treated with endoscopy occurred in 18.6% and 11.9% of TPT and NTPT grafts (p = 0.23). At last follow-up, approximately half of those requiring endoscopy were undergoing a stent-free trial (58.3% TPT; 50.0% NTPT; p = 0.68). IC requiring re-LT in the first year occurred in 0.8% (n = 1) of TPT and 3.0% (n = 2) of NTPT grafts (p = 0.23). There were no differences in patient (hazard ratio [HR], 1.95; 95% confidence interval [CI], 0.76-5.03; p = 0.23) or graft (HR, 1.99; 95% CI, 0.98-4.09; p = 0.10) survival rates. Graft survival at 1 year was 91.5% for TPT grafts and 95.5% for NTPT grafts. Excellent outcomes can be achieved using NTPT for the recovery of DCD livers. There may be an opportunity to expand the use of DCD livers in the United States by increasing the use of NTPT.


Hepatitis A as an etiologic agent of acute liver failure in Latin America.
This prospective, multicenter study examined the importance of hepatitis viruses as etiological agents of acute liver failure (ALF) and the outcome of ALF cases in Latin American children and adolescents. The study was conducted for minimum 12 months in 9 centers in Argentina, Brazil, Chile, Colombia, Costa Rica, and Mexico during 2001-2002. Hospitalized patients aged 1-20 years with a suspected diagnosis of ALF were included in the study and tested for serologic markers for hepatitis A, B, and C viruses. Of the 106 patients enrolled, 88 were included in the analysis. Median age was 5 years, and 55% with ALF were aged 1-5 years. A total of 37 individuals (43%) tested positive for anti-hepatitis A virus (HAV) immunoglobulin M (IgM) as marker of acute HAV infection; one was positive for anti-hepatitis B core antigen IgM and negative for hepatitis B surface antigen. None had markers of hepatitis C virus infection. Mortality rates in the overall study cohort (45%) and for those who tested anti-HAV IgM positive (41%) were similar. Forty-one percent of all patients and 46% of those positive for anti-HAV IgM underwent transplantation. The mortality rate in those with liver transplantation was half of that in patients who were not transplanted (28% versus 57%). HAV was the main etiologic agent of ALF in the population studied.


Co-designing research with Aboriginal and Torres Strait Islander consumers of mental health services, mental health workers, elders and cultural healers.
The disparity in mental health outcomes compared with non-Indigenous Australians means that there is an urgent need to develop an evidence base around how services can better support Aboriginal and Torres Strait Islander communities. A critical first step is to embed cultural safety into research methodologies. Here, we aim to establish the foundation of a research project through co-designing a qualitative interview with Aboriginal and Torres Strait Islander consumers and community members about experiences of cultural safety with mainstream mental health services. Voices of Aboriginal and Torres Strait Islander peoples must be empowered across all stages of research. An Aboriginal-led research team conducted focus groups to understand clear, sensitive, and culturally appropriate ways of asking about experiences in mental health care, to co-design an interview on this topic. Participants were Aboriginal and Torres Strait Islander consumers of mental health services, carers, mental health workers, Elders and Cultural Healers, living in Metropolitan and Regional Western Australia. Results suggest that Indigenous governance, together with investing in ongoing, and meaningful cultural awareness and cultural safety training (cultural awareness being a first step towards safety) for non-Indigenous researchers, together with taking the time to build respectful partnerships with communities through ongoing consultation, were appropriate and comprehensive methods of co-designing an interview. The process of working with Aboriginal and Torres Strait Islander peoples in research is as important as the outcome. Aboriginal and Torres Strait Islander leadership, self-determination, and relationship building with communities are essential. Empowering co-design methodologies are flexible, iterative, and ensure that the experiences and views of participants are valued, leading to more meaningful results.


Involvement of serum amyloid A1 in the rupture of fetal membranes through induction of collagen I degradation.
The de novo synthesis of serum amyloid A1 (SAA1) is augmented in human fetal membranes at parturition. However, its role in parturition remains largely unknown. Here, we investigated whether SAA1 was involved in the rupture of fetal membranes, a crucial event in parturition accompanied with extensive degradation of collagens. Results showed that SAA1 decreased both intracellular and extracellular COL1A1 and COL1A2 abundance, the two subunits of collagen I, without affecting their mRNA levels in human amnion fibroblasts. These reductions were completely blocked only with inhibition of both matrix metalloproteases (MMPs) and autophagy. Consistently, SAA1 increased MMP-2/9 abundance and the markers for autophagic activation including autophagy related (ATG) 7 (ATG7) and the microtubule-associated protein light chain 3 β (LC3B) II/I ratio with the formation of LC3 punctas and autophagic vacuoles in the fibroblasts. Moreover, the autophagic degradation of COL1A1/COL1A2 and activation of MMP-2/9 by SAA1 were blocked by inhibitors for the toll-like receptors 2/4 (TLR2/4) or NF-κB. Finally, reciprocal corresponding changes of SAA1 and collagen I were observed in the amnion following spontaneous rupture of membranes (ROM) at parturition. Conclusively, SAA1 may participate in membrane rupture at parturition by degradating collagen I via both autophagic and MMP pathways. These effects of SAA1 appear to be mediated by the TLR2/4 receptors and the NF-κB pathway.


Parasite fecundity decreases with increasing parasite load in the salmon louse Lepeophtheirus salmonis infecting Atlantic salmon Salmo salar.
Aggregation is common amongst parasites, where a small number of hosts carry a large proportion of parasites. This could result in density-dependent effects on parasite fitness. In a laboratory study, we explored whether parasite load affected parasite fecundity and survival, using ectoparasitic salmon lice (Lepeophtheirus salmonis Krøyer, 1837) infecting Atlantic salmon (Salmo salar) hosts. We found a significant reduction in fecundity with higher parasite load, but no significant effect on survival. Together with previous findings, this suggests that stronger competition amongst female lice under high parasite load is a more likely explanation than increased host immune response.


Multiple mechanisms for the activation of human platelet aggregation by Staphylococcus aureus: roles for the clumping factors ClfA and ClfB, the serine-aspartate repeat protein SdrE and protein A.
The ability of Staphylococcus aureus cells to induce platelet aggregation has long been recognized. However, despite several attempts to identify the mechanisms involved in this interaction, the nature of the bacterial receptors required remains poorly understood. Using genetic manipulation, this study for the first time provides clear evidence that several S. aureus surface proteins participate in the inter-action with platelets. Mutants of S. aureus strain Newman lacking one or more surface proteins were tested for their ability to stimulate platelet aggregation. This approach was complemented by the expression of a number of candidate proteins in the non-aggregating Gram-positive bacterium Lacto-coccus lactis. S. aureus-induced aggregation was monophasic and was dependent on the platelet receptor GPIIb/IIIa. The fibrinogen-binding proteins, clumping factors A and B and the serine-aspartate repeat protein SdrE could each induce aggregation when expressed in L. lactis. Although protein A expressed in L. lactis was not capable of inducing aggregation independently, it enhanced the aggregation response when expressed on the surface of S. aureus. Thus, S. aureus has multiple mechanisms for stimulating platelet aggregation. Such functional redundancy suggests that this phenomenon may be important in the pathogenesis of invasive diseases such as infective endocarditis.


Genome sequence of Lactobacillus helveticus, an organism distinguished by selective gene loss and insertion sequence element expansion.
Mobile genetic elements are major contributing factors to the generation of genetic diversity in prokaryotic organisms. For example, insertion sequence (IS) elements have been shown to specifically contribute to niche adaptation by promoting a variety of genetic rearrangements. The complete genome sequence of the cheese culture Lactobacillus helveticus DPC 4571 was determined and revealed significant conservation compared to three nondairy gut lactobacilli. Despite originating from significantly different environments, 65 to 75% of the genes were conserved between the commensal and dairy lactobacilli, which allowed key niche-specific gene sets to be described. However, the primary distinguishing feature was 213 IS elements in the DPC 4571 genome, 10 times more than for the other lactobacilli. Moreover, genome alignments revealed an unprecedented level of genome stability between these four Lactobacillus species, considering the number of IS elements in the L. helveticus genome. Comparative analysis also indicated that the IS elements were not the primary agents of niche adaptation for the L. helveticus genome. A clear bias toward the loss of genes reported to be important for gut colonization was observed for the cheese culture, but there was no clear evidence of IS-associated gene deletion and decay for the majority of genes lost. Furthermore, an extraordinary level of sequence diversity exists between copies of certain IS elements in the DPC 4571 genome, indicating they may represent an ancient component of the L. helveticus genome. These data suggest a special unobtrusive relationship between the DPC 4571 genome and its mobile DNA complement.


RESISTANCE TO POWDERY MILDEW8.1 boosts pattern-triggered immunity against multiple pathogens in Arabidopsis and rice.
The Arabidopsis gene RESISTANCE TO POWDERY MILDEW8.1 (RPW8.1) confers resistance to virulent fungal and oomycete pathogens that cause powdery mildew and downy mildew, respectively. However, the underlying mechanism remains unclear. Here, we show that ectopic expression of RPW8.1 boosts pattern-triggered immunity (PTI) resulting in enhanced resistance against different pathogens in both Arabidopsis and rice. In Arabidopsis, transcriptome analysis revealed that ectopic expression of RPW8.1-YFP constitutively up-regulates expression of many pathogen-associated molecular pattern (PAMP-)-inducible genes. Consistently, upon PAMP application, the transgenic line expressing RPW8.1-YFP exhibited more pronounced PTI responses such as callose deposition, production of reactive oxygen species, expression of defence-related genes and hypersensitive response-like cell death. Accordingly, the growth of a virulent bacterial pathogen was significantly inhibited in the transgenic lines expressing RPW8.1-YFP. Conversely, impairment of the PTI signalling pathway from PAMP cognition to the immediate downstream relay of phosphorylation abolished or significantly compromised RPW8.1-boosted PTI responses. In rice, heterologous expression of RPW8.1-YFP also led to enhanced resistance to the blast fungus Pyricularia oryzae (syn. Magnaporthe oryzae) and the bacterial pathogen Xanthomonas oryzae pv. oryzae (Xoo). Taken together, our data suggest a surprising mechanistic connection between RPW8.1 function and PTI, and demonstrate the potential of RPW8.1 as a transgene for engineering disease resistance across wide taxonomic lineages of plants.


