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BACKGROUND: Seasonal influenza vaccination is recommended for patients with cardiovascular diseases. Low vaccination coverage among these patients is due to insufficient knowledge about vaccine efficiency and its regular annual use. This work aimed to study the repeated 6-month changes in hemagglutinin antibody titers (AT) for 3 years in patients with cardiac pathology in a comparative study of influenza preventive vaccination. MATERIALS AND METHODS: Analysis of ATs obtained based on the hemagglutination-inhibition test (HAI) was performed in 235 of 817 participants in a prospective follow-up. Blood sampling was performed at baseline, before the vaccination, and 6 months after, and at the same term in unvaccinated patients in the 2012–2013 and 2014–2015 seasons, respectively. The seropositive and seronegative responses to vaccination or acute respiratory or influenza infection were used, according to the reference values of seroconversion and seroprotection and the fact of seroconversion. Multiple regression analysis with a logarithmic scale was used to assess the vaccine effectiveness indices. RESULTS : With vaccination coverage of at least 40% against seasonal influenza within 3 years, the trends of a decrease in seropositive and an increase in seronegative responses in the vaccination group and its reverse nature in the comparison group were determined by traditional analysis. Using logarithmic calculation, an increase in HAI AT seroconversion was revealed over a 3-year follow-up period. It was characterized by a homogeneous serological response at annual vaccination and heterogeneous with a higher serological response in cases without vaccination (p = 0.002 for H1N1 and p = 0.005 for H3N2, respectively). This trend can be determined by a higher and more stable prevaccination level of HAI AT than the same residual level of AT among unvaccinated patients. CONCLUSION : During long-term cardiac studies, the logarithmic calculation in interpreting the results of HAI AT overcomes the limitations of the traditional analysis of assessing the efficiency of the annual influenza vaccine. Further serological programs are required to better understand the role of routine seasonal influenza vaccination in preventing morbidity and mortality of patients with cardiovascular diseases.

Background: Atrial fibrillation (AF) is the most common cardiac arrhythmia. Among patients with AF the subgroup posessing AF without traditional risk factors is differentiated. Such patients are commonly referred as having lone AF and comprise 1020% of all cases. A number of studies have demonstrated that the background of AF, and in particular lone AF, have a substantial genetic component. Aims: To evaluate the influence of gene polymorphism to the development of atrial fibrillation in patients without concomitant valvular pathology and coronary artery disease. Materials and methods: The study included 174 patients with atrial fibrillation and 124 controls without any cardiovascular pathology. All patients were divided into two subgroups: with lone AF (n=94) and with concomitant arterial hypertension (n=80). All patients underwent a complex of clinical, instrumental (ECG, echocardiography, computed tomography of the pulmonary veins) and laboratory tests (thyroid hormones, inflammatory markers, fibrosis), as well as genetic analysis (determination of single nucleotide polymorphisms described as AF risk factors in genes AGXT2, PDE4D, SLN, SCN5A, PITX2, PRRX1, ZFHX3, TBX5, CAV1 и HCN4). Results: For the rare polymorphisms rs12291814 (SLN) and rs137854601 (SCN5A) wasnt found anyone carrier of the minor allele (C and A, respectively). In the both patient subgroups the minor allele T of rs2200733 in PITX2 (OR=3.18, p0.0001), minor allele G of rs3903239 in PRRX1, and minor alleles A of 2 polymorphisms rs2106261 and rs7193343 in ZFHX3 gene were revealed as risk factor of AF (OR=2.96, p0.0001, OR=2.02, p=0.0045, OR=1.64, p=0.04, respectively). We also revealed significant difference between AF and control groups for rs3807989 in CAV1: homozygotic state of minor allele A has a protective effect on the development of arrhythmias (OR=0.39). Conclusions: We revealed the association between the polymorphisms in genes regulating transcription and the development of atrial fibrillation. These polymorphisms have already described but their frequencies have never investigated in Russian population. But the polymorphisms influence to gene functions stays unclear, although attempts to investigate relationship between genotype and gene expression have been made. When the relationship will be discovered it can help us to modify our approach in treatment to patients with atrial fibrillation.

Aim: to study distribution of physiological resources in purposeful activity of patients with epilepsy. Materials and Methods. 70 Healthy individuals and 160 patients with epilepsy were examined. In the examined individuals purposeful activity was modeled with evaluation of its effectiveness using Schulte-Gorbov test. In the dynamics of the modeled activity, parameters of electroencephalogram, variability of heart rhythm were evaluated; in the initial condition (before Schulte-Gorbov test) parameters of evoked visual potentials, P300cognitive potentials and of a conditionally-negative deviation were estimated. Interrelations between parameters of electroencephalography and of variability of heart rhythm were studied using correlation analysis with Spearman rank correlation coefficient; the comparative analysis of variability of heart rhythm and characteristics of respiratory function was performed, and effectiveness of behavior of patients with epilepsy was predicted using logit-regression analysis. Results. Predomination of structural-metabolic forms of epilepsy was found in the group of patients with low effectiveness of the activity. The low-effective group of patients with epilepsy showed a higher correlation of physiological parameters and a high level of characteristics refecting activation of stress-realizing systems. For effective distribution of patients with epilepsy into groups with different effectiveness of behavior, it was necessary to include characteristics of visual evoked potential, P300 cognitive potential and conditionally-negative deviation into logit-regression model, that reflects the role of afferent and association mechanisms in this task. Increase in physiological «cost» and reduction in the effectiveness of activity in the group of patients with epilepsy is associated with prevalence of structural forms of epilepsy. Conclusion. Epileptogenic zones in patients with epilepsy are supposed to play a role not only in reduction of effectiveness of activity, but also in excessive mobilization of physiological resources and in increase in physiological cost of activity, that diminishes its effectiveness.

