Drosophila KASH-domain protein Klarsicht regulates microtubule stability and integrin receptor localization during collective cell migration.

During collective migration of the Drosophila embryonic salivary gland, cells rearrange to form a tube of a distinct shape and size. Here, we report a novel role for the Drosophila Klarsicht-Anc-Syne Homology (KASH) domain protein Klarsicht (Klar) in the regulation of microtubule (MT) stability and integrin receptor localization during salivary gland migration. In wild-type salivary glands, MTs became progressively stabilized as gland migration progressed. In embryos specifically lacking the KASH domain containing isoforms of Klar, salivary gland cells failed to rearrange and migrate, and these defects were accompanied by decreased MT stability and altered integrin receptor localization. In muscles and photoreceptors, KASH isoforms of Klar work together with Klaroid (Koi), a SUN domain protein, to position nuclei; however, loss of Koi had no effect on salivary gland migration, suggesting that Klar controls gland migration through novel interactors. The disrupted cell rearrangement and integrin localization observed in klar mutants could be mimicked by overexpressing Spastin (Spas), a MT severing protein, in otherwise wild-type salivary glands. In turn, promoting MT stability by reducing spas gene dosage in klar mutant embryos rescued the integrin localization, cell rearrangement and gland migration defects. Klar genetically interacts with the Rho1 small GTPase in salivary gland migration and is required for the subcellular localization of Rho1. We also show that Klar binds tubulin directly in vitro. Our studies provide the first evidence that a KASH-domain protein regulates the MT cytoskeleton and integrin localization during collective cell migration.

A cardiovascular simulator tailored for training and clinical uses.

OBJECTIVE: In the present work a cardiovascular simulator designed both for clinical and training use is presented. METHOD: The core of the simulator is a lumped parameter model of the cardiovascular system provided with several modules for the representation of baroreflex control, blood transfusion, ventricular assist device (VAD) therapy and drug infusion. For the training use, a Pre-Set Disease module permits to select one or more cardiovascular diseases with a different level of severity. For the clinical use a Self-Tuning module was implemented. In this case, the user can insert patient's specific data and the simulator will automatically tune its parameters to the desired hemodynamic condition. The simulator can be also interfaced with external systems such as the Specialist Decision Support System (SDSS) devoted to address the choice of the appropriate level of VAD support based on the clinical characteristics of each patient. RESULTS: The Pre-Set Disease module permits to reproduce a wide range of pre-set cardiovascular diseases involving heart, systemic and pulmonary circulation. In addition, the user can test different therapies as drug infusion, VAD therapy and volume transfusion. The Self-Tuning module was tested on six different hemodynamic conditions, including a VAD patient condition. In all cases the simulator permitted to reproduce the desired hemodynamic condition with an error<10%. CONCLUSIONS: The cardiovascular simulator could be of value in clinical arena. Clinicians and students can utilize the Pre-Set Diseases module for training and to get an overall knowledge of the pathophysiology of common cardiovascular diseases. The Self-Tuning module is prospected as a useful tool to visualize patient's status, test different therapies and get more information about specific hemodynamic conditions. In this sense, the simulator, in conjunction with SDSS, constitutes a support to clinical decision - making.

Experimental integration of Autoregulation Unit for left ventricular assist devices in a cardiovascular hybrid simulator.

In this paper, an Autoregulation Unit (ARU) for left ventricular sensorized assist devices (LVAD) has been used with a cardiovascular hybrid simulator mimicking physiological and pathological patient conditions. The functionalities of the ARU have been demonstrating for the successful receiving and visualization of system parameters, sending of commands for LVAD speed changes, and enabling of the autonomous flow control algorithm. Experiments of speed changes and autoregulation are reported, showing the feasibility of the approach for both local and remote control of a LVAD.

Application of a user-friendly comprehensive circulatory model for estimation of hemodynamic and ventricular variables.

