Synthesis of two potential NK<Subscript>1</Subscript>-receptor ligands using [1-<Superscript>11</Superscript>C]ethyl iodide and [1-<Superscript>11</Superscript>C]propyl iodide and initial PET-imaging

Background

The previously validated NK₁-receptor ligand [O-methyl-¹¹C]GR205171 binds with a high affinity to the NK₁-receptor and displays a slow dissociation from the receptor. Hence, it cannot be used in vivo for detecting concentration changes in substance P, the endogenous ligand for the NK₁-receptor. A radioligand used for monitoring these changes has to enable displacement by the endogenous ligand and thus bind reversibly to the receptor. Small changes in the structure of a receptor ligand can lead to changes in binding characteristics and also in the ability to penetrate the blood-brain barrier. The aim of this study was to use carbon-11 labelled ethyl and propyl iodide with high specific radioactivity in the synthesis of two new and potentially reversible NK₁-receptor ligands with chemical structures based on [O-methyl-¹¹C]GR205171.

Methods

[1-¹¹C]Ethyl and [1-¹¹C]propyl iodide with specific radioactivities of 90 GBq/μmol and 270 GBq/μmol, respectively, were used in the synthesis of [O-methyl-¹¹C]GR205171 analogues by alkylation of O-desmethyl GR205171. The brain uptake of the obtained (2S,3S)-N-(1-(2- [1-¹¹C]ethoxy-5-(3-(trifluoromethyl)-4H-1,2,4-triazol-4-yl)phenyl)ethyl)-2-phenylpiperidin-3-amine (I) and (2S,3S)-2-phenyl-N-(1-(2- [1-¹¹C]propoxy-5-(3-(trifluoromethyl)-4H-1,2,4-triazol-4-yl)phenyl)ethyl)piperidin-3-amine (II) was studied with PET in guinea pigs and rhesus monkeys and compared to the uptake of [O-methyl-¹¹C]GR205171.

Results

All ligands had similar uptake distribution in the guinea pig brain. The PET-studies in rhesus monkeys showed that (II) had no specific binding in striatum. Ligand (I) had moderate specific binding compared to the [O-methyl-¹¹C]GR205171. The ethyl analogue (I) displayed reversible binding characteristics contrary to the slow dissociation rate shown by [O-methyl-¹¹C]GR205171.

Conclusion

The propyl-analogue (II) cannot be used for detecting changes in NK₁-ligand levels, while further studies should be performed with the ethyl-analogue (I).

     

Antibacterial activities of the methanol extracts and compounds from <Emphasis Type="Italic">Erythrina sigmoidea against Gram-negative multi-drug resistant phenotypes</Emphasis>

Background

In the present study, the methanol extracts from the leaves, as well as compounds namely sigmoidin I (1), atalantoflavone (2), bidwillon A (3), neocyclomorusin (4), 6α-hydroxyphaseollidin (5) and neobavaisoflavone (6) (from the bark extract) were tested for their activities against a panel of Gram-negative bacteria including multi-drug resistant (MDR) phenotypes.

Methods

Broth microdilution method was used to determine the minimum inhibitory concentrations (MICs) and the minimum bactericidal concentrations (MBCs) of the extracts as well as compounds 1–6.

Results

The MIC results indicated that the crude extracts from the leaves and bark of this plant were able to inhibit the growth of 96.3 % of the 27 tested bacteria. Compounds 2–6 displayed selective activities, their inhibitory effects being obtained on 8.3 %, 41.7 %, 58.3 %, 58.3 % and 66.7 % of tested bacteria respectively for 2, 3, 5, 6 and 4. The lowest MIC value of 8 μg/mL was obtained with 6 against Escherichia coli ATCC8739, Enterobacter cloacae ECCI69, Klebsiella pneumoniae KP55, Providencia stuartii NAE16 and Pseudomonas aeruginosa PA01.