Evaluation of bone mineralization and bone metabolism in children with inflammatory bowel disease (IBD) in accordance with TaqI-polymorphic genotypes of vitamin D receptor (VDR) has been performed in this study. 83 children (38 girls and 54 boys) with IBD (60 with Crohn’s disease, 23 with ulcerative colitis) have been included in the study. Mean age was 13.0 years (11.0; 15.5). All patients have active phase of the disease. Bone mineral density (BMD) of lumbar spine (DEXA) have been assessed in all children. Level of serum osteocalcin (OC) as a marker of osteosynthesis, C-terminal telopeptides (CTT) as a marker of osteoresorbtion, parathyroid hormone (PTH), serum calcium, phosphorus, alkaline phosphatase and 25(OH) vitamin D was measured to evaluate bone metabolism. Molecular-genetic tests: analysis of TaqI (rs731236) polymorphism of vitamin D receptor gene (VDR) was perfomed by polymerase chain reaction with following restriction analysis. Association between molecular markers of VDR gene and bone metabolism and mineralization disturbances have been found. TT genotype of VDR gene was associated with tendency to decrease of linear growth velocity, low linear growth, decrease in osteosynthesis and increase of osteoresorbtion in this group of children. TT-genotype of TaqI-polymorphism of VDR gene can be assessed as a risk factor of bone metabolism disturbances in children with both Crohn’s disease and ulcerative colitis. C allele seems to have protective role in this group.

The structural characterization of the lipopolysaccharide (LPS) from extremophiles has important implications in several biomedical and therapeutic applications. The polyextremophile Gram-negative bacterium Halobacteroides lacunaris TB21, isolated from one of the most extreme habitats on our planet, the deep-sea hypersaline anoxic basin Thetis, represents a fascinating microorganism to investigate in terms of its LPS component. Here we report the elucidation of the full structure of the R-type LPS isolated from H. lacunaris TB21 that was attained through a multi-technique approach comprising chemical analyses, NMR spectroscopy, and Matrix-Assisted Laser Desorption Ionization (MALDI) mass spectrometry. Furthermore, cellular immunology studies were executed on the pure R-LPS revealing a very interesting effect on human innate immunity as an inhibitor of the toxic Escherichia coli LPS.

Background: Discontinuation of gefitinib treatment is often accompanied by a disease flare. Some studies have demonstrated a benefit of the use of epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKI) beyond progression; however, long-term results of these investigations remain limited. Patients and Methods: We observed 70 patients with EGFR-mutated (EGFR-M+) non-small cell lung cancer (NSCLC) receiving single-agent gefitinib in a routine clinical setting; 56 patients were experiencing RECIST progression at the time of the analysis. Results: There was a significant increase (p = 0.00001) in overall survival (OS) in patients continuing on gefitinib beyond progression (n = 21; median duration of continued gefitinib use: 4.2 months; median OS: not reached; expected OS: 29.7 months) as compared to those who stopped gefitinib treatment upon disease progression (n = 35; median OS: 14.0 months). The association between extended gefitinib use and improved OS remained true in multivariate Cox regression analysis (hazard ratio = 4.49, 95% confidence interval 1.25-16.09; p = 0.021). Patient selection bias constitutes an essential limitation of this clinical observational study, given that patients with a more favorable disease course and/or high initial tumor sensitivity to TKI treatment were more likely to be considered for prolonged gefitinib use. Conclusion: This study confirms that continued administration of gefitinib beyond progression is a viable treatment option for some patients with EGFR-M+ NSCLC, in particular those who cannot be rescued by novel EGFR mutation-specific inhibitors such as osimertinib.

Хронические неинфекционные заболевания характеризуются неравномерной заболеваемостью и смертностью в течение года. Цель. Изучить избыточную заболеваемость от сердечно-сосудистых заболеваний (ССЗ) в зимний период в Ивановской области в 2009-2013 гг Материал и методы. Проведен расчет индекса избыточной заболеваемости в зимний период (ИИЗЗП) по специальной формуле. Количество ССЗ определяли по данным медицинского информационно-аналитического центра Ивановской области. Индекс рассчитывали для ССЗ в целом, для артериальной гипертензии (АГ) и для острых форм ССЗ (инфаркт миокарда и инсульт). Результаты. Средний ИИЗЗП для ССЗ за 5 лет составил 77,2%, и оказался у женщин несколько выше, чем у мужчин (80,0% против 69,5%, соответственно). Среди лиц трудоспособного возраста избыточная заболеваемость в зимний период оказалась ниже, чем среди населения старше трудоспособного возраста (53,1 и 88%, соответственно). ИИЗЗП для ССЗ характеризовался значительной вариабельностью и колебался от 29,0% (2013 г.) до 81,1% (2010 г.) у мужчин и от 25,2% до 82,9% у женщин в те же годы. Средний за 5 лет ИИЗЗП для АГ в целом составил 72,7%, выше у мужчин в сравнении с женщинами в большинстве анализируемых лет наблюдения. Выявлена неоднородность показателя ИИЗЗП для АГ в течение анализируемого периода в каждой анализируемой группе (минимальные значения отмечены в 2011 г., вслед за волной жары 2010 г. При анализе ИИЗЗП для инфаркта миокарда выявлена избыточная заболеваемость в зимний период, показатели у мужчин и женщин в целом близки (31,0% и 28,4%, соответственно), но выше у лиц трудоспособного возраста по сравнению с лицами более старших возрастных групп. Этот показатель имел широкий разброс значений. Избыточной заболеваемости инсультом в зимний период не выявлено этот показатель был равномерен в течение года. Заключение. Воздействие на избыточную заболеваемость в зимний период это резерв снижения заболеваемости и смертности от ССЗ за счет снижения воздействия рисков, и важный фактор оптимизации нагрузки на систему здравоохранения, сезонную неравномерность которой необходимо учитывать при планировании работы медицинских учреждений., Chronic non-infectious diseases are characterized by irregular morbidity and mortality throughout a year. Aim. To study excess cardiovascular morbidity during winter in Ivanovo Region in 2009-2013 years. Material and methods. Excess winter morbidity index (EWMI) was calculated using special formula. The incidence of cardiovascular diseases (CVDs) was determined according to the data from the medical information-analytic center of Ivanovo Region. The index was calculated for CVDs in general, for arterial hypertension (HT) and for acute forms of CVDs (acute myocardial infarction and stroke). Results. The mean EWMI for CVDs for 5 years of the follow-up made up to 77.2% and was higher in women than in men (80% vs 69.5%, respectively). People of working age had lower excess winter morbidity than the population older than working age (53.1% and 88%, respectively). EWMI for CVDs had significant variability and ranged from 29.0% (2013 year) to 81.1% (2010 year) in men and from 25.2% to 82.9% in women in the same years. In general, the 5-year mean EWMI for HT was 72.7%; it was higher in men than in women in most of the analyzed years of follow-up. We had revealed the heterogeneity of the EWMI for HT during the analyzed period in each analyzed group (the minimal values were found in 2011 after the heat wave in 2010). EWMI for myocardial infarction demonstrated excess winter morbidity, men and women had in general similar indices (31.0% and 28.4%, respectively), but it was higher in people of working age as compared with the older ones. This index had a wide range of values. In case of stroke we had revealed no excess winter morbidity; this index was equal throughout a year. Conclusion. The influence on excess winter morbidity can result in the reduction of cardiovascular morbidity and mortality due to the decreased effect of risk factors; it would also optimize a load on healthcare system, the season irregularity of which must be taken into account when planning a work of medical centers.