PURPOSE: Application of a comprehensive, user-friendly, digital computer circulatory model to estimate hemodynamic and ventricular variables. METHODS: The closed-loop lumped parameter circulatory model represents the circulation at the level of large vessels. A variable elastance model reproduces ventricular ejection. The circulatory model has been modified embedding an algorithm able to adjust the model parameters reproducing specific circulatory conditions. The algorithm reads input variables: heart rate, aortic pressure, cardiac output, and left atrial pressure. After a preliminary estimate of circulatory parameters and ventricular elastance, it adjusts the amount of circulating blood, the value of the systemic peripheral resistance, left ventricular elastance, and ventricular rest volume. Input variables and the corresponding calculated variables are recursively compared: the procedure is stopped if the difference between input and calculated variables is within the set tolerance. At the procedure end, the model produces an estimate of ventricular volumes and Emaxl along with systemic and pulmonary pressures (output variables). The procedure has been tested using 4 sets of experimental data including left ventricular assist device assistance. RESULTS: The algorithm allows the reproduction of the circulatory conditions defined by all input variable sets, giving as well an estimate of output variables. CONCLUSIONS: The algorithm permits application of the model in environments where the simplicity of use and velocity of execution are of primary importance. Due to its modular structure, the model can be modified adding new circulatory districts or changing the existing ones. The model could also be applied in educational applications.

[Stereotactic biopsy and brachytherapy in the diagnostics and treatment of brain tumors--preliminary report]. / Wczesne wyniki zastosowania biopsji stereotaktycznej i brachyterapii w diagnostyce i leczeniu guzów mózgu.

Promising results have been obtained using brachytherapy in the treatment of brain tumors. Between November 99 and August 2000, 28 patients with brain tumors (15 newly diagnosed gliomas, 11 recurrent gliomas, 2 metastases) underwent implantation of temporary iridium 192 sources with stereotactic technique. This group received external beam radiation therapy (45 Gy) following implantation. Patients were followed-up with CT scans every 3 months. Serious complications occurred in two patients (postradiation brain oedema). Median survival has not been assessed due to short follow-up period and small number of patients. Further clinical assessment is required especially long-term follow-up. Brachytherapy appears to be a useful technique for the treatment of selected brain tumors.

[Clinical and immunohistoenzymatic investigations in patients with vasomotor and perennial allergic rhinitis]. / Badania kliniczne i immunohistoenzymatyczne u chorych z naczynioruchowym i calorocznym alergicznym niezytem nosa.

The aim of our study was evaluation of the results of clinical examination in patients with vasomotor and perennial allergic rhinitis and assessment of number of blood vessels, nerve fibres and mast cells on the basis of immunohistoenzymatic examination. There were 42 patients examined aged from 18 to 50 and divided into three groups: I--16 patients with vasomotor rhinitis and II--14 patients with perennial allergic rhinitis and III--12 patients (control) with nasal septum deviation. On the basis of the patient's history data, clinical otorhinolaryngologic examination and active anterior rhinomanometry the patients were qualified to bilateral inferior turbinectomy. The nasal mucosa removed during surgery underwent immunohistoenzymatic examination using the monoclonal antibody against the tryptase of mast cells (MCT company, DAKO), the endothelin of blood vessels (EC - DAKO) and the neurospecific enolase (NSE - DAKO). In examined groups of patients with vasomotor and perennial allergic rhinitis and control group similar escalation of clinical symptoms expressed by means of points index were stated. In immunohistoenzymatic studies the differences in mean number of blood vessels and nerve fibres between examined groups were not statistically significant, however statistically significant difference concerned higher number of mast cells patients with vasomotor rhinitis in comparison to perennial allergic rhinitis.

[The case of giant pleomorphic adenoma of the parotid gland with the malignancy features]. / Przypadek olbrzymiego gruczolaka wielopostaciowego slinianki przyusznej z cechami zezlosliwienia.

