Complement-mediated inhibition of neovascularization reveals a point of convergence between innate immunity and angiogenesis.
Beyond its role in immunity, complement mediates a wide range of functions in the context of morphogenetic or tissue remodeling processes. Angiogenesis is crucial during tissue remodeling in multiple pathologies; however, the knowledge about the regulation of neovascularization by the complement components is scarce. Here we studied the involvement of complement in pathological angiogenesis. Strikingly, we found that mice deficient in the central complement component C3 displayed increased neovascularization in the model of retinopathy of prematurity (ROP) and in the in vivo Matrigel plug assay. In addition, antibody-mediated blockade of C5, treatment with C5aR antagonist, or C5aR deficiency in mice resulted in enhanced pathological retina angiogenesis. While complement did not directly affect angiogenesis-related endothelial cell functions, we found that macrophages mediated the antiangiogenic activity of complement. In particular, C5a-stimulated macrophages were polarized toward an angiogenesis-inhibitory phenotype, including the up-regulated secretion of the antiangiogenic soluble vascular endothelial growth factor receptor-1. Consistently, macrophage depletion in vivo reversed the increased neovascularization associated with C3- or C5aR deficiency. Taken together, complement and in particular the C5a-C5aR axes are potent inhibitors of angiogenesis.


Dexamethasone implantation in birdshot chorioretinopathy - long-term outcome.
To evaluate the long-term efficacy of the 0.70 mg dexamethasone (DEX) intravitreal implant in patients with birdshot chorioretinopathy (BSCR). Retrospective descriptive case series of BSCR patients treated with DEX implant (DEX implant 0.70 mg, DEX). Patients receiving treatment between September 2013 and November 2016 with a minimum follow-up (FU) of 12 months were included. The outcomes of primary interest were vision-related functioning, Snellen visual acuity, ocular inflammation status, presence or absence of vasculitis, change in central macular thickness, and development of glaucoma and/or cataract. Change in vision-related functioning was evaluated by comparing the National Eye Institute Visual Function Questionnaire-25. The outcomes were assessed at baseline, after DEX implant, at time of relapse, and at last FU. Three patients (six eyes) were included in the study and were followed for 1-3 years. They received 1-4 DEX implants OU. All patients demonstrated improvement in National Eye Institute Visual Function Questionnaire-25 scores. Mean Snellen visual acuity better than or equal to 20/40 was seen in three eyes at baseline and five eyes at last FU. At induction, all of the patients (six eyes) had active vitritis and two (four eyes) had retinal vasculitis. All three patients (six eyes) were quiet at last FU. One patient (two eyes) developed bilateral ocular hypertension requiring topical therapy and discontinuation of DEX implants. Two patients (three eyes) developed posterior subcapsular cataract during therapy. Two patients (four eyes) showed progression of disease while on DEX therapy. All patients were eventually transitioned to systemic immunosuppressive drug therapy. BSCR patients receiving DEX implant experienced clinically meaningful improvements in patient-reported visual function as well as ocular inflammation. However, patients in this study required repeat implantation and were unable to be maintained on DEX implant long term due to development of adverse effects or progression of disease. Eventually, it was necessary to transition to systemic immunosuppressive therapy in all patients.


What Are the Effects of Remplissage on 6-Month Strength and Range of Motion After Arthroscopic Bankart Repair? A Multicenter Cohort Study.
Patients who have undergone shoulder instability surgery are often allowed to return to sports, work, and high-level activity based largely on a time-based criterion of 6 months postoperatively. However, some believe that advancing activity after surgery should be dependent on the return of strength and range of motion (ROM). There will be a significant loss of strength or ROM at 6 months after arthroscopic Bankart repair with remplissage compared with Bankart repair alone. Cohort study; Level of evidence, 2. A total of 38 patients in a prospective multicenter study underwent arthroscopic Bankart repair with remplissage (33 males, 5 females; mean age, 27.0 ± 10.2 years; 82% with ≥2 dislocation events in the past year). Strength and ROM were assessed preoperatively and at 6 months after surgery. Results were compared with 104 matched patients who had undergone Bankart repair without remplissage, although all had radiographic evidence of a Hill-Sachs defect. At 6 months, there were no patients in the remplissage group with anterior apprehension on physical examination. However, 26% had a ≥20° external rotation (ER) deficit with the elbow at the side, 42% had a ≥20° ER deficit with the elbow at 90° of abduction, and 5% had persistent weakness. Compared with matched patients who underwent only arthroscopic Bankart repair, the remplissage group had greater humeral bone loss and had a greater likelihood of a ≥20° ER deficit with the elbow at 90° of abduction (P = .004). Risk factors for a ≥20° ER deficit with the elbow at 90° of abduction were preoperative stiffness in the same plane (P = .02), while risk factors for a ≥20° ER deficit with the elbow at the side were increased number of inferior quadrant glenoid anchors (P = .003), increased patient age (P = .02), and preoperative side-to-side deficits in ER (P = .04). The only risk factor for postoperative ER weakness was preoperative ER weakness (P = .04), with no association with remplissage (P = .26). Arthroscopic Bankart repair with remplissage did not result in significant strength deficits but increased the risk of ER stiffness in abduction compared with Bankart repair without remplissage at short-term follow-up.


Associations of Serum Fatty Acid Proportions with Obesity, Insulin Resistance, Blood Pressure, and Fatty Liver: The Cardiovascular Risk in Young Finns Study.
The links between fatty acids (FAs) and cardiometabolic outcomes are topics of debate. Our aim was to investigate the associations between serum standardized FA percentages and cardiometabolic outcomes. We used cross-sectional (n = 2187-2200 subjects, age 24-39 y, women 54%) and 10-year prospective data (n = 975-1414 subjects) from the Young Finns Study. Outcomes included prevalent and incident obesity, insulin resistance (HOMA-IR index in the upper quintile), elevated blood pressure (BP; taking medication, or diastolic or systolic BP in the upper quintile), and incident nonalcoholic fatty liver. Logistic regression models were used to calculate ORs per SD increase in fatty acids (FAs). The models were adjusted for age and sex, and additionally for other potential confounders. Several cross-sectional findings were also statistically significant in prospective models (Bonferroni corrected P < 0.003). In fully-adjusted models for obesity, these consisted of SFAs (OR: 1.28) and MUFAs (OR: 1.38), including palmitoleic (OR: 1.39) and oleic acids (OR: 1.37). Furthermore, PUFAs (OR: 0.70), including linoleic (OR: 0.67) and docosahexaenoic acids (OR: 0.75), were inversely related with obesity, whereas γ-linolenic acid (OR: 1.32) was positively associated with obesity. In age- and sex-adjusted models for insulin resistance, MUFAs (OR: 1.26) and oleic acid (OR: 1.25) were positively, and PUFAs (OR: 0.81), particularly linoleic acid (OR: 0.78), were inversely associated with HOMA-IR. Similarly with elevated BP, palmitic acid (OR: 1.22), MUFAs (OR: 1.28), and oleic acid (OR: 1.28) were positively associated with elevated BP, whereas PUFAs (OR: 0.77), n-6 (omega-6) PUFAs (OR: 0.79), and linoleic acid (OR: 0.77) were inversely associated. In fully-adjusted models for incident fatty liver, the most consistent predictors were high palmitic (OR: 1.61) and low linoleic acid (OR: 0.63) percentages. The n-6/n-3 (omega-3) PUFA ratio was not linked with any adverse outcomes. High serum percentages of total SFAs and MUFAs and low PUFAs, but also several specific FAs, predict future unfavorable cardiometabolic outcomes in Finnish adults.


Influence of aging of PEEK attachment inserts on the pull-off force of implant-retained overdentures - A laboratory study.
The aim of the current study is to investigate the influence of mechanical stress as well as cleaning agents on the performance of various polyether ether ketone (PEEK) inserts for implant-retained overdentures (IOD). Three different standard PEEK inserts were subjected to rapid artificial aging through storage in chemical denture cleaning agents (acetic acid, sodium hypochlorite, or sparkling denture cleaner) as well as demineralized water. The pre-aged PEEK inserts were then placed in unilateral IOD and subjected to 200,000 chewing loads (5 kg ~ 50 N), with 5000 thermal cycles (5/55°C), and 1100 removal/insertion cycles (vertical movement 2 mm). For all the PEEK inserts, the retention forces decreased significantly with an increasing number of mechanical load cycles and after exposure to all the cleaning agents. PEEK inserts aged by exposure to chemical cleaning agents showed a significantly higher decline in retention force than the inserts stored in water. Confocal laser scanning microscopy indicated that the decline in retention force might be caused by wear on the internal insert surface in contact with the patrix. Within the limitations of this study, it can be concluded that the application of chemical cleaning agents accelerates the decline in the retention forces of PEEK retentive inserts in IODs.


E-cadherin, actin, microtubules and FAK dominate different spheroid formation phases and important elements of tissue integrity.
Spheroids resemble features of tissues and serve as model systems to study cell-cell and cell-ECM interactions in non-adhesive three-dimensional environments. Although it is generally accepted that mature spheroids resemble tissue properties very well, no studies relate different phases in the spheroid formation processes that contribute to tissue integrity. Tissue integrity involves the cellular processes adhesion formation, adhesion reinforcement, rearrangement as well as proliferation. They maintain the structure and function of tissues and, upon dysregulation, contribute to malignancy. We investigated spheroid formation dynamics in cell lines of different metastatic potential. We dissected spheroid formation into phases of aggregation, compaction and growth to identify the respective contributions of E-cadherin, actin, microtubules and FAK. E-cadherin, actin and microtubules drive the first two phases. Microtubules and FAK are involved in the proliferation phase. FAK activity correlates with the metastatic potential of the cells. A robust computational model based on a very large number of experiments reveals the temporal resolution of cell adhesion. Our results provide novel hypotheses to unveil the general mechanisms that contribute to tissue integrity.


Risk of Colorectal and Other Cancers in Patients With Serrated Polyposis.
Patients with serrated polyposis develop multiple colorectal hyperplastic and/or serrated sessile adenomas/polyps. We investigated the risk of colorectal and other cancers by analyzing data from 64 patients with serrated polyposis (mean age at diagnosis, 54 y; 41% men; 92% white) listed in the Johns Hopkins Polyposis Registry. Medical, endoscopic, and histopathology reports were evaluated. Six patients (9.4%) had a history of colorectal cancer, diagnosed at a mean age of 56 years; 6 additional patients (9.4%) had at least 1 advanced colorectal adenoma. Extracolonic cancers were found in 16% of the study population. The standard incidence ratio for colorectal cancer in patients with serrated polyposis was 18.72 (95% confidence interval, 6.87-40.74) and for extracolonic cancer was 31.20 (95% confidence interval, 14.96-57.37), compared with the Surveillance, Epidemiology, and End Results population. Patients with serrated polyposis therefore have a high risk for colorectal cancer and require vigilant colorectal surveillance, starting at the time of diagnosis of serrated polyposis. The risk of extracolonic cancer also appears to be increased, but this requires further evaluation.