Aim: To study the local elastic properties of the arterial wall at the site of atherosclerotic plaque and adjacent areas «healthy» wall of the common carotid artery in patients with arterial hypertension (AH).Materials and methods. The study included 25 patients (3 men, 22 women) with AH 1-2 grad in combination with moderate carotid atherosclerosis and lipid metabolism disorders. All patients underwent conventional ultrasound duplex scanning of the carotid arteries to determine the severity of atherosclerotic carotid arteries and the structure of atherosclerotic plaque. Determination of parameters of the local rigidity - the elasticity of the arterial wall and the atherosclerotic plaque, was carried out on the ultrasound system ALOKA a7. Was scanned and the surroundingarea atherosclerotic plaque sections of the arterial wall in the B/M mode using echo-tracking method.Results. A significant increase in the hardness of the arterial wall in atherosclerotic plaque as in the adventitia and its surface, as compared with the adjacent «healthy» wall of the carotid artery. Atherosclerotic plaque base has significantly greater rigidity and less flexibility than its surface. Conclusion. The formation of atheroma by the action of arterial pulse pressure develops heterogeneity elastic properties carotid wall not only along the artery, but in the atherosclerotic plaque (between its base and the surface). This indicates of different mobility plots artery and plaque and can lead to the formation of the mechanical instability of atherosclerotic plaque in patients with hypertension.

Ferroplasmaceae represent ubiquitous iron-oxidising extreme acidophiles with a number of unique physiological traits. In a genome-based study of Ferroplasma acidiphilum YT, the only species of the genus Ferroplasma with a validly published name, we assessed its central metabolism and genome stability during a long-term cultivation experiment. Consistently with physiology, the genome analysis points to F. acidiphilum YT having an obligate peptidolytic oligotrophic lifestyle alongside with anaplerotic carbon assimilation. This narrow trophic specialisation abridges the sugar uptake, although all genes for glycolysis and gluconeogenesis, including bifunctional unidirectional fructose 1,6-bisphosphate aldolase/phosphatase, have been identified. Pyruvate and 2-oxoglutarate dehydrogenases are substituted by ‘ancient’ CoA-dependent pyruvate and alpha-ketoglutarate ferredoxin oxidoreductases. In the lab culture, after ~550 generations, the strain exhibited the mutation rate of ≥1.3 × 10−8 single nucleotide substitutions per site per generation, which is among the highest values recorded for unicellular organisms. All but one base substitutions were G:C to A:T, their distribution between coding and non-coding regions and synonymous-to-non-synonymous mutation ratios suggest the neutral drift being a prevalent mode in genome evolution in the lab culture. Mutations in nature seem to occur with lower frequencies, as suggested by a remarkable genomic conservation in F. acidiphilum YT variants from geographically distant populations.

In the present study, chemical oxygen demand (COD) removal by coagulation and packed-columns of both fresh and bioregenerated granular activated carbon (GAC) is reported as a feasible treatment for saline and oily wastewaters (slops) generated from marine oil tankers cleaning. The use of Ferric chloride (FeCl3), Aluminium sulphate (Al2(SO4)3) and Polyaluminum chloride (Al2(OH3)Cl3) was evaluated in the pre-treatment by coagulation of a real slop, after a de-oiling phase in a tank skimmer Comparison of coagulation process indicated that Polyaluminum chloride and Aluminium sulphate operate equally well (20–30% of COD removal) when applied at their optimal dose (40 and 90 mg/l respectively) but the latter should be preferred in order to significantly control the sludge production. The results from the column filtration tests indicated the feasibility of using the selected GAC (Filtrasorb 400 -Calgon Carbon Corporation) to achieve the respect of the discharge limits in the slops treatment with a carbon usage rate in the range 0.1–0.3 kg/m3 of treated effluent. Moreover, biological regeneration through Alcalinovorax borkumensis SK2 was proved to be a cost-effective procedure since the reuse of spent GAC through such regeneration process for further treatment could still achieve approximately 90% of the initial sorption capacity, reducing then costs for the use of new sorbents and also the need for waste disposal.

The products of spontaneous and induced proteolysis of human ceruloplasmin (Cp) were studied. Some physico-chemical properties of the six fragments with electrophoretically determined Mr 130,000 (F1), 110,000 (F2), 66,000 (F3), 48,000 (F4) 22,000 (F5) and 18,000 (F6) were compared. The amino acid compositions and N-terminal amino acid sequences coincide in F1-F5, but differ from those of F6. Limited tryptic proteolysis of Cp causes the accumulation of polypeptide fragment with Mr 22,000, the N-terminal primary structure of which is identical to that of F5 produced by spontaneous proteolysis. Electrophoretic fragments of Cp were extracted from polyacrylamide gel, treated with 125I and then studied by peptide mapping with subsequent radioautography. The comparison of the "finger prints" showed the identity of F1 to F2 and F3 and gross similarity between F4 and F1-F3. It also revealed similar peptides in F5 and F6 hydrolyzates and almost perfect matching of the F4 map to the map of F5 + F6 mixture. On the basis of the obtained data general principles of Cp molecular organization are discussed and intramolecular homology is suggested to be a feature of the protein.