Conclusion

The present study demonstrates that Erythrina sigmoidea is a potential source of antibacterial drugs to fight against MDR bacteria. Neobavaisoflavone (6) is the main antibacterial consituents of the bark crude extract.

     

Identification of novel <Emphasis Type="Italic">LEPR</Emphasis> mutations in Pakistani families with morbid childhood obesity

Background

Mutations in the genes encoding leptin (LEP), the leptin receptor (LEPR), and the melanocortin 4 receptor (MC4R) are known to cause severe early-onset childhood obesity. The aim of the current study was to examine the prevalence of damaging LEP, LEPR, and MC4R mutations in Pakistani families having a recessive heritance of early-onset obesity.

Methods

Using targeted resequencing, the presence of rare mutations in LEP, LEPR, and MC4R, was investigated in individuals from 25 families suspected of having autosomal recessive early-onset obesity. Segregation patterns of variants were assessed based on chip-based genotyping.

Results

Homozygous LEPR variants were identified in two probands. One carried a deletion (c.3260AG) resulting in the frameshift mutation p.Ser1090Trpfs*6, and the second carried a substitution (c.2675C?>?G) resulting in the missense mutation p.Pro892Arg. Both mutations were located within regions of homozygosity shared only among affected individuals. Both probands displayed early-onset obesity, hyperphagia and diabetes. No mutations were found in LEP and MC4R.

Conclusions

The current study highlights the implication of LEPR mutations in cases of severe early-onset obesity in consanguineous Pakistani families. Through targeted resequencing, we identified novel damaging mutations, and our approach may therefore be utilized in clinical testing or diagnosis of known forms of monogenic obesity with the aim of optimizing obesity treatment.

     

Quantitative Analysis of Promoter Activity by Green Fluorescent Protein (GFP) Target/Reporter Strategy in a Novel Transgenic <Emphasis Type="Italic">alx/fpr-rs2</Emphasis> Null Mouse

Objective and design: A dual-purpose targeting/reporter vector, incorporating GFP, was utilised to monitor promoter activity in transgenic alx/fpr-rs2 null mice. Materials and Methods: alx/fpr-rs2 null mice were generated by homologous recombination in embryonic stem (ES) cells. A GFP construct was fused inframe within the promoter region allowing representative promoter activity to be monitored by flow cytometry. TNF-α secretion was measured by ELISA. Results: Germline transmission of the alx/fpr-rs2 was confirmed by southern blotting. PCR primer pairs were designed to recognise GFP allowing genotyping. Inducible GFP expression was observed during bone marrow derived macrophage (BMM) differentiation (P<0.01) and following subsequent treatment with LPS (P<0.05), TNF-a secretion mirrored GFP induction. Furthermore GFP positive cell populations were observed in vivo within nai’ve and inflammatory environments. Conclusions: A dual targeting/reporter strategy utilising GFP is a highly effective way of identifying germline transmission in a transgenic colony. In addition the insertion into the promoter region allows a quantitative measure of alx/fpr-rs2 promoter activity and acts as a phenotypic marker in vivo.     

Multidrug-resistant <Emphasis Type="Italic">Mycoplasma genitalium</Emphasis> infections in Europe

In Japan and Australia, multidrug-resistant Mycoplasma genitalium infections are reported with increasing frequency. Although macrolide-resistant M. genitalium strains are common in Europe and North America, fluoroquinolone-resistant strains are still exceptional. However, an increase of multidrug-resistant M. genitalium in Europe and America is to be expected. The aim of this paper is to increase awareness on the rising number of multidrug-resistant M. genitalium strains. Here, one of the first cases of infection with a genetically proven multidrug-resistant M. genitalium strain in Europe is described. The patient was a native Dutch 47-year-old male patient with urethritis. Mycoplasma genitalium was detected, but treatment failed with azithromycin, doxycycline and moxifloxacin. A urogenital sample was used to determine the sequence of the 23S rRNA, gyrA, gyrB and parC genes. The sample contained an A2059G single nucleotide polymorphism (SNP) in the 23S rRNA gene and an SNP in the parC gene, resulting in an amino acid change of Ser83 → Ile, explaining both azithromycin and moxifloxacin treatment failure. The SNPs associated with resistance were probably generated de novo, as a link with high-prevalence areas was not established. It is, thus, predictable that there is going to be an increase of multidrug-resistant M. genitalium strains in Europe. As treatment options for multidrug-resistant M. genitalium are limited, the treatment of M. genitalium infections needs to be carefully considered in order to limit the rapid increase of resistance to macrolides and fluoroquinolones.     