Partial characterization of the C-terminal non-collagenous domain (NC1) of collagen type X.
Collagen type X is composed of three identical alpha 1(X) chains of 59 kDa, each containing a triple-helical region of 45 kDa flanked by a short N-terminal sequence and a larger non-collagenous C-terminal (NC1) domain of approx. 15 kDa. Collagen type X molecules can associate via their C-termini to form a regular hexagonal lattice in vitro, which in vivo may provide a modified extracellular matrix for the events of endochondral ossification. The NC1 domain of chick collagen type X was isolated and purified from a highly purified bacterial collagenase digest of hypertrophic chondrocyte medium proteins. The structure and aggregation properties of the NC1 domain of collagen X were investigated, independently of the triple helix. A trimer, a dimer and a monomer of the individual alpha-chain NC1 polypeptides were identified from a bacterial collagenase digest of cartilage collagens using [14C]tyrosine labelling, N-chlorosuccinimide peptide mapping and N-terminal sequencing. The trimer (50 kDa) remained intact in Laemmli sample buffer unless boiled, upon which it dissociated into the dimer (38 kDa) and the monomer (20 kDa). The dimer persisted even after prolonged periods of heating or reduction with beta-mercaptoethanol, and in preparations obtained from chondrocyte cultures treated with beta-aminoproprionitrile, indicating the presence of non-reducible, non-lysine-derived, covalent cross-links. Hexamers of the individual C-termini were observed in rotary-shadowed preparations of purified NC1 domain, reflecting the ability of collagen type X to self-assemble via its C-termini under appropriate conditions.


Upregulation of RANTES in psoriatic keratinocytes: a possible pathogenic mechanism for psoriasis.
Intraepidermal collections of neutrophils and lymphocytes are unique features of the inflammatory reaction of psoriasis. Migration of leukocytes from dermis to the epidermis suggests a role for chemotactic agent(s). In recent years, increased levels of chemokines such as IL-8 , GRO-a and MCP-1 have been reported in the keratinocytes of psoriatic tissue. IL-8 and GRO-alpha belong to a subfamily (C x C) class and MCP-1 is a beta chemokine. In this study, we investigated RANTES, which is a beta chemokine (C-C class); RANTES has been found to be associated with various cell-mediated hypersensitive disorders. We obtained eight skin biopsies from chronic psoriatic plaques, and five biopsies each from non-lesional psoriatic skin, lichen planus, eczematous dermatitis and skin from healthy controls. Snap-frozen samples were cut into 7 microm cryosections and stained with 6 mg/ml of monoclonal anti-RANTES mouse IgG (DNAX, Palo Alto, CA). Standard immunohistochemistry techniques were applied. RANTES was detected only in the keratinocytes. The number of keratinocytes in per mm2 of epidermis stained for RANTES were 116.79+/-98.42 in psoriatic tissues compared to 32.00+/-46.05 (p<0.05), 6.39+/-3.59 (p<0.01), 2.64 +/-1.15 (p<0.01) and 3.53+/-5.26 (p<0.01), respectively, in the non-lesional, lichen planus, eczematous lesions and normal skin. This is the first study to report that the keratinocytes of psoriatic tissue express high levels of RANTES compared to the controls. IL-8 and related molecules (C x C class) are predominantly chemotactic for neutrophils and MCP-1 is a strong chemotactic factor for monocytes. In contrast, RANTES is chemotactic for memory T cells and activated naive T cells. Increased amounts of RANTES as reported here provide an explanation for migration of the activated T cells to the epidermis of the psoriatic lesions. In addition, RANTES activates T cells. These results suggest that RANTES may have a significant role in the inflammatory process of psoriasis. Our findings further substantiate a regulatory role for keratinocytes in the inflammatory process of psoriasis.


Molecular cloning of cDNA encoding a 55-kDa multifunctional thyroid hormone binding protein of skeletal muscle sarcoplasmic reticulum.
A cDNA clone encoding 55-kDa multifunctional, thyroid hormone binding protein of rabbit skeletal muscle sarcoplasmic reticulum was isolated and sequenced. The cDNA encoded a protein of 509 amino acids, and a comparison of the deduced amino acid sequence with the NH2-terminal amino acid sequence of the purified protein indicates that an 18-residue NH2-terminal signal sequence was removed during synthesis. The deduced amino acid sequence of the rabbit muscle clone suggested that this protein is related to human liver thyroid hormone binding protein, rat liver protein disulfide isomerase, human hepatoma beta-subunit of prolyl 4-hydroxylase and hen oviduct glycosylation site binding protein. The protein contains two repeated sequences Trp-Cys-Gly-His-Cys-Lys proposed to be in the active sites of protein disulfide isomerase. Northern blot analysis showed that the mRNA encoding rabbit skeletal muscle form of the protein is present in liver, kidney, brain, fast- and slow-twitch skeletal muscle, and in the myocardium. In all tissues the cDNA reacts with mRNA of 2.7 kilobases in length. The 55-kDa multifunctional thyroid hormone binding protein was identified in isolated sarcoplasmic reticulum vesicles using a monoclonal antibody specific to the 55-kDa thyroid hormone binding protein from rat liver endoplasmic reticulum. The mature protein of Mr 56,681 contains 95 acidic and 61 basic amino acids. The COOH-terminal amino acid sequence of the protein is highly enriched in acidic residues with 17 of the last 29 amino acids being negatively charged. Analysis of hydropathy of the mature protein suggests that there are no potential transmembrane segments. The COOH-terminal sequence of the protein, Arg-Asp-Glu-Leu (RDEL), is similar to but different from that proposed to be an endoplasmic reticulum retention signal; Lys-Asp-Glu-Leu (KDEL) (Munro, S., and Pelham, H.R.B. (1987) Cell 48, 899-907). This variant of the retention signal may function in a similar manner to the KDEL sequence, to localize the protein to the sarcoplasmic or endoplasmic reticulum. The positively charged amino acids Lys and Arg may thus interchange in this retention signal.


Mobility of Pb, Zn, Ba, As and Cd toward soil pore water and plants (willow and ryegrass) from a mine soil amended with biochar.
Mine soils often contain metal(loid)s that may lead to serious environmental problems. Phytoremediation, consisting in covering the soil with specific plants with the possible addition of amendments, represents an interesting way of enhancing the quality of mine soils by retaining contaminants and reducing soil erosion. In order to study the effect of an assisted phytoremediation (with willow and ryegrass) on the properties of soil pore water (SPW), we investigated the impact of amendment with biochar (BC) combined with the planting of willow and ryegrass on the behavior of several metal(loid)s (Pb, Zn, Ba, As, and Cd) in a mine soil. Data on the physicochemical parameters and concentrations of the different metal(loid)s in both SPW and in plant tissues of willow and ryegrass highlight the importance of BC for SPW properties in terms of reductions in soluble concentrations of Pb and Zn, although there was no effect on the behavior of As and Cd. BC also increased soluble concentrations of Ba, probably related to ion release by the BC. By improving major ions available in mine soil, BC improved the lifetime of plants and enhanced their growth. Plant development did not appear to significantly affect the physicochemical parameters of SPW. Willow and ryegrass growing on soil with BC incorporated Cd and Ba into their tissues. The influence of plants on the behavior of metal(loid)s was noticeable only for ryegrass growing in soil with 2% BC, where it modified the behavior of Pb and Ba.


The non-photosynthetic, pathogenic green alga Helicosporidium sp. has retained a modified, functional plastid genome.
A fragment of the Helicosporidium sp. (Chlorophyta: Trebouxiophyceae) plastid genome has been sequenced. The genome architecture was compared to that of both a non-photosynthetic relative (Prototheca wickerhamii) and a photosynthetic relative (Chlorella vulgaris). Comparative genomic analysis indicated that Helicosporidium and Prototheca are closely related genera. The analyses also revealed that the Helicosporidium sp. plastid genome has been rearranged. In particular, two ribosomal protein-encoding genes (rpl19 and rps23) appeared to have been transposed, or lost from the Helicosporidium sp. plastid genome. RT-PCR reactions demonstrated that the retained plastid genes were transcribed, suggesting that, despite rearrangement(s), the Helicosporidium sp. plastid genome has remained functional. The modified plastid genome architecture is a novel apomorphy that indicates that the Helicosporidia are highly derived green algae, more so than Prototheca spp. As such, they represent a promising model to study organellar genome reorganizations in parasitic protists.


Anti-inflammatory effects of docosahexaenoic acid: Implications for its cancer chemopreventive potential.
The implication of inflammatory tissue damage in pathophysiology of human cancer as well as some metabolic disorders has been under intense investigation. Numerous studies have identified a series of critical signaling molecules involved in cellular responses to inflammatory stimuli. These include nuclear factor κB, peroxisome proliferator-activated receptor γ, nuclear factor erythroid 2 p45-related factor 2 and sterol regulatory element-binding protein 1. The proper regulation of these transcription factors mediating pro- and anti-inflammatory signaling hence provides an important strategy for the chemoprevention of inflammation-associated cancer. There is compelling evidence supporting that dietary supplementation with fish oil-derived ω-3 polyunsaturated fatty acids including docosahexaenoic acid (DHA) ameliorates symptomatic inflammation associated with cancer as well as other divergent human disorders. Acute or physiologic inflammation is an essential body's first line of defence to microbial infection and tissue injuries, but it must be properly completed by a process termed 'resolution'. Failure of resolution mechanisms can result in persistence of inflammation, leading to chronic inflammatory conditions and related malignancies. The phagocytic engulfment of apoptotic neutrophils and clearance of their potentially histotoxic contents by macrophages, called efferocytosis is an essential component in resolving inflammation. Of note, DHA is a precursor of endogenous proresolving lipid mediators which regulate the leukocyte trafficking and recruitment and thereby facilitate efferocytosis. Therefore, DHA and its metabolites may have a preventive potential in the management of human cancer which arises as a consequence of impaired resolution of inflammation as well as chronic inflammation.


