Indocyanine Green tattooing for marking the caudal excision margin of a full-thickness vaginal endometriotic nodule.

Background: The use of Indocyanine Green (ICG) is well-described in oncology and more recently in benign gynaecological surgery. In this article we describe submucosal transvaginal ICG infiltration caudal to a vaginal endometriotic nodule to visualise the lower margin of excision laparoscopically. Objectives: To demonstrates the use of submucosal ICG tattooing to mark and delineate the caudal margin of an ultra-low full thickness vaginal nodule and aid its excision laparoscopically. Materials and Methods: A stepwise approach highlighting the "SOSURE" surgical technique for the excision of endometriosis and the practical use of the ICG to delineate the lowest margin of the full thickness vaginal nodule. Main outcome measures: Laparoscopic complete excision of a 5 cm full-thickness vaginal nodule invading the right parametrium and involving the superficial muscularis layer of the rectum. Result: ICG tattooing was helpful in identifying the lower margin of dissection of the rectovaginal space. Conclusion: ICG tattooing of the margins of full-thickness vaginal nodules could be another use of ICG in benign gynaecology to complement the surgeon's tactile and visual identification of the lower edge of dissection.

Revealing biophysical properties of KfrA-type proteins as a novel class of cytoskeletal, coiled-coil plasmid-encoded proteins.

BACKGROUND: DNA binding KfrA-type proteins of broad-host-range bacterial plasmids belonging to IncP-1 and IncU incompatibility groups are characterized by globular N-terminal head domains and long alpha-helical coiled-coil tails. They have been shown to act as transcriptional auto-regulators. RESULTS: This study was focused on two members of the growing family of KfrA-type proteins encoded by the broad-host-range plasmids, R751 of IncP-1ß and RA3 of IncU groups. Comparative in vitro and in silico studies on KfrAR751 and KfrARA3 confirmed their similar biophysical properties despite low conservation of the amino acid sequences. They form a wide range of oligomeric forms in vitro and, in the presence of their cognate DNA binding sites, they polymerize into the higher order filaments visualized as "threads" by negative staining electron microscopy. The studies revealed also temperature-dependent changes in the coiled-coil segment of KfrA proteins that is involved in the stabilization of dimers required for DNA interactions. CONCLUSION: KfrAR751 and KfrARA3 are structural homologues. We postulate that KfrA type proteins have moonlighting activity. They not only act as transcriptional auto-regulators but form cytoskeletal structures, which might facilitate plasmid DNA delivery and positioning in the cells before cell division, involving thermal energy.

Prophylactic negative-pressure wound therapy after ileostomy reversal for the prevention of wound healing complications in colorectal cancer patients: a randomized controlled trial.

BACKGROUND: The aim of this study was to assess the usefulness of protective negative-pressure wound therapy (NPWT) in the reduction of wound healing complications (WHC) and surgical site infections (SSI) after diverting ileostomy closure in patients who underwent surgery for colorectal cancer. METHODS: In this prospective randomized clinical trial in a tertiary academic surgical center, patients who had colorectal cancer surgery with protective loop ileostomy and were scheduled to undergo ileostomy closure with primary wound closure from January 2016 to December 2018 were randomized to be treated with or without NPWT. The primary endpoint was the incidence of WHC. Secondary endpoints were incidence of SSI, length of postoperative hospital stay (LOS), and length of complete wound healing (CWH) time. RESULTS: We enrolled 35 patients NPWT (24 males [68.6%]; mean age 61.6 ± 11.3 years), with NPWT and 36 patients (20 males [55.6%]; mean age 62.4 ± 11.3 years) with only primary wound closure (control group). WHC was observed in 11 patients (30.6%) in the control group and 3 (8.57%) in the NPWT group (p = 0.020). Patients in the NPWT group had a significantly lower incidence of SSI (2 [5.71%] vs. 8 [22.2%] in the control group; p = 0.046) as well as significantly shorter median CWH (7 [7-7] days vs. 7 [7-15.5] days, p = 0.030). There was no difference in median LOS between groups (3 [2.5-5] days in the control group vs. 4 [2-4] days in the NPWT group; p = 0.072). CONCLUSIONS: Prophylactic postoperative NPWT after diverting ileostomy closure in colorectal cancer patients reduces the incidence of WRC and SSI. CLINICAL TRIAL REGISTRATION: clinicaltrials.gov (NCT04088162).

