[Identifying the Importance of MT-3 Expression for Neuroblastoma Cells]. / Identifikace významu exprese MT-3 pro bunky neuroblastomu.

BACKGROUND: Resistance of cancer cells to cytostatics is caused by a number of mechanisms that are often combined. These include reduced cell entry or increased efflux, increased DNA repair, defects of, apoptotic pathways, increased cytostatic degradation as well as elevated levels of intracellular thiols of glutathione and metallothioneins (MT). It has been reported that high concentrations of thiol groups in the cytoplasm bind platinum alkylation derivatives and chemorezistence is due to the transfer of platinum from the cytostatic to MT, which inactivates them. Because we have shown an increase in MT levels in resistant neuroblastoma (NB) lines, but not in sensitive lines after incubation with platinum cytostatics, we have considered MT-3 for NB cells in our previous studies. METHOD: SiMa NB cell lines transfected with vector containing human MT-3 and GFP or GFP only (control). Expression Microarray Human Cancer 3711 ElectraSense medium density 4 × 2k array slides with 1,609 DNA probes (Custom Array, Bothell, WA, USA), MT-3 expression and most expressed genes validated by real-time polymerase chain reaction. Sensitivity to CDDP (cisplatin) - MTT assay, clonogenicity test, Western blott caspase cleavage and free oxygen radicals fluorescence microscopy after CellROX Deep Red Reagent staining. Levels of MT-3 mRNA in 23 samples of high-risk NB, normal human cortex and bovine adrenal glands were investigated by reverse transcription polymerase chain reaction. RESULTS: Expression microarray showed downregulation 3 and overexpression of 19 genes in MT-3 transfected NB cells. Using gene ontology, over-expressed genes have been shown to drive senescence-induced oncogenes (CDKN2B and ANAPC5), and the genes of glutathione S-transferase M3, caspase 4 and DNAJB6 (chaperone neuronal proteins) were also expressed. We have demonstrated a reduced sensitivity of MT-3 transfected cells to CDDP (24h IC50 of 7.48 ± 0.97 and 19.81 ± 1.2 µg/ml), a higher number of colonies after incubation with CDDP, reduced caspase 3 after incubation with CDDP and lower free oxygen radicals after induction of CDDP. High-grade NB cells expressed MT-3 significantly more than non-tumoral adrenal cells but failed to show a clear relationship to disease course. CONCLUSION: We have demonstrated the relationship between MT-3 and senescence-induced oncogene genes and some other genes relevant to cell fate (glutathione S-transferase M3, caspase 4 and DNAJB6) and a significant proportion of MT-3 on CDDP resistance. High levels of MT-3 in high-risk NB could be one of the causes of frequent relapses in this tumor.Key words: neuroblastoma - metallothionein 3 - chemoresistance The authors declare they have no potential conflicts of interest concerning drugs, products, or services used in the study. The Editorial Board declares that the manuscript met the ICMJE recommendation for biomedical papers.This work was supported by AZV CR grant 15- 28334A. Submitted: 17. 2. 2018Accepted: 16. 4. 2018.

Craniofacial abnormalities and their relevance for sleep apnoea syndrome aetiopathogenesis in acromegaly.

OBJECTIVE: To explain the effect of craniofacial relations on the development of the sleep apnoea syndrome (SAS) in acromegaly, and to elucidate how the activity of acromegaly affects the severity of SAS. DESIGN: Prospective observational study. METHODS: Cephalometry and sleep ventilation measurements were performed in 26 acromegalic men and in 96 men with SAS. RESULTS: SAS was found in 20 acromegalic men. Compared with non-acromegalic men with SAS, patients with acromegaly and SAS were found to have: enlargement of almost all linear dimensions; increased angle indicating mandibular protrusion; increased difference between maxillary and mandibular protrusion; articular angle decrease; soft palate lengthening; and pharyngeal airway space (PAS) enlargement in the palatal and uvular-tip planes. A comparison of acromegalic men with and without SAS revealed no significant difference in the craniofacial skeleton, although there was a narrowing of the minimal PAS (MinPAS) and of PAS in the uvular-tip plane in patients with SAS. SAS was more frequent in the patients with active acromegaly. MinPAS in the patients with active acromegaly was narrower than in those without disease activity. CONCLUSION: Skeletal abnormalities in acromegalic men with SAS were different from those in SAS patients without acromegaly. Upper airway narrowing due to changes in pharyngeal soft tissues takes a more relevant share in the development of SAS in acromegalic men than skeletal anomalies.

[Initial experience with use of cationic propyl gallate as an inducer of thrombocyte aggregation in evaluation of anti-aggregation therapy]. / První zkusenosti s vyuzitím kationického propylgallátu jako induktoru agregace trombocytu v hodnocení úcinnosti antiagregacní terapie.

