Study of Human Fibrinogen Oxidative Modification using Differential Scanning Calorimetry.

For the first time, with the aid of differential scanning calorimetry, the thermal denaturation of fibrinogen under induced oxidation was studied. All fibrinogen structural elements detected by DSC (D region, αC-domain, and E region) are subjected to oxidation. Structural changes in fibrinogen molecule were characterized by the denaturation temperature, denaturation enthalpy, and van't Hoff enthalpy.

Modification of human serum albumin under induced oxidation.

For the first time, by using the complex of physicochemical methods (mass-spectrometry, differential scanning calorimetry, spectrofluorimetry, method of spectral and fluorescent probes, dynamic light scattering, and UV spectrophotometry), the oxidation-mediated modification of chemical and spatial structure of albumin has been studied. All albumin structural regions are subjected to oxidation, methionine and aromatic amino acids primarily involved in oxidation. The albumin melting shows a decrease in thermal stabilization of the structure and changing of aggregation upon oxidation. The change in physical and chemical properties of albumin under different quantity of the oxidizer has been analyzed.

Laparoscopic limited Heller myotomy without anti-reflux procedure does not induce significant long-term gastroesophageal reflux.

BACKGROUND: Laparoscopic Heller myotomy with partial fundoplication is the gold standard treatment for achalasia. Laparoscopic limited Heller myotomy (LLHM) with no anti-reflux procedure is another possible option. METHODS: A review of prospectively collected data was performed on patients who underwent LLHM from January 1998 to December 2012. Evaluation included gastroscopy, esophageal manometry, 24-h pH-metry, and the Short Form(36) Health Survey(SF-36) questionnaire at baseline and 6 months, as well as the global symptom score at baseline, 6 months, and 5 years post-surgery. Comparison between outcomes was performed with a paired t student's test. RESULTS: 126 patients underwent LLHM. Of these, 60 patients had complete pre and post-operative motility studies. 57 % were female, patient mean age was 45.7 years, with a mean follow-up of 10.53 months. Mean operative time was 56.1 min, and the average length of stay was 1.7 days. At 6 months, a significant decrease in the lower esophageal sphincter resting pressure (29.1 vs. 7.1 mmHg; p < 0.001) and nadir (16.4 vs. 4.3 mmHg; p < 0.001) was observed. Normal esophageal acid exposure (total pH < 4 %) was observed in 68.3 % patients. Nevertheless, of the remaining 31.7 % with abnormal pH-metry, only 21.6 % were clinically symptomatic and all were properly controlled with medical treatment without requiring anti-reflux surgery. Significant improvement in all pre-operative symptoms was observed at 6 months and maintained over 5 years. Dysphagia score was reduced from 9.8 pre-operatively to 2.6 at 5 years (p < 0.001), heartburn score from 3.82 to 2 (p < 0.01), and regurgitation score from 7.5 to 0.8 (p < 0.001). Only one patient (0.8 %) presented with recurrent dysphagia requiring reoperation. CONCLUSION: LLHM without anti-reflux procedure is an effective long-term treatment for achalasia and does not cause symptomatic GERD in three quarters of patients. The remaining patients are well controlled on anti-reflux medications. It is believed that similar clinical results would be obtained during a clinical investigation of the POEM procedure.

The recent expansion of Pluto's atmosphere.

Stellar occultations--the passing of a relatively nearby body in front of a background star--can be used to probe the atmosphere of the closer body with a spatial resolution of a few kilometres (ref. 1). Such observations can yield the scale height, temperature profile, and other information about the structure of the occulting atmosphere. Occultation data acquired for Pluto's atmosphere in 1988 revealed a nearly isothermal atmosphere above a radius of approximately 1,215 km. Below this level, the data could be interpreted as indicating either an extinction layer or the onset of a large thermal gradient, calling into question the fundamental structure of this atmosphere. Another question is to what extent Pluto's atmosphere might be collapsing as it recedes from the Sun (passing perihelion in 1989 in its 248-year orbital period), owing to the extreme sensitivity of the equilibrium surface pressure to the surface temperature. Here we report observations at a variety of visible and infrared wavelengths of an occultation of a star by Pluto in August 2002. These data reveal evidence for extinction in Pluto's atmosphere and show that it has indeed changed, having expanded rather than collapsed, since 1988.