D1 dopamine receptors modulate cone ON bipolar cell Nav channels to control daily rhythms in photopic vision.
In amphibians, voltage-gated sodium (Nav) channels in cone ON bipolar cells (ON-CBC) amplify cone signals in the dark and in mesopic background light. However, during light adaptation, dopamine, acting through D1 receptors (D1R), suppresses Nav channels and therefore act as a gain control mechanism. Curiously in rodents, Nav channel contributions to the ON-CBC-generated light-adapted electroretinogram (ERG) b-wave appear to exist even in fully light-adapted conditions. We sought to determine how rodent ON-CBC Nav channels are regulated by dopamine via D1R during light adaptation and during the circadian cycle. We first tested the sensitivity of Nav channels in mouse ON-CBCs to the modulation by dopamine via D1Rs. Although light-adaptation had little effect on Nav channel contributions to the b-wave, these channels were found to be modulated by D1Rs. We pharmacologically isolated the cone to ON-CBC circuit in fully light-adapted retinas to confirm these results. Retinal dopamine release following light adaptation has been previously shown to be increased in mice during circadian night. We first show that circadian fluctuations in ON-CBC function are suppressed in dark-adapted retinas, indicating that circadian fluctuations are a function of light adaptation. Secondly, we show that at night the mouse retina behaves similarly to those of frogs and salamanders with a gain control mechanism utilizing D1R modulation of Nav channels to suppress ON-CBC light responses in light-adapted conditions during circadian night. Taken together, these results suggest that circadian control of ON-CBC function contains an initial phase after approximately 18-30 h of dark adaptation, leading to substantial changes in b-wave amplitude after a relatively short time in free run which are dependent on D1R modulation of Nav channels.


Kinetics and computational analysis of cholinesterase inhibition by REVERC3, a bisdemethoxycurcumin-rich Curcuma longa extract: Relevance to the treatment of Alzheimer's disease.
Objectives: Cholinesterase inhibition is a common strategy to treat Alzheimer's disease. In this study, we have investigated the cholinesterase inhibitory effects of a first-of-its-kind turmeric extract (REVERC3) having enriched content of bisdemethoxycurcumin as major active curcuminoid. Methods: The inhibition studies were performed using Ellman's colorimetric assay. The kinetics of acetylcholinesterase and butyrylcholinesterase was determined in the presence of REVERC3 using the Lineweaver-Burk double reciprocal plots. Furthermore, we used AutoDock tools to predict the binding of bisdemethoxycurcumin with the active sites of cholinesterases. Results: REVERC3 showed 4.8- and 5.39-fold higher inhibitory potential of acetylcholinesterase and butyrylcholinesterase with IC50 values of 29.08 and 33.59 µg/mL, respectively, compared to the regular turmeric extract. The mode of binding of REVERC3 was competitive in the case of acetylcholinesterase while it was uncompetitive for the inhibition of butyrylcholinesterase. Docking analysis revealed that bisdemethoxycurcumin, the major constituent of REVERC3, has different preferences of binding in the active sites of acetylcholinesterase and butyrylcholinesterase. However, the best binding pose predictions are in line with the experimental binding mode of the cholinesterases. Conclusion: These results indicate that bisdemethoxycurcumin-enriched turmeric extract could improve the cholinergic functions via dual inhibition of cholinesterases. However, the predominant role of bisdemethoxycurcumin in REVERC3 must be further validated using preclinical studies and clinical trials.


Induction and characterization of anti-tumor endothelium immunity elicited by ValloVax therapeutic cancer vaccine.
ValloVax is a placental endothelium derived vaccine which induces tissue-nonspecific antitumor immunity by blocking tumor angiogesis. To elucidate mechanisms of action, we showed that production of ValloVax, which involves treating placental endothelial cells with IFN-gamma, results in upregulation of HLA and costimulatory molecules. It was shown that in mixed lymphocyte reaction, ValloVax induces Type I cytokines and allo-proliferative responses. Plasma from ValloVax immunized mice was capable of killing in vitro tumor-like endothelium but not control endothelium. Using defined antigens associated with tumor endothelial cells, specific molecular entities were identified as being targeted by ValloVax induced antibodies. Binding of predominantly IgG antibodies to ValloVax cells was confirmed by flow cytometry. Further suggesting direct killing of tumor endothelial cells was expression of TUNEL positive cells, as well as, reduction in tumor oxygenation. Supporting a role for antibody mediated responses, cell depletion experiments suggested a predominant role of B cells in maintaining an intact anti-tumor endothelial response. Adoptive transfer experiments suggested that infusion of CD3+ T cells from immunized mice was sufficient to transfer tumor protection. Generation of memory T cells selective to tumor endothelial specific markers was observed. Functional confirmation of memory responses was observed in tumor rechallenge experiments. Furthermore, we observed that both PD-1 or CTLA-4 blockade augmented antitumor effects of ValloVax. These data suggest a T cell induced B cell mediated anti-tumor endothelial response and set the framework clinical trials through elucidation of mechanism of action.


Heat inactivation of the neuraminidase and haemagglutinin estimated in the agglutination-separation reactions using red blood cells sensitized with Newcastle disease virus.
The agglutination-separation (AS) reactions estimate the effects of heat on the release of altered Newcastle disease virus (NDV) and HN glycoprotein spikes from red blood cells (RBC) sensitized with NDV (SRBC), the inactivations of the neuraminidase (NA), then the haemagglutinin (HA) in a direct assay. Heating SRBC for 1.5 min at 56 degrees C inactivated the NA by 50%; after 4.5 min no separation occurred indicating 100% inactivation of the NA. Heating a suspension of NDV for 78 min inactivated the NA 50% as assayed by cleavage of fetuin. Comparatively, the AS test was up to 52-fold (78 min/1.5 min = 52) more efficient in detecting NA inactivation than was the basic reference test where cleavage of fetuin was assayed. The HA was 50% inactivated after 18 min of heating and 100% inactivated after 36 min as no agglutination was seen. Free HA on SRBC was agglutinated by and thus was titrated with the sialic acid on NRBC. The large area of RBC increased the efficiency of the AS test when compared with tests using suspensions of NDV. At 51-60 degrees C all NA and HA inactivations were sequential, and invariably the NA was more heat labile than the HA. The release of altered NDV and HN spikes was inhibited with mild heat although the separation of SRBC and NRBC continued. Biological purifications showed that the heat stability of the HA and the lability of the NA were genetically stable.


RNA microarray analysis in prenatal mouse cochlea reveals novel IGF-I target genes: implication of MEF2 and FOXM1 transcription factors.
Insulin-like growth factor-I (IGF-I) provides pivotal cell survival and differentiation signals during inner ear development throughout evolution. Homozygous mutations of human IGF1 cause syndromic sensorineural deafness, decreased intrauterine and postnatal growth rates, and mental retardation. In the mouse, deficits in IGF-I result in profound hearing loss associated with reduced survival, differentiation and maturation of auditory neurons. Nevertheless, little is known about the molecular basis of IGF-I activity in hearing and deafness. A combination of quantitative RT-PCR, subcellular fractionation and Western blotting, along with in situ hybridization studies show IGF-I and its high affinity receptor to be strongly expressed in the embryonic and postnatal mouse cochlea. The expression of both proteins decreases after birth and in the cochlea of E18.5 embryonic Igf1(-/-) null mice, the balance of the main IGF related signalling pathways is altered, with lower activation of Akt and ERK1/2 and stronger activation of p38 kinase. By comparing the Igf1(-/-) and Igf1(+/+) transcriptomes in E18.5 mouse cochleae using RNA microchips and validating their results, we demonstrate the up-regulation of the FoxM1 transcription factor and the misexpression of the neural progenitor transcription factors Six6 and Mash1 associated with the loss of IGF-I. Parallel, in silico promoter analysis of the genes modulated in conjunction with the loss of IGF-I revealed the possible involvement of MEF2 in cochlear development. E18.5 Igf1(+/+) mouse auditory ganglion neurons showed intense MEF2A and MEF2D nuclear staining and MEF2A was also evident in the organ of Corti. At P15, MEF2A and MEF2D expression were shown in neurons and sensory cells. In the absence of IGF-I, nuclear levels of MEF2 were diminished, indicating less transcriptional MEF2 activity. By contrast, there was an increase in the nuclear accumulation of FoxM1 and a corresponding decrease in the nuclear cyclin-dependent kinase inhibitor p27(Kip1). We have defined the spatiotemporal expression of elements involved in IGF signalling during inner ear development and reveal novel regulatory mechanisms that are modulated by IGF-I in promoting sensory cell and neural survival and differentiation. These data will help us to understand the molecular bases of human sensorineural deafness associated to deficits in IGF-I.


[Psychological factors in general practice].
The paper concerns psychological predictors of the pattern of activity of a general practitioner which should be allowed for in collecting data for case records, planning and conduction of examinations, design and discussion of treatment policy. The ability to communicate with the patient has a noticeable psychological effect on the patient's compliance. The analysis of the psychological predictors is important for psychological training of general practitioners whose activity is very specific.


Clinical management of cosmetic extended-wear contact lens failure.
Fifteen patients discontinued use of extended-wear hydrogel contact lenses. The primary cause was neovascularization (60%), although lens soilage, giant papillary conjunctivitis and ulceration were factors in several patients each. All patients were successfully refitted with daily-wear rigid or thin, hydrogel contact lenses following simple clinical techniques.


Case report. Two cases of pulmonary hypertension with Raynaud's phenomenon. Primary pulmonary hypertension and systemic lupus erythematosus.
Two cases of hypertensive pulmonary vascular disease with Raynaud's phenomenon are reported. One is a 32-year-old housewife of primary pulmonary hypertension with some immunological disorders and the other is a 28-year-old female of systemic lupus erythematosus. It may be considered that some immunological process could play a role in the etiology of some cases of primary pulmonary hypertension and the relationship of pathogenesis between these 2 cases was discussed.


Formation of complex extrachromosomal T-DNA structures in Agrobacterium tumefaciens-infected plants.
Agrobacterium tumefaciens is a unique plant pathogenic bacterium renowned for its ability to transform plants. The integration of transferred DNA (T-DNA) and the formation of complex insertions in the genome of transgenic plants during A. tumefaciens-mediated transformation are still poorly understood. Here, we show that complex extrachromosomal T-DNA structures form in A. tumefaciens-infected plants immediately after infection. Furthermore, these extrachromosomal complex DNA molecules can circularize in planta. We recovered circular T-DNA molecules (T-circles) using a novel plasmid-rescue method. Sequencing analysis of the T-circles revealed patterns similar to the insertion patterns commonly found in transgenic plants. The patterns include illegitimate DNA end joining, T-DNA truncations, T-DNA repeats, binary vector sequences, and other unknown "filler" sequences. Our data suggest that prior to T-DNA integration, a transferred single-stranded T-DNA is converted into a double-stranded form. We propose that termini of linear double-stranded T-DNAs are recognized and repaired by the plant's DNA double-strand break-repair machinery. This can lead to circularization, integration, or the formation of extrachromosomal complex T-DNA structures that subsequently may integrate.