4DCT-based evaluation of lung tumour motion during the breathing cycle.

The aim of this work was to quantify the variability in pre-treatment lung tumor motion during a single breathing period for 55 non-small cell lung cancer (NSCLC) targets. The influence of breathing on the volume and position of lung tumor was examined by comparing the information about tumor from respiratory-correlated four-dimensional computed tomography (4DCT) and three-dimensional computed tomography (3DCT) obtained without respiratory monitoring. The impact of age, gender, lung volume changes and immobilization device on tumor respiratory motion was evaluated. Based on the performed analysis, the significant differences were found between tumor volumes on 3DCT and 4DCT, although the comparison of volumes between 4DCT bins showed no statistically significant dependency. The significant differences between tumor centre of mass coordinates in the cranial-caudal (CC) and anterior-posterior (AP) directions were found. According to the results of statistical testing, there was no impact of gender and immobilization device on detected tumor respiratory motion. The impact was found for patient's age, lung volume changes, tumor volume and its location in different lung segments. The dominant lung cancer motion was observed for smaller tumors (up to 20 cc) located in posterior, caudal segments. This effect was also associated with a large variation in the lung volume during one respiratory cycle, observed for older patients. The important finding of the study is connected with the description of different patterns of tumor motion in AP and CC directions.

Sleep after intranasal progesterone vs. zolpidem and placebo in postmenopausal women - A randomized, double-blind cross over study.

CONTEXT: The loss of progesterone during menopause is linked to sleep complaints of the affected women. Previously we demonstrated sleep promoting effects of oral progesterone replacement in postmenopausal women. The oral administration of progesterone, however, is compromised by individual differences in bioavailability and metabolism of the steroid. OBJECTIVE: We compared the sleep-endocrine effects after intranasal progesterone (MPP22), zolpidem and placebo in healthy postmenopausal women. DESIGN: This was a randomized double-blind cross-over study. SETTING: German monocentric study PARTICIPANTS: Participants were 12 healthy postmenopausal women. INTERVENTIONS: Subjects received in randomized order four treatments, 2 doses of intranasal progesterone (4.5 mg and 9 mg of MPP22), 10 mg of zolpidem and placebo. OUTCOME MEASURES: Main outcome were conventional and quantitative sleep-EEG variables. Secondary outcomes were the subjective sleep variables and the sleep related concentrations of cortisol, growth hormone (GH), melatonin and progesterone. RESULTS: Sleep promoting effects were found after the higher dosage of MPP22 and after zolpidem. Zolpidem prompted benzodiazepine-like effects on quantitative sleep EEG as expected, whereas no such changes were found after the two dosages of MP22. Nocturnal progesterone levels increased after 9.0 mg MPP22. No other changes of hormone secretion were found. CONCLUSIONS: Our study shows sleep promoting effects after intranasal progesterone. The spectral signature of intranasal progesterone did not resemble the sleep-EEG alterations induced by GABA active compounds. Progesterone levels were elevated after 9.0 mg MPP22. No other endocrine effects were observed.

Transglutaminase 2 in cartilage homoeostasis: novel links with inflammatory osteoarthritis.

Transglutaminase 2 (TG2) is highly expressed during chondrocyte maturation and contributes to the formation of a mineralised scaffold by introducing crosslinks between extracellular matrix (ECM) proteins. In healthy cartilage, TG2 stabilises integrity of ECM and likely influences cartilage stiffness and mechanistic properties. At the same time, the abnormal accumulation of TG2 in the ECM promotes chondrocyte hypertrophy and cartilage calcification, which might be an important aspect of osteoarthritis (OA) initiation. Although excessive joint loading and injuries are one of the main causes leading to OA development, it is now being recognised that the presence of inflammatory mediators accelerates OA progression. Inflammatory signalling is known to stimulate the extracellular TG2 activity in cartilage and promote TG2-catalysed crosslinking of molecules that promote chondrocyte osteoarthritic differentiation. It is, however, unclear whether TG2 activity aims to resolve or aggravate damages within the arthritic joint. Better understanding of the complex signalling pathways linking inflammation with TG2 activities is needed to identify the role of TG2 in OA and to define possible avenues for therapeutic interventions.