INTRODUCTION AND OBJECTIVE: Acetylsalicylic acid (ASA) is one of the basic preparations which are used in the treatment of cardiovascular diseases. ASA administration leads to irreversible restriction of platelet aggregation. The objective of our work was to test possibilities of monitoring the effectiveness of ASA therapy by measuring the platelet aggregability in vitro after induction with cationic propyl gallate (CPG) which is considered a very potent aggregation inductor. METHOD: We examined a group of 27 healthy volunteers divided into two sub-groups (n = 19, n = 8). In the first sub-group the platelet aggregation was examined before and after 24 hours following ingestion of 400 mg ASA after induction with ADP, collagen, adrenaline and CPG. In the second sub-group the platelet aggregation was examined before and after three-day administration of ASA--100 mg/day. RESULTS AND CONCLUSION: In a group of 27 volunteers we assessed normal values of aggregation after different inductors. A low stability of the methods used was proved (low stability or insignificant correlation of results of the same method before and after ASA ingestion. The most useful parameter by means of which it was possible to monitor the effectiveness of administration of 400 or 100 mg ASA was the C/G slope (paired t-test, p < 0.0000002, and p < 0.001 resp.). In parameter CPG slope we were able to assess in both groups the cut-off value (< 53%/min.) by means of which it is possible to discriminate probands according to ASA therapy (contrary to the other commonly used inductors). From the results ensues that when assessing the thrombocyte aggregation after CPG induction we find a significantly lower percentage of so-called ASA non-respondents than after other inductors. We consider the use of assessment of thrombocyte aggregation after CPG induction when monitoring antiaggregation therapy a very promising procedure.

Prevalence of the sleep apnea syndrome in acromegaly population.

The prevalence of sleep apnea syndrome (SAS) in acromegaly is high. Consequences of SAS are serious and are associated with increased morbidity and mortality. The aim of this study was to assess the relative frequency and predictive factors for SAS in a group of patients with acromegaly (n=55). The presence of SAS was evaluated using the Polymesam device. Hormonal and clinical examination consisted of assessment of growth hormone, insulin-like growth factor I plasma levels, body mass index (BMI), neck circumference, age, sex, treatment modes of acromegaly and ear, nose and throat (ENT) examination. The relative frequency of SAS in our group of patients with acromegaly was 75%. Independent predictors of SAS were: increased activity of acromegaly, higher age and neck circumference. No association between SAS and BMI and ENT findings was observed. The role of gender was controversial.

[Comparison of cephalometric parameters in patients with sleep apnea syndrome and normal individuals]. / Srovnání cefalometrických parametru nemocných se spánkovým apnoickým syndromem s normou.

BACKGROUND: Craniofacial abnormalities which predispose to pharyngeal obstruction can be detected by cephalometry. The aim of the present study was to compare cephalometric findings of the patients with sleep apnoea syndrome (SAS) with those of healthy population. METHODS AND RESULTS: The SAS patients (23 females, 96 males) and normal population (36 females, 50 males) were examined by cephalometry. In the group of females with SAS following significant differences were observed: elongation of the soft palate (p < 0.001), narrowing pharyngeal airway space (p < 0.001), mandibular retrognathism (p < 0.05), micrognathism of the mandible (p < 0.001), dorsocaudal rotation of the mandible (p < 0.01), increased lower gonion angle (p < 0.001) and the articular angle (p < 0.05), increased anterior facial height (p < 0.05), narrowing the depth of bony framework of the nasopharynx (p < 0.001), decreased depth of the upper face (p < 0.01) and decreased length of the cranial base (p < 0.01). In the group of males with SAS, following significant differences were observed: elongation of the soft palate (p < 0.001), narrowing pharyngeal airway space (p < 0.001), decreased depth of the maxilla (p < 0.001), decreased angle of the cranial base (p < 0.01), decrease saddle angle (p < 0.01), decreased depth of the upper face (p < 0.01), increased lower gonion angle (p < 0.01), increased articular angle (p < 0.001) and increased anterior and posterior facial height (p < 0.001). CONCLUSIONS: There are important differences in orofacial skeleton and soft tissue of the nasopharynx measured by cephalometry in the patients with sleep apnoea compared to normal population.

[The sleep apnea syndrome in hypophyseal disorders with the exception of acromegaly]. / Syndrom spánkové apnoe pri hypofyzárních afekcích s výjimkou akromegalie.

