The Optimal Second-Line Systemic Treatment Model for Recurrent and/or Metastatic Head and Neck Squamous Cell Carcinoma: A Bayesian Network Meta-Analysis.
The optimal second-line systemic treatment model for recurrent and/or metastatic head and neck squamous cell carcinoma (R/M HNSCC) remains controversial. A Bayesian network meta-analysis (NMA) was performed to address this issue with regard to efficacy and toxicity. By searching MEDLINE (via PubMed), Embase, the Cochrane Central Register of Controlled Trials and Web of Science, we extracted eligible studies. Efficacy, represented as overall survival (OS) and progression-free survival (PFS), and overall toxicity, represented as ≥ grade 3 severe acute events (sAE), were assessed to compare the following 7 treatment models through an NMA: standard-of-care therapy (SoC), single targeted therapy different from SoC (ST), double targeted therapy (DT), targeted therapy combined with chemotherapy (T+C), single immune checkpoint inhibitor therapy (SI), double immune checkpoint inhibitor therapy (DI) and single chemotherapy different from SoC (SC). Rank probabilities according to the values of the surface under the cumulative ranking curve (SUCRA) were separately determined for efficacy and toxicity. In total, 5285 patients from 24 eligible studies were ultimately screened, with 5184, 4532 and 4026 involved in the NMA of OS, PFS and sAE, respectively. All qualifying studies were absent from first-line immune checkpoint inhibitor therapy. In terms of OS, SI was superior to the other treatments, followed by DI, ST, T+C, SoC, DT and SC. Other than SI and SC, all treatments tended to be consistent, with hazard ratios (HRs) close to 1 between groups. For PFS, ST ranked first, while DT ranked last. For the toxicity profiles, compared with the other models, SI resulted in the lowest incidences of sAE, with statistical significance over SoC (odds ratio [OR] 0.31, 95% credible interval [CrI] 0.11 to 0.90), ST (OR 0.23, 95% CrI 0.06 to 0.86) and DT (OR 0.11, 95% CrI 0.02 to 0.53), while DT was the worst. When the SUCRA values of OS and sAE were combined, a cluster plot illustrated the superiority of SI, which demonstrated the best OS and tolerability toward sAE. For R/M HNSCC patients without immune checkpoint inhibitors in the first-line setting, SI may serve as the optimal second-line systemic treatment model, demonstrating the best OS and least sAE.


Who sells tobacco, who stops? A comparison across different tobacco retailing schemes.
Licensing of tobacco retailers has been proposed as a mechanism to encourage retailers to stop selling tobacco. However, previous studies of tobacco licensing and/or of retailers who have stopped selling have been restricted to one legislative environment. This study examines patterns of tobacco retailing across three legislative environments with three different licensing schemes (an annual fee-based licence, a zero-cost, one-off notification scheme and no notification/licensing scheme). A telephone survey was conducted of 2928 potential tobacco retailers who could personally choose whether or not to sell tobacco (rather than the decision being made at a head office). Unexpectedly, the annual licence fee to sell tobacco was not significantly associated with a lower rate of selling tobacco or a higher rate of stopping. After allowing for other factors, probability of selling, stopping selling and reported importance of tobacco sales varied across outlet types (p<0.001 for all three outcomes), and according to the remoteness of the retailer (p<0.001, p trend=0.041 and p=0.025 respectively). A fee of $A286 was not associated with a lower rate of selling, or a higher rate of stopping. The effect of licensing on retailer numbers will presumably be greater for higher licence fees, but will also depend on the perceived importance of tobacco sales to the retailer. In turn, importance of tobacco sales appears to depend on market factors, including proximity to major urban centres and low-cost competitors. A higher licence fee is likely to have a larger effect on discouraging retailers from selling.


[Projections of fields 5 and 7 in the subdivision of the sensorimotor area of the cat cerebral cortex].
Projections of the parietal cortex (area 5 and 7) to the subdivisions of the sensorimotor cortex have been studied with axonal degeneration method. It was shown that density of distribution of the association fibres is different in the areas of the parietal and sensorimotor cortex. Area 5 projects mainly to the posterolateral part of the cruciate sulcus (areas 4fu, 4 delta); its projections to areas 4y, 4sfu and 6iffu, 6aa, 6ab are less pronounced. Area 7 is connected mostly with the medial part of the lower lip of the cruciate sulcus (areas 6iffu, 6aa, 6ab) and its projection to areas 4fu and 4 delta is less pronounced. Poor interrelation between area 5 and areas SI (2, 3a, 3b) and no projection to SII (second somatosensory area of Wolsey) were observed. No projections were found from area 7 to areas SI and SII.


R115777 (Zarnestra)/Zoledronic acid (Zometa) cooperation on inhibition of prostate cancer proliferation is paralleled by Erk/Akt inactivation and reduced Bcl-2 and bad phosphorylation.
Zoledronic acid (ZOL) has proved activity in bone metastases from prostate cancer through inhibition of mevalonate pathway and of prenylation of intracellular proteins. We have reported that ZOL synergizes with R115777 farnesyltransferase inhibitor (FTI, Zarnestra) in inducing apoptosis and growth inhibition on epidermoid cancer cells. Here, we have studied the effects of the combination of these agents in prostate adenocarcinoma models and, specifically, on androgen-independent (PC3 and DU145) and -dependent (LNCaP) prostate cancer cell lines. We have found that ZOL and R115777 were synergistic in inducing both growth inhibition and apoptosis in prostate adenocarcinoma cells. These effects were paralleled by disruption of Ras-->Erk and Akt survival pathways, consequent decreased phosphorylation of both mitochondrial bcl-2 and bad proteins, and caspase activation. Finally, ZOL/R115777 combination induced cooperative effects also in vivo on tumor growth inhibition of prostate cancer xenografts in nude mice with a significant survival increase. These effects were paralleled by enhanced apoptosis and inactivation of both Erk and Akt. In conclusions, the combination between ZOL and FTI leads to enhanced anti-tumor activity in human prostate adenocarcinoma cells likely through a more efficacious inhibition of ras-dependent survival pathways and consequent bcl-related proteins-dependent apoptosis.


Novel Substrates as Sources of Ancient DNA: Prospects and Hurdles.
Following the discovery in the late 1980s that hard tissues such as bones and teeth preserve genetic information, the field of ancient DNA analysis has typically concentrated upon these substrates. The onset of high-throughput sequencing, combined with optimized DNA recovery methods, has enabled the analysis of a myriad of ancient species and specimens worldwide, dating back to the Middle Pleistocene. Despite the growing sophistication of analytical techniques, the genetic analysis of substrates other than bone and dentine remain comparatively "novel". Here, we review analyses of other biological substrates which offer great potential for elucidating phylogenetic relationships, paleoenvironments, and microbial ecosystems including (1) archaeological artifacts and ecofacts; (2) calcified and/or mineralized biological deposits; and (3) biological and cultural archives. We conclude that there is a pressing need for more refined models of DNA preservation and bespoke tools for DNA extraction and analysis to authenticate and maximize the utility of the data obtained. With such tools in place the potential for neglected or underexploited substrates to provide a unique insight into phylogenetics, microbial evolution and evolutionary processes will be realized.


Integrative taxonomy of the red-finned barb, Enteromius apleurogramma (Cyprininae: Smiliogastrini) from Kenya, supports recognition of E. amboseli as a valid species.
Research on freshwater ecosystems in East Africa is providing a better understanding of the biodiversity in the region. Recent studies of the Kenyan barbs (Cyprininae: Smiliogastrini) revealed diversity within several nominal species from the region. In this study, we examine the molecular and morphological variation in the red-finned barb (Enteromius apleurogramma). The results of this study support the recognition of E. amboseli as a valid species that is endemic to the middle Athi River drainage in southern Kenya. Enteromius amboseli is geographically isolated and distinguished from E. apleurogramma in having longer posterior barbels, a shorter dorsal fin, and generally fewer scales in the longitudinal series. Genetic divergence between E. apleurogramma populations in Lake Victoria and Lake Kanyaboli, provide novel estimates for rates of molecular evolution in the group. Additionally, the biogeography of these barbs and the conservation status of E. amboseli are discussed.


Clinical information systems for primary care: more than just an electronic medical record.
Massachusetts General Hospital Senior Health has integrated a computer-based clinical information system into all aspects of its patient care. This technology includes a longitudinal medical record but has broader functionality that encourages communication, education, and quality improvement activities. Care of geriatric patients is often challenging because of the complex nature of their clinical presentations. The case presented here, a patient at risk for stroke, illustrates how a clinical information system can facilitate quality care. The benefits, as well as difficulties, experienced as the system was integrated into the program's clinical practice are also described.


New and Emerging Treatment Options for Irritable Bowel Syndrome.
Irritable bowel syndrome (IBS) is a functional gastrointestinal disorder associated with abdominal pain, diarrhea, constipation, or a mix of symptoms. The pathophysiology of IBS is not completely understood but appears to involve genetics, the gut microbiome, immune activation, altered intestinal permeability, and brain-gut interactions. There is no gold standard for diagnosis. Several sets of symptom-based guidelines exist. Treatment strategies for IBS may include both nonpharmacologic and pharmacologic approaches. Lifestyle modifications that aim to improve exercise, sleep, diet, and stress may be warranted. Recent data suggest that a gluten-free diet and a diet low in fermentable oligosaccharides, disaccharides, monosaccharides, and polyols (FODMAPs) may benefit some patients. For patients with diarrhea-predominant IBS, treatment options include the synthetic peripheral μ-opioid receptor agonist loperamide, antispasmodic agents, antidepressants, serotonin 5-HT3 antagonists, and the gut-specific antibiotic rifaximin. Ongoing research is evaluating the use of probiotics. For patients with constipation-predominant IBS, therapeutic strategies may include dietary fiber, laxatives, and the prosecretory agents lubiprostone and linaclotide. Research is continuing to optimize the use of available agents and evaluating new approaches to further improve the care of patients with IBS.


Stress factors in modern urban lifestyles: an Indonesian perspective.
An Indonesian perspective of lifestyle in an urban environment is likely to be sharper because of how recent and substantial urbanisation has been. Contrasts between rural and new urban ways of life make the stressors more obvious than they may be to those in societies where the industrial revolution occurred much earlier. Additionally, the post-industrial society is grafted onto a recent industrial society in cities like Jakarta. The impact on nutritional flows through work patterns, eating styles and family life requires attention. There are medical-physiological and psychological approaches to the understanding of stress. In this way, an appreciation of factors other than the food supply itself on nutritional status can be gained. These include fashion, peer pressure, relationships, physical activity patterns and food patterns. Management of these stressors provides additional opportunities for preventive health.