Population genetic data of 74 microhaplotypes in four major U.S. population groups.
Microhaplotypes (microhaps or MHs) are novel forensically relevant genetic markers that demand large and appropriate allele frequency datasets for their implementation in casework. In this study we report on the allele frequency data of 74 microhap loci (230 SNPs) included in a newly developed 74-plex assay. The panel was tested on the Ion S5 system on a total of 347 samples from four main U.S. population groups of African, European, East Asian and Southwest Hispanic descent. Overall, frequencies of individual alleles at each locus varied considerably among the different population groups. An increase in the average value of gene diversity was also observed as the number of SNPs per locus increased. Most microhap markers showed no significant deviation from Hardy-Weinberg ratios within any of the individual population samples displaying an average power of discrimination between 0.74 and 0.81 and an average probability of exclusion between 0.32 and 0.39. Moreover, the four population groups had no clear genetic affinities with the exception of U.S. European and U.S. Southwest Hispanic populations, which showed the lowest FST value. STRUCTURE and principal component analyses (PCA) analysis resulted in effective clustering of the four populations with the U.S. European and Southwest Hispanic showing some overlap. These results support the potential use of this sequence-based 74plex-microhaplotype assay for ancestry inference in addition to previously reported human identification and mixture deconvolution capabilities.


Virtual Hepatic Venous Pressure Gradient with CT Angiography (CHESS 1601): A Prospective Multicenter Study for the Noninvasive Diagnosis of Portal Hypertension.
Purpose To develop and validate a computational model for estimating hepatic venous pressure gradient (HVPG) based on CT angiographic images, termed virtual HVPG, to enable the noninvasive diagnosis of portal hypertension in patients with cirrhosis. Materials and Methods In this prospective multicenter diagnostic trial (ClinicalTrials.gov identifier: NCT02842697), 102 consecutive eligible participants (mean age, 47 years [range, 21-75 years]; 68 men with a mean age of 44 years [range, 21-73 years] and 34 women with a mean age of 52 years [range, 24-75 years]) were recruited from three high-volume liver centers between August 2016 and April 2017. All participants with cirrhosis of various causes underwent transjugular HVPG measurement, Doppler US, and CT angiography. Virtual HVPG was developed with a three-dimensional reconstructed model and computational fluid dynamics. Results In the training cohort (n = 29), the area under the receiver operating characteristic curve (AUC) of virtual HVPG in the prediction of clinically significant portal hypertension (CSPH) was 0.83 (95% confidence interval [CI]: 0.58, 1.00). The diagnostic performance was prospectively confirmed in the validation cohort (n = 73), with an AUC of 0.89 (95% CI: 0.81, 0.96). Inter- and intraobserver agreement was 0.88 and 0.96, respectively, suggesting the good reproducibility of virtual HVPG measurements. There was good correlation between virtual HVPG and invasive HVPG (R = 0.61, P < .001), with a satisfactory performance to rule out (7.3 mm Hg) and rule in (13.0 mm Hg) CSPH. Conclusion The accuracy of a computational model of virtual hepatic venous pressure gradient (HVPG) shows significant correlation with invasive HVPG. The virtual HVPG also showed a good performance in the noninvasive diagnosis of clinically significant portal hypertension in cirrhosis. © RSNA, 2018 Online supplemental material is available for this article. See also the editorial by Malayeri in this issue.


[Hyponatremia with hyperkalemia in the newborn infant].
The electrolyte alterations of hyponatremia/hyperkalemia and their clinical picture are an emergency in the newborn period. Based on a case of pseudohypoaldosteronism the diagnostic work up of these findings and their differential diagnosis are discussed.


The epidemiology of humeral shaft fractures.
We have analysed 249 consecutive fractures of the humeral shaft treated over a three-year period. The fractures were defined by their AO morphology, position, the age and gender of the patient and the mechanism of injury. Open fractures were classified using the Gustilo system and soft-tissue injury, and closed fractures using the Tscherne system. The fractures were classified as AO type A in 63.3%, type B in 26.2% and type C in 10.4%. Most (60%) occurred in the middle third of the diaphysis with 30% in the proximal and 10% in the distal third. The severity of the fracture and soft-tissue injury was greater with increasing injury severity. Less than 10% of the fractures were open. There was a bimodal age distribution with a peak in the third decade as a result of moderate to severe injury in men and a larger peak in the seventh decade after a simple fall in women.


Individual and temporal variability of the retina after chronic bilateral common carotid artery occlusion (BCCAO).
Animal models of disease are an indispensable element in our quest to understand pathophysiology and develop novel therapies. Ex vivo studies have severe limitations, in particular their inability to study individual disease progression over time. In this respect, non-invasive in vivo technologies offer multiple advantages. We here used bilateral common carotid artery occlusion (BCCAO) in mice, an established model for ischemic retinopathy, and performed a multimodal in vivo and ex vivo follow-up. We used scanning laser ophthalmoscopy (SLO), ocular coherence tomography (OCT) and electroretinography (ERG) over 6 weeks followed by ex vivo analyses. BCCAO leads to vascular remodeling with thickening of veins starting at 4 weeks, loss of photoreceptor synapses with concomitant reduced b-waves in the ERG and thinning of the retina. Mononuclear phagocytes showed fluctuation of activity over time. There was large inter-individual variation in the severity of neuronal degeneration and cellular inflammatory responses. Ex vivo analysis confirmed these variable features of vascular remodeling, neurodegeneration and inflammation. In summary, we conclude that multimodal follow-up and subgroup analysis of retinal changes in BCCAO further calls into question the use of ex vivo studies with distinct single end-points. We propose that our approach can foster the understanding of retinal disease as well as the clinical translation of emerging therapeutic strategies.


Extending High-Level Synthesis for Task-Parallel Programs.
C/C++/OpenCL-based high-level synthesis (HLS) becomes more and more popular for field-programmable gate array (FPGA) accelerators in many application domains in recent years, thanks to its competitive quality of results (QoR) and short development cycles compared with the traditional register-transfer level design approach. Yet, limited by the sequential C semantics, it remains challenging to adopt the same highly productive high-level programming approach in many other application domains, where coarse-grained tasks run in parallel and communicate with each other at a fine-grained level. While current HLS tools do support task-parallel programs, the productivity is greatly limited ① in the code development cycle due to the poor programmability, ② in the correctness verification cycle due to restricted software simulation, and ③ in the QoR tuning cycle due to slow code generation. Such limited productivity often defeats the purpose of HLS and hinder programmers from adopting HLS for task-parallel FPGA accelerators. In this paper, we extend the HLS C++ language and present a fully automated framework with programmer-friendly interfaces, unconstrained software simulation, and fast hierarchical code generation to overcome these limitations and demonstrate how task-parallel programs can be productively supported in HLS. Experimental results based on a wide range of real-world task-parallel programs show that, on average, the lines of kernel and host code are reduced by 22% and 51%, respectively, which considerably improves the programmability. The correctness verification and the iterative QoR tuning cycles are both greatly shortened by 3.2× and 6.8×, respectively. Our work is open-source at https://github.com/UCLA-VAST/tapa/.


Tin Oxide Electron-Selective Layers for Efficient, Stable, and Scalable Perovskite Solar Cells.
Perovskite solar cells (PSCs) have become a promising photovoltaic (PV) technology, where the evolution of the electron-selective layers (ESLs), an integral part of any PV device, has played a distinctive role to their progress. To date, the mesoporous titanium dioxide (TiO2 )/compact TiO2 stack has been among the most used ESLs in state-of-the-art PSCs. However, this material requires high-temperature sintering and may induce hysteresis under operational conditions, raising concerns about its use toward commercialization. Recently, tin oxide (SnO2 ) has emerged as an attractive alternative ESL, thanks to its wide bandgap, high optical transmission, high carrier mobility, suitable band alignment with perovskites, and decent chemical stability. Additionally, its low-temperature processability enables compatibility with temperature-sensitive substrates, and thus flexible devices and tandem solar cells. Here, the notable developments of SnO2 as a perovskite-relevant ESL are reviewed with emphasis placed on the various fabrication methods and interfacial passivation routes toward champion solar cells with high stability. Further, a techno-economic analysis of SnO2 materials for large-scale deployment, together with a processing-toxicology assessment, is presented. Finally, a perspective on how SnO2 materials can be instrumental in successful large-scale module and perovskite-based tandem solar cell manufacturing is provided.


[Seroprevalence of Helicobacter pylori infection among children of different socioeconomic levels in Porto Velho, State of Rondônia].
This study investigated the seroprevalence of Helicobacter pylori infection in 200 children in the city of Porto Velho, State of Rondônia, divided in two groups of 100 children. The prevalence of seropositivity varied considerably according to socioeconomic level, such that 51% of the lower-level children and 24% of the middle-class children were positive. The characteristics of the child population relating to sex, ethnicity and diet did not represent risk factors for acquiring the infection. However, most of the infected children were in the age group of five years or older, independent of socioeconomic level. The phenotypic distribution of ABO blood groups among the infected and uninfected individuals showed that the seroreactivity to Helicobacter pylori was greater among the children with the O blood type, thus suggesting that these children have greater genetic susceptibility to infection by Helicobacter pylori.


Multiplexed random peptide library and phospho-specific antibodies facilitate human polo-like kinase 1 inhibitor screen.
One of the challenges to develop time-resolved fluorescence resonance energy transfer (TR-FRET) assay for serine/threonine (Ser/Thr) protein kinase is to select an optimal peptide substrate and a specific phosphor Ser/Thr antibody. This report describes a multiplexed random screen-based development of TR-FRET assay for ultra-high-throughput screening (uHTS) of small molecule inhibitors for a potent cancer drug target polo-like kinase 1 (Plk1). A screen of a diverse peptide library in a 384-well plate format identified several highly potent substrates that share the consensus motif for phosphorylation by Plk1. Their potencies were comparable to FKD peptide, a designed peptide substrate derived from well-described Plk1 substrate Cdc25C. A specific anti-phosphor Ser/Thr antibody p(S/T)F antibody that detects the phosphorylation of FKD peptide was screened out of 87 antibodies with time-resolved fluorometry technology in a 96-well plate format. Using FKD peptide and p(S/T)F antibody, we successfully developed a robust TR-FRET assay in 384-well plate format, and further miniaturized this assay to 1,536-well plate format to perform uHTS. We screened about 1.2 million compounds for Plk1 inhibitors using a Plk1 deletion mutant that only has the kinase domain and subsequently screened the same compound library using a full-length active-mutant Plk1. These uHTSs identified a number of hit compounds, and some of them had selectivity to either the deletion mutant or the full-length protein. Our results prove that a combination of random screen for substrate peptide and phospho-specific antibodies is very powerful strategy to develop TR-FRET assays for protein kinases.