The aim of the study was showed the case of patient aged 41 years, treated in the Otolaryngology Department of Military Medical Academy because of giant pleomorphic adenoma of the parotid gland with the malignancy features. After performed of the BAC of tumour, the CT of neck and the scintigraphy of salivary glands the patient qualified for operation. The tumour resected with capsule and the superficial lobe of the left parotid gland, conservating of function of the facial nerve. The histopathologic examination proved the mucoepidermoid carcinoma in the pleomorphic adenoma didn't infiltrate of capsule. After oncologic consultation the patient had not of complementary radiotherapy, because the operation was radical and the carcinoma had a low radiosensitivity. The above mentioned case confirms of the reports, that the pleomorphic adenoma can undergo of malignancy changes, however the gigantic tumour doesn't write off possibility of operating success.

Comparison of radiometric and histometric features of bone samples from ribs.

OBJECTIVE: To correlate quantitative data gathered from conventional radiograms as well as from quantitative computed tomography (QCT) images of bone samples with histometric coefficients of the same specimens. STUDY DESIGN: Bone samples were obtained from standardized sites of ribs during autopsy of 40 males aged 15-84 years. In the bone samples the following assessments were performed: densitometry of bone using QCT, estimation of radiostructural quantitative coefficients (coefficient of trabecular arrangement anisotropy, trabecular thickness coefficient); histometry (area of bone matrix, trabecular thickness coefficient); relative weights of dry, decalcified bone and bone in ash form; and concentration of calcium in ash. Radiometry and histometry were performed using image analysis methods with IBAS for Windows (Kontron, Eching b. München, Germany) equipment. Radiostructure of bone was evaluated with the use of our own algorithm; concentration of calcium was measured spectrophotometrically. Analysis of variance, cross correlation, regression analysis and others were the statistical tools applied. RESULTS: From the variables analyzed, the following indicates a significant correlation with the histometric area of bone matrix: radiologic trabecular anisotropy, QCT densitometry (especially of cancellous bone) and the ratio of weight of dry sample to fresh sample. The coefficient of radiologic trabecular thickness appeared not to be a valuable feature for estimation of bone histology. CONCLUSION: Quantitative estimation of radiostructure of bone carried out on conventional radiograms may be helpful in the detection of histologic properties of bone, an important factor in assessment of the fragility of bone.

Escape from shock versus escape from shock accompanied by a visual stimulus in rats.

Two groups of 15 rats each were trained in a shuttle box to escape foot-shock either unsignalled or presented in compound with a visual cue: darkness. The visual cue presented in shock compartment amplifies the behavioral tendency actually prevailing in the response repertory of the rat. During the 1st session the compound enhanced the species-specific flight resulting in shortening of the rat's escape latency. Thereafter, during subsequent sessions, darkness exaggerated resistance to enter the other compartment; thus escape latencies were longer under compounded than under unsignalled procedure. The darkness cue reduces the intertrial response rate relative to the unsignalled group. This latter finding supports the discrimination model of the effect, since the compound helps the animals to discriminate the illuminated "safe" period between trials from the aversive shock period. Our data seem to suggest that the darkness presented synchronously with escapable grid-shock acquires aversive properties.

[Delay inhibition of the active avoidance reflex and intertrial responses in rats in the shuttle box]. / Zapazdyvatel'noe tormozhenie reaktsii aktivnogo izbeganiia i mezhsignal'nye reaktsii v chelnochnoi kamere u krys.

A diminished probability of avoidance response in early phases of a warning signal was revealed with salient signals given after short intertrial intervals. The inhibition of the delay in avoidance response is due to an interaction of the safety state conditioning and the excitation elicited by onset of warning signal.

Defensive conditioning-related functional heterogeneity among nuclei of the rat amygdala revealed by c-Fos mapping.