Familial hypercholesterolemia and the atherosclerotic disease.
Familial hypercholesterolemia (FH) is associated with premature atherosclerotic cardiovascular diseases, and is inherited as an autosomal dominant trait. The prevalence of heterozygous FH is one in five hundred people. Owing to dysfunctional low density lipoprotein (LDL) receptors due to genetic mutations, serum low density lipoprotein-cholesterol (LDL-C) levels are considerably increased from birth. FH is clinically diagnosed by confirmation of family history and characteristic findings such as tendon xanthoma or xanthelasma. Thus, clinical concern and suspicion are important for early diagnosis of the disease. Current guidelines recommend lowering LDL-C concentration to at least 50% from baseline. Statins are shown to lower LDL-C levels with high safety, and thus, have been the drug of choice. However, it is difficult to achieve an ideal level of LDL-C with a single statin therapy in the majority of FH patients. Alternatively, lipid lowering combination therapy with the recently-introduced ezetimibe has shown more encouraging results.


What constitutes our understanding of a patient?
A number of approaches to the search for methodology in the human sciences are reviewed. Heidegger's use of Husserl's phenomenology led to his exploration of the background practices on which he believed all our understanding is based. This use of phenomenology was adopted by Sartre and R. D. Laing, and existential psychiatrists and psychoanalysts in their emphasis on the here-and-now encounter between the patient and the therapist, along with various embellishments and emendations on Heidegger's notion of what it meant to live authentically or inauthentically. Foucault maintained that there is a politics embedded in all the human sciences such as psychiatry and psychoanalysis, and this politics revolves around the use of power and its operations, including the actual use of the human sciences in the culture and what the different theories imply in the way of an ideology. Lacan emphasized this when he challenged the prevailing ideology of United States psychoanalysis, in which adaptation was seen as the crucial task of the ego. The existence of background practices and outer horizons required for any understanding in the human sciences leads to a situation that questions the use of empirical methods to establish what is accepted as "truth" in the human sciences. A predominant alternative, that of hermeneutics, has gained acceptance by eminent analysts such as Gill and certain interpersonal theorists. But the term hermeneutics is used in many ways by many authors and runs the danger of a relativism and a nihilism. This leads to a discussion of the value of various philosophical positions in throwing light on our search for a methodology in the human sciences. A review of arguments invoking the "end of philosophy" such as those of Rorty reveals that they are paradoxical and based on premises which in turn represent a philosophy. This debate remains unresolved. In general the postmodern view sees "truth" as more relative and emphasizes the role of social and cultural aspects of prevailing ideologies in constituting and determining what we call truth and understanding, including the "understanding" of a patient in psychotherapy. An example of the postmodern blend of philosophy and psychoanalysis is presented by Arlow's emphasis on unconscious fantasies and his insightful comments on how these unconscious fantasies organize and determine what we experience from the very beginning of life. This I believe is most consistent with Rescher's philosophical notion of "conceptual idealism," also briefly reviewed in this paper.(ABSTRACT TRUNCATED AT 400 WORDS)


[Progressive myoclonic cerebellar ataxia as a manifestation of Creutzfeldt-Jakob disease].
Progressive myoclonic cerebellar ataxia is a clinical entity with an important spectrum of possible diagnoses that requires a complex and exhaustive differential diagnosis. A 53-year-old male patient with no relevant medical history who was admitted to hospital because of an unstable gait, together with mild bilateral dysymmetry and the progressive and insidious widening of the base of support. The patient's symptoms then became more pronounced and included myoclonus and a deterioration of the higher functions. The patient died four months after the onset of the symptoms. The explorations that were conducted included a pathological study of the brain, which confirmed the diagnosis of classical spongiform encephalopathy (Creutzfeldt-Jakob disease). Creutzfeldt-Jakob disease must be included in the differential diagnosis of progressive cerebellar ataxias.


Targeting the renin-angiotensin-aldosterone-system in atrial fibrillation: a shift from electrical to structural therapy?
Despite its increasing incidence and prevalence, treatment options in atrial fibrillation (AF) are far from ideal and often limited. After decades of focus on the electrical aspects of AF with unsatisfactory results, recent research is focusing increasingly on the atrial structural remodelling that underlies the development of AF in different pathological conditions, such as hypertension, heart failure, diabetes mellitus and coronary artery disease. The aim of this review is to provide a comprehensive overview of the role of the renin-angiotensin-aldosterone-system in AF and to highlight the clinical evidence on renin-angiotensin-aldosterone-system blockade as a therapeutic option in AF.


Convection enhanced delivery of cisplatin-loaded brain penetrating nanoparticles cures malignant glioma in rats.
Glioblastoma multiforme (GBM) is highly invasive and uniformly fatal, with median survival<20months after diagnosis even with the most aggressive treatment that includes surgery, radiation, and systemic chemotherapy. Cisplatin is a particularly potent chemotherapeutic agent, but its use to treat GBM is limited by severe systemic toxicity and inefficient penetration of brain tumor tissue even when it is placed directly in the brain within standard delivery systems. We describe the development of cisplatin-loaded nanoparticles that are small enough (70nm in diameter) to move within the porous extracellular matrix between cells and that possess a dense polyethylene glycol (PEG) corona that prevents them from being trapped by adhesion as they move through the brain tumor parenchyma. As a result, these "brain penetrating nanoparticles" penetrate much deeper into brain tumor tissue compared to nanoparticles without a dense PEG corona following local administration by either manual injection or convection enhanced delivery. The nanoparticles also provide controlled release of cisplatin in effective concentrations to kill the tumor cells that they reach without causing toxicity-related deaths that were observed when cisplatin was infused into the brain without a delivery system. Median survival time of rats bearing orthotopic glioma was significantly enhanced when cisplatin was delivered in brain penetrating nanoparticles (median survival not reached; 80% long-term survivors) compared to cisplatin in conventional un-PEGylated particles (median survival=40days), cisplatin alone (median survival=12days) or saline-treated controls (median survival=28days).


A unified view of ligand-protected gold clusters as superatom complexes.
Synthesis, characterization, and functionalization of self-assembled, ligand-stabilized gold nanoparticles are long-standing issues in the chemistry of nanomaterials. Factors driving the thermodynamic stability of well documented discrete sizes are largely unknown. Herein, we provide a unified view of principles that underlie the stability of particles protected by thiolate (SR) or phosphine and halide (PR(3), X) ligands. The picture has emerged from analysis of large-scale density functional theory calculations of structurally characterized compounds, namely Au(102)(SR)(44), Au(39)(PR(3))(14)X(6)(-), Au(11)(PR(3))(7)X(3), and Au(13)(PR(3))(10)X(2)(3+), where X is either a halogen or a thiolate. Attributable to a compact, symmetric core and complete steric protection, each compound has a filled spherical electronic shell and a major energy gap to unoccupied states. Consequently, the exceptional stability is best described by a "noble-gas superatom" analogy. The explanatory power of this concept is shown by its application to many monomeric and oligomeric compounds of precisely known composition and structure, and its predictive power is indicated through suggestions offered for a series of anomalously stable cluster compositions which are still awaiting a precise structure determination.


Power Pattern Sensitivity to Calibration Errors and Mutual Coupling in Linear Arrays through Circular Interval Arithmetics.
The sensitivity to both calibration errors and mutual coupling effects of the power pattern radiated by a linear array is addressed. Starting from the knowledge of the nominal excitations of the array elements and the maximum uncertainty on their amplitudes, the bounds of the pattern deviations from the ideal one are analytically derived by exploiting the Circular Interval Analysis (CIA). A set of representative numerical results is reported and discussed to assess the effectiveness and the reliability of the proposed approach also in comparison with state-of-the-art methods and full-wave simulations.


Spatially dynamic intercellular adhesion junction is coupled to a microtubule-based motility system: evidence from an in vitro binding assay.
During spermatogenesis, spermatids change position in the seminiferous epithelium along an axis that is perpendicular to the seminiferous tubule wall. During this period, spermatids are attached to apical invaginations of Sertoli cells. In areas of this attachment, unique junction plaques occur in Sertoli cells. These plaques consist of regions of the plasma membrane involved with intercellular adhesion, a layer of actin filaments that are hexagonally packed, and an underlying cistern of endoplasmic reticulum (ER). It previously has been proposed that these junction plaques, and therefore the attached spermatids, are translocated, by motor proteins, along microtubule tracts in the Sertoli cell. If this is true, microtubules should bind to the junction plaque in a nucleotide dependent fashion. To verify this prediction, seminiferous epithelia of the rat were separated from tubule walls and then mechanically fragmented. These epithelial preparations were incubated, in both the presence and absence of 10 mM Mg(+)(+)ATP, with exogenous microtubules stabilized with taxol. Then unbound microtubules were separated from microtubules bound to larger epithelial components by centrifuging the samples through a step sucrose gradient. The fraction enriched for elongate spermatids was collected and processed for electron microscopy. The results indicate that the junction plaques remain attached to spermatids, the plaques are intact, and the cytoplasmic face of the ER binds microtubules in a nucleotide dependent fashion. The results are consistent with the presence of motor proteins on the ER component of the junction plaques and with the general hypothesis of microtubule-dependent spermatid translocation.


New Class of Antitrypanosomal Agents Based on Imidazopyridines.
The present work describes the synthesis of 22 new imidazopyridine analogues arising from medicinal chemistry optimization at different sites on the molecule. Seven and 12 compounds exhibited an in vitro EC50 ≤ 1 μM against Trypanosoma cruzi (T. cruzi) and Trypanosoma brucei (T. brucei) parasites, respectively. Based on promising results of in vitro activity (EC50 < 100 nM), cytotoxicity, metabolic stability, protein binding, and pharmacokinetics (PK) properties, compound 20 was selected as a candidate for in vivo efficacy studies. This compound was screened in an acute mouse model against T.cruzi (Tulahuen strain). After established infection, mice were dosed twice a day for 5 days, and then monitored for 6 weeks using an in vivo imaging system (IVIS). Compound 20 demonstrated parasite inhibition comparable to the benznidazole treatment group. Compound 20 represents a potential lead for the development of drugs to treat trypanosomiasis.