From medicinal plant extracts to defined chemical compounds targeting the histamine H<Subscript>4</Subscript> receptor: <Emphasis Type="Italic">Curcuma longa</Emphasis> in the treatment of inflammation

Objectives

The aim was to evaluate the activity of seven medicinal, anti-inflammatory plants at the hH₄R with focus on defined chemical compounds from Curcuma longa.

Materials

Activities were analyzed with membrane preparations from Sf9 cells, transiently expressing the hH₄R, G_αi2 and G_β1γ2 subunits.

Methods

From the methanolic extract of C. longa curcumin (1), demethoxycurcumin (2) and bis(4-hydroxy-cinnamoyl)methane (3) were isolated, purified with HPLC (elution-time 10.20, 9.66, 9.20 min, respectively) and together with six additional extracts, were characterized via radioligand binding studies at the hH₄R.

Results

Compounds from C. longa were the most potent ligands at the hH₄R. They exhibited estimated K _i values of 4.26–6.26 µM (1.57–2.31 µg/mL) (1); 6.66––8.97 µM (2.26–3.04 µg/mL) (2) and 10.24–14.57 µM (3.16–4.49 µg/mL) (3) (95% CI). The estimated K _i value of the crude extract of curcuma was 0.50–0.81 µg/mL. Fractionated curcumin and the crude extract surpassed the effect of pure curcumin with a K _i value of 5.54 µM or 2.04 µg/mL [95% CI (4.47–6.86 µM), (1.65–2.53 µg/mL)].

Conclusion

Within this study, defined compounds of C. longa were recognized as potential ligands and reasonable lead structures at the hH₄R. The mode of anti-inflammatory action of curcumin was further elucidated and the role of extracts in traditional phytomedicine was strengthened.

     

<Emphasis Type="Italic">Mycoplasma hominis</Emphasis> and <Emphasis Type="Italic">Gardnerella vaginalis</Emphasis> display a significant synergistic relationship in bacterial vaginosis

Gardnerella vaginalis plays an important role in bacterial vaginosis (BV,) while the role of genital Mollicutes is less obvious. The diagnosis of BV by use of the current Gram stain Nugent score is also suboptimal for defining the role of Mollicutes that lack a cell wall. Since bacterial load and diversity is an important prerequisite for BV, real-time quantitative polymerase chain reaction (qPCR) assays enable these to be assessed. The purpose of this study was to define the role of genital Mollicutes and potential patterns of synergy with G. vaginalis in women with BV. Vaginal swabs from 130 women categorised by Nugent score as BV (n?=?28), intermediate (n?=?22) and non-BV (n?=?80) were tested against four qPCR TaqMan assays targeting G. vaginalis, Mycoplasma hominis, M. genitalium, Ureaplasma urealyticum and U. parvum. Statistical analyses were used to compare bacterial prevalence and load between the three groups of women. Mycoplasma hominis and G. vaginalis co-infection was significantly more common in BV (60.7 %) compared to intermediate (36.4 %) and non-BV (8.8 %) Nugent scores (p?<?0.001). Significantly higher loads of M. hominis (p?=?0.001) and G. vaginalis (p?<?0.001) were detected in women with BV and the respective loads in M. hominis and G. vaginalis co-infections displayed a significant positive correlation (p?<?0.001; r?=?0.60). No significant associations were seen with the other Mollicutes. The findings strengthen the evidence of a role for M. hominis in BV and a potential synergy with G. vaginalis. This synergy could be an important trigger of the condition and sexual contact the conduit for the transmission of an otherwise commensal bacterium that could initiate it.     

