The activity of selected glycosidases in salivary gland tumors

The monitoring of the patients after salivary gland tumors surgery is an important clinical issue. Still imperfect diagnostic procedures also remain a challenge for searching new sensitive and specific biomarkers of neoplastic processes in salivary glands. The aim of the presented study was an the assessment of the activity of HEX, with its isoforms HEX-A and HEX-B, GLU, GAL, MAN and FUC in salivary gland tumor tissues in comparison to a healthy salivary gland tissues taken during autopsy. A group of 42 patients with benign and malignant salivary gland tumors, aged 25-65 were examined. Fragments of salivary gland tumor tissue, fragments of healthy tissue removed during autopsy, blood serum and saliva were collected from patients with salivary gland tumors and healthy volunteers. In salivary gland tumor tissue the activity of HEX, HEX-A, HEX-B, GAL, FUC was considerably higher than in comparison to healthy salivary gland tissue and ascending trend of activity of GLU, MAN was also noticed. The activity of all lysosomal exoglycosidases in blood serum in patients with salivary gland tumors was considerably higher in comparison to healthy volunteers blood serum. The considerably higher activity of HEX, HEX-A, GLU, GAL, MAN, FUC and descending trend of activity of HEX-B were noticed in saliva of patients with salivary gland tumors in comparison to healthy volunteers. The assessment of HEX in blood serum and saliva of patients with salivary gland tumor can be possibly used in diagnostics and monitoring of salivary glands tumors.     

A psychophysiological assessment battery

This research project was the first step in the development of a psychophysiological assessment battery. The battery consisted of eight tasks that have a history of use within the field of psychophysiology. These tasks were examined on a nonpathological, physically healthy sample. This sample was administered the assessment battery three times over the course of 16 weeks. The response systems of HR, SC, RR, TPA, and FPA were examined. Two major research questions were then examined. The first question was whether a subject would display a stable physiological profile on the assessment battery across the three administrations. The second question was whether there would be individual differences in physiological profiles on the assessment battery. These differences were examined in terms of individual response stereotypy (IRS) and stimulus response specificity (SRS). Depending on the task, from 30 to 100% of the subjects displayed stable physiological profiles across administrations. Twenty-five subjects displayed a high degree of SRS. Five subjects displayed a high degree of IRS.     

A psychophysiological assessment battery

This research project was the first step in the development of a psychophysiological assessment battery. The battery consisted of eight tasks that have a history of use within the field of psychophysiology. These tasks were examined on a nonpathological, physically healthy sample. This sample was administered the assessment battery three times over the course of 16 weeks. The response systems of HR, SC, RR, TPA, and FPA were examined. Two major research questions were then examined. The first question was whether a subject would display a stable physiological profile on the assessment battery across the three administrations. The second question was whether there would be individual differences in physiological profiles on the assessment battery. These differences were examined in terms of individual response stereotypy (IRS) and stimulus response specificity (SRS). Depending on the task, from 30 to 100% of the subjects displayed stable physiological profiles across administrations. Twenty-five subjects displayed a high degree of SRS. Five subjects displayed a high degree of IRS.     

Urinary excretion of factor VIII after renal transplantation.

The urinary excretion of factor-VIII-related antigen (VIIIRAg) was measured in 72 patients with kidney transplants and compared with that of two end-products of fibrin-fibrinogen lysis (fragments D and E) to assess their usefulness in monitoring the onset of rejection episodes. Specific and sensitive radioimmunoassays were used to measure the three proteins. Unconcentrated urine samples of 24-hour collections were obtained from 20 healthy subjects, 48 patients with stable transplants, and 24 patients with recent transplants serially followed up from the day of transplantation. Factor VIIIRAg and fragments E and D were not detectable in the urine from healthy subjects but were present in 39%, 60%, and 100% respectively of samples from patients with stable transplants. During 33 acute rejection episodes in 19 patients with recent transplants factor VIIIRAg and fragments E and D were significantly increased above the values observed in patients with stable transplants in 82%, 73%, and 64% of samples respectively; in patients with recent transplants showing no clinical sign of rejection increased excretion of these proteins was observed in 11%, 26%, and 22% of samples respectively. The presence of factor VIIIRAg in urine from patients with kidney allografts suggests that endothelial cell-factor VIII-platelet interactions might pay a key part in the pathogenesis of acute rejection. The results suggest that the assay of factor VIIIRAg in urine is more useful than assays of fragments D and E as a corroborative index of transplant rejection.     