Addressing the cost of infractions in the online literature and databases.
Myocardial infarction sometimes appears misspelt as myocardial infraction in the cardiovascular research literature. With accurate citation of literature contributions important to advancing the field and its contributors, in this study we aimed to document the frequency and explore the causes and impact of this error specific to the cardiology literature. Literature databases (PubMed, Scopus, Web of Science, WIPO, Google Scholar, Google) were searched to identify the rate of myocardial infraction errors and found an error rate between 0.2% and 1.6%, with substantial differences between search tools used. A Scopus search was used to show changes in errors over time, differences between journals and by specific authors. Myocardial infraction occurred at increasing annual rates over time and at higher rates than other errors. Increasing error rates were associated with increased volume of searchable material rather than quality of the literature. Transcription from article to database is a common source of error and some databases have higher rates of these errors. Simple measures to avoid and to correct these errors in the literature and the databases are also discussed.


[The radiotherapy of advanced breast cancer].
The authors discuss the indications for radiotherapy in late breast cancer, retrospectively analysing the experience of the Radiotherapy Department of the Portuguese Institute of Oncology in Lisbon. Treatment results of locally advanced cancer (T4a-b-c) with megavoltage machines (telecobalt therapy) using two different types of fractionation regimes are discussed. They conclude that conventional fractionation remains the best option. In elderly patients, those with ulcerated and bleeding lesions and those with socio-economic problems, it is acceptable to use non-conventional fractionation regimes with a high dose per fraction for the first part of the treatment. Advantages are overall shortening of treatment time without an increase in morbidity although survival is somewhat lower (30% versus 40% at 5 years). The authors also discuss the role of radiotherapy in disseminated disease which, in combination with systemic treatment, may be extremely useful in improving the patient's quality of life--as a way of stopping haemorrhage in bleeding lesions, for pain relief in secondary deposits, to treat superior vena cava syndrome and spinal cord compression, to prevent pathological fractures and last, but not least, to relieve or prevent symptoms such as those caused by brain metastases. The results presented in this paper are in agreement with current literature. Finally, the authors mention the most frequent complications of radiotherapy in late breast cancer as well as the guidelines for supportive care of the irradiated patient with the aim of a complete re-habilitation.


The Dominant Anterior Thoracic Artery of the Spinal Cord.
Dominant radiculomedullary arteries such as the artery of lumbar enlargement and the artery of cervical enlargement are well-documented. However, variability exists as to the size, number, and location of other radiculomedullary arteries contributing supply to the anterior spinal artery. The aim of this anatomic study was to document the prevalence and characteristics of the dominant anterior thoracic artery in cadaveric specimens. Microsurgical dissection of cadaveric human spinal cord specimens (n = 50) was conducted. The artery of lumbar enlargement was injected with colored latex until the small-caliber arterial vessels were filled. The dominant anterior thoracic artery was identified, injected, and filled with diluted industrial paint. The course, diameter, and location of the dominant anterior thoracic artery, artery of lumbar enlargement, and artery of cervical enlargement were documented. The artery of lumbar enlargement was identified between T3 and L2 in all 50 specimens (100%), and the artery of cervical enlargement was identified in 84% of specimens (42/50). At least 1 dominant anterior thoracic artery distinct from the artery of lumbar enlargement and the artery of cervical enlargement was identified between T1 and T11 in 47 of the 50 specimens (94%). The most frequent origin of the dominant anterior thoracic artery was at the level of T4 on the left. The average size of the dominant anterior thoracic artery was 0.446 mm (range, 0.300-0.759 mm on the left and 0.270-0.569 mm on the right). A dominant anterior thoracic artery is present in 94% of individuals. Variations of the arterial supply to the anterior thoracic cord are of great importance due to their implications for ischemic events as well as surgical and endovascular procedures.


Rapid liposomal formulation for nucleolin targeting to rhabdomyosarcoma cells.
Rhabdomyosarcoma (RMS) is the most common pediatric soft tissue sarcoma. More effective and less toxic therapies are urgently needed for high-risk patients. Peptide-guided targeted drug delivery can increase the therapeutic index of encapsulated drugs and improve patients' well-being. To apply this strategy to RMS, we identified the peptide F3 in a screening for peptides binding to RMS cells surface. F3 binds to nucleolin, which is present on the surface of RMS cells and is abundantly expressed at the mRNA level in RMS patients' biopsies compared to healthy tissues. We developed a rapid microfluidic formulation of F3-decorated PEGylated liposomes and remote loading of the chemotherapeutic drug vincristine. Size, surface charge, drug loading and retention of targeted and control liposomes were studied. Enhanced cellular binding and uptake were observed in three different nucleolin-positive RMS cell lines. Importantly, F3-functionalized liposomes loaded with vincristine were up to 11 times more cytotoxic than non-targeted liposomes for RMS cell lines. These results demonstrate that F3-functionalized liposomes are promising for targeted drug delivery to RMS and warrant further in vivo investigations.


Production of platelet-derived growth factor by interleukin-1 beta and transforming growth factor-beta-stimulated retinal pigment epithelial cells leads to contraction of collagen gels.
In an in vitro model of the later contractile stages of proliferative vitreoretinopathy, interleukin-1 beta (IL-1 beta) and transforming growth factor-beta (TGF-beta) stimulate the contraction of collagen gels by retinal pigment epithelial (RPE) cells. This contraction occurs after a lag period and appears not to be a direct effect of the cytokines but is mediated by another factor produced in the presence of the two cytokines. The nature of this factor has been investigated. Human RPE cells were seeded onto collagen gels in the presence of IL-1 beta and TGF-beta. After 24 hours, the conditioned medium was removed and added to new collagen gels seeded with RPE cells, and the diameter of the collagen gels was measured after various intervals. The ability of the conditioned medium to effect contraction was determined after various treatments, including size fractionation, heating, trypsin digestion, and binding to heparin-Sepharose. The involvement of platelet-derived growth factor (PDGF) as a stimulator of contraction was tested with neutralizing antibodies and by polymerase chain reaction analyses of specific mRNAs. IL-1 beta and TGF-beta cause RPE cells to contract after a delay of up to 24 hours, whereas conditioned medium from cytokine-treated cells results in immediate contraction in a manner similar to that of serum. The factor in the conditioned medium causing immediate contraction was found to be heat-stable, trypsin-sensitive, and resistant to extremes of pH. It has a size of between 30 and 50 kDa and binds heparin. The factor in conditioned medium from cytokine-treated cells does not act in the presence of C-kinase inhibitors or cycloheximide, suggesting that signaling is mediated by way of protein kinase C and new protein synthesis. Stimulation of contraction by conditioned medium is inhibited by anti-PDGF antibodies, and contraction is stimulated by human PDGF. Contraction in the presence of cytokines is mediated by the production of PDGF or a PDGF-like molecule. This factor could have implications in the pathogenesis of proliferative vitreoretinopathy.


Fractionation of single wall carbon nanotubes by length using cross flow filtration method.
A novel system for fractionating single wall carbon nanotubes (SWCNTs) by length via a three-step cross-flow filtration has been developed in which three membrane filters of different pore sizes, 1.0, 0.45, and 0.2 microm, were used. SWCNTs dispersed in water with the help of sodium carboxymethylcellulose (CMC) detergents were successfully sorted into four samples, and the atomic force microscopy (AFM) observation of those samples confirmed that their length distribution peaks are within the expected ranges from pore sizes of used filters. However, the result of the similar filtration process using a different detergent, sodium dodecylbenzenesulfonate (SDBS), showed no pronounced correlation between the length distribution of SWCNTs and the pore size. The observed difference in the sorting phenomena caused by the detergent type suggests that the permeation property depends on the complex structure resulting from the dispersed SWCNTs and detergent molecules.


Retrotransposon insertion into the maize waxy gene results in tissue-specific RNA processing.
We previously reported that three alleles of the maize waxy (wx) gene were alternatively spliced as a result of the insertion of retrotransposons into intronic sequences. In addition, inefficient splicing of element sequences with the surrounding intron produced wild-type transcripts that presumably were responsible for the observed residual gene expression in the endosperm. In this study, we report that one of these alleles, wxG, has a tissue-specific phenotype with 30-fold more WX enzymatic activity in pollen than in the endosperm. Quantification of wxG-encoded transcripts in pollen and the endosperm demonstrates that this difference can be accounted for by tissue-specific differences in RNA processing. Specifically, there is approximately 30-fold more correctly spliced RNA in pollen than in the endosperm. Based on an analogy to similar examples of tissue-specific alternative splicing in animal systems, we hypothesize that the tissue-specific phenotype of the wxG allele may reflect differences in the concentration of splicing factors in these tissues.


Diet quality score is a predictor of type 2 diabetes risk in women: the Australian Longitudinal Study on Women's Health.
The present study aimed to determine the ability of two diet quality scores to predict the incidence of type 2 diabetes in women. The study population comprised a nationally representative sample of 8370 Australian middle-aged (45-50 years) women participating in the ALSWH (Australian Longitudinal Study on Women's Health), who were free of diabetes and completed FFQ at baseline. The associations between the Australian Recommended Food Score (ARFS) and Dietary Guideline Index (DGI) with type 2 diabetes risk were assessed using multiple logistic regression models, adjusting for sociodemographic characteristics, lifestyle factors and energy intake. During 6 years of follow-up, 311 incident cases of type 2 diabetes were reported. The DGI score was inversely associated with type 2 diabetes risk (OR comparing the highest with the lowest quintile of DGI was 0·51; 95% CI 0·35, 0·76; P for trend = 0·01). There was no statistically significant association between the ARFS and type 2 diabetes risk (OR comparing the highest with the lowest quintile of ARFS was 0·99; 95% CI 0·68, 1·43; P for trend = 0·42). The results of the present prospective study indicate that the DGI score, which assesses compliance with established dietary guidelines, is predictive of type 2 diabetes risk in Australian women. The risk of type 2 diabetes among women in the highest quintile of DGI was approximately 50% lower than that in women in the lowest quintile. The ARFS was not significantly predictive of type 2 diabetes.