Frequent <Emphasis Type="Italic">BRAF</Emphasis><Superscript><Emphasis Type="Italic">V600E</Emphasis></Superscript> and Absence of <Emphasis Type="Italic">TERT</Emphasis> Promoter Mutations Characterize Sporadic Pediatric Papillary Thyroid Carcinomas in Japan

Pediatric papillary thyroid carcinoma (PTC) has unique features but requires further genetic investigation. Moreover, there has been increasing concern about the risk for pediatric PTC in Japan after the Fukushima accident. This study aims to evaluate the frequencies of BRAF and TERT promoter mutations and to examine their significance in non-radiation-associated pediatric PTCs in Japan. We enrolled 81 pediatric PTC patients aged ≤20 years. The control group included 91 adult PTCs from patients >20 years old. BRAF and TERT mutations were analyzed by allele-specific-PCR and/or Sanger sequencing. Compared with adult PTCs, pediatric PTCs exhibited larger tumor size, more frequent lymph node metastasis, and less classical histology. The prevalence of BRAF ^V600E in pediatric PTCs was 54% and significantly lower than that in adults of 85%. In the pediatric PTCs, BRAF ^V600E was positively associated with older age, classical histology, and the lymph node metastasis but independent from other clinicopathological factors. TERT mutations were identified in 13% of adults and in none of the pediatric PTCs. In conclusion, pediatric PTCs are characterized by more advanced clinicopathological features, lower BRAF ^V600E frequency, and absence of TERT mutation. The BRAF ^V600E frequency in this study is similar to the reported BRAF ^V600E frequency in the ultrasonographically screened pediatric PTCs in Fukushima.     

Insecticidal activity of essential oils from native medicinal plants of Central Argentina against the house fly, <Emphasis Type="Italic">Musca domestica</Emphasis> (L.)

The insecticidal activity of nine essential oils (EOs) against the house fly (Musca domestica) was evaluated by placing flies in a screw-cap glass jar holding a piece of EO-treated cotton yarn. The dose necessary to kill 50% of flies (LC₅₀) in 30 min was determined at 26?±?1°C. The EOs showed LC₅₀ values ranging from 0.5 to 46.9 mg/dm³. The EO from Minthostachys verticillata was the most potent insecticide (LC₅₀?=?0.5 mg/dm³) followed by EOs from Hedeoma multiflora (LC₅₀?=?1.3 mg/dm³) and Artemisia annua (LC₅₀?=?6.5 mg/dm³). The compositions of the nine EOs, obtained by hydrodistillation of medicinal herbs, were analyzed by gas chromatography/mass spectroscopy. These analyses showed that (4R)(+)-pulegone (69.70%), menthone (12.17%), and limonene (2.75%) were the principal components of M. verticillata EO. (4R)(+)-pulegone was also the main constituent (52.80%) of H. multiflora, while artemisia ketone (22.36%) and 1,8-cineole (16.67%) were the major constituents of A. annua EO. The terpene (4R)(+)-pulegone showed a lower toxicity (LC₅₀?=?1.7 mg/dm³) than M. verticillata or H. multiflora EOs. Dimethyl 2,2-dichlorovinyl phosphate, selected as a positive control, showed an LC₅₀ of 0.5 mg/dm³. EOs from M. verticillata and H. multiflora show promise as natural insecticides against houseflies.     