ERIC-PCR fingerprinting-based community DNA hybridization to pinpoint genome-specific fragments as molecular markers to identify and track populations common to healthy human guts

Bacterial populations common to healthy human guts may play important roles in human health. A new strategy for discovering genomic sequences as markers for these bacteria was developed using Enterobacterial Repetitive Intergenic Consensus (ERIC)-PCR fingerprinting. Structural features within microbial communities are compared with ERIC-PCR followed by DNA hybridization to identify genomic fragments shared by samples from healthy human individuals. ERIC-PCR profiles of fecal samples from 12 diseased or healthy human and piglet subjects demonstrated stable, unique banding patterns for each individual tested. Sequence homology of DNA fragments in bands of identical size was examined between samples by hybridization under high stringency conditions with DIG-labeled ERIC-PCR products derived from the fecal sample of one healthy child. Comparative analysis of the hybridization profiles with the original agarose fingerprints identified three predominant bands as signatures for populations associated with healthy human guts with sizes of 500, 800 and 1000 bp. Clone library profiling of the three bands produced 17 genome fragments, three of which showed high similarity only with regions of the Bacteroides thetaiotaomicron genome, while the remainder were orphan sequences. Association of these sequences with healthy guts was validated by sequence-selective PCR experiments, which showed that a single fragment was present in all 32 healthy humans and 13 healthy piglets tested. Two fragments were present in the healthy human group and in 18 children with non-infectious diarrhea but not in eight children with infectious diarrhea. Genome fragments identified with this novel strategy may be used as genome-specific markers for dynamic monitoring and sequence-guided isolation of functionally important bacterial populations in complex communities such as human gut microflora.     

The influence of laparoscopic and open surgery on the concentration and structural modifications of insulin-like growth factor binding protein 3 in the human circulation

Insulin-like growth factor binding protein 3 (IGFBP-3) is the most abundant insulin-like growth factor binding protein in the circulation. The aim of the present work was to investigate the influence of surgery (laparoscopy and open) on the concentration and carbohydrate content of IGFBP-3. The concentration of IGFBP-3 was measured using an immunoradiometric assay (IRMA), its protein profile was characterised using immunoblotting and its sialic acid content was examined by means of Sambucus nigra agglutinin (SNA) affinity chromatography. The concentration of IGFBP-3 was significantly (p < 0.001) lower in preoperative patients compared with healthy subjects. Only patients that underwent open surgery showed a further significant decrease in the concentration of IGFBP-3. Immunoblotting detected two intact IGFBP-3 isoforms, as well as proteolytic fragments. SNA-affinity chromatography showed that in patients that underwent surgery the ratio between the two IGFBP-3 glycoforms was lower than the ratio in healthy subjects. Patients with gallbladder inflammation that underwent laparoscopy had an increased percentage of specifically bound IGFBP-3 to SNA compared with healthy subjects. Our conclusion is that open surgery decreased the level of IGFBP-3 compared with laparoscopy, whereas patients with gallbladder inflammation that underwent laparoscopy had an increased content of sialic acid in IGFBP-3.     

Spectral and Coherent Analysis of the Relationship between Pulse Wave Forms in the Carotid Artery and Heart Rate

The purpose of this study was the assessment of the parameters of pulse waves in the carotid artery and their relationships with baroreflex-controlled heart rate variability. The early systolic peak (ESP) is closely connected with the filling of the carotid artery, whereas the late systolic peak (LSP) shows the amplitude of the wave reflected from the periphery and affects arterial pressure oscillations activating baroreceptors and changing the heart rate (HR). A cardiac rhythmogram and pulse pressure waves in the carotid artery were continuously recorded for 5 min. ESP and LSP amplitudes (in ml/l) were evaluated in each wave. Patients with coronary artery disease and healthy subjects were examined. Spectrum analysis and assessment of the coherent function between RR intervals, the ESP, and the LSP in the middle frequency band were performed. In healthy subjects, the amplitude of the ESP was higher than that of the LSP, whereas, in patients, the LSP amplitude surpassed that of the ESP. The LSP/ESP ratio was significantly higher in patients, indicating that artery dilation depended more on the reflected wave than on the primary wave. The study revealed that age had the most pronounced effect on the LSP/ESP ratio. Healthy subjects showed the highest age-dependent coherence between the RR interval and the ESP amplitude, whereas in patients this coherence was significantly lower. Measurement of the ESP and LSP amplitudes and their ratio allowed evaluation of vasomotor artery tone. Age and disease produced the maximum effects on the artery tone and decreased the correlation between the changes in the artery tone and the HR in the baroreflex range of frequency regulation.     