Serotonin uptake is largely mediated by platelets versus lymphocytes in peripheral blood cells.
The serotonin transporter (SERT), a primary target for many antidepressants, is expressed in the brain and also in peripheral blood cells. Although platelet SERT function is well accepted, lymphocyte SERT function has not been definitively characterized. Due to their small size, platelets often are found in peripheral blood mononuclear cell preparations aimed at isolating lymphocytes, monocytes, and macrophages. The presence of different cells makes it difficult to assign SERT expression and function to specific cell types. Here, we use flow cytometry and IDT307, a monoamine transporter substrate that fluoresces after uptake into cells, to investigate SERT function in lymphocyte and platelet populations independently, as well as simultaneously without prior isolation. We find that murine lymphocytes exhibit temperature-dependent IDT307 transport but uptake is independent of SERT. Lack of measurable SERT function in lymphocytes was corroborated by chronoamperometry using serotonin as a substrate. When we examined rhesus and human mixed blood cell populations, we found that platelets, and not lymphocytes, were primary contributors to SERT function. Overall, these findings indicate that lymphocyte SERT function is minimal. Moreover, flow cytometry, in conjunction with the fluorescent transporter substrate IDT307, can be widely applied to investigate SERT in platelets from populations of clinical significance.


Liver biopsy analysis has a low level of performance for diagnosis of intermediate stages of fibrosis.
There is controversy about the performance of noninvasive tests such as FibroTest in diagnosing intermediate stages of fibrosis. We investigated whether this controversy results from limitations of biopsy analysis for intermediate-stage fibrosis and inappropriate determination of the standard area under the receiver-operator characteristic curve (AUROC). To determine whether biopsy has a lower diagnostic performance for fibrosis stage F2 (few septa) vs F1 (fibrosis without septa), compared with its performance for F1 vs F0 or F4 vs F3, we determined the fibrotic areas of large surgical samples collected from 20 consecutive patients with chronic liver disease or normal liver tissue that surrounded tumors. We analyzed digitized images of 27,869 virtual biopsies of increasing length and also analyzed data from 6500 patients with interpretable FibroTest results who also underwent biopsy analysis. The overall performance of biopsy analysis (by Obuchowski measure) increased with biopsy length from 0.885 for 5-mm to 0.912 for 30-mm samples (P < .0001). The performance of biopsy was lower for the diagnosis of F2 vs F1 samples (weighted AUROC [wAUROC] = 0.505) than for F1 vs F0 (wAUROC = 0.773; 53% difference; P < .0001) or F4 vs F3 (wAUROC = 0.700; 39% difference; P < .0001), even when 30-mm biopsy samples were used. The performance of FibroTest was also lower for the diagnosis of F2 vs F1 samples (wAUROC = 0.512) than for F1 vs F0 samples (wAUROC = 0.626; 22% difference; P < .0001) or F4 vs F3 (wAUROC = 0.628; 23% difference; P < .0001). However, the FibroTest had smaller percentage differences among wAUROC values than biopsy. Biopsy has a low level of diagnostic performance for fibrosis stages F2 and F1. The recommendation for biopsy analysis, instead of a validated biomarker panel such as FibroTest, for the diagnosis of intermediate stages of fibrosis is therefore misleading.


Structural and kinetic properties of nonglycosylated recombinant Penicillium amagasakiense glucose oxidase expressed in Escherichia coli.
The gene coding for Penicillium amagasakiense glucose oxidase (GOX; beta-D-glucose; oxygen 1-oxidoreductase [EC 184.108.40.206]) has been cloned by PCR amplification with genomic DNA as template with oligonucleotide probes derived from amino acid sequences of N- and C-terminal peptide fragments of the enzyme. Recombinant Escherichia coli expression plasmids have been constructed from the heat-induced pCYTEXP1 expression vector containing the mature GOX coding sequence. When transformed into E. coli TG2, the plasmid directed the synthesis of 0.25 mg of protein in insoluble inclusion bodies per ml of E. coli culture containing more than 60% inactive GOX. Enzyme activity was reconstituted by treatment with 8 M urea and 30 mM dithiothreitol and subsequent 100-fold dilution to a final protein concentration of 0.05 to 0.1 mg ml-1 in a buffer containing reduced glutathione-oxidized glutathione, flavin adenine dinucleotide, and glycerol. Reactivation followed first-order kinetics and was optimal at 10 degrees C. The reactivated recombinant GOX was purified to homogeneity by mild acidification and anion-exchange chromatography. Up to 12 mg of active GOX could be purified from a 1-liter E. coli culture. Circular dichroism demonstrated similar conformations for recombinant and native P. amagasakiense GOXs. The purified enzyme has a specific activity of 968 U mg-1 and exhibits kinetics of glucose oxidation similar to those of, but lower pH and thermal stabilities than, native GOX from P. amagasakiense. In contrast to the native enzyme, recombinant GOX is nonglycosylated and contains a single isoform of pI 4.5. This is the first reported expression of a fully active, nonglycosylated form of a eukaryotic, glycosylated GOX in E. coli.


Effects of nitrate and the pathogenic water mold Saprolegnia on survival of amphibian larvae.
We tested for a synergism between nitrate and Saprolegnia, a pathogenic water mold, using larvae of 3 amphibian species: Ambystoma gracile (northwestern salamander), Hyla regilla (Pacific treefrog) and Rana aurora (red-legged frog). Each species was tested separately, using a 3 x 2 fully factorial experiment with 3 nitrate treatments (none, low and high) and 2 Saprolegnia treatments (Saprolegnia and control). Survival of H. regilla was not affected significantly by either experimental factor. In contrast, survival of R. aurora was affected by a less-than-additive interaction between Saprolegnia and nitrate. Survival of R. aurora was significantly lower in the Saprolegnia compared to the control treatment when nitrate was not added, but there was no significant difference in survival between Saprolegnia and control treatments in the low and high nitrate treatments, consistent with increased nitrate preventing Saprolegnia from causing mortality of R. aurora. Survival of A. gracile followed a similar pattern, but the difference between Saprolegnia and control treatments when nitrate was not added was not significant, nor was the nitrate x Saprolegnia interaction. Our study suggests that Saprolegnia can cause mortality in amphibian larvae, that there are interspecific differences in susceptibility and that the effects of Saprolegnia on amphibians are context-dependent.


Quantitative Analysis of Target Peptides Related to Resistance Against Ascochyta Blight (Peyronellaea pinodes) in Pea.
Peyronellaea pinodes causes Ascochyta blight, one of the major diseases in pea worldwide. Cultivated pea plants have a low resistance to this disease. Although quantitative trait loci (QTLs) involved in the resistance to Ascochyta blight have been identified, the specific genes associated with these QTLs remain unknown, which makes marker-assisted selection difficult. Complex traits alter proteins and their abundance. Quantitative estimation of proteins in pea might therefore be useful in selecting potential markers for breeding. In this work, we developed a strategy using a combination of shotgun proteomics (viz., high performance liquid chromatography-mass spectrometry data-dependent acquisition) and data-independent acquisition (DIA) analysis, to identify putative protein markers associated with resistance to Ascochyta blight and explored its use for breeding selection. For this purpose, an initial list of target peptides based on proteins closely related to resistance to P. pinodes was compiled by using two genotypes with contrasting responses to the disease. Then, targeted data analysis (viz., shotgun proteomics-DIA) was used for constitutive quantification of the target peptides in a representative number of the recombinant inbred line population segregated for resistance as derived from a cross between the two genotypes. Finally, a peptide panel of potential markers for resistance to P. pinodes was built. The results thus obtained are discussed and compared with those of previous gene expression studies using the same parental pea genotypes responding to the pathogen. Also, a molecular defense mechanism against Ascochyta blight in pea is proposed. To the authors' knowledge, this is the first time a targeted proteomics approach based on data analysis has been used to identify peptides associated with resistance to this disease.


Transverse section imaging of mean clearance time.
Transverse sections representing the magnitude of clearance rates or the mean transit time of radionuclide flow through known volumes of interest are computed from angular projections. First the integrated counts for all time intervals at each projection bin for each angle are computed. These modified projection sums are convolved with a filter and backprojected to form an integrated event image. A second image is formed by backprojected the sum over time of the projected values for each angle weighted time. The ratio of the second image to the first image gives a mean clearance time image with high signal to noise and accuracy after thresholding to remove noise in zero-flow regions. The algorithm is verified by computer simulations and has a major benefit in the savings of computational time by a factor of 10 for anticipated clinical studies of brain and kidney blood flow and lung ventilation and perfusion.


Diminished mephobarbital anticonvulsant action following diphenhydramine pretreatment.
Diphenhydramine and other antihistamines produce biphasic effects on drug disposition and lower seizure threshold, thereby potentially diminishing the efficacy of anticonvulsants such as mephobarbital. Accordingly, the influence of diphenhydramine (50 mg/kg, IP) pretreatment on the anticonvulsant activity of mephobarbital (50 mg/kg, IP) was determined in adult female Swiss-Webster mice given pentylenetetrazol (SC). Diphenhydramine lowered the pentylenetetrazol convulsive dose (CD50) by 60%. Administration of diphenhydramine in combination with mephobarbital produced a 65% decrease in the CD50 of pentylenetetrazol in comparison with that of animals given mephobarbital plus pentylenetetrazol. Pharmacokinetic evaluation of mephobarbital blood level data indicates that the mechanism responsible for the observed interaction between diphenhydramine and mephobarbital involves a decrease in mephobarbital uptake from the administration site.