[The functional status of the thyroid system in those irradiated as a consequence of the accident at the Chernobyl Atomic Electric Power Station (based on screening study data)].
A screening study was made of functional indices for the status of the thyroid system in 1097 persons who took part in the elimination of the effects of the Chernobyl accident (PChAEE) during the "iodine" and "non-iodine" periods of irradiation. In 94.6% of the "iodine" period PChAEE, the basal levels of the thyroid-stimulating hormone (TSH) were in the normal range, 4.79% displayed a rise in the parameter. 90.06% of the "non-iodine" period had normal range TSH, 8.77% exhibited augmented levels. 94.68% of the "iodine" period PChAEE showed normal range for the basal levels of free thyroxin, in 3.72% there was a rise, in 1.6% a decrease in the values. Among the examinees of the "non-iodine" period, the levels of free thyroxin were within normal range in 92%, above norm in 5%, decreased in 3%. The level of antibodies to thyroglobulin in 86.38% of the "iodine" period examinees was in the normal range, in 90.99% of the "non-iodine" period it was also normal. There were no statistically significant differences suggesting effects of radiation factors of the "iodine" period of irradiation on the functional indices for the hypophysis-thyroid system during the first 7 years following irradiation.


Fluorouracil, leucovorin, and irinotecan plus either sunitinib or placebo in metastatic colorectal cancer: a randomized, phase III trial.
This double-blind, phase III study aimed to demonstrate that sunitinib plus FOLFIRI (fluorouracil, leucovorin, and irinotecan) was superior to placebo plus FOLFIRI in previously untreated metastatic colorectal cancer (mCRC). Patients were randomly assigned to receive FOLFIRI and either sunitinib (37.5 mg per day) or placebo (4 weeks on treatment, followed by 2 weeks off [schedule 4/2]) until disease progression. The primary end point was progression-free survival (PFS). Secondary end points included overall survival, safety, and patient-reported outcomes. The correlation between genotype and clinical outcomes was also analyzed. In all, 768 patients were randomly assigned to sunitinib plus FOLFIRI (n = 386) or placebo plus FOLFIRI (n = 382). Following a second prespecified interim analysis, the study was stopped because of potential futility of sunitinib plus FOLFIRI. Final results are reported. The PFS hazard ratio was 1.095 (95% CI, 0.892 to 1.344; one-sided stratified log-rank P = .807), indicating a lack of superiority for sunitinib plus FOLFIRI. Median PFS for the sunitinib arm was 7.8 months (95% CI, 7.1 to 8.4 months) versus 8.4 months (95% CI, 7.6 to 9.2 months) for the placebo arm. Sunitinib plus FOLFIRI was associated with more grade ≥ 3 adverse events and laboratory abnormalities than placebo (especially diarrhea, stomatitis/oral syndromes, fatigue, hand-foot syndrome, neutropenia, thrombocytopenia, anemia, and febrile neutropenia). More deaths as a result of toxicity (12 v four) and significantly more dose delays, dose reductions, and treatment discontinuations occurred in the sunitinib arm. Sunitinib 37.5 mg per day (schedule 4/2) plus FOLFIRI is not superior to FOLFIRI alone and has a poorer safety profile. This combination regimen is not recommended for previously untreated mCRC.


Long-Term Tumor-Free Survival in a Patient with Stage IV Epithelial Ovarian Cancer Undergoing High-Dose Chemotherapy and Viscum album Extract Treatment: A Case Report.
Epithelial ovarian cancer (EOC) has a poor prognosis in advanced stages. High-dose chemotherapy (HDC) was pursued in the 1990s but was not found to improve survival of patients with EOC in larger studies. Many patients with cancer use Viscum album extracts (VAE). Also called European mistletoe, Viscum album can lead to improved quality of life and reduced chemotherapy side effects and may have synergistic cytotoxic and proliferation-inhibiting effects when used together with chemotherapy. A high-grade serous epithelial ovarian carcinoma with peritoneal, adrenal, and hepatic metastases (FIGO Stage IV) was diagnosed in a 50-year-old premenopausal woman. Tumor and metastases were surgically removed in cytoreductive surgery, and the patient received adjuvant chemotherapy, without experiencing side effects from treatment. After a second-look surgery revealed lymph node metastases, HDC and autologous hematopoietic stem cell transplantation were performed. Additionally, the patient opted for treatment with VAE, which she continuously received. The patient remained tumor-free in follow-up examinations and has enjoyed good health for 20 years after initial diagnosis. Treatment with VAE in this case might have contributed to the reduction of side effects from HDC and may have acted synergistically with HDC in tumor control. Cases of VAE in EOC should be carefully documented and reported to further illustrate the influence of VAE on this cancer presentation.


Chemopreventive effects of α-tocopherol and its long-chain metabolites α-13'-hydroxy- and α-13'-carboxychromanol in LT97 colon adenoma cells.
Anticancer effects of vitamin E (tocopherols) have been studied extensively. While in vitro and animal studies showed promising results regarding anticancer effects of tocopherols, human intervention studies failed to reproduce these results. In vivo, α-tocopherol (α-TOH) is metabolized to the long-chain metabolites (LCM) 13'-hydroxychromanol (α-13'-OH) and 13'-carboxychromanol (α-13'-COOH), which likely reach the large intestine. The LCM showed antiproliferative effects in different colon cancer cell lines, but the exact mechanism of action remains unclear. To further clarify the chemopreventive action of the LCM, premalignant LT97 colon adenoma cells were treated with α-TOH, α-13'-OH and α-13'-COOH to study their impact on growth, apoptosis, antigenotoxicity, and ROS-scavenging capacity as well as expression of selected genes involved in detoxification and the cell cycle. Growth inhibitory potential was observed for α-13'-OH (IC50: 37.4 μM) and α-13'-COOH (IC50: 5.8 μM) but not for α-TOH in the tested concentrations. Levels of caspase-3 activity and expression of genes regulating the cell cycle and detoxification remained unchanged. However, α-TOH, α-13'-OH and α-13'-COOH exhibited antigenotoxic and partly ROS-scavenging capacity. The results indicate that the LCM exert chemopreventive effects via ROS-scavenging capacity, the protection against DNA damage and the induction of cell death via caspase-independent mechanisms in premalignant colon cells.


[Level of "internal" resistance of E. coli cultures to benyzlpenicillin].
Interaction of benzylpenicillin inactivation by beta-lactamase and the growth of E. coli in the presence of various antibiotic concentrations was studied. It was found that penicillin inactivation began immediately after addition of the culture to the medium in spite of the fact that the number of the viable cells at that time decreased. The culture began to multiply only when benzylpenicillin concentration in the medium lowered up to a definite level (6-7 gamma/ml). Such a concentration must determine the level of the "inner" culture resistance to benzylpenicillin, not connected with the beta-lactamase effect. Moreover, the "inner" resistance proved to be the same for the cultures of E. coli, markedly differing by the amounts of beta-lactamase produced and the resistance levels to benzylpenicillin. Therefore, the routine method for determination of the "inner" resistance of the beta-lactamase-producing cultures to penicillins, using low innoculation doses produced higher results as compared to the real "inner" resistance. Significant differences observed between the values of the "inner" resistance (6-7 gamma/ml) and the MIC of benzylpenicillin at high innoculation doses with respect to E. coli 2 (129 gamma/ml) and E. coli 82 (15000 gamma/ml) may probably be explained by destruction of the drug by beta-lactamase.


Low-grade myofibroblastic sarcoma of the larynx.
Low-grade myofibroblastic sarcoma is an uncommon sarcoma with myofibroblastic differentiation. It occurs in a wide variety of sites and has a predilection for the head and neck region. Biologically, low-grade myofibroblastic sarcoma has a propensity for local recurrence and is associated with a low risk of metastatic spread. Histologically, it can mimic a variety of different types of benign and malignant processes and often requires immunohistochemical analysis for its accurate identification. This report describes a case and discusses the differential diagnosis of a low-grade myofibroblastic sarcoma that arose in the larynx of a 69-year-old woman with a history of metastatic skin melanoma. To the best of the authors' knowledge this is the first description in the English literature of low-grade myofibroblastic sarcoma originating in the larynx.


The effect of carbohydrate and caffeine mouth rinsing on kicking performance in competitive Taekwondo athletes during Ramadan.
Mouth rinsing has been proposed as a strategy to minimize performance decrements during Ramadan. We investigated the effect of 4 weeks of Ramadan on kicking performance in 27 Taekwondo athletes performing weekly Taekwondo Anaerobic Intermittent Kick Tests (TAIKT). The effects of a placebo, 6.4% glucose and 6-mg/kg caffeine mouth rinses on TAIKT performance and perceived exertion were investigated before, during weekly training sessions, and after Ramadan in a counterbalanced, crossover design. Ramadan had a significant negative impact on the percentage of successful kicks in Week 1 of Ramadan (pre: 76.7±0.4%, Week 1: 69.9±3.2%). The percentage of successful kicks was significantly greater in the caffeine mouth rinse condition compared to the glucose and placebo conditions during the first 3 weeks of Ramadan (caffeine: 38.3±6.8%, glucose: 36.4±6.9%, placebo: 36.0±6.5%). Caffeine decreased perceived exertion during Ramadan (0.74-1.15 AU, p>0.05). Our results showed that Ramadan had a significant negative effect on repeated high-intensity kicking efforts that should be considered when training and competing. Additionally, there were significant positive effects of a caffeine mouth rinse in a sport-specific test. These data suggest that athletes can consider mouth rinsing as a strategy to enhance performance when undertaking training or competition during a period of privation.


Combining Dietary Intervention with Metformin Treatment Enhances Non-Alcoholic Steatohepatitis Remission in Mice Fed a High-Fat High-Sucrose Diet.
Non-alcoholic fatty liver disease (NAFLD) and non-alcoholic steatohepatitis (NASH) are serious health concerns for which lifestyle interventions are the only effective first-line treatment. Dietary interventions are effective in body weight reduction, but not in improving insulin sensitivity and hepatic lipid mobilization. Conversely, metformin increases insulin sensitivity and promotes the inhibition of de novo hepatic lipogenesis. In this study, we evaluated the metformin effectiveness in NASH prevention and treatment, when combined with dietary intervention in male mice fed a high-fat high-sucrose diet (HFHSD). Eighty 5-week-old C57BL/6J male mice were fed a chow or HFHSD diet and sacrificed at 20 or 40 weeks. The HFHSD-fed mice developed NASH after 20 weeks. Lipoprotein and lipidomic analyses showed that the changes associated with diet were not prevented by metformin administration. HFHSD-fed mice subject to dietary intervention combined with metformin showed a 19.6% body weight reduction compared to 9.8% in those mice subjected to dietary intervention alone. Lower hepatic steatosis scores were induced. We conclude that metformin should not be considered a preventive option for NAFLD, but it is effective in the treatment of this disorder when combined with dietary intervention.