Presence and location of endothelin and endothelin-binding sites in human nasal mucosa under normal and metaplastic conditions

The presence and site of production of endothelin-1 (ET-1) was investigated in biopsies obtained from the nasal mucosa of 10 healthy human subjects and 10 patients affected by chronic rhinitis. The presence and localization of receptors for ET-1 was also investigated. Bioptic fragments were examined by scanning electron microscopy. ET-1 was present in the vessels and in the respiratory epithelium of normal subjects, whereas in patients affected by epithelial metaplasia induced by chronic rhinitis, it was absent in the metaplastic epithelium and present in the endothelium and vascular wall. Receptors for ET (A- and B-receptor subtypes) were localized in the vessels of the nasal mucosa, both in normal and in pathological subjects. In particular, A-receptors were identified in the vascular wall, whereas B-receptors were mainly distributed in the endothelium. We suggest that ET-1 is involved in the homeostasis of nasal blood flow (shunting the blood toward the deep cavernous plexus and inducing mucosal swelling) by an autocrine and/or paracrine mechanism. Normal epithelium seems to be important in this mechanism, since it is able to produce ET. However, when pathologic conditions induce squamous or cuboidal metaplasia, the epithelium is no longer able to play this role.     

Reflection of the Process of Emotional Perception of Short Musical Fragments in the Parameters of Long-Latency Auditory Evoked Potentials

The amplitude–temporal parameters of long-latency components of evoked potentials (recorded in symmetrical leads of the parietal regions) were studied during listening to short fragments of classical musical works, which had different emotional expressiveness, presented to healthy subjects by different modes. In random order of presentation, the maximal markedness of the N1-P2 complex and the P3 wave was found to be observed for fragments causing predominantly negative emotional experiences, while the minimal amplitude values and the highest values of the latencies were observed in response to presentation of a musical fragment assessed as emotionally neutral. Significantly higher amplitude values of these oscillations were recorded in the right parietal lead as compared to the left. The data obtained are interpreted in the context of the specific features of perception of short musical fragments in a human with the use of data concerning functional interhemispheric asymmetry when opposite-sign emotional experiences occur.     

Method for Qualitative and Quantitative Assessment of Proprioceptive Perception of Single-joint Arm Movements

The phenomenon of reproduction of the series of passive single-joint movements in the tested arm by the contralateral arm just in the course of passive movements with no visual control was studied in 35 healthy subjects and 13 post-stroke patients in order to develop a new method for objective assessment of sense of the arm motion for the detection of proprioceptive deficit and for monitoring of the changes in proprioception during rehabilitation. We examined the reproduction of flexion–extension at the elbow and wrist joints, abduction–adduction at the wrist joint and the forearm pronation–supination in both right and left arms in healthy subjects and in the affected arm in post-stroke patients. Displacements of the angles in the tested joint and a homonymous joint of the other arm were acquired by means of video recording system, goniometers, or 9-DoF inertional-magnetometric sensors. Qualitative and quantitative indicators were evaluated to assess the similarity of the passive and active movements. It has been found that the healthy subjects are able to actively reproduce the repeated passive movements at different joints of either the left or right tested arm almost simultaneously and with quite accurate reproduction of an amplitude and shape of movement. At the same time, most of post-stroke patients reproduce movements either with qualitative errors demonstrating incorrect location or wrong estimation of direction or number of repeated test movements, or with significant reduction of accuracy (increased latency or shape distortion). We proposed a method for the assessment of movement proprioception at individual joints. The procedure is easy and convenient for both physicians and patients. It does not require special heavy equipment and can easily be performed under different conditions in a wide range of patients.     

Migration inhibition test with thyroid membrane antigen in Graves-Basedow disease and nodular goiter]

The evaluation of the migration inhibition test with thyroid membrane antigen was carried out in 20 patients with Graves' disease, 13 patients with neutral nodular goiter and 13 healthy subjects. A significant inhibition of migration by thyroid antigen as compared to the control group was demonstrated only in the patients with Graves' disease (the value of the migration index was 0.57 +/- 0.07). The respective value was 0.79 +/- 0.16 in patients with nodular goiter and 0.85 +/- 0.11 in healthy subjects. The results obtained indicate the practical value of the migration inhibition test in diagnosing the thyroid diseases having an autoimmune background.     