In order to study the effect of a long-term stress and/or starvation on a natural microbial community we have analysed both the quantitative and the qualitative changes in the composition of a bacterial community after 30 months incubation in a seawater microcosm system. Measures of the bacterial density showed a general decrement of values of living, total and heterotrophic cultivable bacteria. Nevertheless, 16S rRNA taxonomical analysis revealed a microbial population characterised principally by the persistence of sequences related toAlteromonas genus. The application of the theory of stochastic Boolean network on biological systems has given an important speculative tool to the development of the hypothesis for the explication of the dynamics of microbial populations during a long-term stress and/or starvation conditions.

Aims This study focused on the influence of different amounts of NaCl in the medium in Vibrio anguillarum EmpA protease production at both the transcriptional and post-transcriptional levels. Methods and results Vibrio anguillarum 975/I was cultivated in cM9 medium with varying concentrations of NaCl: 0·5, 1·5, 3·0%. EmpA protease was monitored in the supernatants by the skim milk test, azocasein assay and Western blot analysis. The empA gene expression was measured by real-time PCR. A mutant strain 975/I defective for the empA gene confirmed the specificity of the response for EmpA protease. Active protease production was induced by 0·5 and 1·5% NaCl-amended media; however, the strain cultivated in 3·0% NaCl was unable to secrete EmpA protease. The quantitative expression of the empA gene was very similar in all tested conditions. Conclusions The NaCl concentration in the medium modulates the secretion of active EmpA protease in V. anguillarum at a post-transcriptional level. Significance and impact of the study EmpA protease is one of the most important virulence factors in V. anguillarum. We demonstrated the influence of osmotic changes in the regulation of EmpA protease in the V. anguillarum 975/I strain. This finding has an important impact on the evaluation of factors determining the onset of disease in fish.

The cpn60 and cpn10 genes from psychrophilic bacterium, Oleispira antarctica RB8, showed a positive effect in Escherichia coli growth at low temperature, shifting its theoretical minimal growth temperature from +7.5 degrees C to -13.7 degrees C [Ferrer, M., Chernikova, T.N., Yakimov, M., Golyshin, P.N., and Timmis, K.N. (2003) Nature Biotechnol 21: 1266-1267]. To provide experimental support for this finding, Cpn60 and 10 were overproduced in E. coli and purified to apparent homogeneity. Recombinant O.Cpn60 was identical to the native protein based on tetradecameric structure, and it dissociates during native PAGE. Gel filtration and native PAGE revealed that, in vivo and in vitro, (O.Cpn60)(7) was the active oligomer at 4-10 degrees C, whereas at > 10 degrees C, this complex was converted to (O.Cpn60)(14). The dissociation reduces the ATP consumption (energy-saving mechanism) and increases the refolding capacity at low temperatures. In order for this transition to occur, we demonstrated that K468 and S471 may play a key role in conforming the more advantageous oligomeric state in O.Cpn60. We have proved this hypothesis by showing that single and double mutations in K468 and S471 for T and G, as in E.GroEL, produced a more stable double-ring oligomer. The optimum temperature for ATPase and chaperone activity for the wild-type chaperonin was 24-28 degrees C and 4-18 degrees C, whereas that for the mutants was 45-55 degrees C and 14-36 degrees C respectively. The temperature inducing unfolding (T(M)) increased from 45 degrees C to more than 65 degrees C. In contrast, a single ring mutant, O.Cpn60(SR), with three amino acid substitutions (E461A, S463A and V464A) was as stable as the wild type but possessed refolding activity below 10 degrees C. Above 10 degrees C, this complex lost refolding capacity to the detriment of the double ring, which was not an efficient chaperone at 4 degrees C as the single ring variant. We demonstrated that expression of O.Cpn60(WT) and O.Cpn60(SR) leads to a higher growth of E. coli at 4 degrees C ( micro (max), 0.22 and 0.36 h(-1) respectively), whereas at 10-15 degrees C, only E. coli cells expressing O.Cpn60 or O.Cpn60(DR) grew better than parental cells (-cpn). These results clearly indicate that the single-to-double ring transition in Oleispira chaperonin is a wild-type mechanism for its thermal acclimation. Although previous studies have also reported single-to-double ring transitions under many circumstances, this is the first clear indication that single-ring chaperonins are necessary to support growth when the temperature falls from 37 degrees C to 4 degrees C.

Objectives. To determine the effectiveness of tormentil root extract (TRE) for treatment of rotavirus diarrhea in children. Background. Rotavirus, one of the most widely spread pathogens of acute, dehydrating diarrhea in children, is estimated to cause >800 000 annual deaths of young children in developing countries. Currently no rotavirus vaccine is available. Management involves rehydration therapy. Available antiperistaltic or antisecretory drugs to reduce the severity of diarrhea can cause serious side effects in children. Methods. A randomized, double blinded, placebo-controlled trial was conducted at Children's Hospital for Infectious Diseases #3, St. Petersburg, Russia in 40 children ranging in age from 3 months to 7 years with rotavirus diarrhea. We constructed 2 groups for comparison: a treatment group that consisted of 20 children treated with tormentil root extract; and a control group of 20 children who received a placebo. All patients received 3 drops of tormentil root extract or placebo per year of life, three times daily until discontinuation of diarrhea, or a maximum of 5 days. An objective method was used to evaluate diarrhea, and physical examination was used to assess degree of dehydration in children. Results. The duration of diarrhea in the tormentil root extract treatment group was 3 days, compared with 5 days in the control group (P < 0.0001). In the treatment group 8 of 20 (40%) children were diarrhea-free 48 h after admission to the hospital, compared with 1 of 20 (5%) in the control group (P < 0.0001). Subjects in the treatment group received smaller volumes of parenteral fluids than subjects in the control group. Conclusions. The administration of tormentil root extract in controlled doses shortened the duration of rotavirus diarrhea and decreased the requirement for rehydration solutions. Tormentil root extract appears to be an effective measure to treat rotavirus diarrhea in children.