Fourier-transform infrared spectroscopy for characterization of protein chain reductions in enzymatic reactions.
The potential of dry-film Fourier-transform infrared (FTIR) measurements as a monitoring tool for enzymatic hydrolysis of protein-based substrates is explored in this study. As a proof-of-concept, the enzymatic digestion of bovine serum albumin using Alcalase was monitored. To evaluate the analytical approach on complex substrates with industrial relevance, salmon- and chicken-based substrates were digested for 80 minutes using Alcalase and a total of 12 FTIR spectra were acquired during the course of the hydrolysis. The observed changes in the IR spectral features as a function of hydrolysis time were found to be in agreement with the breakdown of the amide backbone and formation of amino and carboxylate terminals. Some of the most consistent markers for hydrolysis time were the bands at 1516 cm-1 (-NH3+) and ∼1400 cm-1 (-COO-). Moreover, principal component analysis (PCA) of the FTIR spectra was used to demonstrate the systematic relationship of the hydrolysis time with key variables (wavelengths) in the protein backbone region (800-1800 cm-1). Scores in the first principal component versus the hydrolysis time have been shown to provide an overview of the process dynamics related to protein structural changes. The herein presented results suggest that dry-film FTIR measurements have potential as a rapid tool for monitoring industrial protein hydrolysis processes.


Effects of higher dietary protein intake on energy balance and metabolic control in children with long-chain 3-hydroxy acyl-CoA dehydrogenase (LCHAD) or trifunctional protein (TFP) deficiency.
The incidence of overweight and obesity is increasing among children with long-chain 3-hydroxyacyl-CoA dehydrogenase (LCHAD) or mitochondrial trifunctional (TFP) deficiency. Traditional treatment includes fasting avoidance and consumption of a low-fat, high-carbohydrate diet. A diet higher in protein and lower in carbohydrate may help to lower total energy intake while maintaining good metabolic control. To determine the short-term safety and efficacy of a high protein diet, subjects were admitted to the General Clinical Research Center and fed an ad-libitum high-protein diet and a high-carbohydrate diet for 6 days each using a randomized, crossover design. Nine subjects with LCHAD or TFP deficiency, age 7-14 were enrolled. Body composition was determined by DEXA. Total energy intake was evaluated daily. Resting energy expenditure and substrate utilization were determined by indirect calorimetry. Post-prandial metabolic responses of plasma glucose, insulin, leptin, ghrelin, acylcarnitines, and triglyceride were determined in response to a liquid meal. Subjects had a higher fat mass, lower lean mass and higher plasma leptin levels compared to reference values. While on the high protein diet energy consumption was an average of 50 kcals/day lower (p = 0.02) and resting energy expenditure was an average of 170 kcals/day higher (p = 0.05) compared to the high carbohydrate diet. Short-term higher protein diets were safe, well tolerated, and resulted in lowered energy intake and increased energy expenditure than the standard high-carbohydrate diet. Long-term studies are needed to determine whether higher protein diets will reduce the risk of overweight and obesity in children with LCHAD or TFP deficiency.


[Atypical femoral fractures associated with long-term treatment with bisphosphonates].
Long-term treatment with bisphosphonates has been associated to atypical femoral fractures whose features are now clearly defined. We present two cases of female patients under bisphosphonate treatment for over 10 years who were admitted to our institution for subtrochanteric and femoral shaft fractures after low-energy trauma. They presented, respectively, a transverse and a short oblique femoral fracture, with thickening of the lateral cortex. They underwent surgical treatment obtaining good functional and imaging result. These cases fulfill the established criteria for atypical femoral fracture, thereby illustrating a serious adverse event of long-term treatment with bisphosphonates. Such relationship has still not been clearly established by scientific evidence. However, its effectiveness in preventing osteoporotic fractures is well proven. The atypical femoral fractures are possibly a serious adverse effect of the long-term treatment with bisphosphonates. Scientific evidence still supports its use, however, the physician must be aware of these events and closely follow-up these patients.


Genetic alterations of the RET proto-oncogene in familial and sporadic pheochromocytomas.
The identification of RET proto-oncogene mutations associated with multiple endocrine neoplasia type 2 (MEN-2) has provided a convenient screening test for MEN-2 in patients with pheochromocytoma (PH). In 120 patients with apparently sporadic PH, we analyzed RET exons 10, 11, 13 and 16 using denaturing gradient gel electrophoresis and found a Leu to Phe missense mutation at codon 790 (exon 13) in 1 case. A TaqI polymorphism located at exon 13 and an AluI polymorphism at exon 14 were present with a similar frequency in the 120 sporadic PH and in 94 unaffected normotensive Caucasian subjects. In 60 patients with PH, including 14 with documented MEN-2, we compared genetic testing with the pentagastrin stimulation test. The latter was 100% sensitive and 92% specific, whereas genetic testing was 88% sensitive and 100% specific. Additional somatic mutations were sought in 35 sporadic PH. Two missense mutations affecting RET exons 11 (C634R) and 16 (M918T) and three neutral mutations at codon 836 of exon 14 associated with the AluI polymorphism were detected. Detection of RET mutations in patients with PH is safe, specific and convenient. Tumoral mutations of the RET gene may play a role in medullary tumorigenesis but seem to be less frequent than previously reported.


[Expression and localization of merlin in vestibular schwannoma].
To clarify the expression and subcellular localization of merlin in vestibular schwannoma. Fifty four paraffin embedded vestibular schwannoma samples confirmed by pathology after resection were included in the study. The expression of merlin in vestibular schwannoma was analyzed by immunohistochemistry. Nerve tissues that were resected during surgical treatment for trigeminal neuralgia and Meniere's disease were used as control. Western blotting was used to analyze the electrophoresis migration of merlin in the acoustic neuroma. Image analysis was used to calculate the positive expression percentage of merlin in each individual. The expression percentage of merlin in the tumor tissue was compared with age and gender of the patients, clinical course of the tumor, tumor growth index, tumor diameter and clinical stage. Merlin was expressed in 0 to 87.5% of the cells in vestibular schwannoma tissue with a mean of (46.66 +/- 5.75)%. There was a negative correlation between merlin expression percentage and tumor growth index. There were no correlations between merlin expression percentage and the age, gender, tumor diameter and clinical stage. There exists a difference for the location of merlin, mainly in the nucleus and perinucleus. There was also a cytoplasmic location. Merlin in the tumor tissue was shown by western blot to be in 65000 and 125000 positions. Merlin was expressed in vestibular schwannoma tissue, with a different intra-cellular location. Merlin might also exist as a complex with other proteins in the tumor tissue.


Bimetallic complexes from amphoteric group 13/15 ligands: syntheses and X-ray crystal structures.
Bimetallic, pentel-bridged complexes of the type [(dmap)Me2M-E(SiMe3)2-M'(CO)n] (M=Al, Ga; E=P, As, Sb; M'=Cr, Fe, Ni; DMAP=4-(dimethylamino)pyridine) are formed by reactions of DMAP-coordinated monomeric Group 13/15 compounds [(dmap)Me2M-E(SiMe3)2] with the transition metal complexes [(Me3N)Cr(CO)5], [Fe3(CO)12], and [Ni(CO)4]. For the first time, this reaction offers a general pathway to compounds containing a Group 13 metal and a transition metal bridged by a pentel atom. Complexes prepared in this way were characterized by IR and multinuclear NMR spectroscopy and by single-crystal X-ray structure analysis. Their electronic and structural properties are discussed in detail. The Group 13/15 ligands are very weak acceptors, which is likely to be due to the electropositive Group 13 metal, and the complexes feature comparatively long pentel-transition metal bonds. In addition, the synthesis and structural characterization of the parent DMAP-coordinated gallanes [(dmap)Me2Ga-E(SiMe3)2] (E=P, As) are reported.


Myosin light chain kinase in endothelium: molecular cloning and regulation.
The phosphorylation of myosin light chains by myosin light chain kinase (MLCK) is a key event in agonist-mediated endothelial cell gap formation and vascular permeability. We now report the cloning and expression of a nonmuscle MLCK isoform in cultured endothelium. Screening of a human endothelial cell cDNA library identified a 7.7 kb cDNA with substantial (> 95%) homology to the coding region of the rabbit and bovine smooth muscle (SM) MLCK (amino acid #923-1913) as well as with the reported avian nonmuscle MLCK (65-70% homology). Sequence analysis also identified, however, a 5' stretch of novel sequence (amino acids #1-922) which is not contained in the open reading frame of mammalian SM MLCK, and is only 58% homologous to the avian fibroblast MLCK sequence. Immunoprecipitation with NH2-specific antisera revealed a 214 kD high molecular weight MLCK in bovine and human endothelium which exhibits MLC phosphorylation properties. Amino acid sequence analysis revealed endothelial MLCK consensus sequences for a variety of protein kinases including highly conserved potential phosphorylation sites for cAMP-dependent protein kinase A (PKA) in the CaM-binding region. Augmentation of intracellular cAMP levels markedly enhanced MLCK phosphorylation (2.5-fold increase) and reduced kinase activity in MLCK immunoprecipitates (4-fold decrease). These data suggest potentially novel mechanisms of endothelial cell contraction and barrier regulation.


Single-incision percutaneous pericardial ICD lead placement in a piglet model.
Our group has demonstrated the feasibility of percutaneous pericardial ICD lead placement in a piglet model utilizing direct visualization from a lateral thoracoscopic approach. Development of a novel delivery tool that incorporates visualization allows for the procedure to be performed with a 1 cm subxiphoid incision. A 1 cm incision is made in the subxiphoid area and a novel self-anchoring delivery tool is inserted. A rigid thoracoscope and needle are inserted into two crossed working channels of the tool. After needle visualization, pericardial needle access, followed by sheath access is obtained. A modified side-biting ICD lead is inserted and fixated to the ventricular epicardial surface. The lead is connected to an ICD generator and lead testing followed by defibrillation threshold testing (DFT) is performed. Single-incision ICD lead placement was performed in 6 piglets without acute complications. Median time from subxiphoid incision to DFT testing was 64 minutes; median time from thoracoscope insertion to lead fixation was 16.5 minutes (range 9-30). All had adequate ventricular sensing and pacing at implant and underwent successful defibrillation (range 3-5 J). Survival period ranged from 1 to 16 weeks. Two piglets had survival periods of 12 and 16 weeks with mean weight gain of 7 kg; both had successful repeat DFT at 10 J. All survival animals had stable lead impedances and R-wave amplitudes throughout the survival period. Percutaneous pericardial placement of an ICD lead using our novel access tool can be safely performed through a 1 cm incision without complications.


Fabrication of CaFe2O4/TaON heterojunction photoanode for photoelectrochemical water oxidation.
Tantalum oxynitride photoanode is fabricated and modified with calcium ferrite to form a heterojunction anode for a photoelectrochemical water splitting cell. The synthesized powders are loaded sequentially to the transparent conducting glass by electrophoretic deposition, which is advantageous to form a uniform layer and a junction structure. X-ray diffraction, UV-vis diffuse reflectance spectroscopy, scanning electron microscopy, and impedance spectroscopy analysis are conducted to investigate the structural, morphological, and electrochemical characteristics of the anode. The introduction of CaFe2O4 overlayer onto TaON electrode increases the photocurrent density about five times at 1.23 V vs reversible hydrogen electrode without any co-catalyst. Impedance spectroscopy analysis indicates that the junction formation increased photocurrent density by reducing the resistance to the transport of charge carriers and thereby enhancing the electron-hole separation. This photocurrent generation is a result of the overall water splitting as confirmed by evolution of hydrogen and oxygen in a stoichiometric ratio. From the study of different junction configurations, it is established that the intimate contact between TaON and CaFe2O4 is critical for enhanced performance of the heterojunction anode for photoelectrochemical water oxidation under simulated sun light.