A study on the immunological basis of the dissociation between type I-hypersensitivity skin reactions to <Emphasis Type="Italic">Blomia tropicalis</Emphasis> antigens and serum anti-<Emphasis Type="Italic">B. tropicalis</Emphasis> IgE antibodies

Background

Two conditions are used as markers of atopy: the presence of circulating anti-allergen IgE antibodies and the presence of positive skin prick test (SPT) reactions to allergenic extracts. The correlation between these conditions is not absolute. This study aimed at investigating immunological parameters that may mediate this lack of correlation. Individuals whose sera contained anti-B. tropicalis extract IgE antibodies (α-Bt E IgE) were divided into two groups, according to the presence or absence of skin reactivity to B. tropicalis extract (Bt E). The following parameters were investigated: total IgE levels; α-Bt E IgE levels; an arbitrary α-Bt E IgE/total IgE ratio; the proportion of carbohydrate-reactive α-Bt E IgE; the proportion of α-Bt E IgE that reacted with Ascaris lumbricoides extract (Al E); the production of IL-10 by Bt E- and Al E-stimulated peripheral blood cells (PBMC).

Results

Total IgE levels were similar in the two groups, but α-Bt E IgE was significantly higher in the SPT-positive group (SPT+). A large overlap of α-Bt E IgE levels was found in individuals of both groups, indicating that these levels alone cannot account for the differences in SPT outcome. Individuals of the two groups did not differ, statistically, in the proportion of α-Bt E IgE that reacted with carbohydrate and in the production of IL-10 by Bt E- and Al E-stimulated PBMC. Both groups had part of α-Bt E IgE activity absorbed out by Al E, indicating the existence of cross-reactive IgE antibodies. However, the α-Bt E IgE from the SPT-negative individuals (SPT-) was more absorbed with AlE than the α-Bt E IgE from the SPT+ individuals. This finding may be ascribed to avidity differences of the α-Bt E IgE that is present in the two groups of individuals, and could occur if at least part of the α-Bt E IgE from the SPT- individuals were elicited by A. lumbricoides infection.

Conclusion

The present results suggest that a low ratio of specific IgE to total IgE levels (in a minority of individuals), and differences in α-Bt E IgE avidities (which would have high affinities for A. lumbricoides antigens in SPT- than in SPT+ individuals) may play a role in the down-modulation of type-I hypersensitivity reaction against aeroallergens described in helminth-infected individuals.

     

Distinct Early Inflammatory Events during Ear Tissue Regeneration in Mice Selected for High Inflammation Bearing S<Emphasis Type="Italic">lc11a1 R</Emphasis> and <Emphasis Type="Italic">S</Emphasis> Alleles

High inflammatory AIRmax mice homozygous for Slc11a1 R and S alleles were produced. AIRmax ^SS mice showed faster ear tissue regeneration than AIRmax ^RR mice, suggesting that the S allele favored tissue restoration. Here, we investigated the gene expression profiles and the inflammatory reactions of AIRmax ^RR and AIRmax ^SS mice during the initial phase of ear tissue regeneration. We observed superior levels of analysis of wound myeloperoxidase and edema in AIRmax ^SS mice, although similar cell influx was verified in both lines. Of the genes, 794 were up- and 674 down-regulated in AIRmax ^RR , while 735 genes were found to be up- and 1616 down-regulated in AIRmax ^SS mice 48 h after punch. Both mouse lines showed significant over-represented genes related to cell proliferation; however AIRmax ^SS displayed up-regulation of inflammatory response genes. Quantitative PCR experiments showed higher expressions of Tgfb1, Dap12 and Trem1 genes in AIRmax ^SS mice. These results indicate that Slc11a1 gene modulated the early inflammatory events of ear tissue regeneration.     

Selective β-adrenergic Receptor Expression on Human Memory CD8+ T Lymphocyte Subsets Regulates Mobilization and INF-y Production