Propionic acid and butyric acid inhibit lipolysis and de novo lipogenesis and increase insulin-stimulated glucose uptake in primary rat adipocytes.
Fermentation of dietary fibers by colonic microbiota generates short-chain fatty acids (SCFAs), e.g., propionic acid and butyric acid, which have been described to have "anti-obesity properties" by ameliorating fasting glycaemia, body weight and insulin tolerance in animal models. In the present study, we therefore investigate if propionic acid and butyric acid have effects on lipolysis, de novo lipogenesis and glucose uptake in primary rat adipocytes. We show that both propionic acid and butyric acid inhibit isoproterenol- and adenosine deaminase-stimulated lipolysis as well as isoproterenol-stimulated lipolysis in the presence of a phosphodiesterase (PDE3) inhibitor. In addition, we show that propionic acid and butyric acid inhibit basal and insulin-stimulated de novo lipogenesis, which is associated with increased phosphorylation and thus inhibition of acetyl CoA carboxylase, a rate-limiting enzyme in fatty acid synthesis. Furthermore, we show that propionic acid and butyric acid increase insulin-stimulated glucose uptake. To conclude, our study shows that SCFAs have effects on fat storage and mobilization as well as glucose uptake in rat primary adipocytes. Thus, the SCFAs might contribute to healthier adipocytes and subsequently also to improved energy metabolism with for example less circulating free fatty acids, which is beneficial in the context of obesity and type 2 diabetes.


Overcoming barriers to the recruitment of nurses as participants in health care research.
Adequate participant recruitment is critical for the successful execution of research studies involving human subjects. Participant recruitment can be particularly challenging when the intended study participants are health care providers as opposed to patients. Discussions of the challenges and successful strategies associated with provider recruitment are limited, particularly regarding providers other than physicians and settings outside of primary care. Using a case study format, we present our experiences recruiting inpatient nurses into a study which involved focus group methodology at a United States Veterans Health Administration facility. We describe the initial logistical and ethical issues involved, various challenges we encountered, and five successful action strategies used during a second recruitment wave addressing logistical issues; on-site recruitment; increased scheduling flexibility and peer-to-peer recruitment; attention to the sensitive nature of the research topic; and increased involvement of stakeholders. In conclusion, we advocate for multimodal recruitment strategies that facilitate ongoing investigator-clinician partnerships.


Does Complexity Always Correlate With Compensation? An Analysis of Work Relative Value Units in 3 Common Nerve Decompression Syndromes.
Work relative value units (wRVUs) are part of Resource Based Relative Value Scale system. It is expected that a more difficult and time-consuming procedure would yield higher wRVUs. Brachial plexus nerve decompression surgery is a more time-consuming procedure compared with carpal and cubital tunnel procedures. The aim of this study was to analyze physician reimbursement in upper limb decompression procedures by comparing mean operative times, wRVUs per minute, and dollars per minute. A retrospective cohort study was conducted from June 2016 to June 2019, including all patients who underwent carpal tunnel, cubital tunnel, and brachial plexus release procedures. Operating time was collected, and calculations of mean operative time, wRVUs per minute, and dollars per minute were performed and compared between groups. A total of 209 cases were included. Carpal tunnel accounted for 75.1% of the cases, followed by cubital tunnel and brachial plexus releases. Brachial plexus release had the highest median operative time (147 minutes), followed by cubital tunnel (57 minutes) and carpal tunnel release (16 minutes, P < 0.0001). Carpal tunnel release procedures had a significantly higher wRVUs per minute (0.310) when compared with cubital tunnel and brachial pleaxus release procedures, 0.127 and 0.077, respectively (P < 0.0001). Same was true for dollars per minute; carpal tunnel procedures yielded significantly more compensation than cubital tunnel and thoracic outlet procedures (P < 0.0001). More complex and time-consuming procedures yielded a lower reimbursement for physicians. The current work relative unit system does not account adequately for the time spent in each procedure.


Structural Basis of Oligomerization of N-Terminal Domain of Spider Aciniform Silk Protein.
Spider silk is self-assembled from water-soluble silk proteins through changes in the environment, including pH, salt concentrations, and shear force. The N-terminal domains of major and minor ampullate silk proteins have been found to play an important role in the assembly process through salt- and pH-dependent dimerization. Here, we identified the sequences of the N-terminal domains of aciniform silk protein (AcSpN) and major ampullate silk protein (MaSpN) from Nephila antipodiana (NA). Different from MaSpN, our biophysical characterization indicated that AcSpN assembles to form large oligomers, instead of a dimer, upon condition changes from neutral to acidic pH and/or from a high to low salt concentration. Our structural studies, by nuclear magnetic resonance spectroscopy and homology modelling, revealed that AcSpN and MaSpN monomers adopt similar overall structures, but have very different charge distributions contributing to the differential self-association features. The intermolecular interaction interfaces for AcSp oligomers were identified using hydrogen-deuterium exchange mass spectrometry and mutagenesis. On the basis of the monomeric structure and identified interfaces, the oligomeric structures of AcSpN were modelled. The structural information obtained will facilitate an understanding of silk fiber formation mechanisms for aciniform silk protein.


Phases and Phenotypes of Multiple Sclerosis.
This article describes the dynamic evolution of multiple sclerosis (MS) through its phases and the impact of this understanding on treatment decisions. MS consists of three phases: (1) the high-risk phase, (2) the relapsing-remitting phase, and (3) the progressive phase. Increasingly, subclinical disease activity is becoming an integral part of our definition of disease course in MS. In many patients, the relapsing-remitting phase starts as subclinical activity, likely long before they present with a clinically isolated syndrome. Differentiating progressive MS subgroups is also becoming less relevant. This is illustrated by comparing progressive MS that evolves from an asymptomatic state in individuals with radiologically isolated syndrome (primary progressive MS) and symptomatic individuals with relapsing-remitting MS (secondary progressive MS). In each case, the background disease activity and pathology can be indistinguishable. These phases evolve on a continuum and largely follow the aging process with little influence by the preceding clinical activity level. Recently, it also became evident that one or a few poorly recovered relapses at the beginning of clinical manifestations of MS predict much earlier progressive MS onset. These findings suggest that interventions to prevent progressive MS, when they become available for clinical practice, may need to be considered as early as when the asymptomatic radiologically isolated syndrome is detected. This early treatment approach is being evaluated with ongoing trials with available disease-modifying therapies. In contrast, continuing the use of disease-modifying therapy beyond a certain age may have little benefit. However, being in the progressive phase of MS is not, in itself, an argument against disease-modifying therapy use in active disease in younger patients.


Predicting treatment-seeking behavior in Guatemala: a comparison of the health services research and decision-theoretic approaches.
This study attempts to identify and describe factors associated with the choice of a health care source in rural Guatemala. Because of limited choice options, rural Guatemala makes an excellent location for studying the factors that affect utilization patterns. Illness case histories were collected from a random sample of 270 households in six villages. Then, two different methodological approaches were used to predict treatment actions. First, a sociobehavioral model, which encompasses enabling, predisposing, and need factors, was used to predict treatment choices. Using discriminant analysis we identified factors associated with the use of home remedies, a pharmacy, the health post, a physician, or folk healer. In a second, parallel study, descriptive interviews were used to identify important factors in choosing a treatment strategy. From these interviews, and from responses to hypothetical illness cases, we developed a decision model of treatment actions. Both models were tested against the set of illness cases. Results indicate that both approaches identify similar variables (especially, severity), although selection of variables through the multivariate analysis was much more successful in predicting treatment actions.


Adrenergic mechanisms for the effects of epinephrine on glucose production and clearance in man.
THE PRESENT STUDIES WERE UNDERTAKEN TO ASSESS THE ADRENERGIC MECHANISMS BY WHICH EPINEPHRINE STIMULATES GLUCOSE PRODUCTION AND SUPPRESSES GLUCOSE CLEARANCE IN MAN: epinephrine (50 ng/kg per min) was infused for 180 min alone and during either alpha (phentolamine) or beta (propranolol)-adrenergic blockade in normal subjects under conditions in which plasma insulin, glucagon, and glucose were maintained at comparable levels by infusion of somatostatin (100 mug/h), insulin (0.2 mU/kg per min), and variable amounts of glucose. In additional experiments, to control for the effects of the hyperglycemia caused by epinephrine, variable amounts of glucose without epinephrine were infused along with somatostatin and insulin to produce hyperglycemia comparable with that observed during infusion of epinephrine. This glucose infusion suppressed glucose production from basal rates of 1.8+/-0.1 to 0.0+/-0.1 mg/kg per min (P < 0.01), but did not alter glucose clearance. During infusion of epinephrine, glucose production increased transiently from a basal rate of 1.8+/-0.1 to a maximum of 3.0+/-0.2 mg/kg per min (P < 0.01) at min 30, and returned to near basal rates at min 180 (1.9+/-0.1 mg/kg per min). Glucose clearance decreased from a basal rate of 2.0+/-0.1 to 1.5+/-0.2 ml/kg per min at the end of the epinephrine infusion (P < 0.01). Infusion of phentolamine did not alter these effects of epinephrine on glucose production and clearance. In contrast, infusion of propranolol completely prevented the suppression of glucose clearance by epinephrine, and inhibited the stimulation of glucose production by epinephrine by 80+/-6% (P < 0.001). These results indicate that, under conditions in which plasma glucose, insulin, and glucagon are maintained constant, epinephrine stimulates glucose production and inhibits glucose clearance in man predominantly by beta adrenergic mechanisms.


Stability of Calcium Ion Battery Electrolytes: Predictions from Ab Initio Molecular Dynamics Simulations.
Multivalent batteries, such as magnesium-ion, calcium-ion, and zinc-ion batteries, have attracted significant attention as next-generation electrochemical energy storage devices to complement conventional lithium-ion batteries (LIBs). Among them, calcium-ion batteries (CIBs) are the least explored due to difficult reversible Ca deposition-dissolution. In this work, we examined the stability of four different Ca salts with weakly coordinating anions and three different solvents commonly employed in existing battery technologies to identify suitable candidates for CIBs. By employing Born-Oppenheimer molecular dynamics (BOMD) simulations on salt-Ca and solvent-Ca interfaces, we find that the tetraglyme solvent and carborane salt are promising candidates for CIBs. Due to the strong reducing nature of the calcium surface, the other salts and solvents readily decompose. We explain the microscopic mechanisms of salt/solvent decomposition on the Ca surface using time-dependent projected density of states, time-dependent charge-transfer plots, and climbing-image nudged elastic band calculations. Collectively, this work presents the first mechanistic assessment of the dynamical stability of candidate salts and solvents on a Ca surface using BOMD simulations, and provides a predictive path toward designing stable electrolytes for CIBs.