Proteoglycan mediated lipoprotein retention: a mechanism of diabetic atherosclerosis.
The response to retention hypothesis outlines the initial stages of atherosclerotic lesion formation. The central theme of the hypothesis is that proteoglycan mediated lipoprotein retention plays a critical step in the initiation of atherosclerosis development. Recent research using human arterial specimens, transgenic mouse models and molecular biology techniques have added to our understanding of atherosclerosis development, and provided experimental data in support of the response to retention hypothesis. In this review we summarize the recent data, in particular that which addresses mechanisms by which diabetes can accelerate atherosclerosis formation, with a focus on proteoglycan-mediated LDL retention.


A call to action for nurses: declining enrollment and the nursing shortage.
Once again, nursing and health care face a crisis related to the continued decline in nursing school enrollment and the nursing shortage. This particular shortage seems to be especially problematic because of the increased number of aging baby boomers and the dwindling professional nurse pool. Nurses must work together to increase understanding and awareness of the opportunities in the profession and to resolve this crisis. There are specific actions nurses can take to improve the perception of nursing and renew interest in a dynamic field that is so crucial to society.


Investigating mixotrophic metabolism in the model diatom Phaeodactylum tricornutum.
Diatoms are prominent marine microalgae, interesting not only from an ecological point of view, but also for their possible use in biotechnology applications. They can be cultivated in phototrophic conditions, using sunlight as the sole energy source. Some diatoms, however, can also grow in a mixotrophic mode, wherein both light and external reduced carbon contribute to biomass accumulation. In this study, we investigated the consequences of mixotrophy on the growth and metabolism of the pennate diatom Phaeodactylum tricornutum, using glycerol as the source of reduced carbon. Transcriptomics, metabolomics, metabolic modelling and physiological data combine to indicate that glycerol affects the central-carbon, carbon-storage and lipid metabolism of the diatom. In particular, provision of glycerol mimics typical responses of nitrogen limitation on lipid metabolism at the level of triacylglycerol accumulation and fatty acid composition. The presence of glycerol, despite provoking features reminiscent of nutrient limitation, neither diminishes photosynthetic activity nor cell growth, revealing essential aspects of the metabolic flexibility of these microalgae and suggesting possible biotechnological applications of mixotrophy.This article is part of the themed issue 'The peculiar carbon metabolism in diatoms'.


Safety evaluation of COVID-19 vaccination during early pregnancy: A single-center prospective cohort study of Chinese pregnant women.
This prospective cohort study aimed to evaluate the safety of an inactivated coronavirus disease 2019 (COVID-19) vaccine in pregnant women at a tertiary hospital in Hubei, China. Pregnancy outcomes were compared between pregnant vaccinated and unvaccinated women. Composite adverse pregnancy outcomes were defined as one or more of maternal adverse outcomes (prenatal pyrexia, postpartum hemorrhage, maternal intensive care unit admission, and a prethrombotic state) and adverse neonatal outcomes (premature delivery, intrauterine fetal death or induction of labor, fetal macrosomia, fetal growth restriction, small-for-gestational age, fetal abnormalities, neonatal admission to the neonatal intensive care unit, and birth asphyxia). Of a total of 845 participants in the delivery cohort, 41.2% (348/845) received at least one dose of the COVID-19 vaccination, and 33.6% (284/845) received two doses. In total, 25.3% (88/348) of the vaccinated group were vaccinated at 0-4 weeks of pregnancy. No significant difference was found in the composite adverse outcomes between the vaccinated and unvaccinated participants, regardless of whether vaccination occurred before (44/243 [18.1%] vs. 71/497 [14.3%]; P = .17) or during early pregnancy (14/105 [13.3%] vs. 71/497[14.3%]; P = .79). These findings indicate that the pregnancy outcomes of women who received the COVID-19 vaccination, even if they were vaccinated early in pregnancy, were comparable to those of unvaccinated women. This study was registered with the Chinese Clinical Trial Center (ChiCTR2100051756).


Amino acid variants in the human leptin receptor: lack of association to juvenile onset obesity.
The recently described putative lipostat system mediated in part by leptin and its hypothalamic receptor provides logical candidate genes for the molecular basis of inherited obesity in humans on the basis of the occurrence of profound obesity observed in obese and diabetic mice, in which the genes for leptin or its receptor, respectively, are mutated. In this study we tested the hypothesis that juvenile onset obesity in humans may be caused by leptin resistance mediated through genetic variations in isoforms of the hypothalamic leptin receptor. One hundred and fifty-six obese Danish men with a history of juvenile onset obesity were selected at the draft board examination with a body mass index (BMI) > or = 31 kg/m2. From the same study population a control group of 205 control subjects (mean BMI = 21,5 kg/m2) were randomly selected. Single strand conformational polymorphism scanning of genomic DNA from 56 obese subjects revealed a total of four amino acid variants located in coding exons 2, (Lys109Arg), 4 (Lys204Arg and Gln223Arg), and 12 (Lys656Asn), respectively. The codons 109, 223, and 656 variants were common, but their prevalence was not significantly different between obese and lean carriers with regard to allele or carrier frequency (p > 0.1 in each case). The codon 204 mutation was only found in one obese subject. In conclusion, it is unlikely that mutations in the coding region of the long isoform of the leptin receptor are a common cause of juvenile onset obesity.


Isometric training with maximal co-contraction instruction does not increase co-activation during exercises against external resistances.
The present investigation verified that strength is improved by a training programme consisting of repetitions of maximal isometric voluntary co-contractions without increasing co-activations during contractions against external resistances. Ten participants performed 12 training sessions (four sets of 6 × 4 second maximal isometric co-contraction of the elbow flexor and extensors, 3 days a week for 4 weeks). Surface electromyograms of triceps and biceps brachii were collected during maximal voluntary isometric elbow flexion and extension against a force transducer. Maximal voluntary isometric force increased significantly after training, by 13.8 ± 6.0% (extension) and 9.6 ± 9.5% (flexion), but the observed increases in EMG of agonist muscles during maximal voluntary contraction were not significant. No significant changes in the levels of co-activation of the elbow flexors and extensors were observed. No significant change was observed for all the parameters in a control group of ten participants. These results indicated that the strength improvements after co-contraction training occur without increases in co-activation level.


Whole-brain mapping of socially isolated zebrafish reveals that lonely fish are not loners.
The zebrafish was used to assess the impact of social isolation on behaviour and brain function. As in humans and other social species, early social deprivation reduced social preference in juvenile zebrafish. Whole-brain functional maps of anti-social isolated (lonely) fish were distinct from anti-social (loner) fish found in the normal population. These isolation-induced activity changes revealed profound disruption of neural activity in brain areas linked to social behaviour, social cue processing, and anxiety/stress. Several of the affected regions are modulated by serotonin, and we found that social preference in isolated fish could be rescued by acutely reducing serotonin levels.


Normalizing cDNA libraries.
The characterization of rare messages in cDNA libraries is complicated by the substantial variations that exist in the abundance levels of different transcripts in cells and tissues. The equalization (normalization) of cDNA is a helpful approach for decreasing the prevalence of abundant transcripts, thereby facilitating the assessment of rare transcripts. This unit provides a method for duplex-specific nuclease (DSN)-based normalization, which allows for the fast and reliable equalization of cDNA, thereby facilitating the generation of normalized, full-length-enriched cDNA libraries, and enabling efficient RNA analyses.


Tunable birefringence and elliptical polarization eigenmodes in a biplate of two quarter-wave plates.
Birefringence of elliptical polarization eigenmodes can be conceptualized as a composite system comprising two distinct media: one with linear polarization eigenmodes and the other with circular polarization eigenmodes. However, the practical realization of such a system often involves the combination of two birefringent quarter-wave plates (QWPs). In this study, our objective is to characterize the variable retardation and variable elliptical polarization eigenmodes exhibited by a biplate consisting of two quarter-wave plates. Additionally, we aim to analyze the geometric properties of the transformation of one state of polarization on the Poincaré sphere, employing the emerging state's curve. This curve corresponds to the intersection between the Poincaré sphere and a cone. The outcomes of our study are presented as a function of the angle between the fast axes of the two QWPs. The findings have the potential to contribute to the configuration of q-plates and facilitate the development of quantum communication protocols.


A Concise Review on the Classification and Nomenclature of Stem Cells.
Stem cell biology and regenerative medicine is a relatively young field. However, in recent years there has been a tremendous interest in stem cells possibly due to their therapeutic potential in disease states. As a classical definition, a stem cell is an undifferentiated cell that can produce daughter cells that can either remain a stem cell in a process called self-renew¬al, or commit to a specific cell type via the initiation of a differentiation pathway leading to the production of mature progeny cells. Despite this acknowledged definition, the classification of stem cells has been a perplexing notion that may often raise misconception even among stem cell biologists. Therefore, the aim of this brief review is to give a conceptual approach to classifying the stem cells beginning from the early morula stage totipotent embryonic stem cells to the unipotent tissue-resident adult stem cells, also called tissue-specific stem cells.


Resistance to Bacillus sphaericus involves different mechanisms in Culex pipiens (Diptera:Culicidae) larvae.
Field Culex pipiens pipiens (L.) mosquitoes that were collected after a control failure with Spherimos in southern France developed high resistance (> 10,000-fold) to Bacillus sphaericus crystal toxin after < 8 generations of laboratory selection. We show that this resistance is encoded by a single major recessive gene on linkage group I at 22.1 recombination units from the sex locus, and that it is not associated with any loss of binding affinity between brush border membrane fractions and the B. sphaericus radiolabeled toxin. Thus, in Southern France, resistance differs from the high B. sphaericus resistance developed after laboratory selection of Californian C. p. quinquefasciatus. This demonstrates that at least 2 different mechanisms may confer high levels of resistance to B. sphaericus crystal toxin in mosquitoes of the C. pipiens complex. These results have important implications for mosquito control strategies.