Lower Extremity Peripheral Artery Disease Without Chronic Limb-Threatening Ischemia: A Review.
Lower extremity peripheral artery disease (PAD) affects approximately 8.5 million people in the US and approximately 230 million worldwide. Peripheral artery disease is uncommon before aged 50 years but affects up to 20% of people aged 80 years and older. It can be noninvasively diagnosed with the ankle-brachial index (ABI), a ratio of Doppler-recorded pressures in the dorsalis pedis and/or posterior tibial artery in each leg to brachial artery pressures. An ABI value less than 0.90 is 57% to 79% sensitive and 83% to 99% specific for arterial stenosis of at least 50%. Intermittent claudication, consisting of exertional calf pain that does not begin at rest and that resolves within 10 minutes of rest, is considered the classic symptom of PAD. However, 70% to 90% of people with an ABI value less than 0.90 either report no exertional leg symptoms (ie, asymptomatic) or report leg symptoms with walking that are not consistent with classic claudication. Over time, people with PAD restrict walking activity or slow walking speed to avoid leg symptoms. Thus, although approximately 75% of people with PAD report no change in leg symptoms over time, those with PAD have significantly greater annual declines in 6-minute walk performance compared with those without it. Approximately 11% of people with PAD develop chronic limb-threatening ischemia, the most severe form of PAD. Compared with people without PAD, those with the disease have approximately twice the rate of all-cause mortality, cardiovascular mortality, and major coronary events at 10-year follow-up. High-dose statins and antiplatelet therapy with or without antithrombotic therapy reduced rates of coronary events and stroke in people with PAD. Supervised treadmill exercise improved 6-minute walk distance by 30 to 35 m, consistent with a clinically meaningful change, whereas effective home-based walking exercise interventions improved 6-minute walk by 42 to 53 m. Effective home-based exercise programs require behavioral methods, including monitoring by a coach. Peripheral artery disease affects approximately 230 million people worldwide and is associated with increased rates of cardiovascular events, lower extremity events, and functional decline compared with that of people without PAD. People with PAD should be treated with the highest dose of statin tolerated, antithrombotic and/or antiplatelet therapy, and exercise.


GTRAC: fast retrieval from compressed collections of genomic variants.
The dramatic decrease in the cost of sequencing has resulted in the generation of huge amounts of genomic data, as evidenced by projects such as the UK10K and the Million Veteran Project, with the number of sequenced genomes ranging in the order of 10 K to 1 M. Due to the large redundancies among genomic sequences of individuals from the same species, most of the medical research deals with the variants in the sequences as compared with a reference sequence, rather than with the complete genomic sequences. Consequently, millions of genomes represented as variants are stored in databases. These databases are constantly updated and queried to extract information such as the common variants among individuals or groups of individuals. Previous algorithms for compression of this type of databases lack efficient random access capabilities, rendering querying the database for particular variants and/or individuals extremely inefficient, to the point where compression is often relinquished altogether. We present a new algorithm for this task, called GTRAC, that achieves significant compression ratios while allowing fast random access over the compressed database. For example, GTRAC is able to compress a Homo sapiens dataset containing 1092 samples in 1.1 GB (compression ratio of 160), while allowing for decompression of specific samples in less than a second and decompression of specific variants in 17 ms. GTRAC uses and adapts techniques from information theory, such as a specialized Lempel-Ziv compressor, and tailored succinct data structures. The GTRAC algorithm is available for download at: https://github.com/kedartatwawadi/GTRAC CONTACT: : firstname.lastname@example.com Supplementary data are available at Bioinformatics online.


The effects of remnant-preserving anterior cruciate ligament reconstruction on proprioception: A prospective comparative study.
The aim of this study was to investigate the effects of anterior cruciate ligament reconstruction performed by preserving remnant tissue on proprioception and to assess the effects it has on isokinetic quadriceps and hamstring muscle strength, as well as on range of motion and functional scores. A prospective study was conducted with 44 patients who underwent either anterior cruciate ligament reconstruction with remnant preservation (study group, n=22) or with remnant excision (control group, n=22) with the use of a 4-strand hamstring allograft. The mean follow-up time was 20.2 ± 1.4 months after surgery. Using an isokinetic dynamometer, proprioception was evaluated with passive joint position perception at 150, 450, and 600, and quadriceps femoris, and hamstring muscle strength were evaluated at speeds of 900, 1800, and 2400 per second. Range of motion was measured using a goniometer. Functional outcomes were assessed using International Knee Documentation Committee subjective knee evaluation score and Lysholm knee scoring questionnaires. It was only at 15° of knee flexion that there was a statistically significant difference in proprioception; the median of the difference in the amount of deviation from the target angle between the healthy knee and the operated side was 1.7 (range, 0.7-20.7) in those with remnant preserved, and 2.7 (range, 1-26) in those with remnant excised (P=.016). At 2400/s speed, the mean quadriceps femoris strength was 77.2 ± 24.3 Nm in those with remnant preserved and 67.6 ± 24.2 Nm in those with remnant excised. (P=.048) There was no difference between the 2 groups in terms of range of motion, International Knee Documentation Committee, and Lysholm knee scoring. (P > .05) Conclusion: The present study has demonstrated that better proprioception and higher quadriceps femoris muscle strength can be obtained by remnant-preserving anatomical single-bundle anterior cruciate ligament reconstruction using a hamstring autograft. Level II, Therapeutic study.


How can patient preferences be used and communicated in the regulatory evaluation of medicinal products? Findings and recommendations from IMI PREFER and call to action.
Objective: Patients have unique insights and are (in-)directly affected by each decision taken throughout the life cycle of medicinal products. Patient preference studies (PPS) assess what matters most to patients, how much, and what trade-offs patients are willing to make. IMI PREFER was a six-year European public-private partnership under the Innovative Medicines Initiative that developed recommendations on how to assess and use PPS in medical product decision-making, including in the regulatory evaluation of medicinal products. This paper aims to summarize findings and recommendations from IMI PREFER regarding i) PPS applications in regulatory evaluation, ii) when and how to consult with regulators on PPS, iii) how to reflect PPS in regulatory communication and iv) barriers and open questions for PPS in regulatory decision-making. Methods: PREFER performed six literature reviews, 143 interviews and eight focus group discussions with regulators, patient representatives, industry representatives, Health Technology Assessment bodies, payers, academics, and clincians between October 2016 and May 2022. Results: i) With respect to PPS applications, prior to the conduct of clinical trials of medicinal products, PPS could inform regulators' understanding of patients' unmet needs and relevant endpoints during horizon scanning activities and scientific advice. During the evaluation of a marketing authorization application, PPS could inform: a) the assessment of whether a product meets an unmet need, b) whether patient-relevant clinical trial endpoints and outcomes were studied, c) the understanding of patient-relevant effect sizes and acceptable trade-offs, and d) the identification of key (un-)favorable effects and uncertainties. ii) With respect to consulting with regulators on PPS, PPS researchers should ideally have early discussions with regulators (e.g., during scientific advice) on the PPS design and research questions. iii) Regarding external PPS communication, PPS could be reflected in the assessment report and product information (e.g., the European Public Assessment Report and the Summary of Product Characteristics). iv) Barriers relevant to the use of PPS in regulatory evaluation include a lack of PPS use cases and demonstrated impact on regulatory decision-making, and need for (financial) incentives, guidance and quality criteria for implementing PPS results in regulatory decision-making. Open questions concerning regulatory PPS use include: a) should a product independent broad approach to the design of PPS be taken and/or a product-specific one, b) who should optimally be financing, designing, conducting, and coordinating PPS, c) when (within and/or outside clinical trials) to perform PPS, and d) how can PPS use best be operationalized in regulatory decisions. Conclusion: PPS have high potential to inform regulators on key unmet needs, endpoints, benefits, and risks that matter most to patients and their acceptable trade-offs. Regulatory guidelines, templates and checklists, together with incentives are needed to foster structural and transparent PPS submission and evaluation in regulatory decision-making. More PPS case studies should be conducted and submitted for regulatory assessment to enable regulatory discussion and increase regulators' experience with PPS implementation and communication in regulatory evaluations.


Oleanolic acid exerts bone protective effects in ovariectomized mice by inhibiting osteoclastogenesis.
Postmenopausal osteoporosis (POP) is quite prevalent and many new drugs are under development to obtain better therapeutic outcomes. Oleanolic acid (OA) has been reported to prevent bone loss in ovariectomized (OVX) rats by stimulating osteoblastogenesis. One previous study has demonstrated that acetate of OA suppressed lipopolysaccharides (LPS)-induced bone loss in mice. However, the role of OA in the receptor activator of nuclear factor kappa-B ligand (RANKL)-mediated osteoclastogenesis is still not elucidated. Here we show that OA dose-dependently inhibits RANKL-mediated osteoclastogenesis and the formation of functional osteoclasts without impairing the viability and osteoclastic potential in bone marrow macrophages (BMMs). Moreover, OA administration attenuates bone loss in OVX mice by inhibiting osteoclast's densities. Mechanistically, OA does not affect RANKL-induced activation of the NF-кB, JNK, p38, ERK and Akt pathways, but inhibits the expression of the nuclear factor of activated T-cells c1(NFATc1) and c-Fos. Moreover, OA significantly suppresses the expression of RANKL-activated osteoclast genes encoding matrix metalloproteinase 9 (MMP9), Cathepsin K(Ctsk), tartrate-resistant acid phosphatase (TRAP) and carbonic anhydrase II (Car2). This work has elucidated the molecular mechanism of OA in RANKL-mediated osteoclastogenesis and revealed the promising potential of OA to be further developed as a new drug to prevent and treat POP.


Claudins play a role in normal and tumor cell motility.
Claudins are key integral proteins of the tight junction. Although they play an essential role in controlling paracellular diffusion in epithelia, increasing evidence supports a role for these proteins in non-barrier forming activities. To elucidate a potential function for claudins outside of their traditional role in tight junctions, subcellular localization of claudin-4 was determined in normal mammary epithelial cells as well as breast and ovarian cancer cell lines and the effects of a claudin mimic peptide on cell motility were determined. Immunofluorescence revealed that claudin-4 was localized along cellular projections. Using a fluorescent peptide that mimics a conserved sequence in the second extracellular loop of a set of claudin subtypes, that includes claudin-4, exposure of this loop to the extracellular environment was confirmed in non-polarized cells. This peptide inhibited cell motility when normal mammary epithelial cells as well as breast and ovarian tumor cells were subjected to a wound healing assay. Knockdown of claudin-4 also inhibited cell motility and the mimic peptide had no effect on motility in the claudin-4 knockdown cells. This effect on motility was seen when cells were grown on collagen, but not when cells were grown on non-physiological cell adhesive or fibronectin. The second extracellular loop of claudins is able to interact with the extracellular environment to promote normal and tumor cell motility when it is not associated with tight junction structures.