Jovian Atmosphere: Structure and Composition between the Turbopause and the Mesopause.

The occultation of the star Beta Scorpii by Jupiter was observed at high time resolution in three wavelength channels. The results imply a temperature of 220 degrees K at an altitude in the Jovian atmosphere corresponding to 10(14) molecules per cubic centimeter, and temperature fluctuations of 2 degrees to 10 degrees K over vertical scales of 2 to 10 kilometers. They suggest that the vertical eddy diffusion coefficient near the turbopause has a lower limit of 7 x 10(5)K square centimeters per second, and that the turbopause lies above the altitude where the density is 5 x 10(13) molecules per cubic centimeter. Below the turbopause, the ratio of hydrogen to helium is consistent with cosmic abundances.

Diamond-Blackfan syndrome: lymphocyte-mediated suppression of erythropoiesis.

Peripheral blood lymphocytes from six patients with congenital hypoplastic anemia suppressed erythroid cell formation by normal human bone marrow cells in response to erythropoietin in vitro. The results suggest that the anemia in these children has an immunologic basis.

Resistance of alpha-crystallin quaternary structure to UV irradiation.

The damaging effect of UV radiation (lambda > 260 nm) on bovine alpha-crystallin in solution was studied by small-angle X-ray scattering, gel permeation chromatography, electrophoresis, absorption and fluorescence spectroscopy, and differential scanning calorimetry. The results obtained show that damage to even a large number of subunits within an alpha-crystallin oligomer does not cause significant rearrangement of its quaternary structure, aggregation of oligomers, or the loss of their solubility. Due to the high resistance of its quaternary structure, alpha-crystallin is able to prevent aggregation of destabilized proteins (especially of gamma- and beta-crystallins) and so to maintain lens transparency throughout the life of an animal (the chaperone-like function of alpha-crystallin).

Some physico-chemical and thermodynamic characteristics of maize starches hydrolyzed by glucoamylase.

Glucoamylolysis of maize starch at 55 °C has been studied by means of scanning electron microscopy (SEM), wide-angle X-ray diffraction spectroscopy (WAXD), and differential scanning calorimetry (DSC). It was found that hydrolysis is accompanied by changes in thermodynamic parameters of diluted aqueous dispersions of partially hydrolyzed starches. Such changes are ensured by two processes directly from hydrolysis and accompanying annealing. At relatively low degrees of hydrolysis (less than 30%), changes in thermodynamic parameters are mainly controlled by annealing. At the same time, at high degrees of hydrolysis (more than 40%), the main contribution to changes in thermodynamic parameters of partially hydrolyzed starch granules is due to the hydrolysis itself. It has been established that the main controlling parameter is the thickness of crystalline lamellae Lcrl, which, when annealed, increases, but tends to decrease at deeper glucoamylolisis. It has been established that the thickness Lcrl of crystalline lamellae, which increases with annealing, but shows a tendency to decrease with deeper glucoamylolysis is the most representative parameter of changes in maize starch after hydrolysis.

[Psychological assessment of visual hemispatial neglect: standardization and approbation of the modified digit cancellation test]. / Diagnostika odnostoronnego zritel'no-prostranstvennogo nevnimaniia: standartizatsiia i aprobatsiia modifitsirovannoi tsifrovoi korrekturnoi proby.

AIM: To study the phenomena of visual-hemispatial neglect in healthy people and patients with brain diseases of different genesis. MATERIAL AND METHODS: Eighty-eight patients with schizophrenia spectrum disorders, 68 patients with exogenous organic brain diseases and 240 healthy adults of different age were included in the study. The digit cancellation test modified by the authors was used. RESULTS AND CONCLUSION: The validity of the modified digit cancellation test was approved and its age standards were obtained. In healthy right-handed people, there was the bias of attention focus to the left, the decrease of asymmetry intensity of visual-spatial inattention during physiological aging and the presence of some clinical peculiarities of neglect in schizophrenia spectrum disorders and lateralized organic damages of the brain. This variant of the test can be recommended for practical use as the sensitive psychometric tool.

Erythroid colony formation by polycythemia vera bone marrow in vitro. Dependence on erythropoietin.