Sociocultural and community factors influencing the use of Native Hawaiian healers and healing practices among adolescents in Hawai'i.
Recently, there has been much emphasis placed on both alternative approaches to health care and the provision of culturally competent care. Despite these trends, few studies have examined the use of alternative therapies on the part of adolescents. Even fewer studies have been done focusing on traditional Hawaiian approaches to health care. This is essential, especially in Hawai'i, where Hawaiians have not attained health parity with other majority ethnic populations, despite significant efforts and funding to achieve this end. This study examines the sociocultural and community factors influencing the use of traditional Native Hawaiian healers and healing practices by adolescents in Hawai'i. The Hawaiian High Schools Health Survey was administered at five high schools on three islands in Hawai'i during the 1993-1994 school year. The sample included 1,321 high school students who preferred either an allopathic or alternative practitioner. Ethnicity, gender, community access, healer preference, health status, level of education, and health insurance status were used to predict healer use and participation in healing practices. Community access and healer preference predicted both healer use and participation in native healing practices. Mental health predicted healer use (i.e., seeing a Native Hawaiian healer in the past six months), but did not predict taking part in native healing practices, such as ho'oponopono and lomilomi. Hawaiian ethnicity, female gender, and a measure of health insurance predicted participation in native healing practices, but not healer use. These results suggest that native healing practices and traditional healers are being used in Native Hawaiian communities and this is perhaps not due to a lack of health insurance. Given the general separation between Western and Native Hawaiian health services, traditional healing practices should be made available in Native Hawaiian communities to see whether a collaboration between Western practitioners and traditional healers can have a greater positive impact on the health of Native Hawaiians, particularly for adolescents.


A SERIES OF FIVE PATIENTS WITH FOVEAL HYPOPLASIA DEMONSTRATING GOOD VISUAL ACUITY.
Foveal hypoplasia is associated with low vision in a variety of conditions. We report and show imaging studies for a series of five patients with foveal hypoplasia who had visual acuity ranging from 20/25 to 20/60. Observational case series. This series includes five patients aged 5 to 18 years who presented to the Greater Baltimore Medical Center, Hoover Low Vision Clinic. All patients showed a partial or complete absence of the fovea in both eyes on examination. Eight eyes exhibited absence of foveal depression on spectral domain optical coherence tomography (Cirrus high-definition OCT; Carl Zeiss Meditec). Four eyes exhibited Grade 3 foveal hypoplasia, two eyes exhibited Grade 4 foveal hypoplasia, and two eyes exhibited Grade 1 foveal hypoplasia. For two eyes (one patient), the optical coherence tomography was no longer available for grading, but was interpreted as foveal hypoplasia in both eyes in the chart. Visual acuities ranged from 20/25 to 20/60. Only two of five patients showed signs of albinism. Despite the presence of foveal hypoplasia, patients can retain a high level of visual acuity, which suggests that a foveal depression is not essential for the development of good visual acuity.


The c-erbB-2 protein in primary and metastatic breast carcinomas.
Material from 41 patients with primary breast carcinoma and lymph node metastases at the time of primary surgical intervention was immunostained for c-erbB-2 protein, neuron-specific enolase (NSE), and estrogen receptors. Thirty of the primary breast carcinomas were of ductal type. Six were classified as infiltrating lobular carcinomas, 2 were apocrine, 1 was mucinous, and 1 was a tubular carcinoma. One tumor could not be classified as ductal or lobular by light microscopic examination alone. The number of lymph node metastases available varied from 1 to 14 per case (median, 3.9). Nine (22%) of the primary breast carcinomas (8 ductal and 1 apocrine) expressed c-erbB-2 protein and showed c-erbB-2 gene amplification; 12 expressed NSE immunoreactivity. None expressed both markers. Estrogen receptor immunoreactivity was present in 23 of the 41 cases, including 9 of the NSE-positive cases. C-erbB2- protein-positive metastases were present in 18 cases (44%), and in 13 cases all metastases were immunostained. In 5 cases the expression of c-erbB-2 protein varied from metastasis to metastasis. NSE immunoreactivity was expressed in 10 cases, and in 3 cases with minor NSE-positive cell populations the metastatic lesions expressed c-erbB-2 protein as well. All 9 primary breast carcinomas expressing c-erbB-2 protein had lymph node metastases with c-erbB-2-immunoreactive tumor cells. Eight of the 9 c-erbB-2 protein-negative primary tumors with metastases expressing c-erbB-2 protein showed no amplification of the c-erbB-2 gene.(ABSTRACT TRUNCATED AT 250 WORDS)


Vitreous surgery. XII. New instrumentation for vitrectomy.
An improved instrument for removing vitreous tissue consists of a replaceable cutting tip and a micromotor that powers the cutting tip. The cutting tip consists of a helical cutter and a metallic tube housing the cutter. An automated power-control unit is also available to control evacuation, infusion, and speed of rotation of the instrument. Several devices for monitoring the outflow of evacuation material and pressure in the eye and in the evacuation line are included. The cutting tip is illuminated by a fiber-optic system, which provides an additional factor of safety. The instrument contains a cleaning mechanism that eliminates the need for disassembling the cutting tip. When an automated control unit is not available, the vitreous nibbler may be used with manual suction, gravity-controlled infusion, and a simplified control unit.


Constructing an efficient electrocatalyst for water oxidation: an Fe-doped CoO/Co catalyst enabled by in situ MOF growth and a solvent-free strategy.
The development of non-precious metal electrocatalysts with high activity for the oxygen evolution reaction (OER) is a crucial and challenging task. In this work, we proposed a solvent-free in situ metal-organic framework (MOF) growth strategy for the fabrication of an Fe-doped CoO/Co electrocatalyst. This approach not only partially granted the MOF's porous structure to the catalyst but also resulted in a tighter combination between the Co metal and CoO, thereby enhancing its electrical conductivity. Furthermore, this method enabled the Fe species to be more uniformly dispersed on CoO/Co, which significantly exposed more active sites for efficient electrocatalysis. The entire synthesis process was solvent-free, except for a small amount of water and ethanol used during catalyst washing. The as-synthesized Fe-CoO/Co electrocatalyst exhibited superior OER activity on a glass carbon electrode, with η = 276 mV at a current density of 10 mA cm-2, even higher than that of the commercial precious IrO2/C catalyst. Additionally, it was also extended to prepare a Ni-doped CoO/Co electrocatalyst by the same procedure with satisfactory OER performance. This work presents a new preparation approach for MOF-derived catalysts with potential applications in energy conversion and beyond.


Lorentz Force Electrical-Impedance Tomography Using Linearly Frequency-Modulated Ultrasound Pulse.
Lorentz force electrical-impedance tomography (LFEIT) combines ultrasound stimulation and electromagnetic field detection with the goal of creating a high-contrast and high-resolution hybrid imaging modality. To reduce the peak stimulation power to the ultrasound transducer in LFEIT, linearly frequency-modulated (LFM) ultrasound pulse was investigated in this paper. First, the coherency between LFM ultrasound excitation and the resulting local current density was established theoretically. Then, experiments were done using different agar phantoms of conductivity ranging from 0.2 to 0.5 S/m. The results showed: 1) using electrical signal of peak instantaneous power of 39.54 dBm to the ultrasound transducer, which was 25.5 dB lower than the peak instantaneous power of the high-voltage narrow pulse adopted in traditional LFEIT (65.05 dBm), the LFM ultrasound pulse-based LFEIT can detect the electrical conductivity discontinuity positions precisely; 2) the reconstructed B-scan image of the electrical conductivity discontinuity distribution is comparable to that obtained through LFEIT with high-voltage narrow pulse; and 3) axial resolution of 1 mm was achieved with the experimental setup. The method of LFM ultrasound pulse stimulation and coherent detection initiates an alternative scheme toward the clinical application of LFEIT.


Mitochondria transfer enhances proliferation, migration, and osteogenic differentiation of bone marrow mesenchymal stem cell and promotes bone defect healing.
Bone marrow-derived mesenchymal stem cell (BMSC) transplantation is considered a promising therapeutic approach for bone defect repair. However, during the transplantation procedure, the functions and viability of BMSCs may be impaired due to extended durations of in vitro culture, aging, and disease conditions of patients. Inspired by spontaneous intercellular mitochondria transfer that naturally occurs within injured tissues to rescue cellular or tissue function, we investigated whether artificial mitochondria transfer into pre-transplant BMSCs in vitro could improve cellular function and enhance their therapeutic effects on bone defect repair in situ. Mitochondria were isolated from donor BMSCs and transferred into recipient BMSCs of the same batch and passage. Subsequently, changes in proliferative capacity and cell senescence were evaluated by live cell imaging, Cell Counting Kit-8 assay, cell cycle analysis, Ki67 staining, qPCR and Western blot analysis of c-Myc expression, and β-galactosidase staining. Migration ability was evaluated by the transwell migration assay, wound scratch healing, and cell motility tests. Alkaline phosphatase (ALP) staining, Alizarin Red staining, and combined with qPCR and Western blot analyses of Runx2 and BMP2 were performed to elucidate the effects of mitochondria transfer on the osteogenic potential of BMSCs in vitro. After that, in vivo experiments were performed by transplanting mitochondria-recipient BMSCs into a rat cranial critical-size bone defect model. Micro CT scanning and histological analysis were conducted at 4 and 8 weeks after transplantation to evaluate osteogenesis in situ. Finally, in order to establish the correlation between cellular behavioral changes and aerobic metabolism, OXPHOS (oxidative phosphorylation) and ATP production were assessed and inhibition of aerobic respiration by oligomycin was performed. Mitochondria-recipient BMSCs exhibited significantly enhanced proliferation and migration, and increased osteogenesis upon osteogenic induction. The in vivo results showed more new bone formation after transplantation of mitochondria-recipient BMSCs in situ. Increased OXPHOS activity and ATP production were observed, which upon inhibition by oligomycin attenuated the enhancement of proliferation, migration, and osteogenic differentiation induced by mitochondria transfer. Mitochondria transfer is a feasible technique to enhance BMSC function in vitro and promote bone defect repair in situ through the upregulation of aerobic metabolism. The results indicated that mitochondria transfer may be a novel promising technique for optimizing stem cell therapeutic function.


[Cutaneous neoplasms].
The skin is the organ most commonly affected by malignancies. Various cancers of the skin show a dramatic increase in incidence over the last decades. Epithelial skin tumors are most frequently, e.g., basal cell carcinoma and the squamous cell carcinoma with its precursors, the actinic keratoses. Melanoma, which is extremely difficult to treat in advanced tumor stages, is dreaded. Besides that, there are other epithelial malignant diseases, e.g. Morbus Bowen and adnexal tumors originating from the skin appendices. Mesenchymal malignant neoplasias such as Morbus Kaposi, angiosarcomas and other dermal sarcomas, are rare. Since the majority of malignant neoplasms is removable and curable by a simple surgical intervention, the knowledge of the different skin tumors is essential for non-dermatologist.