Concentration response functions for ultrafine particles and all-cause mortality and hospital admissions: results of a European expert panel elicitation.
Toxicological studies have provided evidence of the toxicity of ultrafine particles (UFP), but epidemiological evidence for health effects of ultrafines is limited. No quantitative summary currently exists of concentration-response functions for ultrafine particles that can be used in health impact assessment. The goal was to specify concentration-response functions for ultrafine particles in urban air including their uncertainty through an expert panel elicitation. Eleven European experts from the disciplines of epidemiology, toxicology, and clinical medicine selected using a systematic peer-nomination procedure participated. Using individual ratings supplemented with group discussion, probability distributions of effect estimates were obtained for all-cause mortality and cardiovascular and respiratory hospital admissions. Experts judged the small database of epidemiological studies supplemented with experimental studies sufficient to quantify effects of UFP on all-cause mortality and to a lesser extent hospital admissions. Substantial differences in the estimated UFP health effect and its uncertainty were found between experts. The lack of studies on long-term exposure to UFP was rated as the most important source of uncertainty. Effects on hospital admissions were considered more uncertain. This expert elicitation provides the first quantitative evaluation of estimates of concentration response functions between urban air ultrafine particles and all-cause mortality and hospital admissions.


Stretchable Semiconducting Polymers with Hydrogen-Bonding-Capable Conjugation Breakers: Synthesis and Application in Organic Thin-Film Transistors.
Diketopyrrolopyrrole (DPP)-based conjugated copolymers are important organic semiconductors for applications in high-efficiency organic thin-film transistors (OTFTs). However, the direct application of these polymers with rigid backbones in stretchable devices has limitations. In this study, we designed and synthesized three kinds of DPPBT-based copolymers, DPPBT-A1, DPPBT-A3, and DPPBT-A5, which have amide-coupled alkylene conjugation breakers capable of hydrogen bonding. Linkers with different segment lengths were copolymerized with DPP and bithiophene (BT) backbone units. A DPP-based copolymer with alternating BT moieties, DPPBT, was synthesized as a reference fully conjugated copolymer. The synthesized polymers with freely rotational backbone linkers have sufficient flexibility to develop ordered phase domains, even in thin films, in comparison to the reference copolymer. However, the introduction of the conjugation breakers, which disconnect the intramolecular π-π overlapping, tends to decrease the hole mobility (μ) from 0.76 to 0.20 cm2 V-1 s-1 in the corresponding OTFT devices. The TFT fabricated using DPPBT-A3 showed a mobility of 0.50 cm2 V-1 s-1, and the mobility value did not show a significant change even when elongated by more than 50%. Therefore, the molecular design strategy of introducing amide-coupled alkylene conjugation breakers into conjugated polymer chains can contribute significantly to the development of high-mobility stretchable conjugated polymers in future.


Site-targeted mutagenesis for stabilization of recombinant monoclonal antibody expressed in tobacco (Nicotiana tabacum) plants.
This study examined the degradation pattern of a murine IgG1κ monoclonal antibody expressed in and extracted from transformedNicotiana tabacum Gel electrophoresis of leaf extracts revealed a consistent pattern of recombinant immunoglobulin bands, including intact and full-length antibody, as well as smaller antibody fragments. N-terminal sequencing revealed these smaller fragments to be proteolytic cleavage products and identified a limited number of protease-sensitive sites in the antibody light and heavy chain sequences. No strictly conserved target sequence was evident, although the peptide bonds that were susceptible to proteolysis were predominantly and consistently located within or near to the interdomain or solvent-exposed regions in the antibody structure. Amino acids surrounding identified cleavage sites were mutated in an attempt to increase resistance. Different Guy's 13 antibody heavy and light chain mutant combinations were expressed transiently inN. tabacumand demonstrated intensity shifts in the fragmentation pattern, resulting in alterations to the full-length antibody-to-fragment ratio. The work strengthens the understanding of proteolytic cleavage of antibodies expressed in plants and presents a novel approach to stabilize full-length antibody by site-directed mutagenesis.-Hehle, V. K., Paul, M. J., Roberts, V. A., van Dolleweerd, C. J., Ma, J. K.-C. Site-targeted mutagenesis for stabilization of recombinant monoclonal antibody expressed in tobacco (Nicotiana tabacum) plants.


Quinolone derivative, flumequine, as short-term treatment for gonorrhoea.
The antigonococcal activity of the quinolone derivative flumequine was evaluated. Of 246 strains examined, 240 (97.5%) strains showed minimum inhibitory concentrations (MICs) of flumequine of less than or equal to 0.4 microgram/ml, including three beta-lactamase-producing strains. The six remaining strains showed MICs from 3.2 to 9.6 micrograms/ml. By disc diffusion tests using 3-micrograms discs of flumequine the zones of growth inhibition correlated well with the MICs of flumequine. The effect of treatment with flumequine was compared in 239 patients with uncomplicated gonorrhoea. A single dose regimen of 1200 mg flumequine orally, a two-dose regimen of 1200 and 800 mg, and a three-dose regimen of 1200, 800, and 800 mg (six hours apart) were given. With a single dose of flumequine the failure rate was 26%. The two-dose and three-dose regimens were equally effective with an overall cure rate of 95.4%. In patients harbouring beta-lactamase-producing gonococci the infection was cured. The failures (10 men) included all of the six patients infected with flumequine-resistant gonococci. Side effects were noted by 14.6% of the patients and were mostly described as dizziness.


Preparation of ribonucleic acid probes specific for Bacteroides fragilis.
Nucleic acid probes promise to be very useful in the detection of infectious agents. Several RNA probes have been developed by random cloning of chromosomal DNA for the detection of Bacteroides fragilis. Four such probes prepared by transcription of inserts of 400 bp, 800 bp, 2.4 kbp, and 6 kbp all hybridized to denatured B. fragilis DNA on nitrocellulose, but did not bind to DNA from a variety of other Bacteroides spp. or from selected aerobic strains. Each probe readily detected five clinical isolates of B. fragilis and did not react with 11 clinical isolates representing four other species in the "B. fragilis group".


X-DC: Explainable Deep Clustering Based on Learnable Spectrogram Templates.
Deep neural networks (DNNs) have achieved substantial predictive performance in various speech processing tasks. Particularly, it has been shown that a monaural speech separation task can be successfully solved with a DNN-based method called deep clustering (DC), which uses a DNN to describe the process of assigning a continuous vector to each time-frequency (TF) bin and measure how likely each pair of TF bins is to be dominated by the same speaker. In DC, the DNN is trained so that the embedding vectors for the TF bins dominated by the same speaker are forced to get close to each other. One concern regarding DC is that the embedding process described by a DNN has a black-box structure, which is usually very hard to interpret. The potential weakness owing to the noninterpretable black box structure is that it lacks the flexibility of addressing the mismatch between training and test conditions (caused by reverberation, for instance). To overcome this limitation, in this letter, we propose the concept of explainable deep clustering (X-DC), whose network architecture can be interpreted as a process of fitting learnable spectrogram templates to an input spectrogram followed by Wiener filtering. During training, the elements of the spectrogram templates and their activations are constrained to be nonnegative, which facilitates the sparsity of their values and thus improves interpretability. The main advantage of this framework is that it naturally allows us to incorporate a model adaptation mechanism into the network thanks to its physically interpretable structure. We experimentally show that the proposed X-DC enables us to visualize and understand the clues for the model to determine the embedding vectors while achieving speech separation performance comparable to that of the original DC models.


Reading skills in children and adults with albinism: the role of visual impairment.
To evaluate whether visual impairment in albinism contributes significantly to the acquisition of normal reading skills. The authors administered standardized reading tests to 41 children and 18 adults with albinism. The Young Children's Achievement Test was used for children between 4 and 6 years old and the Woodcock-Johnson III was used for children 7 years and older and adults. Parents of children and adult subjects also completed a questionnaire to document developmental, academic, and/or work experiences. The Spearman test was used to evaluate the relationship between binocular best-corrected visual acuity (BCVA) and reading test results. Standardized reading tests in both children and adults with albinism showed a normal distribution of scores. BCVA did not appear to play a significant role in the development of normal reading ability in these individuals who were visually impaired except for a mild correlation of decreased reading fluency on the Woodcock-Johnson III with decreased BCVA (r = 0.287, P = .046). Many young children with albinism had superior reading skills despite having a BCVA of 20/200 or worse (legal blindness). This study of cognitively normal children and adults with albinism demonstrates that impaired vision during childhood does not by itself significantly impede a child's ability to acquire normal reading skills. However, the lower reading fluency that occurs in the more visually impaired individuals suggests they would benefit, both in the school system and workplace, with an accommodation involving more time to complete reading tasks.


A comparison of the efficacy of new monopyridinium oximes with the oxime HI-6 against mevinphos in mice.
1. The therapeutic efficacy of three new monopyridinium oximes (2,4-PAEtM, 2,5-PAEtM, 2,5-PAAM) and the bispyridinium oxime HI-6 was evaluated in combination with benactyzine against acute poisoning with the organophosphorus insecticide mevinphos in mice. 2. When mice were treated two min after mevinphos poisoning, no significant differences in the therapeutic effectiveness of tested oximes were observed. They increased the 24h LD50 values of mevinphos about three times in comparison with non-treated intoxicated animals. 3. On the other hand, there were significant differences in their therapeutic efficacy when they were administered 30 sec following mevinphos challenge. The monopyridinium oxime 2,5-PAEtM seems to be the most efficacious against mevinphos toxicity. 4. Use of new monopyridinium oxime 2,5-PAEtM appears to be the improvement in the antidotal treatment of poisoning with organophosphorus insecticide mevinphos in comparison with HI-6.


Six autoantibodies associated with autoimmune encephalitis are not detectable in the cerebrospinal fluid of suicide attempters.
Previous findings suggest a link between neuroinflammatory processes and suicidality. Despite several lines of evidence supporting this link, including increased pro-inflammatory markers in blood-, cerebrospinal fluid (CSF)- and in post-mortem brain samples from suicidal individuals, the underlying mechanisms remain poorly understood. In this pilot study, we explored the possibility that autoimmune encephalopathies might be found among suicide attempters. We analysed the presence of six different autoantibodies (N-methyl-D-aspartate receptor, the α-amino-3-hydroxy-5-methyl-4-isoxazol-propionic acid receptor, the γ-amino-butyric acid B-receptor, the leucine-rich, glioma-inactivated 1, the contactin-associated protein-like 2, and the dipeptidyl-peptidase-like protein-6), all previously associated with psychopathology, in CSF samples from 29 unmedicated suicide attempters. Five of these subjects had high CSF/serum albumin ratio, indicative of increased blood-brain-barrier permeability. We were not able to detect any of these autoantibodies in the CSF samples. These pilot data do not support a role for autoimmune encephalopathies in suicidal behaviour, although the presence of lower levels of these autoantibodies cannot be ruled out in these patients.