Evidence-based rehabilitation following anterior cruciate ligament reconstruction.
Following a bone-patellar tendon-bone autograft (BPTB) or four-stranded semitendinosus/gracilis tendons autograft (ST/G) anterior cruciate ligament (ACL) reconstruction, the speed and safety with which an athlete returns to sports (or regains the pre-injury level of function) depends on the rehabilitation protocol. Considering the large differences in clinical and outpatient protocols, there is no consensus regarding the content of such a rehabilitation program. Therefore, we conducted a systematic review to develop an optimal evidence-based rehabilitation protocol to enable unambiguous, practical and useful treatment after ACL reconstruction. The systematic literature search identified 1,096 citations published between January 1995 and December 2006. Thirty-two soundly based rehabilitation programs, randomized clinical trials (RCT's) and reviews were included in which common physical therapy modalities (instruction, bracing, cryotherapy, joint mobility training, muscle-strength training, gait re-education, training of neuromuscular function/balance and proprioception) or rehabilitation programs were evaluated following ACL reconstruction with a BPTB or ST/G graft. Two reviews were excluded because of poor quality. Finally, the extracted data were combined with information from background literature to develop an optimal evidence-based rehabilitation protocol. The results clearly indicated that an accelerated protocol without postoperative bracing, in which reduction of pain, swelling and inflammation, regaining range of motion, strength and neuromuscular control are the most important aims, has important advantages and does not lead to stability problems. Preclinical sessions, clear starting times and control of the rehabilitation aims with objective and subjective tests facilitate an uncomplicated rehabilitation course. Consensus about this evidence-based accelerated protocol will not only enhance the speed and safety with which an athlete returns to sports, but a standardized method of outcome measurement and reporting will also increase the evidential value of future articles.


Multiple in vivo Effects of Cadmium on Photosynthetic Electron Transport in Pea Plants.
The inhibitory effects of cadmium (CdSO4 ) on the primary photosynthetic processes were studied in vivo in Pisum sativum by using Multi-function Plant Efficiency Analyser (M-PEA-2). Photosynthetic parameters related to photosystem (PS) II, PS I and intersystem electron carriers were calculated from the light-induced kinetics of prompt chlorophyll a fluorescence (OJIP transient), delayed chlorophyll a fluorescence (DF), and 820 nm modulated reflection (MR). Low-dose exposure to cadmium (20 μm CdSO4 for 48 h) reduced probability of electron transfer from plastoquinones to the terminal electron acceptors of PSI (δRo ) accompanied by a decrease in the rate of P700 + and PC reduction (Vred ) and the magnitude of the I2 step on the DF kinetics. Electron transport through PSI remained unaltered. The obtained results allowed us to propose existence of the potential site of inhibition of photosynthetic electron flow by cadmium between PSII and PSI. We propose to use parameters δRo , Vred , and I2 /I1 as sensitive indicators of an early contamination by heavy metals.


Blood group, red cell, and serum protein variation in the Cabecar and Huetar, two Chibchan Amerindian tribes of Costa Rica.
Genetic variation, using blood groups and red cell and serum proteins, was surveyed in the Cabecar of Chirripo and the Huetar of Quitirrisi, Costa Rica. Thirty-nine loci were screened in a sample of 91 Cabecars and 40 loci in 45 Huetars. Twenty-seven loci were monomorphic in the Cabecar and 30 in the Huetar. The proportions of polymorphic loci (P), out of 34 studied by electrophoresis, were 0.235 and 0.177, respectively. Estimated gene diversities (H) of the polymorphic loci were 0.050 in the Cabecar and 0.053 in the Huetar. Two polymorphisms, reported until now in Costa Rican and Panamanian Chibchan groups only, occurred at very high frequencies: TF*DGUA = 0.357 in the Cabecar, the highest frequency ever reported, and 0.033 in the Huetar; and PEPA*F, which reached 0.26 in the Cabecar and 0.29 in the Huetar. Nei's genetic distances and trees (two methods) were used to compare them to seven other Chibchan tribes of Costa Rica. The results placed both the Cabecar of Chirripo and the Huetar closer to the Talamancan Tribes (Bribri and Cabecar). This was an unexpected result for the Huetar, since linguistic studies suggested a closer relationship to the Guatuso. GST, DST, RST, and Dm for three Cabecar subpopulations (Atlantic, Chirripo, and Pacific) doubled their values compared to estimates based on comparison of only two subpopulations: Atlantic and Pacific. Total genetic diversity considering just the three Cabecar subpopulations resembled that obtained including them plus six other Chibchan populations of Costa Rica.


Cytoplasmic acidification is not an effector mechanism of VP16 or DEX-induced apoptosis in CEM T leukaemia cells.
The role of intracellular acidification in the execution phase of apoptosis is not well understood. Here we examine the effect of Bcl-2 over-expression on intracellular acidification occurring during apoptosis. We found, that in CEM cells, neither DEX nor VP16-induced apoptosis lead to a significant change in intracellular pH (pHi). Furthermore, we found that shifting pHi away from physiological values was unable to induce chromatin condensation or poly(ADP-ribose) polymerase (PARP) cleavage in the presence of Bcl-2 over-expression. However, it was found that maximum chromatin condensation and PARP cleavage occurred at near physiological pHi values. Taken together these data suggest that intracellular acidification does not trigger the effector phase of CEM apoptosis.


Professional satisfaction of physicians.
Nineteen physicians were interviewed to identify how they defined and solved the problems facing their profession. Problems cited by the physicians included loss of control over medical decisions, loss of control of the referral process, the threat of malpractice suits, ethical issues facing health maintenance organization physicians, and reduction of income. The attitudes and problem-solving approaches of physicians who remained satisfied with their profession are presented and compared with those of physicians who are essentially dissatisfied with the practice of medicine. Among satisfied physicians, happiness in medicine was attributed more to attitudinal adjustments than to success at the "business of medicine."


Clinical Significance of Ubiquitin Specific Protease 7 (USP7) in Predicting Prognosis of Hepatocellular Carcinoma and its Functional Mechanisms.
BACKGROUND Hepatocellular carcinoma (HCC) accounts for one of the most prevalent cancer types in the world. The ubiquitin specific protease 7 (USP7), a kind of deubiquitylating enzyme, has been reported to play multifaceted roles in different tumor types. The aim of this study was to investigate the expression and function of USP7 in HCC. MATERIAL AND METHODS Immunohistochemical staining and quantitative PCR were performed to explore the expression of USP7 in both HCC tissues and adjacent normal liver tissues. Chi-square test, univariate analysis, and multivariate analysis were conducted to statistically evaluate the clinical significance of USP7 in HCC. Proliferation, migration, and invasion capacities of HCC cells were assessed after overexpressing or silencing USP7. RESULTS Both the RNA and protein levels of USP7 were upregulated in HCC tissues compared to normal liver tissues. High expression of USP7 was correlated with advanced tumor stage and poor overall survival. Moreover, USP7 was identified as a novel independent prognostic factor for HCC patients. Cellular studies showed that USP7 could enhance the proliferation, migration, and invasion capacities of HCC cells, thereby promoting tumor progression. CONCLUSIONS High expression of USP7 is frequent in HCC tissues, which promotes tumor proliferation and invasion, and is correlated with a poor overall survival. Targeting USP7 may be a novel direction for the drug development of HCC therapy.


Taste perception in kidney disease and relationship to dietary sodium intake.
Taste abnormalities are prevalent in Chronic Kidney Disease (CKD) potentially affecting food palatability and intake, and nutrition status. The TASTE CKD study aimed to assess taste and explore the relationship of dietary sodium intake with taste disturbance in CKD subjects. This was a cross-sectional study of 91 adult stage 3-5 CKD participants (78% male) aged 65.9 ± 13.5 years with mean estimated glomerular filtration rate of 33.1 ± 12.7 ml/min/1.73 m(2), and 30 controls (47% male) aged 55.2 ± 7.4 years without kidney dysfunction. Taste assessment was performed in both groups, presenting five basic tastes (sweet, sour, salty, umami and bitter) in blinded 2 ml solution which the participants tasted, identified (identification) and rated perceived strength (intensity) on a 10 cm visual analogue scale. Sodium intake was measured in the CKD group using validated food frequency questionnaire to determine high or low sodium intake (cut-off 100 mmol sodium/day). Differences between groups (CKD vs controls; high vs low sodium intake) were analysed using chi-square for identification and t-test for intensity. Multivariate analysis was used to adjust for age and gender differences between CKD and controls. The control group identified mean 3.9 ± 1.0 tastants correctly compared with 3.0 ± 1.2 for CKD group (p < 0.001), which remained significant after adjustment for age and gender. After adjustment for age and gender, sour identification and intensity and salty and umami intensity were impaired in CKD compared with controls. Participants with low sodium intake were more likely to correctly identify salty and umami, and rated intensity of umami and bitter significantly higher than those with high sodium intake. These findings add to the body of evidence suggesting that taste changes occur with CKD, independent of age and gender differences, with specific impairment in sour, umami and salty tastes. Our finding that sodium intake is related to umami and bitter disturbance as well as salty taste warrants further investigation.


Genetic markers of inflammation and their role in cardiovascular disease.
Atherosclerosis and associated cardiovascular diseases (CVD) are multifaceted disorders, influenced by environmental and heritable risk factors. Inflammation plays a significant role in each stage of atherosclerosis and as such, discovery and characterization of inflammatory biomarkers associated with risk of CVD is an active area of research. Because of the strong predicted genetic components of both CVD and inflammatory biomarkers, there is interest in identifying genetic determinants of inflammatory markers and characterizing their role in CVD. Recent developments in the methodological approaches of genetic epidemiology, especially genome-wide association studies and Mendelian randomization studies, have been effective in identifying novel gene associations and determining the causality of these genes with CVD. In this review, we will summarize the current understanding of the genetic architecture of inflammatory markers. The markers selected for this review include C-reactive protein, soluble intercellular adhesion molecule-1, interleukin-6, and P-selectin.


[The rehabilitation of older people].
Although age itself is not a disease, older people are more likely to contact diseases. These diseases and the loss of facilities in old age can develop slowly and almost unnoticed or can result from an acute illness. It is always important to intervene in good time as for older people spontaneous healing is often impossible without rehabilitation. This article deals with the development of geriatrics in Germany and introduces the institutions for geriatric rehabilitation. Taking the example of the geriatric department, it will describe in-patient and partial in-patient rehabilitation, the geriatric day clinic with out-patient rehabilitation and the rehabilitation team. The article ends by formulating some demands of governmental health policies and points out that nowadays the success of geriatric work is measurable.