[Efficacy of the energy-modifier cytoflavin in the treatment of patients with hypertensive encephalopathy].
To evaluate the efficacy of cytoflavin in the treatment of patients with hypertensive encephalopathy (HE). One hundred and forty patients aged 39 to 73 years, diagnosed with HE, were examined and randomized to 2 groups. A study group (n = 74) received cytoflavin in a dose of 2 tablets b.i.d. on days 1 to 25 days inclusive during standard basic therapy. A comparison group (n = 66 persons) had standard basic therapy only. A control group consisted of 30 apparently healthy individuals. The investigators studied the frequency of headache, dizziness, and other complaints and the intensity of cephalalgic syndrome, by using a visual analog scale, the quality of life by the Medical Outcomes Study 36-Item Short-Form Health Survey (MOS SF-36) questionnaire, that of sleep by the subjective sleep characteristics questionnaire elaborated at the Moscow City Somnological Center, the level of asthenia by a subjective asthenia rating scale (Multidimensional Fatigue Inventory (MFI-20), and autonomic status, by applying objective and subjective scales on days 1 and 25 of therapy. The study has shown that cytoflavin used in the above dose for 25 days reduces the frequency and magnitude of complaints of headache, dizziness, "venous" complaints, the degree of autonomic and asthenic disorders, and impairments in the quality of sleep and life in the patients with HE at all disease stages. A stepwise discriminant analysis has indicated that the degree of cephalgic syndrome, and autonomic disorders, and worse sleep quality are the most effective points for using the energy-modifier cytoflavin. HE treatment based on the current pathogenetic principles may have a preventive impact on the development of HE or slow down the rate of its progression.


Nuclear-Electronic Orbital Quantum Mechanical/Molecular Mechanical Real-Time Dynamics.
Simulating the nuclear-electronic quantum dynamics of large-scale molecular systems in the condensed phase is key for studying biologically and chemically important processes such as proton transfer and proton-coupled electron transfer reactions. Herein, the real-time nuclear-electronic orbital time-dependent density functional theory (RT-NEO-TDDFT) approach is combined with a hybrid quantum mechanical/molecular mechanical (QM/MM) strategy to enable the accurate description of coupled nuclear-electronic quantum dynamics in the presence of heterogeneous environments such as solvent or proteins. The densities of the electrons and quantum protons are propagated in real time, while the other nuclei are propagated classically on the instantaneous electron-proton vibronic surface. This approach is applied to phenol bound to lysozyme, intramolecular proton transfer in malonaldehyde, and nonequilibrium excited-state intramolecular proton transfer in o-hydroxybenzaldehyde. These examples illustrate that the RT-NEO-TDDFT framework, coupled with an atomistic representation of the environment, allows the simulation of condensed-phase systems that exhibit significant nuclear quantum effects.


Uterine contractility and regional blood flow responses to oxytocin and prostaglandin E2 in pregnant rhesus monkeys.
The effects of oxytocin and prostaglandin E2 (PGE2) infusions on regional blood flow (measured by radioactive microspheres) and myometrial contractility were studied in 18 pregnant rhesus monkeys near term. We observed no significant differences between PGE2 and oxytocin in cardiac output, hemodynamics, or uterine activity. Their effects on the regional distribution of systemic blood flow were similar although the gastrointestinal tract received an increased percentage of cardiac output after PGE2. A relative placental ischemia, together with a myometrial hyperemia, was observed during labor. Uterine contraction produced a large reduction (average 73 per cent) in placental blood flow whereas myometrial blood flow was maintained or sometimes increased. A significant negative correlation was observed between intra-amniotic pressure and placental blood flow. During uterine relaxation, placental flow partially recovered whereas myometrial flow nearly doubled the prelabor values. We conclude that (1) in the dose ranges studied, oxytocin and PGE2 produce similar effects on myometrial contractility and uteroplacental blood flow, and (2) the vascular beds of the placenta and myometrium respond differently to labor of moderate intensity.


Modeling intracranial electrodes. A simulation platform for the evaluation of localization algorithms.
Intracranial electrodes are implanted in patients with drug-resistant epilepsy as part of their pre-surgical evaluation. This allows the investigation of normal and pathological brain functions with excellent spatial and temporal resolution. The spatial resolution relies on methods that precisely localize the implanted electrodes in the cerebral cortex, which is critical for drawing valid inferences about the anatomical localization of brain function. Multiple methods have been developed to localize the electrodes, mainly relying on pre-implantation MRI and post-implantation computer tomography (CT) images. However, they are hard to validate because there is no ground truth data to test them and there is no standard approach to systematically quantify their performance. In other words, their validation lacks standardization. Our work aimed to model intracranial electrode arrays and simulate realistic implantation scenarios, thereby providing localization algorithms with new ways to evaluate and optimize their performance. We implemented novel methods to model the coordinates of implanted grids, strips, and depth electrodes, as well as the CT artifacts produced by these. We successfully modeled realistic implantation scenarios, including different sizes, inter-electrode distances, and brain areas. In total, ∼3,300 grids and strips were fitted over the brain surface, and ∼850 depth electrode arrays penetrating the cortical tissue were modeled. Realistic CT artifacts were simulated at the electrode locations under 12 different noise levels. Altogether, ∼50,000 thresholded CT artifact arrays were simulated in these scenarios, and validated with real data from 17 patients regarding the coordinates' spatial deformation, and the CT artifacts' shape, intensity distribution, and noise level. Finally, we provide an example of how the simulation platform is used to characterize the performance of two cluster-based localization methods. We successfully developed the first platform to model implanted intracranial grids, strips, and depth electrodes and realistically simulate thresholded CT artifacts and their noise. These methods provide a basis for developing more complex models, while simulations allow systematic evaluation of the performance of electrode localization techniques. The methods described in this article, and the results obtained from the simulations, are freely available via open repositories. A graphical user interface implementation is also accessible via the open-source iElectrodes toolbox.


Recent Advances in Perovskite Photodetectors for Image Sensing.
In recent years, metal halide perovskites have been widely investigated to fabricate photodetectors for image sensing due to the excellent photoelectric performance, tunable bandgap, and low-cost solution preparation process. In this review, a comprehensive overview of the recent advances in perovskite photodetectors for image sensing is provided. First, the key performance parameters and the basic device types of photodetectors are briefly introduced. Then, the recent developments of image sensors on the basis of different dimensional perovskite materials, including 0D, 1D, 2D, and 3D perovskite materials, are highlighted. Besides the device structures and photoelectric properties of perovskite image sensors, the preparation methods of perovskite photodetector arrays are also analyzed. Subsequently, the single-pixel imaging of perovskite photodetectors and the strategies to fabricate narrowband perovskite photodetectors for color discrimination are discussed. Finally, the potential challenges and possible solutions for the future development of perovskite image sensors are presented.


Priming of autoreactive CD8(+) T cells is inhibited by immunogenic peptides which are competitive for major histocompatibility complex class I binding.
In the present study, we investigated if priming of autoreactive CD8(+) T cells would be inhibited by competitive peptides for major histocompatibility complex (MHC) class I binding. We used a mouse model of vitiligo which is induced by immunization of K(b)-binding tyrosinase-related protein 2 (TRP2)-180 peptide. Competitive peptides for K(b) binding inhibited IFN-γ production and proliferation of TRP2-180-specific CD8(+) T cells upon ex vivo peptide restimulation, while other MHC class I-binding peptides did not. In mice, the capability of inhibition was influenced by T-cell immunogenicity of the competitive peptides. The competitive peptide with a high T-cell immunogenicity efficiently inhibited priming of TRP2-180-specific CD8(+) T cells in vivo, whereas the competitive peptide with a low T-cell immunogenicity did not. Taken together, the inhibition of priming of autoreactive CD8(+) T cells depends on not only competition of peptides for MHC class I binding but also competitive peptide-specific CD8(+) T cells, suggesting that clonal expansion of autoreactive T cells would be affected by expansion of competitive peptide-specific T cells. This result provides new insights into the development of competitive peptides-based therapy for the treatment of autoimmune diseases.


Angelica sinensis extract inhibits RANKL-mediated osteoclastogenesis by down-regulated the expression of NFATc1 in mouse bone marrow cells.
Destructive erosion of bone or osteolysis is a major complication of inflammatory conditions such as rheumatoid arthritis (RA), periodontal disease, and periprosthetic osteolysis. Natural plant-derived products have received recent attention as potential therapeutic and preventative drugs in human disease. The effect of Angelica sinensis (AS) extract on RANKL-induced osteoclast differentiation was examined in this study. The osteoclast precursor cell line bone marrow macrophages (BMMs) was cultured and stimulated with RANKL followed by treatment with AS at several doses. Gene expression profiles of c-Fos, c-Jun, NFATc1, TRAP, and OSCAR were sequentially evaluated. AS extract inhibited RANKL-mediated osteoclast differentiation in BMMs in a dose-dependent manner without any evidence of cytotoxicity. AS extract strongly inhibited p38, ERK, JNK, p65 phosphorylation and I-κB degradation in RANKL-stimulated BMMs. AS extract also inhibited the mRNA expression of c-Fos, c-Jun, NFATc1, TRAP, and OSCAR in RANKL-treated BMMs. Moreover, RANKL-induced c-Fos, c-Jun and NFATc1 protein expression was suppressed by AS extract. These results collectively suggested that AS extract demonstrated inhibitory effects on RANKL-mediated osteoclast differentiation in bone marrow macrophages in vitro, indicating that AS may therefore serve as a useful drug in the prevention of bone loss.


Making a world of difference.
Volunteering on hospital ships that take health care to the world's poorest people has given Elaine Gordon extraordinary new experiences, as well as skills and experience to bring back to the NHS.


Cutaneous Disseminated and Extracutaneous Sporotrichosis: Current Status of a Complex Disease.
Sporotrichosis is an implantation or inoculation mycosis caused by species of Sporothrix schenckii complex; its main manifestations are limited to skin; however, cutaneous-disseminated, disseminated (visceral) and extracutaneous variants of sporotrichosis can be associated with immunosuppression, including HIV-AIDS, chronic alcoholism or more virulent strains. The most common extracutaneous form of sporotrichosis includes pulmonary, osteoarticular and meningeal. The laboratory diagnosis requires observing yeast forms and isolating the fungus; the two main causative agents are Sporothrix schenckii (ss) and Sporothrix brasiliensis. Antibody levels and species recognition by Polimerase Chain Reaction using biological samples or cultures are also useful. The treatment of choice for most cases is amphotericin B and subsequent itraconazole for maintenance therapy.