Vulvar melanoma presenting as postmenopausal bleeding: a case report.

Primary melanomas of the vulva are extremely rare, creating obstacles in the differential diagnosis of other epithelial and non-epithelial malignancies. Due to their rarity, there are only approximately 250 cases reported in the current literature. Vulvar melanomas tend to relapse locally, as well as develop locoregional and distant metastasis through lymph node and haematic dissemination. The authors describe a case of an 84-year-old Caucasian female patient, presenting with postmenopausal bleeding, consistent with primary vulvar melanoma cause, which was successfully diagnosed and treated accordingly.

Impact of the Intra- and Inter-observer Variability in the Delineation of Parotid Glands on the Dose Calculation During Head and Neck Helical Tomotherapy.

The intra- and inter-observer variability in delineation of the parotids on the kilo-voltage computed tomography (kVCT) and mega-voltage computed tomography (MVCT) were examined to establish their impact on the dose calculation during adaptive head and neck helical tomotherapy (HT). Three observers delineated left and right parotids for ten randomly selected patients with oropharynx cancer treated on HT. The pre-treatment kVCT and the MVCT from the first fraction of irradiation were selected to delineation. The delineation procedure was repeated three times by each observer. The parotids were delineated according to the institutional protocol. The analyses included intra-observer reproducibility and inter-structure, -observer and -modality variability of the volume and dose. The differences between the left and right parotid outlines were not statistically significant (p > 0.3). The reproducibility of the delineation was confirmed for each observer on the kVCT (p > 0.2) and on the MVCT (p > 0.1). The inter-observer variability of the outlines was significant (p < 0.001) as well as the inter-modality variability (p < 0.006). The parotids delineated on the MVCT were 10% smaller than on the kVCT. The inter-observer variability of the parotids delineation did not affect the average dose (p = 0.096 on the kVCT and p = 0.176 on the MVCT). The dose calculated on the MVCT was higher by 3.3% than dose from the kVCT (p = 0.009). Usage of the institutional protocols for the parotids delineation reduces intra-observer variability and increases reproducibility of the outlines. These protocols do not eliminate delineation differences between the observers, but these differences are not clinically significant and do not affect average doses in the parotids. The volumes of the parotids delineated on the MVCT are smaller than on the kVCT, which affects the differences in the calculated doses.

Genetics of rapid eye movement sleep in humans.

The trait-like nature of electroencephalogram (EEG) is well established. Furthermore, EEG of wake and non-rapid eye movement (non-REM) sleep has been shown to be highly heritable. However, the genetic effects on REM sleep EEG microstructure are as yet unknown. REM sleep is of special interest since animal and human data suggest a connection between REM sleep abnormalities and the pathophysiology of psychiatric and neurological diseases. Here we report the results of a study in monozygotic (MZ) and dizygotic (DZ) twins examining the heritability of REM sleep EEG. We studied the architecture, spectral composition and phasic parameters of REM sleep and identified genetic effects on whole investigated EEG frequency spectrum as well as phasic REM parameters (REM density, REM activity and organization of REMs in bursts). In addition, cluster analysis based on the morphology of the EEG frequency spectrum revealed that the similarity among MZ twins is close to intra-individual stability. The observed strong genetic effects on REM sleep characteristics establish REM sleep as an important source of endophenotypes for psychiatric and neurological diseases.

Effect of temperature on calibration quality of structured-light three-dimensional scanners.

This paper presents the outcome of research into the effects of ambient temperature changes on structured-light three-dimensional (3D) scanners. The tests were conducted in a thermal chamber and consisted of a comparison of the 3D measurement of a special reference unit (made of a carbon composite) performed at different temperatures, with measurements performed at the calibration temperature. A contact measuring arm with temperature compensation was used as a reference. Based on the results of these experiments, we propose a method that allows us to extend the existing scanner calibration method by using a temperature-correction procedure that is based on linear and nonlinear mathematical models. An exemplary application of this procedure has shown that the range of temperatures in which scanner accuracy is within declared limits can be increased 11-fold.

Impact of the Intra- and Inter-observer Variability in the Delineation of Parotid Glands on the Dose Calculation During Head and Neck Helical Tomotherapy.