Do the Different Sensory Areas Within the Cat Anterior Ectosylvian Sulcal Cortex Collectively Represent a Network Multisensory Hub?
Current theory supports that the numerous functional areas of the cerebral cortex are organized and function as a network. Using connectional databases and computational approaches, the cerebral network has been demonstrated to exhibit a hierarchical structure composed of areas, clusters and, ultimately, hubs. Hubs are highly connected, higher-order regions that also facilitate communication between different sensory modalities. One region computationally identified network hub is the visual area of the Anterior Ectosylvian Sulcal cortex (AESc) of the cat. The Anterior Ectosylvian Visual area (AEV) is but one component of the AESc that also includes the auditory (Field of the Anterior Ectosylvian Sulcus - FAES) and somatosensory (Fourth somatosensory representation - SIV). To better understand the nature of cortical network hubs, the present report reviews the biological features of the AESc. Within the AESc, each area has extensive external cortical connections as well as among one another. Each of these core representations is separated by a transition zone characterized by bimodal neurons that share sensory properties of both adjoining core areas. Finally, core and transition zones are underlain by a continuous sheet of layer 5 neurons that project to common output structures. Altogether, these shared properties suggest that the collective AESc region represents a multiple sensory/multisensory cortical network hub. Ultimately, such an interconnected, composite structure adds complexity and biological detail to the understanding of cortical network hubs and their function in cortical processing.


No effect of HMB or α-HICA supplementation on training-induced changes in body composition.
β-hydroxy-β-methylbutyrate (calcium: HMB-Ca and free acid: HMB-FA) and α-hydroxyisocaproic acid (α-HICA) are leucine metabolites that have been proposed to improve body composition and strength when combined with resistance exercise training (RET). In this double-blind randomized controlled pragmatic trial, we evaluated the effects of off-the-shelf supplements: α-HICA, HMB-FA and HMB-Ca, on RET-induced changes in body composition and performance. Forty men were blocked randomized to receive α-HICA (n = 10, fat-free mass [FFM] = 62.0 ± 7.1 kg), HMB-FA (n = 11, FFM = 62.7 ± 10.5 kg), HMB-Ca (n = 9, FFM = 65.6 ± 10.1 kg) or placebo (PLA; n = 10, FFM = 64.2 ± 5.7 kg). The training protocol consisted of a whole-body resistance training routine, thrice weekly for 8 weeks. Body composition was assessed by dual-energy x-ray absorptiometry (DXA) and total body water (TBW) by whole-body bioimpedance spectroscopy (BIS), both at baseline and at the end of weeks 4 and 8. Time-dependent changes were observed for increase in trunk FFM (p < 0.05). No statistically significant between-group or group-by-time interactions were observed. Supplementation with HMB (FA and Ca) or α-HICA failed to enhance body composition to a greater extent than placebo. We do not recommend these leucine metabolites for improving body composition changes with RET in young adult resistance trained men.


A 1-year community-based health economic study of ciprofloxacin vs usual antibiotic treatment in acute exacerbations of chronic bronchitis: the Canadian Ciprofloxacin Health Economic Study Group.
To evaluate the costs, consequences, effectiveness, and safety of ciprofloxacin vs standard antibiotic care in patients with an initial acute exacerbation of chronic bronchitis (AECB) as well as recurrent AECBs over a 1-year period. Randomized, multicenter, parallel-group, open-label study. Outpatient general practice. A total of 240 patients, 18 years or older with chronic bronchitis, with a history of frequent exacerbations (three or more in the past year) presenting with a type 1 or 2 AECB (two or more of increased dyspnea, increased sputum volume, or sputum purulence). The assessment included AECB symptoms, antibiotics prescribed, concomitant medications, adverse events, hospitalizations, emergency department visits, outpatient resources such as diagnostic tests, procedures, and patient and caregiver out-of-pocket expenses. Patients completed the Nottingham Health Profile, St. George's Respiratory Questionnaire, and the Health Utilities Index. The parameters were recorded with each AECB and at regular quarterly intervals for 1 year. These variables were compared between the ciprofloxacin-treated group and the usual-care-treated group. Patients receiving ciprofloxacin experienced a median of two AECBs per patient compared to a median of three AECBs per patient receiving usual care. The mean annualized total number of AECB-symptom days was 42.9+/-2.8 in the ciprofloxacin arm compared to 45.6+/-3.0 days in the usual-care arm (p=0.50). The overall duration of the average AECB was 15.2+/-0.6 days for the ciprofloxacin arm compared to 16.3+/-0.6 days for the usual-care arm. Treatment with ciprofloxacin tended to accelerate the resolution of all AECBs compared to usual care (relative risk=1.20; 95% confidence interval [CI], 0.91 to 1.58; p=0.19). Treatment assignment did not affect the interexacerbation period but a history of severe bronchitis, prolonged chronic bronchitis, and an increased number of AECBs in the past year were associated with shorter exacerbations-free periods. There was a slight, but not statistically significant, improvement in all quality of life measures with ciprofloxacin over usual care. The only factors predictive of hospitalization were duration of chronic bronchitis (odds ratio=4.6; 95% CI, 1.6, 13.0) and severity of chronic bronchitis (odds ratio=4.3; 95% CI, 0.8, 24.6). The incremental cost difference of $578 Canadian in favor of usual care was not significant (95% CI, -$778, $1,932). The cost for the ciprofloxacin arm over the usual care arm was $18,588 Canadian per quality-adjusted life year gained. When the simple base case analysis was expanded to examine the effect of risk stratification, the presence of moderate or severe bronchitis and at least four AECBs in the previous year changed the economic and clinical analysis to one favorable to ciprofloxacin with the ciprofloxacin-treated group having a better clinical outcome at lower cost ("win-win" scenario). Treatment with ciprofloxacin tended to accelerate the resolution of all AECBs compared to usual care; however, the difference was not statistically significant. Further, usual care was found to be more reflective of best available care rather than usual first-line agents such as amoxicillin, tetracycline, or trimethoprim-sulfamethoxazole as originally expected. Despite the similar antimicrobial activities and broad-spectrum coverage of both ciprofloxacin and usual care, the trends in clinical outcomes and all quality of life measurements favor ciprofloxacin. In patients suffering from an AECB with a history of moderate to severe chronic bronchitis and at least four AECBs in the previous year, ciprofloxacin treatment offered substantial clinical and economic benefits. In these patients, ciprofloxacin may be the preferred first antimicrobial choice.


Improved prediction of peptide detectability for targeted proteomics using a rank-based algorithm and organism-specific data.
The in silico prediction of the best-observable "proteotypic" peptides in mass spectrometry-based workflows is a challenging problem. Being able to accurately predict such peptides would enable the informed selection of proteotypic peptides for targeted quantification of previously observed and non-observed proteins for any organism, with a significant impact for clinical proteomics and systems biology studies. Current prediction algorithms rely on physicochemical parameters in combination with positive and negative training sets to identify those peptide properties that most profoundly affect their general detectability. Here we present PeptideRank, an approach that uses learning to rank algorithm for peptide detectability prediction from shotgun proteomics data, and that eliminates the need to select a negative dataset for the training step. A large number of different peptide properties are used to train ranking models in order to predict a ranking of the best-observable peptides within a protein. Empirical evaluation with rank accuracy metrics showed that PeptideRank complements existing prediction algorithms. Our results indicate that the best performance is achieved when it is trained on organism-specific shotgun proteomics data, and that PeptideRank is most accurate for short to medium-sized and abundant proteins, without any loss in prediction accuracy for the important class of membrane proteins. Targeted proteomics approaches have been gaining a lot of momentum and hold immense potential for systems biology studies and clinical proteomics. However, since only very few complete proteomes have been reported to date, for a considerable fraction of a proteome there is no experimental proteomics evidence that would allow to guide the selection of the best-suited proteotypic peptides (PTPs), i.e. peptides that are specific to a given proteoform and that are repeatedly observed in a mass spectrometer. We describe a novel, rank-based approach for the prediction of the best-suited PTPs for targeted proteomics applications. By building on methods developed in the field of information retrieval (e.g. web search engines like Google's PageRank), we circumvent the delicate step of selecting positive and negative training sets and at the same time also more closely reflect the experimentalist´s need for selecting e.g. the 5 most promising peptides for targeting a protein of interest. This approach allows to predict PTPs for not yet observed proteins or for organisms without prior experimental proteomics data such as many non-model organisms.


[Vacuum assisted closure in the treatment of pleural empyema -- first experiences with intra-thoracal application].
Meanwhile vacuum-assisted closure (V.A.C.) therapy belongs to the standard procedures in cardiovascular surgery. Its intra-thoracal application is mentioned in one unique case-report in literature. We like to show our first experiences with this technique demonstrating three cases with pleural infection. One patient with bronchial stump insufficiency, one with pleural empyema and one with penetrating abscess of the chest was treated with V.A.C.-therapy. We applied a combination of microporous silicon fleece and polyurethane foam in direct neighbourhood to ventilated lung areas inside the thoracic cavity. One patient died of metastasis of his cancer disease, while wound situation and bronchial stump insufficiency could be treated well. Two patients could be treated successfully and be discharged out of the hospital. No complications due to V.A.C.-therapy could be seen, such as fistulas of the lung or bleeding of the mediastinal organs. Intra-thoracal V.A-C.-therapy can be performed as a good therapeutic option in the treatment of septic diseases inside the chest.


Retroviral stem cell gene therapy.
Long-term in vivo gene transfer studies in mice have shown that recombinant murine retroviruses are able to infect murine hemopoietic stem cells with high efficiency. Taken together the results indicated that the proviral structure was present at high frequency in circulating hemopoietic cells resulting in significant expression levels. Because of the success of these murine studies, it was believed that gene therapy would soon be applicable to treat a wide variety of congenital or acquired human diseases associated with the hemopoietic system. However, results from gene transfer studies in nonhuman primates and first human clinical trails have indicated that murine retrovirus infection of primate hemopoietic stem cells is inefficient. Although there are essential differences between the murine and primate gene therapy studies with respect to the recombinant viruses and transduction protocols used, these differences cannot solely account for the differences observed in infection efficiency. Therefore, in recent years effort has been spent on the identification of factors limiting retroviral transduction of primate hemopoietic stem cells. Increasing knowledge concerning hemopoiesis and retroviral infection has helped in identifying a number of limiting factors. Novel transduction strategies and tools have been generated which attempt to circumvent these limiting factors. These factors as well as the strategies that showed increased retroviral infection of primate hemopoietic stem cells will be discussed.