Endoscopic fetal surgery may reduce preterm labor associated with open hysterotomy but is partially limited by current visualization technology. We investigated a three-dimensional (3D) imaging system coupled to a head-mounted display (3D-HMD) and also employed a computer-controlled zoom endoscope for noninsufflated amnioscopy. Pregnant sheep were prepared in aseptic fashion for general anesthesia. Uterine access was obtained following maternal laparoscopy. A 10-mm zoom endoscope (Vista Medical Technologies, Carlsbad, CA) was used to examine the fetus and uterine contents. Fetal limbs were exteriorized for microsurgery. A new system (Vista Medical Technologies) was attached to an operative microscope, permitting projection of a 3D image via an HMD. The fetus and umbilical cord were inspected using the zoom endoscope, which changes the depth of focus under computer control. Basic manipulations of the fetus and cord were easily completed. Real-time 3D fetal imaging was accomplished. The added depth perception enabled detailed fetal and placental examination, fostering manipulation of the fetus and cord. The HMD was adjusted to fit several surgeons, permitting a natural operative posture. This unit has the capacity to display any video, CT, MR, or ultrasound image as a picture-in-picture. The success of minimally invasive fetal surgery is in part dependent on the development of video technologies capable of providing both magnification and optimal resolution. The zoom endoscope affords excellent visibility of multiple surgical targets without instrument repositioning. A 3D HMD system such as this provides greater anatomic detail and an appreciation of fetal movements that may make intrauterine procedures more feasible.

Certain Bacillus licheniformis strains isolated from oil wells have been shown to produce a very effective biosurfactant, lichenysin A, which is structurally similar to another less active lipopeptide, surfactin. Surfactin, like many small peptides in prokaryotes and lower eukaryotes, is synthesized non-ribosomally by multi-enzyme peptide synthetase complex. Analysis of several peptide synthetases of bacterial and fungal origin has revealed a high degree of sequence conservation. Two 35-mer oligonucleotides derived from highly conserved motifs ('core I' and 'core II') of surfactin synthetase were used to identify the cloned putative operon of lichenysin A synthetase lchA from B. licheniformis BNP29, a strain not amenable to genetic manipulation in a BAC system (F-plasmid-based bacterial artificial chromosome) based on Escherichia coli and its single-copy plasmid F-factor. A 32.4 kb fragment containing lichenysin A biosynthesis locus was sequenced and analysed. The structural architecture of putative lichenysin A synthetase protein containing seven amino acid (aa) activation-thiolation, two epimerization and one thioesterase domains is discussed in terms of its similarity to surfactin and other peptide synthetases. The 100 aa peptide chain situated between the highly conserved signature sequences FDXX and NXYGPTE(IV)X within amino acid binding domains of peptide synthetases is proposed to be a minimal block dictating the substrate specificity of the enzymes. A new operon-type structure has been localized directly upstream from the lichenysin A synthetase genes which, on the basis of sequence determination, potentially encode a four-member ABC-type transport system involved in product secretion.

This experimental study investigates the effect of mitomycin C (MMC) on sinus mucosal healing. MMC has an antiproliferative action on fibroblasts. It is used in glaucoma surgery to prevent restenosis of fistulas. Antrostomies were drilled in rabbit maxillary sinuses. One side was used as a control and the other treated with MMC at a concentration of 0.04, 0.4, or 1 mg/mL. Two animals from each group were sacrificed at 1, 2, 4, and 12 weeks. The antrostomies in the control and 0.04-mg/mL groups had closed by 1 week; in the 0.4-mg/mL group by 4 weeks, and in the 1.0-mg/mL group by 12 weeks. Ciliary function was initially impaired but normalized within 1 week. Both light and scanning electron microscopy showed no permanent damage to the cilia. These results suggest that MMC can be used to delay closure of antrostomies in sinus surgery.

Background The fluorescence characteristics of tissues depend upon their biochemical composition and histomorphological architecture, both of which undergo a change during malignant transformation. These changes are detectable as an alteration in the fluorescence spectral profile of the tissues. Methods Biopsy specimens from clinically suspicious lesions and normal-appearing oral mucosa were obtained from patients. Fluorescence spectroscopic measurements were obtained to study the differences between normal and dysplastic tissues and to determine the most appropriate excitation wavelength(s) for exploiting these differences. Results Fluorescence spectra from a total of 12 histologically normal (healthy mucosa or benign lesions) and ten abnormal (dysplastic or malignant) tissue samples were compared. Significant spectral differences were seen between the two groups. These differences were most marked at the excitation wavelength of 410 nm. Using this wavelength, fluorescence correctly diagnosed 20 of 22 samples studied. Conclusions This technique accurately differentiates normal from abnormal tissues in vitro and has the potential applications for in vivo use as a noninvasive diagnostic tool. © 1997 John Wiley & Sons, Inc. Head Neck 19: 27–32, 1997.

Endoscopic laser resection usually has been discouraged for anterior commissure (AC) carcinoma because of inadequate exposure and close proximity to underlying cartilage. A new technique combining endoscopic CO 2 laser incision and an external approach, creating a window in the thyroid cartilage, was tested in this in vivo study of six dogs. An en bloc specimen including adjacent cartilage was excised while preserving the thyroid framework. The glottic reconstruction was accomplished with external placement of a sternohyoid muscle flap, by either using a bipedicled muscle flap with overlying skin or a unipedicled muscle flap with a graft of free mucosa. The results showed a satisfactory anatomic reconstruction and acceptable functions. The bipedicled muscle flap was superior to the unipedicled muscle flap due to a better AC reconstruction. It is believed that this new combined technique may overcome limited access with endoscopic technique and excessive cartilage resection with the external partial laryngectomy.