In the plasma clot culture system both normal and polycythemia vera (PV) bone marrow cells respond to erythropoietin (Ep), giving rise to large numbers of colonies of erythroid cells. In PV, but not in normal individuals, the marrow produced endogenous erythroid colonies (EED) in the absence of exogenous Ep. The number of EEC formed varied from patient to patient comprising anywhere from 6 to 29% of the total number of colonies formed in the presence of Ep. Exposure, before use in culture, of fetal calf serum and citrated bovine plasma to the gammaglobulin fraction of rabbit anti-Ep serum followed by treatment with goat anti-rabbit gamma-globulin re sulted in a significant decrease in EEC formation. Addition of anti-Ep directly to the culture medium produced similar results. In addition, the production of EEC in response to added Ep was inhibited in the presence of anti-Ep. Addition of very small doses of highly purified Ep to anti-Ep-treated cultures resulted in the reappearance of a significantnumber of EEC formation in PV may be due to a population of erythroid-committed precursors that are abnormally sensitive to small concentrations of Ep which may be present in fetal calf serum and citrated plasma. Although the mechanism of formation of these cells is not known, it appears that the final steps in the formation of red cells derived from this clone of precursors is subject to the usual Ep control.

Dimethylthiourea inhibition of melanoma cell growth in vitro and in vivo.

The effect of dimethylthiourea (DMTU), an agent known as a hydroxyl radical scavenger, was determined on growth and differentiation of the B16 murine melanoma cell line. DMTU inhibited melanoma cell growth in vitro and induced changes in the morphology of melanoma cells. Prolonged treatment of cells with DMTU resulted in an increase in melanin content. DMTU-treated melanoma cells had a decreased capacity to form tumors in syngeneic mice. Systemic administration of DMT to C57BL/6J mice inoculated with melanoma cells resulted in a delay in tumor appearance and a prolongation of survival. The doses of DMTU used did not cause any apparent toxic effects. A potential therapeutic role for DMTU in the treatment of melanoma is suggested.

Characterization of purine nucleotide metabolism in primary rat cardiomyocyte cultures.

Primary rat cardiomyocyte cultures were utilized as a model for the study of purine nucleotide metabolism in the heart muscle, especially in connection with the mechanisms operating for the conservation of adenine nucleotides. The cultures exhibited capacity to produce purine nucleotides from nonpurine molecules (de novo synthesis), as well as from preformed purines (salvage synthesis). The conversion of adenosine to AMP, catalyzed by adenosine kinase, appears to be the most important physiological salvage pathway of adenine nucleotide synthesis in the cardiomyocytes. The study of the metabolic fate of IMP formed from [14C]formate or [14C]hypoxanthine and that of AMP formed from [14C]adenine or [14C]adenosine revealed that in the cardiomyocyte the main flow in the nucleotide interconversion pathways is from IMP to AMP, whereas the flux from AMP to IMP appeared to be markedly slower. Following synthesis from labeled precursors by either de novo or salvage pathways, most of the radioactivity in purine nucleotides accumulated in adenine nucleotides, and only a small proportion of it resided in IMP. The results suggest that the main pathway of AMP degradation in the cardiomyocyte proceeds through adenosine rather than through IMP. About 90% of the total radioactivity in purines effluxed from the cells during de novo synthesis from [14C]formate or following prelabeling of adenine nucleotides with [14C]adenine were found to reside in hypoxanthine. The activities in cell extracts of AMP 5'-nucleotidase and IMP 5'-nucleotidase, which catalyze nucleotide degradation, and of AMP deaminase, a key enzyme in the purine nucleotide cycle, were low. The nucleotidase activity resembles, and that of the AMP deaminase contrasts the respective enzyme activities in extracts of cultured skeletal-muscle myotubes. The results indicate that in the cardiomyocyte, in contrast to the myotube, the main mechanism operating for conservation of nucleotides is prompt phosphorylation of AMP, rather than operation of the purine nucleotide cycle. The primary cardiomyocyte cultures are a plausible model for the study of purine nucleotide metabolism in the heart muscle.

The customer approach to patienthood. Attending to patient requests in a walk-in clinic.