The amygdala is a complex forebrain structure proposed to play a pivotal role in fear conditioning circuitry. In this study, c-Fos immunomapping was applied to investigate the functional activation of particular amygdalar nuclei following a 50-trial training session of two-way active avoidance reaction. To dissect distinctive responses displayed by the animals and to cluster them into groups of correlated behaviors, factor analysis was employed. The training procedure resulted in an increase of c-Fos expression within the cortical, medial, lateral and basolateral, but not central, nuclei. The expression in the cortical nucleus correlated negatively with grooming behavior, whereas c-Fos immunolabeling of the other three subdivisions of the amygdala could be associated with the number of intertrial responses. No correlation was observed between c-Fos expression and avoidance reactions performed or the amount of shock received by the animal. The results obtained with c-Fos mapping of various regions of rat amygdala, combined with a fine dissection of behavioral repertoire, imply that there are specific functional links between particular parts of the structure and distinctive behaviors that reflect various emotional states of the animal.

How do rats cope with the two-way escape problem in a homogeneous shuttle box?

The behavior of 25 rats trained in a homogeneous shuttle box to escape unsignalled grid-shock was analyzed. Three categories of escape were distinguished: (1) species-specific fly away from the charged grid, (2) long-latency crossing preceded and accompanied by other behaviors that compete with the escape response, and (3) short-latency escape which followed an anticipatory postural pose. The animals displayed species-specific fly away only during the initial trials of a session. Subsequently long-latency crossings develops, reflecting a resistance to enter the opposite compartment. A measure based on a comparison of escape latency distributions in the two halves of the 1st session discriminates between good and poor learners. Subgroups of good and poor learners differed in performance efficiency in all five training sessions. Good learners were able to overcome the resistance to enter the opposite compartment and recall the learned short-latency escape.

Effects of octreotide on propylotiouracil-induced goiter in rats: a quantitative evaluation.

To evaluate the possible antigoitrogenic effect of somatostatin, the influence of long-acting somatostatin analog--octreotide--on experimental goiter developed in rats treated with propylthiouracil was examined. Goiter formation was assessed by measurement of the main histological compartments of the thyroid as well as by morphometric analysis of the vascularization and blood supply of the gland. Although treatment with octreotide did not prevent the goiter formation, it clearly reduced blood supply and vascularization of the thyroid and counteracted propylthiouracil-induced increase in the relative volume of follicular epithelium. To conclude, the somatostatin analog--octreotide--is effective in reduction of goiter vascularisation. This finding provides a rationale for the clinical trials of the treatment of hypervascular goiter by somatostatin analogs.

CS modality transfer of two-way avoidance in rats with central and basolateral amygdala lesions.

Post-lesion acquisition of two-way avoidance and subsequent transfer to two warning signals (conditioned stimulus, CS) of different modality were investigated in 60 rats. In Experiment I the animals were originally trained with less salient (darkness) CS, then transferred to more salient compound (darkness and white noise), and finally to white noise CS. The opposite arrangement of the conditioned stimuli (CSi) during the subsequent stages was employed in Experiment II. In control animals, avoidance acquisition was faster and the intertrial responding (ITR) rate lower with the auditory than with the visual CS. Lesioned rats learned avoidance responses more slowly, independently of CS modality. The transfer to other CSi revealed dramatic between-group difference in the level and consistency of avoidance response, shuttle-box latencies and ITR rate. In control animals, transfer to more salient CSi enhanced avoidance performance, whereas change to less salient CS decreased it. Rather small changes in shuttle-box performance and consistency of avoidance response due to CS modality were seen in rats with the basolateral lesions. In contrast, central nucleus injury caused a strong deterioration in the avoidance transfer, especially when the visual CS followed the acoustic one. The results indicate differential involvement of the basolateral and central amygdala nuclei in stimulus-processing mechanisms of instrumental defensive behavior.

[The dependence of learning an active avoidance reaction on overcoming a problem-solving situation in a shuttle box]. / Zavisimost' obucheniia reaktsii aktivnogo izbeganiia ot preodoleniia problemnoi situatsii v chelnochnoi kamere.