The intra- and inter-observer variability in delineation of the parotids on the kilo-voltage computed tomography (kVCT) and mega-voltage computed tomography (MVCT) were examined to establish their impact on the dose calculation during adaptive head and neck helical tomotherapy (HT). Three observers delineated left and right parotids for ten randomly selected patients with oropharynx cancer treated on HT. The pre-treatment kVCT and the MVCT from the first fraction of irradiation were selected to delineation. The delineation procedure was repeated three times by each observer. The parotids were delineated according to the institutional protocol. The analyses included intra-observer reproducibility and inter-structure, -observer and -modality variability of the volume and dose. The differences between the left and right parotid outlines were not statistically significant (p > 0.3). The reproducibility of the delineation was confirmed for each observer on the kVCT (p > 0.2) and on the MVCT (p > 0.1). The inter-observer variability of the outlines was significant (p < 0.001) as well as the inter-modality variability (p < 0.006). The parotids delineated on the MVCT were 10% smaller than on the kVCT. The inter-observer variability of the parotids delineation did not affect the average dose (p = 0.096 on the kVCT and p = 0.176 on the MVCT). The dose calculated on the MVCT was higher by 3.3% than dose from the kVCT (p = 0.009). Usage of the institutional protocols for the parotids delineation reduces intra-observer variability and increases reproducibility of the outlines. These protocols do not eliminate delineation differences between the observers, but these differences are not clinically significant and do not affect average doses in the parotids. The volumes of the parotids delineated on the MVCT are smaller than on the kVCT, which affects the differences in the calculated doses.

Tomotherapy: implications on daily workload and scheduling patients based on three years' institutional experience.

Helical tomotherapy (HT) was introduced at the Greater Poland Cancer Centre (GPCC) in April 2009. Retrospective analysis included data from the treatments performed for the first 656 patients treated with HT between May 2009 and May 2012 at the GPCC. In order to evaluate the implications on daily workload and scheduling of patients, stepwise regression and time analysis for each component of the overall treatment time, such as positioning, imaging, registration, and irradiation were performed. A detailed analysis included: (1) learning curves and optimized time needed for positioning and registration; (2) relation between irradiation time and parameters used for plan creation; and (3) average time of daily imaging. The irradiation component has the highest influence on the overall treatment time (R = 0.911). The lowest influence was observed for the imaging (R = 0.670). The learning curve for positioning was 7 months while the reduction of the average daily time needed for registration was observed even after two years. The irradiation time strongly depends on the planning parameters. Changing the pitch from 0.215 to 0.287 for pelvic cancer cases decreased the average daily beam-on time per patient by about 2 minutes. Similar changes for head and neck reduced this time by 1.3 minutes. The limitation in the usage of 1 cm field width only for complex cases, lower than 10 cm in the cranio-caudal direction, reduced the beam-on time per patient by 2 minutes. The average overall treatment time decreased from 21.5 minutes per patient in the first year of the HT usage to 13.8 minutes per patient in current practice. Our current practice shows that for a group of patients including mainly those with pelvis and head and neck cancers, the HT treatment takes approximately 15 minutes per patient allowing 40 patients to be treated within 10 hours.

Electric permittivity anomaly close to the critical consolute point of a nitrobenzene + octane liquid mixture.

Electric permittivity and density were measured in a nitrobenzene and octane mixture in the vicinity of the upper critical consolute point. Measurements were conducted in the one-phase region, at the critical concentration. The possibility of stirring in the course of measurements allowed us to check if the density and concentration gradients had any influence on the obtained results. No signs of the presence of the gradients mentioned above were found. Using the data obtained in the reported measurements, different methods of the fitting of the equation describing the permittivity anomaly were tested. The calculation of a reliable value of the critical amplitude, used to estimate the critical temperature shift under the influence of the electric field, was of particular interest. The derivative (∂T(c)/∂E(2)) was found to be (-3.9 ± 0.3) × 10(-16) K m(2) V(-2).

Variation in fat content and fatty-acid composition of the Baltic herring Clupea harengus membras.