Safety and efficacy of 2.5-mm sirolimus-eluting stent implantation at lower deployment pressures in very small vessels (<2.5 mm).
Percutaneous coronary interventions for very small vessels are common in clinical practice despite an unavailability of the 2.25-mm sirolimus-eluting stent (SES) in some countries. We sought to evaluate the clinical and angiographic outcomes of 2.5-mm SES implantation at lower deployment pressures in very small coronary arteries. Between June 2004 and March 2007, a total of 244 patients underwent percutaneous coronary interventions in vessels with reference diameters less than 2.5 mm at our centers: outcomes in 126 consecutive patients undergoing 2.5-mm SES implantation at lower deployment pressures (< or =10 atmospheres) with predilatation and postdilatation were compared with those in 118 patients who received bare-metal stents (BMS). In the SES group, rates of predilatation and postdilatation were 73.8 and 81% respectively, and mean deployment pressure was 8.3+/-1.2 atmospheres. At follow-up, in-segment late loss was markedly lower in SES versus BMS (0.21+/-0.41 vs. 0.48+/-0.63 mm, P=0.001), resulting in significantly lower rates of restenosis (14.7 vs. 37.5%, P<0.001). At 1 year, SES versus BMS use was associated with similar rates of stent thrombosis (0.8 vs. 0.8%, P>0.999), but significantly lower rates of major adverse cardiac events (MACE) (11.9 vs. 27.1%, P=0.003), mainly driven by a significantly lower need for target-lesion revascularization (9.5 vs. 26.3%, P=0.001). Multivariable analysis identified the SES use as independently associated with a reduced 1-year MACE risk (hazard ratio: 0.32; 95% confidence interval: 0.15-0.66; P=0.002). Implantation of 2.5-mm SES in vessels with reference diameters less than 2.5 mm using lower deployment pressures and predilatation and postdilatation may lead to reduced risks of restenosis and MACE without an increased risk of stent thrombosis up to 1 year.


Obesity as a predictor for a poor prognosis of COVID-19: A systematic review.
COVID-19 is an emerging pandemic due to droplet infection of 2019-novel coronavirus (2019-nCoV). Due to its rapid transmission and high case-fatality rate, recognition of its risk and prognostic factor is important. Obesity has been associated with impaired immune system, increasing the susceptibility for 2019-nCoV infection. We aimed to study the impact of obesity to the prognosis and disease severity of COVID-19. A systematic search and handsearching was conducted in four databases: Cochrane, MEDLINE, EMBASE, and PubMed. The identified articles were screened using the chosen eligibility criteria. We obtained three retrospective cohort studies (Wu J et al., Lighter J et al., and Simonnet A et al.) to be critically appraised using Newcastle Ottawa Scale. The findings of all included studies were consistent in stating the contribution of obesity as a risk factor to increase the requirement for advanced medical care. Study with the highest quality, Simonnet A et al., reported an increase need of invasive mechanical ventilation in COVID-19 patients with body mass index higher than 35 kg/m2, OR: 7.36 (1.63-33.14; p = 0.021). This is associated with a higher mortality rate in obese population infected with COVID-19. Obesity is an independent risk and prognostic factor for the disease severity and the requirement of advanced medical care in COVID-19. This systematic review highlights a particularly vulnerable group - obese, and emphasises on the importance of treatment aggression and disease prevention in this population group.


The corpus luteum of the guinea pig. III. Cytochemical studies on the Golgi complex and GERL during normal postpartum regression of luteal cells, emphasizing the origin of lysosomes and autophagic vacuoles.
The postpartum involution of corpora lutea was examined by electron microscope cytochemistry of guinea pig ovaries previously fixed by vascular perfusion, a method which produces optimal preservation of steroid-secreting cells and yet maintains enzyme activity. The intracellular digestive apparatus was identified through the localization of two acid hydrolases, acid phosphatase (ACPase) and arylsulfatase. Other marker enzymes localized were thiamine pyrophosphatase (in Golgi cisternae) and alkaline phosphatase (along plasma membranes). Prolonged osmication was used to mark the outer Golgi cisterna. The results demonstrate that luteal cell regression is characterized by a striking increase in the number of lysosomes and the appearance of numerous, double-walled autophagic vacuoles. Both lysosomes and the space between the double walls of autophagic vacuoles exhibit ACPase and arylsulfatase activity. In contrast to earlier periods, just before and during regression, Golgi complex-endoplasmic reticulum-lysosomes (GERL) is markedly hypertrophied, displaying intense acid hydrolase activity. On the basis of various criteria, GERL is proposed to function in the formation of lysosomes and autophagic vacuoles. Lysosomes seem to develop from GERL as focal protuberances of varying size and shape, which detach from the parent structure. Double-walled autophagic vacuoles, often large and complex in structure, initially are produced as GERL cisternae envelop small areas of cytoplasm. Lytic enzymes, perhaps furnished by the engulfing membranes and trapped lysosomes, presumably bring about digestion of the contents of these vacuoles, producing first aggregate-type inclusions, then, as the contents are further degraded, myelin figure-filled residual bodies. ACPase activity occasionally appears within smooth endoplasmic reticulum tubules and cisternae in advanced regression, possibly suggesting that lytic enzymes utilize this membrane system as an access route to GERL. These data indicate that cellular autophagy is a prominent mechanism underlying luteal cell involution during normal postpartum degeneration of guinea pig corpora lutea. Furthermore they suggest that in regressing luteal cells GERL is responsible for packaging acid hydrolases into lytic bodies.


[Participation of the adenyl cyclase system in estradiol-activated growth of tumors in rat mammary glands].
Early effects of estradiol on the adenylate cyclase system in target tissues were investigated. The proliferative processes in the estradiol-dependent mammary tumours and uterus of the rat were arrested by ovariectomy. Subsequent hormonal treatment resulted in adenylate cyclase activation and the increase of the cAMP level in these tissues, probably to be connected with the initiation of the cell growth. These events were followed by a decrease of the cAMP content necessary for active proliferation of tumour cells. cAMP phosphodiesterase is not responsible for the increase in the cyclic nucleotide content, but provides for its further hydrolysis. The stimulation of the adenylate cyclase system at early steps of estradiol action is typical both for normal and transformed estradiol-dependent tissues.


Impact of age, gender, and race on circulating γδ T cells.
A major subset of human peripheral blood γδ T cells expresses the Vγ2Vδ2 T cell receptor and responds to malignant or infectious diseases. We noted significant differences in the numbers of Vγ2Vδ2 T cells in blood samples from healthy Caucasian CA or African American (AA) donors. On average, CA donors had 3.71% ± 4.37% Vδ2 cells (as a percentage of total lymphocytes) compared with 1.18% ± 2.14% Vδ2 cells for AA donors (p < 0.0001). Age and race had the greatest impact on Vδ2 cell levels; the effect of age was similar for both racial groups. The Vδ2 cell population was dominated, for both donor groups, by cells expressing the Vγ2-Jγ1.2 Vδ2 T cell receptor, an apparent result of strong positive selection and there was substantial overlap in the public Vγ2 clonotypes from both racial groups. Mechanisms for selection and amplification of Vδ2 cells are nearly identical for both groups, despite the significant difference in baseline levels. These data show that appropriate controls, matched for age and race, may be required for clinical studies of Vγ2Vδ2 T cells in infectious disease or cancer and raise important questions about the mechanisms regulating the levels of circulating Vδ2 cells.


The 3' end formation in small RNAs.
Small RNAs are a major class of RNAs along with transfer RNAs, ribosomal RNAs, and messenger RNAs. They vary in size from less than 100 nucleotides to several thousand nucleotides and have been identified and characterized both in prokaryotes and eukaryotes. Small RNAs participate in a variety of cellular functions including regulating RNA synthesis, RNA processing, guiding modifications in RNA, and in transport of proteins. Small RNAs are generated by a series of posttranscriptional processing steps following transcription. While RNA 5' end structure, 5' cap formation, and RNA processing mechanisms have been fairly well characterized, the 3' end processing is poorly understood. Recent data point to an emerging theme in small RNAs metabolism in which the 3' end processing is mediated by the exosome, a large multienzyme complex. In addition to removal of nucleotides by the exosome, there is simultaneous rebuilding of the 3' end of some small RNA by adenylation and/or uridylation. This review presents a picture of both degradative and rebuilding reactions operative on the 3' end of some small RNA molecules in prokaryotes and eukaryotes.


Vocal and Electric Fish: Revisiting a Comparison of Two Teleost Models in the Neuroethology of Social Behavior.
The communication behaviors of vocal fish and electric fish are among the vertebrate social behaviors best understood at the level of neural circuits. Both forms of signaling rely on midbrain inputs to hindbrain pattern generators that activate peripheral effectors (sonic muscles and electrocytes) to produce pulsatile signals that are modulated by frequency/repetition rate, amplitude and call duration. To generate signals that vary by sex, male phenotype, and social context, these circuits are responsive to a wide range of hormones and neuromodulators acting on different timescales at multiple loci. Bass and Zakon (2005) reviewed the behavioral neuroendocrinology of these two teleost groups, comparing how the regulation of their communication systems have both converged and diverged during their parallel evolution. Here, we revisit this comparison and review the complementary developments over the past 16 years. We (a) summarize recent work that expands our knowledge of the neural circuits underlying these two communication systems, (b) review parallel studies on the action of neuromodulators (e.g., serotonin, AVT, melatonin), brain steroidogenesis (via aromatase), and social stimuli on the output of these circuits, (c) highlight recent transcriptomic studies that illustrate how contemporary molecular methods have elucidated the genetic regulation of social behavior in these fish, and (d) describe recent studies of mochokid catfish, which use both vocal and electric communication, and that use both vocal and electric communication and consider how these two systems are spliced together in the same species. Finally, we offer avenues for future research to further probe how similarities and differences between these two communication systems emerge over ontogeny and evolution.