Estimation of short musical fragments in normal subjects and patients with chronic depression

Emotional estimation of short fragments of classical music was analyzed in healthy subjects and patients with depressive neurosis and retarded depression (the depressive phase of maniac-depressive psychosis). The values and stability of point estimates varied between the studied groups. Narrowing of the range of emotional estimates was found in depressed patients. Standard deviation of estimates was related to musical key and tempo in normal subjects and patients with retarded depression.     

Novel parameters of surface EMG in patients with Parkinson’s disease and healthy young and old controls

The aim of this study was to evaluate a variety of traditional and novel surface electromyography (SEMG) characteristics of biceps brachii muscle in patients with Parkinson’s disease (PD) and compare the results with the healthy old and young control subjects. Furthermore, the aim was to define the optimal biceps brachii loading level that would most likely differentiate patients from controls. The results indicated that such nonlinear SEMG parameters as %Recurrence, %Determinism and SEMG distribution kurtosis, correlation dimension and sample entropy were significantly different between the PD patients and healthy controls. These novel nonlinear parameters, unlike traditional spectral or amplitude parameters, correlated with the Unified Parkinson’s Disease Rating Scale (UPDRS) and finger tapping scores. The most significant between group differences were found in the loading condition where no additional weights were applied in isometric elbow flexion. No major difference of SEMG characteristics was detected between old and young control subjects. In conclusion, the novel SEMG parameters can differentiate the patients with PD from healthy control subjects and these parameters may have potential in the assessment of the severity of PD.     

Dynamic Stability and Risk of Tripping during the Timed Up and Go Test in Hemiparetic and Healthy Subjects

Background

The Timed Up and Go (TUG) test is often used to estimate risk of falls. Foot clearance and displacement of the center of mass (COM), which are related to risk of tripping and dynamic stability have never been evaluated during the TUG. Accurate assessment of these parameters using instrumented measurements would provide a comprehensive assessment of risk of falls in hemiparetic patients. The aims of this study were to analyze correlations between TUG performance time and displacement of the COM and foot clearance in patients with stroke-related hemiparesis and healthy subjects during the walking and turning sub-tasks of the TUG and to compare these parameters between fallers and non-fallers.

Methods

29 hemiparetic patients and 25 healthy subjects underwent three-dimensional gait analysis during the TUG test. COM and foot clearance were analyzed during the walking and turning sub-tasks of the TUG.

Results

Lateral displacement of the COM was greater and faster during the walking sub-tasks and vertical displacement of the COM was greater during the turn in the patients compared to the healthy subjects (respectively p<0.01 and p<0.05). Paretic foot clearance was greater during walking and displacement of the COM was slower during the turn in the patients (p<0.01). COM displacement and velocity during the turn were correlated with TUG performance in the patients, however, vertical COM displacement was not. These correlations were significant in the healthy subjects. There were no differences between COM parameters or foot clearance in fallers and non-fallers.

Discussion and Conclusion

Hemiparetic patients are less stable than healthy subjects, but compensate with a cautious gait to avoid tripping. Instrumented analysis of the TUG test appears relevant for the assessment of dynamic stability in hemiparetic patients, providing more information than straight-line gait.     

Microflora of the mucous membrane of human tonsils in the normal state and in pathology

A total of 50 healthy persons and 50 chronic tonsillitis patients were examined with the use of the bacteriological method. The microflora of tonsillar mucous membrane was represented by 8 genera of facultative anaerobic microorganisms. In the biocenosis of the tonsillar mucosa of the healthy subjects the occurrence of Streptococcus was 1.5-fold greater than in that of the patients; microorganisms of the genus Staphylococcus prevailed on the tonsillar mucosa of chronic tonsillitis patients 1.3-fold prevailed more often and Klebsiella and Neisseria prevailed 4-fold more often. Among the microorganisms isolated from the patients, cultures simultaneously having two or more pathogenicity markers occurred twice as often as among the representatives of the microbiocenosis obtained from the healthy subjects. The microflora of the tonsillar mucosa of the chronic tonsillitis patients was characterized by resistance to penicillins, macrolides and aminoglygosides, but was simultaneously sensitive to lincosamides and glycopeptides.     