Repair of anterior tracheal wall collapse is a common and troublesome problem encountered by the head and neck surgeon. The standard treatment calls for an open procedure with or without stenting, depending on the extent of the damage. To avoid the morbidity of the open procedure, a new concept of endoscopic cartilage reshaping was investigated in a laboratory animal study. It involved the application of a 1.44-μm pulsed neodymium:yttrium-aluminum-garnet (Nd: YAG) laser at relatively low power to restructure without devitalizing cartilage. An in vivo study was done in six dogs to determine appropriate laser dosimetry in a model of tracheal wall collapse created by a tracheotomy. The deformed cartilage was treated endoscopically with a noncontact 1.44-μm Nd: YAG laser, at 2 to 4 W of power with a repetition rate of 20 Hz, in three animals. As a control, three animals had endoscopic cartilage incisions followed by stent placement. Six weeks postoperatively, both groups had an adequate airway lined by healthy mucosa. In the animals with stenting, however, there was stenosis formation due to scarring at both ends of the stent, with significant inflammatory response in the local area. This study shows that it is possible to use low-power laser energy to reshape cartilage without destroying its viability, and to restore the tracheal wall to a normal contour without ablation or vaporization. The reshaped cartilage will tend to retain its shape with functional elastic force, as seen in in vitro studies. These preliminary results are encouraging, and it seems reasonable to consider using the technique in selected clinical cases as an alternative to conventional open surgery.

Objective: To determine the effectiveness of the holmium: YAG (Ho:YAG) laser in otolaryngologic procedures that necessitate the ablation of osseous and soft tissue. Design: Case series. Setting: Lahey Clinic, Burlington, Mass. Patients: Consecutive series of 37 patients; 29 with chronic sinusitis, five with chronic dacryocystitis, one with recurrent choanal stenosis, one with tracheopathia osteoplastica, and one with a sphenoid sinus mucocele. Intervention: The Ho:YAG laser was used to assist in 37 procedures, including endoscopic sinus surgery, dacryocystorhinostomy, treatment of choanal stenosis, ablation of obstructive tracheopathia osteoplastica, and removal of a sphenoid sinus mucocele. Main Outcome Measures: Postsurgical success and complications, satisfaction of the patients, and the abiliity of the laser to remove tissue. Results: Complications occurred in eight patients: intranasal or ethmoid scarring (four), persistent polyps (one), bleeding (one), stent dislodgment (one), and tracheitis (one). Three patients required revision surgery. None of the complications were related to use of the laser, although the laser may produce increased scarring. The laser was effective for osseous and soft-tissue ablation, but its usefulness was limited for hemostasis. Conclusions: The Ho:YAG laser can be used in otolaryngologic procedures when surgical access is difficult or when controlled, precise ablation of osseous tissue is necessary. (Arch Otolaryngol Head Neck Surg. 1995;121:1162-1166)

Objective: To investigate (1) the possibility of survival of free mucosa "stamp" grafts fixed in the airway with a new technique using indocyanine green—dyed albumin solder activated with a diode laser and (2) the degree of improvement of wound healing in the airway by applying modified microskin transplantation techniques from burn surgery to cover a relatively large wound with a few small pieces of mucosa anchored in place with the previously mentioned technique. Design: Three (one control and two experimental) rectangular (10×8 mm) wounds in tracheal mucosa were produced in four experimental animals (dogs) using a carbon dioxide laser. The control wound was left uncovered. In the first experimental wound, a mucosal flap was raised and then fixed in place by a trapdoor flap method. In the second experimental wound, two small (each 2×3 mm) autogenous mucosa grafts were anchored onto the surface with indocyanine green—dyed albumin activated with an 810-nm diode laser. Histomorphologically, the postoperative results from three wounds were compared. Results: The experimental wounds were completely covered by regenerated squamous cells in 1 week and by ciliated epithelium in 2 weeks after the operation despite the discrepancy in size of the graft to wound area (1:6.7) covered with the stamp mucosa. No thermal damage from the diode laser was noted in the second experimental wounds. In the control wounds, no coverage was observed at 1 week, and only squamous cells were noted 2 weeks postoperatively. All the wounds had normal ciliated epithelium coverage at 4 weeks. Conclusions: Transplanted stamp grafts provided similar or better healing than trapdoor flap transplants. This new technique made endoscopic mucosal grafting possible and offers a potential breakthrough in the management of laryngotracheal stenosis. (Arch Otolaryngol Head Neck Surg. 1995;121:773-777)

Epithelial coverage of a laryngotracheal wound is an important factor in preventing stenosis, and endoscopic transplantation of a free mucosal graft without stents or sutures would be a significant therapeutic advance. In vitro and in vivo canine studies were performed to explore the feasibility of transplantation with a low-power diode laser (400 mW) enhanced by indocyanine green dye-doped albumin. The tensile strength of graft adherence in 10 cadaver larynges was strong (35.25 +/- 10.39 g). Survival studies in live canine models with a specially designed endoscopic instrument set showed excellent healing at 6, 14, and 28 days. Healing was documented with photography and by histologic examination. Successful endoscopic transplantation of a free mucosal graft should improve results of treatment for laryngotracheal stenosis and laryngeal reconstructive surgery.

A 22-year-old patient was admitted because of abdominal pain and vomiting. Computed tomography diagnosed small intestinal malignancy. Ileal resection was performed, and the histological findings were consistent with sclerosing mesenteritis. The patient was treated with enteral nutrition, corticosteroids, azathioprine and methotrexate, but died 2 years later. Dig Liver Dis

Cyclooxygenases (COXs) are the key enzymes in the production of prostaglandins (PGs) and exist in two isoforms. Isoform 1 (COX-1) is constitutively expressed in most tissues, whereas cyclooxygenase-2 (COX-2) is rapidly induced by a variety of different stimuli. In this study, we have quantitatively analyzed mRNA expression of COX-1 and COX-2 and protein distribution during corneal reparative processes after wound. Total RNA was isolated from cornea samples of New Zealand rabbits that had been subjected to corneal wound by mechanical brush scraping. Quantification of RT-PCR results was made by using a DNA mimic approach. The localization and expression of the enzymes was studied by immunocytochemistry and Western blotting. In normal corneas COX-1 is expressed throughout the cornea in the whole tissue, while COX-2 is strongly expressed in stromal keratocytes. Following injury, COX-2 levels drastically increase and, at least in the epithelium, COX-2 becomes the predominant isoform of cyclooxygenases at an early stage of healing. Moreover, in the epithelium COX-2 is expressed predominantly by those cells close to the wound. These cells become migratory and move toward the injured area. In contrast, COX-1 levels remain unaffected in all corneal tissues. The system returns to the pre-injury state in about 24 h. Thus, the expression of COX-2 in the corneal epithelium during wound repair is tightly regulated both temporally and spatially.