Introduction: CD8⁺ T lymphocytes (CD8TLs) express β-adrenergic receptors (βAR), which bind the neurotransmitter norepinephrine and stress hormone epinephrine released during inflammation, trauma, and psychological stress. Little is known about the functions of this βAR expression on CD8TLs. Methods: Volunteers were exposed to a psychological stressor (N=24). Flow cytometry identified CD8TL subsets by CCR7, CD27, CD28 and CD45RA expression. Adrenergic receptor subtype expression was determined by micro-array. The effects of βPAR stimulation on IFN-γ production in activated CD8TLs was tested in vitro using PMA/Ionomycin. Results: Stress caused selective migration of effector-memory (CCR7^?CD27^?CD28^?) CD8TLs into the blood (+148%, p<.001). An 8-fold up-regulation of the β₂AR was demonstrated in effector-memory cells as compared to naïve CD8TLs. Stimulation of the β₂AR subtype completely inhibited IFN-γ production. Conclusion: These results show that β₂AR stimulation enhances peripheral immune surveillance in a highly selective manner, and might protect against excessive cytokine release during inflammation and stress.     

Opening–closing pattern of four pericardial prostheses: results from an in vitro study of leaflet kinematics

Pericardial and porcine stented aortic valves have different leaflet kinematics. To study the biomechanics of a prosthesis thoroughly, the in vitro setting is the most appropriate. The aim of our study was to find out whether the prosthesis design in which the pericardial sheet is outside the stent post might influence the opening and closing patterns of the leaflets. Four pericardial prostheses (Magna Ease [MG] 21, Trifecta [TRI] 21, Soprano-Armonia [SA] 20 and Mitroflow [MF] 23) that fitted aortic roots with a native annulus diameter of 2.1 cm were implanted and their leaflet kinematics was studied by a high-speed digital camera. In the opening phase, MG showed the shortest RVOT and the highest RVOVI, with values of 12 ± 2 and 209 ± 17 ms, respectively. The RVOT of MG was significantly shorter than that of MF (p < 0.01), but not than that of TRI (p = 0.286). Both TRI and SA showed similar opening patterns (TRI: RVOT of 15 ± 3 ms and RVOVI of 132 ± 25 ms; SA: 17 ± 2 ms and 126 ± 19 ms), without statistically significant difference. Conversely, MF showed the slowest profile, with an RVOT of 23 ± 3 ms and an RVOVI of 94 ± 8 ms (Table 1; Fig. 3). The opening/closing profile is not influenced by the position of the pericardial leaflets, but depends on other intrinsic structural characteristics related to the material used for the stent and leaflets. Moreover, the kinematics does not affect the valve performance.

Table 1 Kinematics and hydrodynamic results, reported as means and standard deviations, evaluated over the tested heart samples

	TRI	SA	MG	MF	ANOVA	TRI versus SA	TRI versus MG	TRI versus MF	SA versus MG	SA versus MF	MG versus MF
					p Value	p Value	p Value	p Value	p Value	p Value	p Value
ET (ms)						1.0	1.0	1.0	1.0
RVOT (ms)	15 ± 3	17 ± 2	12 ± 2	23 ± 3	<0.01	1.0	0.286	<0.01	0.03	<0.01	<0.01
SVCT (ms)	247 ± 14	231 ± 15	256 ± 26	241 ± 11	0.170	0.463	0.853	0.931	0.213	1.0	1.0
RVCT (ms)	35 ± 19	52 ± 13	32 ± 17	52 ± 4	0.07	0.474	1.0	0.494	0.236	1.0	0.247
TVCT (ms)	283 ± 10	283 ± 19	289 ± 10	293 ± 11	0.584	1.00	1.0	1.0	1.0	1.0	1.0
RVOVI (ms?1)	132 ± 25	126 ± 19	209 ± 17	94 ± 8	<0.01	0.959	<0.01	0.02	<0.01	0.07	<0.01
SVCVI (ms?1)	?0.9 ± 0.3	?1.1 ± 0.4	?0.57 ± 0.1	?0.55 ± 0.1	<0.01	1.0	0.353	0.292	0.045	0.04	1.0
RVCVI (ms?1)	?16 ± 4	?10 ± 2	?18 ± 6	?10 ± 1	<0.01	0.396	1.0	0.513	0.025	1.0	0.03
Δp (mmHg)	6.7 ± 3.6	10.6 ± 5.5	15.2 ± 7.9	10.7 ± 6.1	<0.01	0.01	<0.01	0.01	0.04	1.0	<0.01
EOA (cm2)	2.2 ± 1.2	1.7 ± 0.9	1.5 ± 0.8	1.7 ± 0.9	<0.01	0.03	<0.01	0.01	0.261	0.617	0.11
El  %	7.3 ± 1	11.9 ± 1	15.4 ± 2	11.8 ± 3	<0.01	<0.01	<0.01	<0.01	0.04	1.00	0.03
CO (L/min)	3.1 ± 0.4	2.8 ± 0.5	3.1 ± 0.3	3.0 ± 0.5	0.534	0.282	0.792	0.702	0.106	0.552	0.559