Can Caffeine Intake Improve Neuromuscular and Technical-Tactical Performance During Judo Matches?
Athayde, MSdS, Lima Kons, R, and Detanico, D. Can caffeine intake improve neuromuscular and technical-tactical performance during judo matches? J Strength Cond Res 32(11): 3095-3102, 2018-This study aimed to verify the effect of caffeine intake on blood lactate concentrations and neuromuscular and technical-tactical performance during simulated judo matches. Fourteen male judo athletes received capsules containing caffeine or placebo and were evaluated in both conditions. Before the baseline and 1 hour after the caffeine/placebo intake, athletes performed the countermovement jump (CMJ) test, handgrip strength, judogi grip strength test (JGST), and blood samples were collected. Subsequently, the athletes were submitted to three 5-minute matches with 15-minute intervals. All neuromuscular tests were performed and blood samples were taken during the matches' intervals. Significant interaction between condition and time (p = 0.01) was found for blood lactate concentrations. No interaction between condition and time was verified for CMJ performance (p > 0.05), handgrip strength (p > 0.05), JGST performance (p = 0.32), and number of attacks (p = 0.97). However, for all variables, a significant decrease was observed throughout time (p ≤ 0.05). According to clinical results (magnitude-based inference), we identified that post-match 2 and post-match 3 showed higher effect (most likely positive effect) regarding delta caffeine-placebo than other moments (pre-match and post-match 1) for blood lactate. Caffeine increased the estimated glycolytic contribution over the matches; however, it did not induce improvements in neuromuscular performance or number of attacks.


Parylene-based implantable platinum-black coated wire microelectrode for orbicularis oculi muscle electrical stimulation.
A novel and simple process was proposed to fabricate a parylene-based platinum-black coated wire microelectrode for orbicularis oculi muscle electrical stimulation. Compared with conventional microelectrodes, wire microelectrodes would enable smaller wounds, increased ease of implantation, and improved cosmesis. Meanwhile, the circumferential electrode sites of this wire microelectrode fabricated by lift-off process would contribute to fully contact with tissue and reduction of electrode-tissue interface impedance. The width and the amount of electrode sites could be decided by the thickness and the amount of sacrificial layer, respectively. The platinum-black coatings were electroplated on electrode sites by applying a current pulse train in chloroplatinic acid solution with ultrasonic bath for further electrode-tissue interface impedance reduction and good mechanical stability of coatings. Electrode impedance at 1 kHz has been significantly reduced by 90%, and the cathodic charge storage capacity (CSCc) has been increased by 13 times. In addition, hematoxylin-eosin (HE) staining section of muscle demonstrated the good biocompatibility of this electroplated platinum-black. By applying a charge imbalanced biphasic stimulation waveform for orbicularis oculi muscle stimulation, the rabbits with facial paralysis rehabilitated the function of closing eyes. This kind of microelectrode will be promising for neuromuscular applications.


Investigating and Modeling the Regulation of Extracellular Antibiotic Resistance Gene Bioavailability by Naturally Occurring Nanoparticles.
Extracellular antibiotic resistance genes (eARGs) are widespread in the environment and can genetically transform bacteria. This work examined the role of environmentally relevant nanoparticles (NPs) in regulating eARG bioavailability. eARGs extracted from antibiotic-resistant B. subtilis were incubated with nonresistant recipient B. subtilis cells. In the mixture, particle type (either humic acid coated nanoparticles (HASNPs) or their micron-sized counterpart (HASPs)), DNase I concentration, and eARG type were systematically varied. Transformants were counted on selective media. Particles decreased bacterial growth and eARG bioavailability in systems without nuclease. When DNase I was present (≥5 μg/mL), particles increased transformation via chromosomal (but not plasmid-borne) eARGs. HASNPs increased transformation more than HASPs, indicating that the smaller nanoparticle with greater surface area per volume is more effective in increasing eARG bioavailability. These results were also modeled via particle aggregation theory, which represented eARG-bacteria interactions as transport leading to collision, followed by attachment. Using attachment efficiency as a fitting factor, the model predicted transformant concentrations within 35% of experimental data. These results confirm the ability of NPs to increase eARG bioavailability and suggest that particle aggregation theory may be a simplified and suitable framework to broadly predict eARG uptake.


Extended wear soft contact lenses: to market or not to market.
1. Today, cosmetic extended wear lenses have numerous aliases; continuous wear, provisional extended wear, and flexible wear, all referring to the same products, but on a relative scale of wearing time. Cosmetic extended wear lenses are elective and differ from the medical necessity of extended wear that few patients require. Refractive surgery has made significant advances in the cosmetic extended wear lens market. 2. Despite growing conservatism, positive marketing kept growing the EW philosophy for almost a decade until the FDA laid down some new rules, including a 1-week maximum for overnight wear. Coincidentally, this mandate came with the advent of the disposable lens in 1989. Since then, 1-week EW with disposable contacts has proved to be a safer modality for cosmetic EW; however, the concern of corneal complications continues to decline the market. 3. Patients risk a microbial ulcerative keratitis secondary to cosmetic EW. If daily wear contact lens options or occasional part-time EW is not amenable to the patient, refer the patient to an experienced refractive surgeon. Today, patients have more healthful vision choices than ever before.


Social Isolation and Breast Cancer.
Although the role of life stressors in breast cancer remains unclear, social isolation is consistently associated with increased breast cancer risk and mortality. Social isolation can be defined as loneliness or an absence of perceived social connections. In female mice and rats, social isolation is mimicked by housing animals 1 per cage. Social isolation causes many biological changes, of which an increase in inflammatory markers and disruptions in mitochondrial and cellular metabolism are commonly reported. It is not clear how the 2 traditional stress-induced pathways, namely, the hypothalamic-pituitary-adrenocortical axis (HPA), resulting in a release of glucocorticoids from the adrenal cortex, and autonomic nervous system (ANS), resulting in a release of catecholamines from the adrenal medulla and postganglionic neurons, could explain the increased breast cancer risk in socially isolated individuals. For instance, glucocorticoid receptor activation in estrogen receptor positive breast cancer cells inhibits their proliferation, and activation of β-adrenergic receptor in immature immune cells promotes their differentiation toward antitumorigenic T cells. However, activation of HPA and ANS pathways may cause a disruption in the brain-gut-microbiome axis, resulting in gut dysbiosis. Gut dysbiosis, in turn, leads to an alteration in the production of bacterial metabolites, such as short chain fatty acids, causing a systemic low-grade inflammation and inducing dysfunction in mitochondrial and cellular metabolism. A possible causal link between social isolation-induced increased breast cancer risk and mortality and gut dysbiosis should be investigated, as it offers new tools to prevent breast cancer.


Investigating the structural and functional effects of mutating Asn glycosylation sites of horseradish peroxidase to Asp.
Horseradish peroxidase (HRP) has long attracted intense research interest and is used in many biotechnological fields, including diagnostics, biosensors, and biocatalysis. Enhancement of HRP catalytic activity and/or stability would further increase its applications. One of the problems with heterologus expression of HRP especially in prokaryotic host is lack of glycosylation that affects it's stability toward H(2)O(2) and thermal inactivation. In this study, two asparagine residues which constitute two of the eight glycosylation sites in native HRP (Asn 13 and 268) with respectively 83% and 65% surface accessibility were substituted with aspartic acid in recombinant HRP. Both mutant proteins expressed in Escherichia coli showed increased stabilities against heat (increase in t (1/2) from 20 min in native rHRP to 32 and 67 min in N13D and N268D) and H(2)O(2) (up to threefold). Unexpectedly, despite the distance of the mutated positions from the active site, notable alterations in steady-state k (cat) and K (m) values occurred with phenol/4-aminoantipyrine as reducing substrate which might be due to conformational changes. No significant alteration in flexibility was detected by acrylamide quenching analyses, but ANS binding experiments purposed lesser binding of ANS to hydrophobic patches in mutated HRPs. Double mutation was non-additive and non-synergistic.


[An enzymatic sensor for estimation of the content of nitro and nitroso compounds in biological objects].
Nitric oxide (NO) generation in the biological cells and tissues is a cause of the production of some nitro and nitroso compounds that differ in their physiological significance and toxicity. The resultant problem in the estimation of the content of all nitro and nitroso compounds totally and singly in complex systems, such as biological objects, remains to be very urgent since simple, highly sensitive and highly selective methods have not been proposed so far to determine all these NO derivatives. The method based on some specific biochemical property of NO metabolites, which manifests itself under physiological conditions and permits the fixation of all nitro and nitroso compounds without their prior modification fraught with unpredictable artifacts, seems to be optimal. The authors have designed an enzymatic sensor based on the previously established ability to inhibit the enzyme catalase in the presence of halide ions. The level of dinitrosyl iron complex (DNIC) was measured, by using its property to lose its ability to inhibit catalase on addition of a trap of NO and an iron chelator to a reaction medium. S-nitrosothiols are detected as substances that are able to produce DNIC after addition of ferrous iron and thiols, unlike nitrite that has not this property. The total content of nitro compounds was estimated, by reducing to nitroso compounds by vanadium (III) chloride. The nitro compounds showing the properties of NO donors (RNO2) were determined as the substances that acquire the properties of DNIC in the presence of ferrous iron and thiols. The content of nitrates was estimated as a difference between the total level of nitro compounds and the content of RNO2. The sensitivity of this method was as high as 50 nM. That is it is more than an order higher in sensitivity than the classical methods based on the Griess reaction. By keeping its high sensitivity in mind, the proposed catalase method as an enzymatic detector of nitro and nitroso compounds allows one to detect these compounds in the neutral medium, without pre-purifying the object, thereby preventing the influence of the factors that contribute to the uncontrolled modification of the compounds under study.


Cervical injuries scored according to the Subaxial Injury Classification system: An analysis of the literature.
The Subaxial Injury Classification (SLIC) system and severity score has been developed to help surgeons in the decision-making process of treatment of subaxial cervical spine injuries. A detailed description of all potential scored injures of the SLIC is lacking. We performed a systematic review in the PubMed database from 2007 to 2014 to describe the relationship between the scored injuries in the SLIC and their eventual treatment according to the system score. Patients with an SLIC of 1-3 points (conservative treatment) are neurologically intact with the spinous process, laminar or small facet fractures. Patients with compression and burst fractures who are neurologically intact are also treated nonsurgically. Patients with an SLIC of 4 points may have an incomplete spinal cord injury such as a central cord syndrome, compression injuries with incomplete neurologic deficits and burst fractures with complete neurologic deficits. SLIC of 5-10 points includes distraction and rotational injuries, traumatic disc herniation in the setting of a neurological deficit and burst fractures with an incomplete neurologic deficit. The SLIC injury severity score can help surgeons guide fracture treatment. Knowledge of the potential scored injures and their relationships with the SLIC are of paramount importance for spine surgeons who treated subaxial cervical spine injuries.