Flow cytometry application in the assessment of sperm DNA integrity of men with asthenozoospermia

Sperm genomic integrity and ultrastructural features of ejaculated spermatozoa contributing to the assessment of gamete fertility potential in patients with asthenozoospermia are discussed. The proportion of TUNEL-positive cells was significantly higher in the semen of patients with low sperm motility (n=40; p<0.01) as compared to men with normal sperm motility (n=54). Sperm DNA fragmentation negatively correlated (n=94) with sperm motility, sperm concentration, and integrity of the sperm cellular membrane (HOS-test). Two categories of patients were distinguished: (1) patients (23 out of 94 subjects) with < or = 4% of TUNEL-positive cells and (2) patients (71 subjects) with 4% of TUNEL-positive cells. A significant difference was noted in the sperm motility and HOS-test results between patients from both groups. Large numbers of immature spermatozoa with extensive cytoplasmic retention, ultrastructural chromatin and midpiece abnormalities, and conglomerates containing sperm fragments were present more frequently in the semen of asthenozoospermic subjects with >4% of TUNEL-positive sperm cells. Low sperm motility seems to be accompanied by serious defects of gamete chromatin expressed as diminished sperm genomic integrity and abnormal DNA condensation and by defects of sperm midpiece. These abnormalities may reflect developmental failure during the spermatogenic remodeling process. The DNA fragmentation test may be considered as an additional assay for the evaluation of spermatozoa beside standard analysis and taken together with electron microscopy may help to determine the actual number of "healthy" spermatozoa thereby playing an important role during diagnosis and treatment of male infertility.     

Porphyromonas gingivalis prevalence related to other micro-organisms in adult refractory periodontitis.

Forty-six adult periodontal patients, selected on the basis of clinical examination, and 46 adult healthy subjects were examined. The subgingival plaque samples from one inflammatory and one non-inflammatory site of each periodontal patient were studied to determine Porphyromonas gingivalis prevalence related to other periodontal micro-organisms and to periodontal tissue destruction. The results showed Porphyromonas gingivalis as the main pathogenic micro-organism isolated in the inflammatory sites together with Bacteroides forsythus. Peptostreptococcus sp., Actinomyces sp. and Prevotella sp. were found as a normal oral flora in the healthy subjects. Fusobacterium nucleatum, Prevotella intermedia, Campylobacter rectus and Eikenella corrodens were detected both in inflammatory and in non-inflammatory sites of periodontal patients as well as in the healthy subjects.     

The Insular Cortex: Relationship to Skin Conductance Responses to Facial Expression of Emotion in Temporal Lobe Epilepsy

The insula plays an important role both in emotion processing and in the generation of epileptic seizures. In the current study we examined thickness of insular cortices and bilateral skin conductance responses (SCR) in healthy subjects in addition to a small number of patients with temporal lobe epilepsy. SCR measures arousal and is used to assess non-conscious responses to emotional stimuli. We used two emotion tasks, one explicitly about emotion and the other implicit. The explicit task required judgments about emotions being expressed in photographs of faces, while the implicit one required judgments about the age of the people in the photographs. Patients and healthy differed in labeling neutral faces, but not other emotions. They also differed in their SCR to emotions, though the profile depended on which hand the recordings were from. Finally, we found relationships between the thickness of the insula and SCR to each task: in the healthy group the thickness of the left insula was related to SCR to the emotion-labeling task; in the patient group it was between the thickness of the right insula and SCR in the age-labeling task. These patterns were evident only for the right hand recordings, thus underscoring the importance of bilateral recordings.     

The use of emotional evaluations of short musical fragments for analyzing the functional states of students

Emotional evaluations of short fragments of classical music were compared in students (boys and girls) in the state of relative rest and during examinations. Substantial gender differences were found with respect to the sign and intensity of the emotional evaluations. The functional states of subjects affect the numbers of positive, neutral, and negative evaluations of musical fragments.     

Changes in the EEG spectral power during perception of neutral and emotionally salient words in schizophrenic patients, their relatives and healthy individuals from the general population

To search for EEG-correlates of emotional processing that might be indicators of genetic predisposition to schizophrenia, changes in EEG spectral power during perception of neutral and emotionally salient words were examined in 36 schizophrenic patients, 50 of their unaffected first-degree relatives, and 47 healthy individuals without any family history of psychoses. In healthy persons, passive listening to neutral words induced minimum changes in cortical rhythmical activity, predominantly in the form of synchronization of slow and fast waves, whereas perception of emotional words was followed by a generalized depression of the alpha and beta1 activity and a locally specific decrease in the power of theta and beta2 frequency bands. The patients and their relatives showed a decrease in the alpha and beta1 activity simultaneously with an increase in the power of delta activity in response to both groups of words. Thus, in the patients and their relatives, reactions to neutral and emotional words were ulterior as a result of augmented reactions to the neutral words. These findings suggest that the EEG changes reflect familial and possibly hereditable abnormal involuntary attention. No prominent decrease in reactivity to emotional stimuli was revealed in schizophrenic families.