Phthalate esters reveal skin-sensitizing activity of phenethyl isothiocyanate in mice.
Phenethyl isothiocyanate (PEITC) is a constituent of edible cruciferous vegetables and has received attention due to its potential cancer chemopreventive activity. Although its protein-binding activity is known, PEITC has not been studied as a contact sensitizer. We previously demonstrated that phthalate esters, including dibutyl phthalate (DBP) and di-n-propyl phthalate (DPP), enhance skin sensitization in a fluorescein isothiocyanate (FITC)-induced contact hypersensitivity mouse model. In this study, we examined whether DBP and DPP enhance skin sensitization to haptens other than FITC. During this study, we tested PEITC as a contact sensitizer. BALB/c mice were epicutaneously sensitized with a suboptimal dose of a hapten with or without a phthalate ester. Sensitization was evaluated as the ear swelling response after a challenge with the respective hapten. DBP and DPP did not enhance the sensitization to two T-helper 1-type (2,4-dinitrofluorobenzene and oxazolone) or three T-helper 2-type (trimellitic anhydride, methylenediphenyl 4,4'-diisocyanate, and tolulene 2,4-diisocyanate) haptens. In contrast, DBP and DPP enhanced the sensitization to two FITC analogues (eosin 5-isothiocyanate and rhodamine B isothiocyanate) as well as to PEITC. Adjuvant effects of DBP and DPP were observed in contact hypersensitivity to haptens other than FITC, including a food ingredient PEITC.


Genotype GI.6 norovirus, United States, 2010-2012.
We report an increase in the proportion of genotype GI.6 norovirus outbreaks in the United States from 1.4% in 2010 to 7.7% in 2012 (p<0.001). Compared with non-GI.6 outbreaks, GI.6 outbreaks were characterized by summer seasonality, foodborne transmission, and non-health care settings.


ACLS update. What you must know, now!
The latest resuscitation guidelines from the American Heart Association expand the scope of emergency care and emphasize interventions with proven safety and usefulness. Here's what new therapies are in, and what gold standards are out because of a lack of strong supporting evidence.


Orthodontic examination of 2,301 Danish children aged 9-10 years in a community dental service.
Orthodontic therapy in the Danish Child Dental Health Service is offered according to patient need. In general, systematic screenings of children in specific age groups are carried out to insure uniformity in selection of patients for orthodontic treatment. The screening described in this investigation was carried out among 2,301 3rd grade schoolchildren in order to diagnose symptoms of malocclusion at an early stage with regard to tooth formation, the aim being to evaluate the need for orthodontic observation, prophylactic measures or therapy. Each examination took approximately 3-4 min.


A formula for the profile of voltammogram spikes in the quasistatic regime.
A phase transition occurring at electrode-electrolyte interfaces is reflected in voltammograms (the current versus voltage plots) as a sharp spike. We derive a general formula fitting the profile of the spikes due to the first-order phase transitions that can be microscopically modeled by classical two-dimensional lattice gases. The quasistatic (near equilibrium) regime is required. The profile is especially essential when interpreting generic voltammograms, where two or more close or overlapping spikes usually appear. Simple direct links between the microscopics of a phase transition and the macroscopic properties of the associated spike are explicitly given. We demonstrate our results on the voltammograms for the underpotential deposition of copper on platinum (111) and (100) and on gold (111) and achieve very good agreement with experiment.


[In vivo study on antisense-micro ribonucleic acid-21 oligonucleotide inhibiting tongue squamous cell carcinoma growth].
To detect antisense-micro ribonucleic acid-21 oligonucleotide (AS-miR-21)'s inhibiting effect to tongue squamous cell carcinoma. Living image and TUNEL experiments were performed, based upon the xenograft animal models set up by introduction of Tca8113-luc cells which were stably transfected with pGL6 luciferase report gene plasmid into nude mice, while the tumors were injected with AS-miR-21. Tca8113-luc cell line which steadily expressed luciferase activity was constructed by transfecting pGL6 report gene plasmid. The subcutaneous tumor formation rate was much higher in nude mice introduced with the cells, and the tumors grew well. After injection of AS-miR-21 into mice tumors, it was obviously viewed that tumors grew slower, the volume of the tumors was smaller, the photon number in live body imaging was getting less, the necrosis in the tumor specimens was rare, cell nuclei was getting smaller, dyeing color was lighter, heteromorphism and new vessels were decreased, micro ribonucleic acid-21 expression in tumor cells was considerably lower, and apoptotic index was increased. All the results indicate that the injection of AS-miR-21 can inhibit growth of tongue squamous cell carcinoma in nude mice model, and effectively promote cell apoptosis of tongue squamous cell carcinoma.


Antemortem Prediction of Braak Stage.
We examined the extent to which tauopathy distribution, as determined by Braak staging, might be predicted by various risk factors in older individuals. The Swedish Twin Registry provided extensive information on neuropsychological function, lifestyle, and cardiovascular risk factors of 128 patients for whom autopsy data including Braak staging were available. Logistic regression was used to develop a prognostic model that targeted discrimination between Braak stages 0 to II and III to VI. The analysis showed that Braak stages III to VI were significantly predicted by having 1 or more APOE ε4 alleles, older age, high total cholesterol, absence of diabetes and cardiovascular disease, and poorer scores on the Wechsler Adult Intelligence Score Information test, verbal fluency, and recognition memory but better verbal recall. The algorithm predicted Braak stages III to VI well (receiver-operating characteristic area under curve, 0.897; 95% confidence interval, 0.842-0.951). Using a cutoff of 50% risk or more, the sensitivity was 85%, the specificity was 70%, and the negative predictive value was 69%. This study demonstrates that tauopathy distribution can be accurately predicted using a combination of antemortem patient data. These results provide further insight into tauopathy development and AD-related disease mechanisms and suggest a prognostic model that predicts the spread of neurofibrillary tangles above the transentorhinal stage.


Using expanded polytetrafluoroethylene for bridging arterial microvascular defects: an experimental study with evaluation in the scanning electron microscopy.
In an experimental investigation the replacement of arterial microvessels with microporous expanded polytetrafluoroethylene was studied. The infrarenal segment of the rat aorta was used as the scientific model. The inside diameter of the micrograft was 1 mm. Using microvascular techniques a total of 18 prostheses were inserted. The animals were harvested at time intervals from 3 days up to 3 months. Patency was determined by angiography and at autopsy. The aorta implant junction, the characteristics of the neointima development and the differentiation of the interstitial cells were studied by scanning electron microscopy. Fourteen of the implanted micrografts remained patent at the time of sacrifice, yielding an early patency rate of 77,7%. As shown by scanning electron microscopy, the development of the neointima took place in 2 stages. The initial fibrin/platelet layer was reduced after one month and replaced by a definite neointima with mononuclear cells, connected with interstitial cells. Investigation of the aorta-implant junction showed the prosthesis functionally embedded in the structure of the aorta. Expanded polytetrafluoroethylene as a prosthetic material for microvascular prostheses was easy to handle and emerged compatible with other tissues. It can be used as a substitute for bridging arterial microvascular defects.


Purification of Pyruvate Dehydrogenase Complex from an Extreme Thermophile, Bacillus caldolyticus, and Its Thermal Stability.
Pyruvate dehydrogenase multienzyme complex was purified from Bacillus caldolyticus. The complex was composed of four polypeptides with molecular masses of 39.8, 41.7, 53.7, and 57.5kDa estimated by SDS-PAGE and they were presumed to be pyruvate decarboxylase (E1, dimeric), lipoate acetyltransferase (E2), and lipoamide dehydrogenase (E3) on the analogy of those from Bacillus stearothermophilus. E1 and E3 were stable at pH 5.7-10.2 and 4.5-11.3, respectively. Halves of E1 and E3 activity were abolished by incubation for 30min at 65°C and 85°C, respectively. Loss of overall activity was principally due to inactivation of E1. Structural changes in the complex incubated at high temperature were studied by fluorescence spectroscopy. The results suggested that the thermal denaturation of the complex proceeded through at least two different steps: inactivations of E1 and E3, and the former process is accompanied by a reduction of the complex size.


The use of milk as a saliva substitute.
Xerostomia, a clinical manifestation of salivary gland dysfunction, affects many people. These individuals frequently sip liquids to alleviate the discomforts associated with hyposalivation. Milk appears to have many of the chemical and physical properties of a good saliva substitute. Besides the obvious benefit of providing moisture and lubrication for the dehydrated mucosa, milk buffers oral acids, reduces enamel solubility, and contributes to enamel remineralization. These anticariogenic factors are generally attributed to the high calcium and phosphate content along with the milk phosphoproteins that strongly adsorb to enamel. Patients with xerostomia frequently have difficulty in obtaining proper nutrition due to problems associated with lubricating, masticating, tasting, and swallowing food. Milk is a food with high nutritional quality that would certainly benefit most patients with xerostomia. Because of the nutritional, anticariogenic, and moisturizing properties of milk, patients with xerostomia may find milk of value as a saliva substitute to help reduce the oral health problems associated with hyposalivation.


Wells Syndrome with Generalized Oedema.
Wells syndrome or eosinophilic cellulitis is a rare and relapsing skin disease which lacks systemic involvement. A skin biopsy is needed to establish a diagnosis. Several precipitating factors have been proposed but no proven causative link has been found. On the other hand, Churg-Strauss syndrome, also known as eosinophilic granulomatosis with polyangiitis (EGPA), an auto-immune disease, is associated with multiorgan, including cutaneous manifestations. We report a case with overlapping features of Wells and Churg-Strauss syndrome, suggesting that these syndromes could be part of the same nosological entity. Biopsies are essential to establish a diagnosis in unusual cases of cellulitis.COVID-19 vaccination is a putative trigger for Wells syndrome.There is significant overlap between Wells and Churg-Strauss syndromes.


"Chain-like" trimetallic ruthenium complexes with C7 carbon-rich bridges: experimental and theoretical investigations of electronic communication tuning in five distinct oxidation states.
In this work, we report the synthesis and the electronic properties of the unique highly conjugated molecular wires trans-[Cl-(dppe)(2)Ru=C=C=(Ph)C-CH=(CH(3))C-C[triple bond]C-(X)(2)Ru-C=C-C(CH(3))=CH-C(Ph)=C=C=Ru(dppe)(2)Cl](n+) (n = 2, X = dppe ([3a](OTf)(2)) and dppm ([3b](OTf)(2)) with three similar metal centers spanned by two odd-numbered unsaturated C(7) chains providing a 28 A long conjugated path and displaying five well-separated redox states (n = 0-4). Successive one-electron transfer steps were studied by means of cyclic voltammetry, EPR and UV-vis-NIR-IR spectroelectrochemistry. The electronic and physical properties of the different states were further rationalized with the help of DFT-based calculations. Upon one-electron reduction (n = 1), the single electron is delocalized over the two carbon chains through the central metal atom to an extent driven by the rotations within and between the chains. The second reduction (n = 0) involves the whole carbon-rich conjugated path of the molecule in a spin polarized scheme: one electron is delocalized over each chain, and the two electrons are antiferromagnetically coupled with a coupling on the order of kT. Interestingly, oxidation processes strongly involve both the metal atoms and the bridging ligands. The combined investigations reveal that the mono-oxidized system (n = 3) presents a spin density uniformly distributed between the metal atoms and the carbon atoms of the chains, whereas in the second oxidation state (n = 4) the compounds show a strong antiferromagnetic coupling on the order of 4 kT between the two single electrons localized in two distinct delocalized spin orbitals implying all the carbon atoms of the bridges and the three metal atoms. Thus, for the first time, electronic communication was fully evidenced and tuned in homonuclear trimetallic oligomeric carbon-rich systems in either an oxidation or a reduction process.