In the report the variant of the operative dynamical tomography is considered for the opticallytransparent biological tissues investigations based upon the linear moving of the emitting-receiving unitrelative to the investigated tissue section. The radiation is sent and detected simultaneously at five directions , specified by condition = arctan (A.H)/i x zR, where i=1.. .5, tiR and EiH are spatialresolution at the tissue surface and penetration depth into the tissue, respectively. The interpretation ofthe obtained scattering signals is carried out at varying integration limit, corresponding to the emitting- receiving unit moving. Keywords: dynamical optical tomography, biological tissues, optical scattering 1. INTRODUCTION The realization of optical tomography principle requires the irradiation of the object in differentaspects and registration of the radiation left the object. The recorded radiation, transformed into the numerical codes, is processed relative to the quantitative values of specified physical characteristics. The

Background: A laser technique has been developed in the laboratory using a noncontact 810-nm diode laser. Results: Mucosal intact laser tonsillar ablation (MILTA) coagulates tonsillar lymphoid tissue while preserving the overlying mucosa. A canine model was used to evaluate this new method. The canine tonsils were exposed to laser energy for 5 to 6 minutes each at 8 to 10 W of power. This technique was compared with standard "cold" knife tonsillectomy. The dogs were observed carefully during the postoperative period. When comparing parameters such as onset of eating solid food, amount of food ingested per day, onset of normal activity, and degree of weight loss, MILTA showed significant benefit over the standard technique. The operative sites were evaluated and compared at 1.5 hours and at 7, 21, and 45 days after treatment. Conclusions: None of the tonsils treated by MILTA showed mucosal ulceration. At the end of 45 days, histopathological examination showed complete absence of tonsillar tissue with normal mucosa in both the MILTA and standard groups. Traditional tonsillectomy techniques are associated with moderate morbidity and the potential for serious hemorrhage. With the preservation of the overlying mucosa, the possible clinical advantages of this approach to tonsillectomy are the absence of bleeding, avoidance of general anesthesia in older patients, less operating time, and decreased postoperative morbidity. The potential for office treatment exists. Additional laboratory studies, as well as careful clinical trials, are indicated based on these encouraging results. Arch Otolaryngol Head Neck Surg. 1996;122:1355-1359

Excimer lasers with stimulated Raman scatteiing cell and Nd YAG laser with frequency doubler were employed to study the effect oflaser radiation both on intact and atheroscierotically damaged aortic wall. After inadiation,, samples of aorta were subjected to detailed morphologic study and sizes of craters, necrosis and tissue destruction zones were determined. Lipid content and spectra of the extracts affected by laser beam were under consideration as well. Key words: laser angioplasty, tissue ablation, photodissociation ofbiological molecules

Advancements in minimally invasive surgery, in areas such as otology, require that the surgeon operates through a small opening, often with indirect access to the surgical field. Conventional carbon-dioxide laser delivery systems, such as the micromanipulator and the semi-flexible waveguide limit the surgeon's ability to perform such procedures. The ablation properties of a small, flexible, hollow carbon-dioxide laser waveguide (Medical Optics, Carlsbad, CA) were compared to a conventional surgical carbon-dioxide laser micromanipulator on chinchilla calvarium bone. This waveguide has a small diameter (O.D./I.D. equals 500/300 micrometer) and is therefore potentially useful for minimally invasive middle ear surgical procedures, such as carbon-dioxide laser stapedotomy. The beam diameter and the half angle divergence of the waveguide were estimated using a standard knife-edge measurement technique. Lesions were created at clinically relevant power settings with each delivery system on fresh harvested chinchilla bone. The crater size and the thermal damage width were then measured under a light microscope. The size of the lesions were found to be comparable between the two delivery systems. This preliminary study demonstrates that the two delivery systems have comparable effects on bone. However, further work is necessary to compare their effects on healing before the waveguide can be used clinically for surgical procedures such as carbon-dioxide laser stapedotomy.© (1996) COPYRIGHT SPIE--The International Society for Optical Engineering. Downloading of the abstract is permitted for personal use only.

Cadaveric human temporal bones, cadaveric rabbits, and live rats were used to demonstrate the utility and safety of the erbium:yttrium-scandium-gallium-garnet (Er:YSGG) laser for otologic applications. The shallow penetration in water of this wave-length and its ability to ablate bony tissue with minimal collateral thermal effects spare underlying and adjacent structures and make it appealing for stapedotomy. The authors were able to satisfactorily perform small fenestra stapedotomy, atticotomy facial nerve decompression, and mastoidectomy. Temperature measurements from the round window area during Er:YSGG stapedotomy recorded an elevation of less than 2 degrees C, which is well within acceptable limits. An acoustic shock produced at the impact site is the major disadvantage and requires further in vivo investigation of the transient and sustained deleterious effects away from the application site. This work supports further investigation into potential applications of the Er:YSGG laser in otology.