The fat content and fatty-acid profiles of herring, Clupea harengus membras, from the southern Baltic Sea varied depending on when (fishing season) and where (fishing grounds) the fish were caught as well as on their size and sex. The fat, protein and dry matter content and the fatty-acid profiles were assayed in C. h. membras muscle tissue. The changes observed in fatty-acid profiles were determined by factors such as specimen mass and fat content, which, in turn, depended on fishing season. This is explained by dietary differences between juvenile and older fish. Gonad maturation and spawning in the latter are also factors. The study results provide confirmation of the hypothesis that polyunsaturated fatty acids (PUFA), in particular docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA), play vital roles in the sexual maturation of C. h. membras.

Influence of cigarette smoking on the level of mRNA of somatostatin receptor 3 (SSTR3) in the gastric mucosa of patients with functional dyspepsia.

PURPOSE: Helicobacter pylori (H. pylori) infection and smoking of cigarettes increase individual risk to gastric carcinoma. In stomach tumors, an expression of somatostatin receptor 3 (SSTR3) is diminished or completely lost. The purpose of these studies was to determine the influence of smoking cigarettes and H. pylori infection on the expression of SSTR3 in patients with functional dyspepsia. MATERIALS AND METHODS: The study comprised 109 patients with functional dyspepsia in the age range 28-61 years. The total 218 biopsies used for analysis were divided into two groups: group I - 176 biopsies from non-smokers (72 from H. pylori positive ones), and group II - 42 biopsies from cigarette smokers (28 from H. pylori positive patients). The SSTR3 mRNA amount in the gastric mucosa (1 biopsy from the antrum and 1 biopsy from the corpus) was determined by real time RT-PCR. The presence of H. pylori colonization in the stomach tissue was evaluated by multiplex PCR. RESULTS: In the H. pylori negative samples the amount of the SSTR3 mRNA was significantly lower for smokers than for non-smokers (by 40%, p < 0.010). Infection with H. pylori caused reduction of the level of SSTR3 mRNA in non-smoking patients by ca. 30% (p < 0.01), while in samples from smokers the SSTR3 mRNA level was similar regardless of H. pylori infection. CONCLUSIONS: The cigarettes smoking and H. pylori infection are independent factors leading to decreasing of the SSTR3 mRNA level in gastric mucosa of patients with functional dyspepsia.

Complete nucleotide sequence of the pCTX-M3 plasmid and its involvement in spread of the extended-spectrum beta-lactamase gene blaCTX-M-3.

Here we report the nucleotide sequence of pCTX-M3, a highly conjugative plasmid that is responsible for the extensive spread of the gene coding for the CTX-M-3 extended-spectrum beta-lactamase in clinical populations of the family Enterobacteriaceae in Poland. The plasmid belongs to the IncL/M incompatibility group, is 89,468 bp in size, and carries 103 putative genes. Besides bla(CTX-M-3), it also bears the bla(TEM-1), aacC2, and armA genes, as well as integronic aadA2, dfrA12, and sul1, which altogether confer resistance to the majority of beta-lactams and aminoglycosides and to trimethoprim-sulfamethoxazole. The conjugal transfer genes are organized in two blocks, tra and trb, separated by a spacer sequence where almost all antibiotic resistance genes and multiple mobile genetic elements are located. Only bla(CTX-M-3), accompanied by an ISEcp1 element, is placed separately, in a DNA fragment previously identified as a fragment of the Kluyvera ascorbata chromosome. On the basis of sequence analysis, we speculate that pCTX-M3 might have arisen from plasmid pEL60 from plant pathogen Erwinia amylovora by acquiring mobile elements with resistance genes. This suggests that plasmids of environmental bacterial strains could be the source of those plasmids now observed in bacteria pathogenic for humans.

Comparison of the efficacy of rosuvastatin versus atorvastatin, simvastatin, and pravastatin in achieving lipid goals: results from the STELLAR trial.