This report conceptualizes the initial psychiatric interview as a process of negotiation between the clinician and patient. Patients are conceived of as appearing with one or more requests, many of which represent legitimate needs. It is the clinician's task to elicit the request, collect the relevant clinical data, and enter into a "negotiation" that should foster a relationship of mutual influence between patient and clinician. We have attempted to show that this approach to patienthood not only improves patient care and patient satisfaction but also leads to improved staff morale. The "customer approach" has special relevance to those clinical settings (walk-in clinics and community mental health centers) in which clinicians see a broad range of patients with a broad definition of psychiatric problems and requests.

Thallium 201 myocardial scintigraphy. Advantages and limitations.

Myocardial scintigraphy with thallium 201 is a simple, safe, and valuable noninvasive technique in evaluating the condition of patients with cardiac disorders. Images obtained at rest appear to have limited usefulness at the present time. However, rest imaging may prove to have advantages in the future for early diagnosis of myocardial infarction, thus aiding in the selection of patients to be admitted to the coronary care unit. Exercise imaging, on the other hand, has a high degree of sensitivity and specificity in detecting ischemic heart disease; when combined with treadmill exercise testing, imaging improves the diagnostic accuracy even further.

[Cognitive impairments in depression: neuropsychological and MRI-studies]. / Kognitivnye rasstroistva pri depressiiakh: neiropsikhologicheskoe i MRT-issledovanie.

OBJECTIVE: A complex neuropsychological and neuroimaging study of deep brain structures in depression with cognitive impairment. MATERIAL AND METHODS: We studied 73 patients with endogenous depression and 86 patients with depressive syndrome in temporal epilepsy. MRI and neuropsychological methods were used to study brain structures. Results and сonclusion. Neurocognitive impairment was more severe in patients with depressive syndrome in the structure of temporal epilepsy. The differences between the patients with endogenous and organic (temporal epilepsy) affective disturbances were determined by the more marked memory and spatial-constructive impairments in patients with organic disorders. Deficit of executive functions, planning functions and cognitive organization were more typical for the patients with endogenous affective disorders. The MRI study revealed the decrease in the left hippocampus due to sclerotic processes in patients with temporal epilepsy and the increase in the right amygdale in patients with endogenous depressive disorders. The results demonstrate the significant similarity between characteristics of the cognitive profile in patients with endo- and exogenous depressions.

Therapeutic recommendations in polycythemia vera based on Polycythemia Vera Study Group protocols.

The PVSG was organized in 1967 to establish effective diagnostic criteria for polycythemia vera, to study the natural history of the disease and to define the optimal treatment. Although polycythemia vera and the other myeloproliferative diseases are relatively uncommon, the PVSG was able to accumulate well over 1,000 patients with these various disorders and to study them according to a total of 15 different protocols. PVSG-01, a long-term randomized controlled study of phlebotomy alone compared with the myelosuppressive agents, 32P or chlorambucil supplemented by phlebotomy, continues to receive follow-up data on 93% of surviving patients 18 years after initiation of the study. During its lifetime, PVSG has developed a widely accepted and highly effective set of criteria for the specific diagnosis of polycythemia vera as well as useful criteria for the diagnosis of essential thrombocythemia. It has gathered an enormous volume of data on the natural history of the myeloproliferative diseases and in particular on the nature of the prevalent complications, such as thrombotic events and hematologic and nonhematologic malignancies. With respect to the final question, the optimal treatment for polycythemia vera, it is apparent that the expectation of a single optimal therapy that would apply to all patients at all ages and stages of the disease was naive. Nevertheless considerable progress has been made. Moreover, the group has defined more precisely than ever before the nature of the complications of the disease and the association of the risks of specific complications with specific forms of therapy. It thus has made it possible to pose the next series of therapeutic questions that must be addressed in this disorder with a greater degree of sophistication than was previously possible.

Anti-proliferative effects and phenotypic alterations induced by 8-hydroxyquinoline in melanoma cell lines.