ET ejection time, RVOT rapid valve-opening time, SVCT slow valve-closing time, RVCT rapid valve-closing time, TVCT total valve-closing time, RVOVI rapid valve-opening velocity index, SVCVI slow valve-closing velocity index, RVCVI rapid valve-closing velocity index, Δp mean pressure drop, EOA effective orifice area, El % energy loss, CO cardiac output

     

Predicting Cumulative and Maximum Brain Strain Measures From HybridIII Head Kinematics: A Combined Laboratory Study and <Emphasis Type="Italic">Post-Hoc</Emphasis> Regression Analysis

Due to growing concern on brain injury in sport, and the role that helmets could play in preventing brain injury caused by impact, biomechanics researchers and helmet certification organizations are discussing how helmet assessment methods might change to assess helmets based on impact parameters relevant to brain injury. To understand the relationship between kinematic measures and brain strain, we completed hundreds of impacts using a 50th percentile Hybrid III head-neck wearing an ice hockey helmet and input three-dimensional impact kinematics to a finite element brain model called the Simulated Injury Monitor (SIMon) (n = 267). Impacts to the helmet front, back and side included impact speeds from 1.2 to 5.8 ms^?1. Linear regression models, compared through multiple regression techniques, calculating adjusted R ² and the F-statistic, determined the most efficient set of kinematics capable of predicting SIMon-computed brain strain, including the cumulative strain damage measure (specifically CSDM-15) and maximum principal strain (MPS). Resultant change in angular velocity, Δω _R, better predicted CSDM-15 and MPS than the current helmet certification metric, peak g, and was the most efficient model for predicting strain, regardless of impact location. In nearly all cases, the best two-variable model included peak resultant angular acceleration, α _R, and Δω _R.     

Clinical and microbiological characteristics of peritoneal dialysis-related peritonitis caused by <Emphasis Type="Italic">Escherichia coli</Emphasis> in southern Taiwan

Peritonitis is a serious complication and major cause of treatment failure in patients undergoing peritoneal dialysis (PD). Escherichia coli is the major pathogen in extraintestinal Gram-negative infections, including PD-related peritonitis. The outcomes of E. coli peritonitis in PD varied from relatively favorable outcomes to a higher incidence of treatment failure. The aim of this study was to investigate the impact of bacterial virulence and host characteristics on the outcomes of PD-related peritonitis caused by E. coli. From January 2000 to June 2016, a total of 47 episodes of monomicrobial and 10 episodes of polymicrobial E. coli PD-related peritonitis, as well as 89 episodes of monomicrobial Gram-positive (56 Staphylococcus spp. and 33 Streptococcus spp.) PD-related peritonitis cases, were retrospectively enrolled. Clinical features, E. coli bacterial virulence, and outcomes were analyzed. Compared to Streptococcus spp. peritonitis, E. coli peritonitis had a higher peritoneal catheter removal rate (38 versus 12%; P =?0.0115). Compared to the monomicrobial group, patients in polymicrobial group were older and had higher peritoneal catheter removal rate (80 versus 38%; P =?0.0324). Treatment failure of E. coli peritonitis was associated with more polymicrobial peritonitis and immunocompromised comorbidity, longer duration of PD therapy, and more antimicrobial resistance. E. coli isolates with more iron-related genes had higher prevalence of phylogenetic group B2 and papG II, iha, ompT, and usp genes. This study demonstrates the important roles of clinical and bacterial characteristics in the outcomes of monomicrobial and polymicrobial E. coli PD-related peritonitis.     