In the Statin Therapies for Elevated Lipid Levels compared Across doses to Rosuvastatin (STELLAR) trial, the efficacy of rosuvastatin calcium (Crestor) was compared with that of atorvastatin (Lipitor), simvastatin (Zocor), and pravastatin (Pravachol) for lowering plasma low-density lipoprotein cholesterol (LDL-C) after 6 weeks of treatment. In this multicenter, parallel-group, open-label trial, adults with hypercholesterolemia were randomized to treatments with rosuvastatin 10, 20, 40, or 80 mg, atorvastatin 10, 20, 40, or 80 mg, simvastatin 10, 20, 40, or 80 mg, or pravastatin 10, 20, or 40 mg. Efficacy and safety results from this trial have been previously published. The additional analyses included in this report show that 53% (83/156) to 80% (125/157) of patients in the rosuvastatin 10- to 40-mg groups achieved LDL-C levels < 100 mg/dl (< 2.6 mmol/l), compared with 18% (28/158) to 70% (115/165) of patients who received atorvastatin, 8% (13/165) to 53% (86/163) of patients who received simvastatin, and 1% (1/160) to 8% (13/161) of patients who received pravastatin. Other additional analyses showed that more patients in the rosuvastatin 10- to 40-mg groups than in the comparator groups who were at high risk of coronary heart disease according to National Cholesterol Education Program Adult Treatment Panel (ATP) III, Joint European Societies, or Canadian guidelines achieved the LDL-C goals of < 100 mg/dl (< 2.6 mmol/l) (55% to 77% compared with 0 to 64%), < 3.0 mmol/l (< 116 mg/dl) (76% to 94% compared with 6% to 81%), and < 2.5 mmol/l (< 97 mg/dl) (47% to 69% compared with 0 to 53%), respectively. Results favoring rosuvastatin versus the comparators were also reported for patients: (a) who had triglycerides > or = 200mg/dl (> or = 2.3 mmol/l), and achieved both ATP III LDL-C and non-high-density lipoprotein cholesterol (non-HDL-C) goals (80% to 84% versus 15% to 84%); (b) overall who achieved the Canadian LDL-C goals of < 2.5 (< 97 mg/dl) to < 5.0 mmol/l (< 193 mg/dl) (85% to 91% versus 44% to 86%); and (c) who achieved all 3 Canadian goals for LDL-C, triglycerides (< 3.0 mmol/l [< 266 mg/dl] to < 2.0 mmol/l [< 177 mg/dl]), and the total cholesterol/high-density lipoproteincholesterol ratio (< 4 to < 7) (70% to 83% versus 35% to 79%).

Enantioselective synthesis of (2-pyridyl)alanines via catalytic hydrogenation and application to the synthesis of L-azatyrosine.

[reaction: see text] A novel method for the synthesis of (2-pyridyl)alanines 2a-b was developed by converting (2-pyridyl)dehydroamino acid derivatives 1a-b to the corresponding N-oxides 3a-b followed by asymmetric hydrogenation using (R,R)-[Rh(Et-DUPHOS)(COD)]BF(4) [(R,R)-6] catalyst and subsequent N-oxide reduction in 80-83% ee. This methodology was applied to the total synthesis of L-azatyrosine [(+)-12], an antitumor antibiotic, starting from (5-benzyloxy)-2-pyridylmethanol (7), in >96% enantiomeric purity.

Selective analysis of phosphopeptides within a protein mixture by chemical modification, reversible biotinylation and mass spectrometry.

A new method combining chemical modification and affinity purification is described for the characterization of serine and threonine phosphopeptides in proteins. The method is based on the conversion of phosphoserine and phosphothreonine residues to S-(2-mercaptoethyl)cysteinyl or beta-methyl-S-(2-mercaptoethyl)cysteinyl residues by beta-elimination/1,2-ethanedithiol addition, followed by reversible biotinylation of the modified proteins. After trypsin digestion, the biotinylated peptides were affinity-isolated and enriched, and subsequently subjected to structural characterization by liquid chromatography/tandem mass spectrometry (LC/MS/MS). Database searching allowed for automated identification of modified residues that were originally phosphorylated. The applicability of the method is demonstrated by the identification of all known phosphorylation sites in a mixture of alpha-casein, beta-casein, and ovalbumin. The technique has potential for adaptations to proteome-wide analysis of protein phosphorylation.

Quenching of biotinylated aequorin bioluminescence by dye-labeled avidin conjugates: application to homogeneous bioluminescence resonance energy transfer assays.

[see reaction]. Avidin conjugates containing the covalently attached dyes QSY-7 and dabcyl were prepared and shown to quench the bioluminescence of biotinylated aequorin. Quenching efficiency was shown to be dependent on both the label-to-avidin ratio and the concentration of the avidin conjugate. These properties were exploited to develop a homogeneous bioluminescence resonance energy transfer (BRET) assay for biotin.