The aim of the study was to ascertain the occurrence of the sleep apnoea syndrome (SAS) in patients after pituitary afunctional adenoma operation or in patients with other affection in sella turcica region except acromegaly patients. Eighty-one patients were addressed. Sixteen patients (eight men, eight women) underwent the examination; the rest of them had no interest to be explored. Afunctional pituitary adenoma was diagnosed in twelve patients, afunctional pituitary adenoma and subsequently optic glioma in one patient, craniopharyngioma in one patient, prolactinoma in one patient and undifferentiated cystic pituitary tumour in one patient. Neurological examination was done in all patients (standardized sleep questionnaire was included). SAS was quantified using PolyMESAM. Severity of SAS were set up from the value of M index (M = ODI.delta saturation; ODI--oxygen desaturation index). Mild SAS: 40 < M < 100; middle SAS: 100 < M < 210; severe SAS: M > 210. SAS was diagnosed in 13 patients: mild SAS in three of them (average value of M index: 81.0 +/- 12.7), moderate SAS in seven patients (average value of M index: 129.0 +/- 18.7) and severe SAS in three patients (average value of M index: 790.0 +/- 563.0). Therapy by continuous positive airway pressure (CPAP) was recommended in six patients. Five of them continue in using of CPAP--four patients with good compliance. The occurrence of SAS was 16.05% in measured group, higher than in normal population.

Seizures in sleep apnea patients: occurrence and time distribution.

A retrospective survey of consecutive 480 adult patients (394 men, 96 women, mean age 50.3, SD +/- 10.4 years) with sleep apnea syndrome (SAS) was made in attempt to find out among them the patients with epilepsy. In total 19 subjects (17 men, 2 women, average age 55.8 +/- 8.7 years) representing 4.0% experienced at least two seizures in adult age. The average age at the first seizure was 48.4 +/- 16.1 years. Ten subjects suffered from primarily generalized tonic-clonic seizures, seven from secondarily generalized tonic-clonic seizures, one had complex partial seizures and one simple partial seizures. In 15 subjects (78.9%) the seizures occurred exclusively during sleep. Two patients (11.1%) had seizures exclusively during vigilance and in remaining two subjects the seizures occurred during sleep and vigilance. The findings of high proportion of subjects with epilepsy among SAS subjects, of late seizure onset and of predominantly sleep occurrence of seizures suggest that sleep apnea and hypopnea can facilitate epileptic seizures.

[Sleep apnea syndrome in acromegaly. Treatment and development-- retrospective analysis]. / Spánkový apnoický syndrom (SAS) u akromegalie. Lécení a vývoj--retrospektivní analýza.

In total 56 subjects with acromegaly (37 men and 19 females) were examined in attempt to find sleep apnoea syndrome (SAS). The examination consisted of clinical examination and of all-night monitoring MESAM4 or polyMESAM. The diagnosis of SAS was established in 76.4% of subjects: 31 men and 11 females, average age 51.8 (SD +/- 9.6) years. The average ODI (oxygen desaturation index--number of oxygen saturation drops per 1 hour of sleep) of SAS patients was 29.2 (+/- 20.7). The therapy of SAS was recommended to 27 patients: sleeping position on the side (3 patients), reduction of the weight (8 patients), change of hypnotic drug (1 patient) and CPAP--continuous positive airway pressure (24 patients). CPAP titration was performed in 18 patients (in one subject the titration was repeated 4 years later). CPAP was titrated within all-night polysomnography in 10 subjects and in 9 subjects using self adjusting CPAP. Primary acceptance of CPAP was 94.4%. The average CPAP pressure was 7.8 (+/- 2.1) mbar. The theoretical duration of CPAP use was 546.6 (+/- 533.7) days. Long term compliance was considered as sufficient (weekly 25 hours or more) in 66.7% of patients. Ten patients underwent important acromegaly therapy or its change during the follow up and the improvement or the disappearance of SAS symptoms occurred in 6 of them.

Craniofacial abnormalities in sleep apnoea syndrome.

One hundred and four men with the suspected diagnosis of sleep apnoea syndrome (SAS) not suffering from neuromuscular diseases or acromegaly were examined by X-ray cephalometry. Subjects suffering from bronchial obstruction, laryngeal carcinoma, suspected Treacher Collins syndrome as well as subjects in whom SAS was not verified were excluded. The remaining 81 men were divided into two groups depending upon disease severity as expressed by the mean index of oxygen desaturations per hour (less than 30-group A; more than 30-group B). In group B, the following parameters were significantly altered as compared with group A: dorsocaudal rotation of the mandible, elongation of the soft palate, and an increase of the saddle angle, increase of the angle of the cranial base, the lower gonion angle and the angle of the inclination of the mandible. Differences in the size of the soft palate, the rotation of the mandible and the size of the lower gonion angle can be found not only between healthy and subjects with SAS but also between subjects with mild and severe apnoea.