Recent studies have indicated the existence of an extensive trans-genomic trans-mural co-metabolism between gut microbes and animal hosts that is diet-, host phylogeny- and provenance-influenced. Here, we analyzed the biodiversity at the level of small subunit rRNA gene sequence and the metabolic composition of 18 Mbp of consensus metagenome sequences and activity characteristics of bacterial intra-cellular extracts, in wild Iberian lynx (Lynx pardinus) fecal samples. Bacterial signatures (14.43% of all of the Firmicutes reads and 6.36% of total reads) related to the uncultured anaerobic commensals Anaeroplasma spp., which are typically found in ovine and bovine rumen, were first identified. The lynx gut was further characterized by an over-representation of 'presumptive' aquaporin aqpZ genes and genes encoding 'active' lysosomal-like digestive enzymes that are possibly needed to acquire glycerol, sugars and amino acids from glycoproteins, glyco(amino)lipids, glyco(amino)glycans and nucleoside diphosphate sugars. Lynx gut was highly enriched (28% of the total glycosidases) in genes encoding α-amylase and related enzymes, although it exhibited low rate of enzymatic activity indicative of starch degradation. The preponderance of β-xylosidase activity in protein extracts further suggests lynx gut microbes being most active for the metabolism of β-xylose containing plant N-glycans, although β-xylosidases sequences constituted only 1.5% of total glycosidases. These collective and unique bacterial, genetic and enzymatic activity signatures suggest that the wild lynx gut microbiota not only harbors gene sets underpinning sugar uptake from primary animal tissues (with the monotypic dietary profile of the wild lynx consisting of 80-100% wild rabbits) but also for the hydrolysis of prey-derived plant biomass. Although, the present investigation corresponds to a single sample and some of the statements should be considered qualitative, the data most likely suggests a tighter, more coordinated and complex evolutionary and nutritional ecology scenario of carnivore gut microbial communities than has been previously assumed.

Autofluorescence characteristics of normal, dysplastic, and malignant squamous tissues from the oral cavity were measured with a spectrofluorometer in the excitation range of 250 - 500 nm and emission range of 350 - 750 nm. Fluorescence excitation-emission matrices (EEM) were obtained from samples collected from patients in the clinic and in the operating room. The same samples were submitted for histopathological examination following spectroscopic measurements. The contour plots obtained from the EEMs of the samples showed consistent differences between normal and abnormal tissues. All the abnormal samples showed enhanced red region (> 600 nm) fluorescence with a prominent peak at 635 nm, when excited by 410 nm light. A ratio contour plot (abnormal/normal) enhanced spectral differences in the red region. A fiber-optic based spectrofluorometer for EEM measurements is being developed for further investigations.

Vascular ectasias (dilatation) and vascular lesions of the larynx are difficult to treat with exciting modalities. Varix (enlarged vessel) of the vocal folds, vocal fold hemorrhage, vascular polyp, hemangioma, intubation or contact granuloma are common problems which disturb voice. Current applications of CO2 laser and cautery often damage the delicate vocal fold cover. The 585 nm dermatologic pulsed dye laser may be an ideal substitute. Two adult canines were examined under anesthesia via microlaryngoscopy technique. Pulsed dye laser (SPTL-1a, Candela Laser Corp., Wayland, MA) energy was delivered via the micromanipulator with the 3.1-mm spot size in single pulses of 6, 8, and 10 Joules/cm2 and applied to the vessels of the vocal folds, epiglottis, and arytenoid cartilage. Endoscopic examination was carried out immediately after the treatment and at 4 weeks postoperatively. The animals were sacrificed at 3 weeks, larynges excised, and whole organ laryngeal section were prepared for histology. Pulsed dye laser thrombosed vessels of the vocal fold using 6 or 8 Joules/cm2. Vascular break and leakage occurred at 10 Joules/cm2. Follow up examination showed excellent vessel obliteration or thrombosis without scarring or injury to the overlying tissues. Histologic examination shows vascular thrombosis without inflammation and fibrosis in the vocal fold cover. Pulsed dye laser may have promise in treatment of vascular lesions of the larynx and upper airway.© (1995) COPYRIGHT SPIE--The International Society for Optical Engineering. Downloading of the abstract is permitted for personal use only.

Correction of cartilaginous defects in the head and neck area remains a challenge for the surgeon. This study investigated a new technique for laser-assisted cartilage reshaping. The pulsed 1.44 micrometers Nd:YAG laser was used in vitro and in vivo experiments to irradiate cartilage to change it's shape without carbonization or vaporization of tissue. Two watts of average power in non contact manner was used to irradiate and reshape the cartilage. The extracted reshaped cartilage specimens underwent testing of elastic force with a computer assisted measurement system that recorded the changes in elastic force in the specimens from 1 hr to 11 days post-irradiation. An animal model of defective tracheal cartilage (collapsed tracheal wall) was created, allowed to heal for 6 weeks and then corrected endoscopically with the laser-assisted technique. The results of the in vitro and in vivo investigations demonstrated that it was possible to alter the cartilage and that cartilage would retain its new shape. The clinical significance of the technique is evident and warrants further animal studies and clinical trials.

The Nd:YAG laser 1.44 micrometers wavelength is a relatively new wavelength with water absorption coefficient higher even than 1.06 micrometers , permitting the creation of a more precise lesion with less damage to surrounding tissue. Very little, however, is known about its effects in live experimental model. The non-contact 1.44 micrometers Nd:YAG laser was used in four animals (dogs) to perform a resection of middle turbinate and to create a 1 cm lesion on the septal mucosa and inferior turbinate in the left side through a sinuscope. The same procedure was performed on the opposite side using microsurgical forceps as a control. The animals were followed-up, examined endoscopically, and sacrificed after 48 hours (1 animal), 1 month (1 animal) and 2 months (2 animals). Moderate-to-severe bleeding was commonly encountered on the control side, which obscured vision, and required packing in 2 dogs. In contrast, the laser had a satisfactory hemostatic result with easy control for endoscopic resection. Normal healing was observed under gross and by histologic examination 48 hours and 1 month after the surgery and healing was complete by 2 months. There was more scaring and adhesions produced by the forceps on the control side. Histologically, tissue thermal damage produced by the laser was superficial and confined to the lesion. The epithelial regeneration has been nearly completed in 1 month, with normal mucosa coverage in 2 months after the surgery. This study suggests that the 1.44 micrometers wavelength of the Nd:YAG should be considered as safe and reliable for intranasal application.