Decline in telomere length with increasing age across nonhuman vertebrates: A meta-analysis.
The prediction that telomere length (TL) shortens with increasing age is a major element in considering the role of telomeres as a key player in evolution. While telomere attrition is found in humans both in vitro and in vivo, the increasing number of studies reporting diverse age-specific patterns of TL challenges the hypothesis of a universal decline of TL with increasing age. Here, we performed a meta-analysis to estimate the relationship between TL and age across 175 estimates encompassing 98 species of vertebrates. We found that, on average, TL does decline with increasing age during adulthood. However, this decline was weak and variable across vertebrate classes, and we also found evidence for a publication bias that might weaken our current evidence of decreasing TL with increasing age. We found no evidence for a faster decline in TL with increasing age when considering the juvenile stage (from birth to age at first reproduction) compared to the adult stage. Heterogeneity in TL ageing rates was explained by the method used to measure telomeres: detectable TL declines with increasing age were found only among studies using TRF with in-gel hybridisation and qFISH methods, but not in studies using qPCR and Southern blot-based TRF methods. While we confirmed that TL declines with increasing age in most adult vertebrates, our results identify an influence of telomere measurement methodology, which highlights the need to examine more thoroughly the effect of the method of measurement on TL estimates.


Monopylocystis visvesvarai n. gen., n. sp. and Sawyeria marylandensis n. gen., n. sp.: two new amitochondrial heterolobosean amoebae from anoxic environments.
Two new species of heterolobosean amoebae from anoxic environments, Monopylocystis visvesvarai and Sawyeria marylandensis, are described on the basis of light microscopy, electron microscopy, and their phylogenetic affiliation based on analyses of nuclear small-subunit ribosomal RNA gene sequences. Both species lack mitochondria but have organelles provisionally interpreted as hydrogenosomes, and neither can tolerate aerobic conditions. As their conditions of culture do not exclude all oxygen, they may be microaerophiles rather than strict anaerobes. Both species have unusual nucleolar morphologies. Monopylocystis visvesvarai, from a marine sediment, has nucleolar material distributed around the nuclear periphery. It is the first non-aerobic heterolobosean protist for which a cyst is known; the cyst is unmineralized and unornamented except for a single, raised, plugged pore. Sawyeria marylandensis, from an iron-rich freshwater stream, has nucleolar material distributed in one or two parietal masses, which persist during mitosis. In phylogenetic analyses of small-subunit rRNA gene sequences, Monopylocystis visvesvarai, Sawyeria marylandensis and Psalteriomonas lanterna converge to form a single clade of non-aerobic (anaerobic/microaerophilic) heteroloboseans.


A new method for measuring auricular inflammation in the mouse.
The croton oil ear test is widely used to identify prospective topical antiinflammatory drugs. Ear inflammation is produced by applying a 2% solution of croton oil on the ears of mice or rats. The effectiveness of the drug that is dissolved in the croton oil solution can be gauged by comparing the croton oil treated ears with the croton oil plus drug treated ears. The effect is measured following sacrifice of the animal by weighing either the excised ear (Tonelli et al., 1965; Glenn et al., 1978; Swingle et al., 1981; Soliman et al., 1983; Mantione and Rodriguez 1990) or a plug taken from the ear (Tubaro et al., 1985; Davis et al., 1989a; Davis et al., 1989b). Use of this technique for the generation of a time-course evaluation of antiinflammatory activity requires a large amount of the chemical to be tested and the sacrifice of many animals. In other assays, ear thickness has been measured by caliper (Carlson et al., 1985; Maloff et al., 1989) or by dial micrometer (Griswold et al., 1987), which allow multiple measurements to be made, but the pressure on the ear was not reported. In a recent review of pharmacological methods, Chang and Lewis (1989) caution that using calipers to measure ear thickness is subject to operator error and bias. Furthermore, they emphasize care must be taken to not leave the calipers in contact with the ear too long, as it is possible to squeeze substantial amounts of edema fluid out of the ear tissue.(ABSTRACT TRUNCATED AT 250 WORDS)


Calf deep venous thrombosis. A wolf in sheep's clothing?
To determine the natural history of calf deep venous thrombosis (C-DVT), an analytic review of the 20 relevant English-language papers published since 1942 was performed. Remarkably little methodologically sound research on this subject was found. However, available evidence suggests that C-DVT propagates to the thigh in up to 20% of cases and that propagation invariably occurs before embolization. No fatal emboli were reported in patients presenting with isolated C-DVT. Traditional anticoagulation treatment with heparin sodium and warfarin sodium of symptomatic patients with C-DVT appears to prevent extension, embolization, and early recurrence. There is no convincing evidence that C-DVT leads to chronic venous insufficiency or whether the risks of anticoagulation exceed the risks of no treatment. As an option to anticoagulation, physicians may choose to follow patients with C-DVT with serial impedance plethysmography, treating only if there is evidence of proximal extension.


First Female-to-Male Facial Confirmation Surgery with Description of a New Procedure for Masculinization of the Thyroid Cartilage (Adam's Apple).
Although male-to-female transgender patients commonly seek facial feminization surgery, facial masculinization surgery in the female-to-male transgender population is unreported in the literature. This report documents the first known female-to-male facial masculinization surgery, including a new technique for creating an "Adam's apple" to enhance the facial masculine appearance of a natal female. The authors "reversed" the methods typically used to feminize male facial features, and modified the forehead, nose, and chin to masculinize the patient's natal female facial features. The authors devised a novel technique to augment the thyroid cartilage using autologous rib cartilage to create a visible Adam's apple. Initially, masculinization of the chin was accomplished with a multisegment chin osteotomy with grafts to vertically expand and widen the chin along with correcting pronounced microgenia. Subsequently, a second facial masculinization procedure was performed to masculinize the forehead, nose, and thyroid cartilage. Rib cartilage was harvested and carved into an appropriately shaped thyroid cartilage onlay graft and then attached and integrated with the native thyroid cartilage, creating a fully mobile cartilage that translocates up and down with swallowing and a visible Adam's apple. Previously described techniques to masculinize the facial features of natal male patients were adapted to masculinize the female-to-male patient. Those procedures were combined with the novel technique to create a visually perceptible and naturally mobile Adam's apple in the female-to-male transsexual patient. Collectively, these described procedures can now provide most female-to-male transsexual patients with a satisfying transformation of their facial features. Therapeutic, V.


Is Breast Conserving Therapy a Safe Modality for Early-Stage Male Breast Cancer?
Male breast cancer (MBC) is a rare disease and lacks data-based treatment guidelines. Most men are currently treated with modified radical mastectomy (MRM) or simple mastectomy (SM). We compared the oncologic treatment outcomes of early-stage MBC to determine whether breast conservation therapy (BCT) is appropriate. We searched the Surveillance, Epidemiology, and End Results database for MBC cases. That cohort was narrowed to cases of stage I-II, T1-T2N0 MBC with surgical and radiation therapy (RT) data available. The patients had undergone MRM, SM, or breast conservation surgery (BCS) with or without postoperative RT. We calculated the actuarial 5-year cause-specific survival (CSS). We identified 6263 MBC cases and included 1777 men with stage I or II, T1-T2, node-negative disease, who had the required treatment information available. MRM without RT was the most common treatment (43%). Only 17% underwent BCS. Of the BCS patients, 46% received adjuvant RT to complete the traditional BCT. No deaths were recorded in the BCT group, regardless of stage, or in the 3 stage I surgical groups if the men had received RT. The actuarial 5-year CSS was 100% in each BCT group. MRM alone resulted in an actuarial 5-year CSS of 97.3% for stage 1% and 91.2% for stage 2. The results from our study suggest that BCT for early-stage MBC yields comparable survival compared with more invasive treatment modalities (ie, MRM or SM alone). This could shift the treatment paradigm to less-invasive interventions and might have the added benefit of increased functional and psychological outcomes. Further prospective studies are needed to confirm our conclusions.


Role of Multidrug Resistance Proteins in Nonresponders to Immunomodulatory Therapy for Noninfectious Uveitis.
Nearly a third to half of patients with noninfectious uveitis (NIU) fail to achieve control with immunomodulatory therapy (IMT). Multidrug resistance (MDR) proteins are transmembrane proteins that allow efflux of intracellular drugs, leading to drug resistance. The aim of our study was to compare MDR protein function in blood CD4+ cells between responders and nonresponders to IMT. We included NIU patients on IMT for ≥6 months and corticosteroid dose ≤10 mg/d. Nonresponders to treatment were those with worsening (two or more steps) of inflammation in the past 3 months on full-dose immunosuppressive therapy. MDR function was assessed by Rhodamine-123 dye retention in blood CD4+ cells. Three nonresponders were treated with adjunctive oral cyclosporine A (CSA, MDR inhibitor) therapy for 2 months and reevaluated. Fourteen NIU patients were recruited. Most (n = 8) had Vogt-Koyanagi-Harada disease. These included nine nonresponders and five responders to IMT. Nonresponders produced significantly higher MDR function and proinflammatory cytokines (interferon γ, tumor necrosis factor α, interleukin 17, and Granulocyte Macrophage Colony Stimulating Factor (GM-CSF)) than responders. In vitro CSA treatment of CD4+ cells inhibited MDR expression and proinflammatory cytokine production while increasing Foxp3. Finally, adjunctive oral CSA therapy led to improvement in clinical inflammatory scores with a concurrent decrease in MDR function and proinflammatory cytokine secretion. MDR function is significantly higher in CD4+ T cells of nonresponders to IMT. Adjunctive CSA therapy may decrease MDR function and allow improvement in treatment response to IMT. Our study highlights the need for MDR inhibition strategies in NIU patients not responding to IMT for improving the efficacy of anti-inflammatory therapy.


PEGylated NiPAM microgels: synthesis, characterization and colloidal stability.
The objective of this work is to synthesize highly stable thermoresponsive microgels that could be used in diverse applications. To achieve this, N-isopropylacrylamide (NiPAM) based microgels were first synthesized by surfactant-free precipitation polymerization of NiPAM in the presence of poly(ethylene glycol)methacrylate (PEG) as a macro-comonomer and methylenebisacrylamide (MBA) as a chemical crosslinker. By combining a complete set of techniques such as dynamic light scattering (DLS), scanning electron microscopy (SEM), zetametry, 1H NMR and micro-differential scanning calorimetry (μDSC), we clearly demonstrate that (i) the incorporation of the PEG chains controls the size and the polydispersity of the NiPAM-based microgels, whereas the thermal behavior in solution (enthalpy, volume phase transition temperature (VPTT)) remains almost the same as for pure NiPAM microgels; (ii) the PEG chains are mainly located on the microgel periphery; and (iii) the presence of the PEG chains strongly increases the colloidal stability of microgels in electrolyte solutions at high temperatures.