Eukaryotic initiation factor 5A-1 (eIF5A-1) as a diagnostic marker for aberrant proliferation in intraepithelial neoplasia of the vulva.
The mature eukaryotic translation initiation factor 5A contains the unusual amino acid hypusine, formed post-translationally from a specific lysine residue and essential for proliferation of eukaryotic cells. We hypothesized that the major eIF5A isoform, eIF5A-1, is an in situ biomarker for proliferation. NIH-353, a polyclonal immunoreagent specific for hypusine-containing eIF5A-1, was used to test this proposal in biopsies of vulvar high-grade intraepithelial neoplasia (VIN), characterized by the presence of proliferating cells throughout the thickness of the epithelium. Methods. Formalin-fixed and paraffin-embedded archival samples with an independently established diagnosis of VIN 3 were stained immunohistochemically after antigen retrieval, employing NIH-353 and, for comparison, the standard Ki-67 antibody. NIH-353 labeled neoplastic keratinocytes throughout the thickness of the epithelium in all VIN 3 samples. Malignant cells in a case of focally invasive squamous cell carcinoma also stained strongly for mature, hypusine-containing eIF5A-1. Epithelium adjacent to these lesions, though still of apparently normal morphology, was immunoreactive throughout its full thickness. At inflammatory foci of lesional sites, solitary reactive lymphocytes were positive, as were individual proliferating cells within dermal appendages. The submucosal stroma lacked reactive cells. NIH-353 identifies mature eIF5A-1 as an in situ biomarker for proliferation. Like Ki-67, this immunoreagent promises broad applicability in histopathological diagnosis and may be helpful in outcome prediction. In contrast to Ki-67, NIH-353 visualizes a molecular target for antineoplastic therapy, and thus may guide the development and clinical testing of drugs that, like the fungicide ciclopirox, inhibit hypusine formation and cell proliferation.


Antimicrobial polyketide furanoterpenoids from seaweed-associated heterotrophic bacterium Bacillus subtilis MTCC 10403.
Brown seaweed Anthophycus longifolius (Turner) Kützing (family Sargassaceae) associated heterotrophic bacterium Bacillus subtilis MTCC 10403 was found to be a potent isolate with broad range of antibacterial activity against important perceptive food pathogens Vibrio parahaemolyticus, V. vulnificus, and Aeromonas hydrophila. This bacterium was positive for polyketide synthetase gene (KC589397), and therefore, was selected to bioprospect specialized metabolites bearing polyketide backbone. Bioactivity-guided chromatographic fractionation of the ethyl acetate extract of the seaweed-associated bacterium segregated four homologous polyketide furanoterpenoids with potential antibacterial activities against clinically important pathogens. The minimum inhibitory concentration (MIC) assay showed that the referral antibiotics tetracycline and ampicillin were active at 25 μg/mL against the test pathogens, whereas the previously undescribed (4E)-methyl 13-((16-(furan-2-yl) ethyl)-octahydro-7-hydroxy-4-((E)-23-methylbut-21-enyl)-2H-chromen-6-yl)-4-methylpent-4-enoate (compound 1) and methyl 3-(hexahydro-9-((E)-3-methylpent-1-enyl)-4H-furo[3,2-g]isochromen-6-yl) propanoate (compound 3) displayed antibacterial activities against the test pathogens at a lesser concentration (MIC < 7 μg/mL). The title compounds were characterized by comprehensive nuclear magnetic resonance and mass spectroscopic experiments. Polyketide synthase catalyzed putative biosynthetic mechanism additionally corroborated the structural ascriptions of the hitherto undescribed furanoterpenoids from seaweed-associated bacterial symbiont. The electronic and hydrophobic parameters appeared to hold a conspicuous part in directing the antibacterial properties of the compounds. Seaweed-associated B. subtilis MTCC 10403 demonstrated to represent a potential source of antimicrobial polyketides for pharmaceutical applications.


Key role of effector memory CD4+ T lymphocytes in a short-incubation heparin-binding hemagglutinin gamma interferon release assay for the detection of latent tuberculosis.
The treatment of latent tuberculosis infection (LTBI) in target populations is one of the current WHO strategies for preventing active tuberculosis (TB) infection and reducing the Mycobacterium tuberculosis reservoir. Therefore, powerful LTBI screening tools are indispensable. A gamma interferon release assay (IGRA) in response to the stimulation of peripheral blood mononuclear cells by the latency antigen native heparin-binding hemagglutinin (nHBHA-IGRA) has proven its potential for this purpose. We have evaluated its possible optimization through a reduction of incubation time from 96 to 24 h, while compensating for this by adding interleukin 7 (IL-7) to the medium. We have also investigated the phenotypes of the gamma interferon (IFN-γ)-producing cells after both short and long incubation times. One hundred thirty-one nonimmunocompromised patients were recruited from 3 Brussels-based university hospitals. They were divided into 1 of 4 subgroups according to their M. tuberculosis infection status (LTBI, TB infection, undetermined M. tuberculosis infection status, and noninfected controls). The novel 24-h nHBHA-IGRA was performed for all subjects, and a simultaneous 96-h classical HBHA-IGRA was performed for 79 individuals. The results showed a good correlation between the two tests, and the novel 24-h nHBHA-IGRA maintained the principal advantages of the classical test, namely, a high specificity for LTBI diagnosis, an absence of interference of Mycobacterium bovis BCG vaccination during infancy, and a relative discrimination between LTBI and TB infection. Whereas the commercialized IGRAs show a greater sensitivity for recent than for remote M. tuberculosis infections, the 24-h nHBHA-IGRA appears to have comparable diagnostic powers for recent and remote LTBI. The IFN-γ detected by the 24-h nHBHA-IGRA was mainly secreted by effector memory CD4(+) T lymphocytes, a finding suggestive of continuous HBHA presentation during latency.


Feasibility and accuracy of coronary artery calcium score on virtual non-contrast images derived from a dual-layer spectral detector CT: A retrospective multicenter study.
This retrospective study was to evaluate the feasibility and accuracy of coronary artery calcium score (CACS) from virtual non-contrast (VNC) images in comparison with that from true non-contrast (TNC) images. A total of 540 patients with suspected of coronary artery disease (CAD) who underwent a dual-layer spectral detector CT (SDCT) in three hospitals were eligible for this study and 233 patients were retrospectively enrolled for further analysis. The CACS was calculated from both TNC and VNC images and compared. Linear regression analysis of the CACS was performed between TNC and VNC images. The correlation of overall CACS from VNC and TNC images was very strong (r = 0.923, p < 0.001). The CACS from VNC images were lower than that from TNC images (221 versus. 69, p < 0.001). When the regression equation of the overall coronary artery was applied, the mean calibrated CACS-VNC was 221 which had a significant difference from the CACS-TNC (p = 0.017). When the regression equation of each coronary branch artery was applied, the mean calibrated CACS-VNC was 221, which had a significant difference from the CACS-TNC (p = 0.003). But the mean difference between the CACS-TNC and the calibrated CACS-VNC in either way was less than 1. The agreement on risk stratification with CACS-TNC and CCACS-VNC was almost perfect. This multicenter study with dual-layer spectral detector CT showed that it was feasible to calculate CACS from the VNC images derived from the spectral coronary artery CT angiography scan, and the results were in good accordance with the TNC images after correction. Therefore, the TNC scan could be omitted, reducing the radiation dose to patients and saving examination time while using dual-layer spectral detector CT.


Large-area 2D bismuth antimonide with enhanced thermoelectric properties via multiscale electron-phonon decoupling.
It is a challenge to obtain high thermoelectric efficiency owing to the conflicting parameters of the materials that are required. In this work, the composition-adjustable 2D bismuth antimonide (Bi100-xSbx) is synthesized using an e-beam evaporation system with homemade targets. Engineering multiscale defects is done to optimize the thermoelectric performance in the compound. Sb alloying introduces atomic defects, lattice distortion and increased grain boundary. They drastically decrease the thermal conductivity, with an ultralow value of ∼0.23 W m-1 K-1 obtained for the composition with x = 18. It is noticed that the atomic and nanoscale defects do not deteriorate the electrical conductivity (105 S m-1), and the value is even comparable to the bulk counterpart over a wide composition range (0 ≤ x ≤ 35). Annealing induces pore structure with microscale defects, which increase the Seebeck coefficient by 84% due to the energy barrier. The resultant ZT of 0.13 is enhanced by 420% in the annealed Bi82Sb18 when compared with the as-grown Bi. This work demonstrates a cost-effective and controllable way to decouple electrons and phonons in the thermoelectric field.


Targeted gene inactivation for the elucidation of deoxysugar biosynthesis in the erythromycin producer Saccharopolyspora erythraea.
The production of erythromycin A by Saccharopolyspora erythraea requires the synthesis of dTDP-D-desosamine and dTDP-L-mycarose, which serve as substrates for the transfer of the two sugar residues onto the macrolactone ring. The enzymatic activities involved in this process are largely encoded within the ery gene cluster, by two sets of genes flanking the eryA locus that encodes the polyketide synthase. We report here the nucleotide sequence of three such ORFs located immediately downstream of eryA, ORFs 7, 8 and 9. Chromosomal mutants carrying a deletion either in ORF7 or in one of the previously sequenced ORFs 13 and 14 have been constructed and shown to accumulate erythronolide B, as expected for eryB mutants. Similarly, chromosomal mutants carrying a deletion in either ORF8, ORF9, or one of the previously sequenced ORFs 17 and 18 have been constructed and shown to accumulate 3-alpha-mycarosyl erythronolide B, as expected for eryC mutants. The ORF13 (eryBIV), ORF17 (eryCIV) and ORF7 (eryBII) mutants also synthesised small amounts of macrolide shunt metabolites, as shown by mass spectrometry. These results considerably strengthen previous tentative proposals for the pathways for the biosynthesis of dTDP-D-desosamine and dTDP-L-mycarose in Sac. erythraea and reveal that at least some of these enzymes can accommodate alternative substrates.