The effect of the transition metal chelator, 8-hydroxyquinoline (8-HQ), was examined on the growth and phenotype expression of B16 mouse melanoma cells. Micromolar concentrations of 8-HQ inhibited the growth of B16 cells as well as human melanoma cell lines. Removal of 8-HQ from the culture medium restored normal cell growth. Growth inhibition by 8-HQ was accompanied by phenotypic alterations that included changes in cell morphology, increased production of melanin and enhanced activities of the enzymes gamma-glutamyl transpeptidase and NADPH cytochrome c reductase. These changes might be associated with a better differentiated phenotype.

Adenosine acts as an inhibitor of lymphoma cell growth: a major role for the A3 adenosine receptor.

In this study, we demonstrated several mechanisms exploring the inhibitory effect of low-dose adenosine on lymphoma cell growth. Adenosine, a purine nucleoside present in plasma and other extracellular fluids, acts as a regulatory molecule, by binding to G-protein associated cell-surface receptors, A1, A2 and A3. Recently we showed that low-dose adenosine released by muscle cells, inhibits tumour cell growth and thus attributes to the rarity of muscle metastases. In the present work, a cytostatic effect of adenosine on the proliferation of the Nb2-11C rat lymphoma cell line was demonstrated. This effect was mediated through the induction of cell cycle arrest in the G0/G1 phase and by decreasing the telomeric signal in these cells. Adenosine was found to exert its antiproliferative effect mainly through binding to its A3 receptor. The cytostatic anticancer activity, mediated through the A3 adenosine receptor, turns it into a potential target for the development of anticancer therapies.

A cell line with unusual characteristics from an ovarian carcinoma patient: modulation of sensitivity to antitumour drugs.

A cell line (GZL-8) was established by cloning from ascitic fluid of an untreated ovarian carcinoma patient. The cells grew rapidly, accumulated lipids and showed chromosomal alterations. One of the marker chromosomes showed characteristics of a Y-like chromosome. This unusual finding was confirmed by DNA hybridisation using specific probes to the Y chromosome. The cells stained with fluorescent antibodies to desmoplakin and cytokeratins 8, 18, 19, and weakly with vimentin but not with desmin. The presence of epithelial membrane antigen, human milk fat globulin, alpha-lactalbumin, alpha-fetoprotein, placental alkaline phosphatase and oestrogen receptor-related antigen was demonstrated by indirect immunoperoxidase staining, but no CA-125 antigen could be detected. The cells showed positive reaction with antibodies to P-glycoprotein. The function of the P-glycoprotein transport system was demonstrated by the rhodamine-123 release test. The cells were initially responsive to doxorubicin, and to high concentrations of cisplatin. Growth inhibition by doxorubicin, especially at low doses was enhanced by the addition of verapamil or tamoxifen. This was shown by the soft agar clonogenic assay, by direct cell counting and by the MTT reducing test. Our results show that combination between drug and sensitivity modulators may be of potential clinical value in ovarian cancer.

Left ventricular perfusion and performance from a single radiopharmaceutical and one camera.

To test the hypothesis that a small field of view portable multicrystal scintillation camera can perform stress/rest combined LV function by first-pass and perfusion studies using 99mTc-teboroxime, 26 patients with positive stress thallium studies within 2 wk and 8 healthy volunteers were studied. A 241Am point source marker over the sternum was used for motion correction. Dynamic dual-isotope (99mTc/241Am) acquisition was performed following injection of 15.6 +/- 2.3 mCi of 99mTc-teboroxime at peak treadmill exercise. Two minutes later (blood-pool clearance), while still standing on the flat treadmill, 3-4 40-sec planar images were acquired. One hour later patients were reinjected with 22.7 +/- 3.4 mCi of 99mTc-teboroxime while standing in front of the camera and the same dynamic/static acquisition protocol repeated. The planar images were interpolated from a 20 x 20 matrix to a 160 x 160 matrix, a sharpening filter and an interpolative background subtraction algorithm applied. The scans were divided into segments, each scored as normal, reversible and fixed. The agreement with thallium imaging for identifying an abnormal scan was 24/26 (92%) and for identifying abnormal vascular territories was 43/52, (83%). Fourteen patients had exercise LVEF less than 50% and all had either prior myocardial infarction, myocardial infarction plus ischemia or LAD ischemia. Diagnostic planar perfusion images and exercise LVEF can be acquired in less than 4 min using 99mTc-teboroxime and a portable multicrystal scintillation camera.