Analysis of <Emphasis Type="Italic">KRAS</Emphasis> and <Emphasis Type="Italic">BRAF</Emphasis> genes mutation in the central nervous system metastases of non-small cell lung cancer

KRAS mutations are associated with tumor resistance to EGFR TKIs (erlotinib, gefitinib) and to monoclonal antibody against EGFR (cetuximab). Targeted treatment of mutated RAS patients is still considered as a challenge. Inhibitors of c-Met (onartuzumab or tiwantinib) and MEK (selumetinib—a dual inhibitor of MEK1 and MEK2) signaling pathways showed activity in patients with mutations in KRAS that can became an effective approach in carriers of such disorders. BRAF mutation is very rare in patients with NSCLC, and its presence is associated with sensitivity of tumor cells to BRAF inhibitors (vemurafenib, dabrafenib). In the present study, the frequency and type of KRAS and BRAF mutation were assessed in 145 FFPE tissue samples from CNS metastases of NSCLC. In 30 patients, material from the primary tumor was simultaneously available. Real-time PCR technique with allele-specific molecular probe (KRAS/BRAF Mutation Analysis Kit, Entrogen, USA) was used for molecular tests. KRAS mutations were detected in 21.4 % of CNS metastatic lesions and in 23.3 % of corresponding primary tumors. Five mutations were identified both in primary and in metastatic lesions, while one mutation only in primary tumor and one mutation only in the metastatic tumor. Most of mutations were observed in codon 12 of KRAS; however, an individual patient had diagnosed a rare G13D and Q61R substitutions. KRAS mutations were significantly more frequent in adenocarcinoma patients and smokers. Additional analysis indicated one patient with rare coexistence of KRAS and DDR2 mutations. BRAF mutation was not detected in the examined materials. KRAS frequency appears to be similar in primary and CNS.     

Prevalence and antimicrobial resistance of <Emphasis Type="Italic">Ureaplasma</Emphasis> spp. and <Emphasis Type="Italic">Mycoplasma hominis</Emphasis> isolated from semen samples of infertile men in Shanghai,China from 2011 to 2016

This study was conducted to estimate the prevalence and antimicrobial resistance rate of Ureaplasma spp. and Mycoplasma hominis that were isolated from the semen samples of infertile males in Shanghai, China from 2011 to 2016. A total of 5016 infertile males and 412 healthy male controls were examined. The cultivation, identification, and antimicrobial susceptibilities of Ureaplasma spp. and M. hominis were assessed by using a Mycoplasma IST kit that was performed in parallel to selective solid agar cultivation. The positive rate of genital Mycoplasma infections in infertile men from 2011 to 2016 was 30–55%, which initially decreased during the first four years and then increased in the last two. Two distinct high-risk age ranges of Mycoplasma infections were observed: 26–30 years (37.8%) and 31–35 years (30.7%). Semisynthetic tetracyclines and macrolide antibiotics were the most effective agents against Ureaplasma spp. Among the fluoroquinolones, sparfloxacin and levofloxacin were also effective. Antibiotic resistance of Ureaplasma spp. against tetracyclines and macrolide antibiotics in the last six years did not vary significantly. However, the rate of resistance to fluoroquinolones (except norfloxacin) and spectinomycin decreased in the last two years. The rate of genital Mycoplasma presence in infertile patients between the ages of 26 and 35 years in Shanghai was high. The prevalence of genital Mycoplasma decreased during the first four years and then increased, with a peak in 2016. Doxycycline, minocycline, josamycin, and sparfloxacin can be recommended for first-line empirical treatment of Mycoplasma infections in infertile men in Shanghai, China.