The two-way avoidance procedure dramatically differs from the one-way procedure in rate of learning. The present study was conducted to prove that retardation of the two-way avoidance acquisition resulted from development of the behavioral conflict tendency not to reenter the previous shock compartment. Cluster analysis of avoidance response indices divided rats into three distinctive classes. The occurrence of avoidance, escape, and freezing responses in the first session was analyzed in these three groups. Freezing during the shock action reflected the conflict tendency and showed a negative correlation with avoidance response indices. Only the rats which overcame the conflict and performed at least one avoidance reaction toward the end of the first session significantly improved their avoidance score in the following session. Discriminant analysis of indices of the three reaction types in the first session revealed sufficiency of these indices for prediction of the success in avoidance learning in subsequent sessions. Our results proved the hypothesis formulated previously that the conflict situation inherent to the two-way shuttle box procedure retarded the active avoidance acquisition. A number of trials have to be reserved in the first session for solving the conflict situation (direct effect on the rate of learning). The conflict intensity which directly effects the avoidance performance in the first session, presumably, influences learning in subsequent sessions (secondary effect on learning).

[The study of individual differences as a method for dividing into stages the acquisition of a complex reflex]. / Izuchenie individual'nykh razlichii kak metod vychleneniia étapov vyrabotki slozhnogo refleksa.

Two-way active avoidance learning is a complex task characterized by a high level of interindividual variability. We have demonstrated in our previous paper that rats with different rate of learning vary in expression of freezing responses, which reflects a conflict tendency not to re-enter the previous chock compartment [4]. In the present work we analyzed as main indices the proportions of avoidance, rapid escape, and freezing responses and their distribution within a session. Sequences of occurrence of these three types of responses were examined in groups of rats with different success of avoidance learning. The obtained results confirmed a hypothesis that overcoming the conflict tendency was a separate stage of learning which preceded avoidance response appearance and subsequent stabilization. Only in rats which failed to learn and develop avoidance responses within the first session, freezing responses were observed in successive session being indicative of reappearance of the conflict tendency. The intensity of the conflict tendency did not depend on the type of the first response of an animal (fleeing or freezing) to an unexpected aversive stimulus.

[Macroscopic estimation of the placenta using a morphometric grid. Part I: pre-pregnancy and post-pregnancy diabetes mellitus]. / Ocena makroskopowa lozyska przy zastosowaniu siatki morfometrycznej. Czesc I: Cukrzyca przedciazowa i ciazowa.

Kurt Benirschke has once written that the placenta provides the most accurate records of prenatal history of foetus. Our study included 104 placentas obtained from pregnancy complicated by diabetes. An obstetrician did the planimetric estimation directly after delivery. A measurement of lesions was conducted with using a special own idea's morphometric grid that allowed us to evaluate a placental surface and a relative size of change statement in a very quick and convenient way. Our method is very useful for making a quantitative estimation of pathological changes in the chorionic plate of placenta.

[Macroscopic assessment of the placenta using a morphometric grid. Part II: other selected pathologies]. / Ocena makroskopowa lozyska przy zastosowaniu siatki morfometrycznej. Czesc II: Inne wybrane patologie.

Kurt Benirschke has once written that the placenta provides the most accurate records of prenatal history of foetus. Our study included 2498 placentas obtained from pregnancy complicated by pregnancy-induced hypertension (PIH), intrauterine growth retardation (IUGR) and epilepsy. An obstetrician did the planimetric estimation directly after delivery. A measurement of lesions was conducted with using a special own idea's morphometric grid that allowed us to evaluate a placental surface and a relative size of change statement in a very quick and convenient way. Our method is very useful for making a quantitative estimation of pathological changes in the chorionic plate of placenta.

[Macroscopic assessment of the placenta using a morphometric grid. Part III: preterm delivery]. / Ocena makroskopowa lozyska przy zastosowaniu siatki morfometrycznej. Czesc III: Poród przedwczesny.

Kurt Benirschke has once written that the placenta provides the most accurate records of prenatal history of foetus. Our study included 198 placentas obtained from pregnancy complicated by preterm delivery. An obstetrician did the planimetric estimation directly after delivery. A measurement of lesions was conducted with using a special own idea's morphometric gird that allowed us to evaluate a placental surface and a relative size of change statement in a very quick and convenient way. Our method is very useful for making a quantitative estimation of pathological changes in the chorionic plate of placenta.