Preventable Hazards from in Vitro Fertilization - A Case Series of CF Patients from Bulgaria.

Pre-implantation genetic diagnosis (PGD) is not often performed when donor gametes are used, due to its high cost. This is with the presumption that the donors are healthy. We report on five cases of babies with confirmed cystic fibrosis (CF), being the result from in vitro fertilization (IVF) with donor (4 cases) or own gametes (one case). There has been no family history for CF in any of the families affected. The clinical presentation in the children ranged from meconium ileus to recurrent respiratory infections and severe nasal polyposis. The age of diagnosis also varied from birth until 9 years. Since one of the presented cases was discovered in a very renowned private IVF clinic, the clinic changed their own protocol, and currently they test every donor for CF carriership. The percentage of CF carriers in the donor population is roughly the same as the one predicted in the general population of Bulgaria - 1/33. Although PGD is costly, the costs for proper care for a CF patient are currently much higher. The more economical option would to screen every donor for CF carriership. IVF requires a lot of physical and psychological stamina. The couples that go through this procedure also require a great deal of hope. It is essential to be more preconscious for possible congenital diseases. We advocate every IVF center to test the donors for CF carriership or to provide PGD for their clients.

Predominance of influenza B/Yamagata lineage viruses in Bulgaria during the 2017/2018 season.

In this study, we investigated the antigenic and genetic characteristics of influenza viruses circulating in Bulgaria during the 2017/2018 season. The detection and typing/subtyping of influenza viruses were performed using real-time RT-PCR. Results of antigenic characterisation, phylogenetic and amino acid sequence analyses of representative influenza strains are presented. The season was characterised by the predominance of B/Yamagata viruses, accounting for 77% of detected influenza viruses, followed by A(H1N1)pdm09 (17%), B/Victoria (3.7%) and A(H3N2) (2.4%). The sequenced B/Yamagata, B/Victoria, A(H1N1)pdm09 and A(H3N2) viruses belonged to the genetic groups 3, 1A, 6B.1 and 3C.2a1, respectively. Amino acid analysis of B/Yamagata isolates revealed the presence of three changes in haemagglutinin (HA), eight changes in neuraminidase (NA) and a number of substitutions in internal proteins compared with the B/Phucket/3073/2013 vaccine virus. Despite the amino acid changes, B/Yamagata viruses remained antigenically related to the vaccine strain. B/Victoria isolates fell into a group of viruses with double deletion (Δ162-163) in HA1. Substitutions in HA and NA sequences of B/Victoria, A(H1N1)pdm09 and A(H3N2) viruses were also identified compared with the vaccine strains, including in antigenic sites. The results of this study confirm the genetic variability of circulating influenza viruses and the need for continual antigenic and molecular surveillance.

Middle Term Results of Simple Open Hip Reduction of Irreducible DDH - What Is the Cut-off Age to Safely Perform It with Lower Complications?

PURPOSE OF THE STUDY When developmental dysplasia of the hip (DDH) is irreducible by conservative means, then surgical open hip reduction is the choice. When done before walking age simple open hip reduction (SOHR) is most often enough to stabilize the hip. We tested the hypothesis that simple open hip reduction gives enough stability even in severe Tönnis 4 grades of dislocation. We tried to find what is the cut-off age to safely perform SOHR with lower complications. MATERIAL AND METHODS From 193 open hip reductions (OHR) of irreducible DDH in 123 children for the period 1995-2010 year with X-ray follow up of at least 8 years and full documentation we investigated 75 SOHR. Mean age at follow-up was 13 years and 5 months for the whole group and 8 years and 3 months for the SOHR patients. Age at operation - 7 months to 7.5 years for all patients with OHR, with average OR age for SOHR - 18 months. The traditional surgical technique of open hip reduction was performed through lateral Murphy's approach but with sparing the attachment of the piriformis muscle with the underlying blood supply to the epiphysis during circumferential capsulotomy. We used McKay clinical criteria, radiological classifications of Tönnis, Severin, Herring-Mose and Kalamchi & MacEwen. RESULTS When analyzing the results, there came up a strong statistical correlation between bilaterality and Tönnis grade 4. The final CE angle of Wiberg was on average 28º. According to Severin classification: 74% were excellent and 10% good or 84 % successful results. But according to Herring-Mose sphericity scoring - 60% were good and 30% fair. Mose's fair is a potential cam-type femoro-acetabular impingement (FAI). Clinically according to McKay criteria - 10% excellent and 54% good, or 64% of the operated hips were clinically acceptable, which correlates more with the radiological results according to Mose than with Severin. Avascular necrosis (AVN) of the femoral head (FH) according to Kalamchi was: Type I - 6%, Type II - 12 %, Type III - 0%, Type IV - 8 %. Types II and IV represent 20 % of the operated hips. The presence of ossific nucleus in the femoral epiphysis on initial X-ray didn't have a statistical impact on AVN frequency. When analyzing the impact of severity of dislocation and trying to find the cut-off age for fewer complications, there came up a strong statistical significance between AVN IV type appearance in Tonnis grade 4 hips when age at operation was above 9 months. CONCLUSIONS The thorough capsuloplasty after FH reduction in the acetabular socket is enough for stability even in Tonnis 4. When OR age is below 9 months severe AVN is lower and Herring-Mose FH sphericity scoring is higher. Key words: DDH, simple open hip reduction, avascular necrosis of femoral head.

Neonatal muscle hypotonia--an early manifestation of cerebral palsy.

UNLABELLED: Cerebral palsy causes a number of diagnostic, therapeutic, organisational, and social problems. A timely diagnosis reduces the unfavourable effects of the disease and the resulting disability of the children. AIM: The aim of the present study was to evaluate the role of the neonatal muscle hypotonia as an early indicative symptom of cerebral palsy (CP). MATERIAL AND METHODS: Forty-two children with clinical and anamnestic data of muscle hypotonia (from the neonatal period to the age of 3) were enrolled in the study. An active and dynamic follow-up including specialized neurologic examinations was realised. RESULTS: The etiologic factors of utmost importance for CP were prematurity (40.5%) and perinatal asphyxia (26.2%). On the background of differently expressed muscle hypotonia, persisting from the neonatal period, the abnormalities in the psychoneurologic development during the first year after birth were characterized mainly by psychoemotional retardation. Motor deficit was marked from the 1st through 2nd year, and speech disorders appeared after the age of 2. CONCLUSIONS: The presence of partial or generalized muscle hypotonia in the neonatal age and its persisting in the subsequent stages of the child's life is an early indicative symptom of CP. The diagnosis is possible within the first two years of life, especially in children with perinatal pathology, by dynamic neurologic follow-up and prompt evaluation of the abnormalities in the psychoneurologic development.

[Effect of bombesin on the frequency of slow potentials of the smooth muscles of the gastrointestinal tract and migrating myoelectric complex]. / Vliianie bombezina na chastotu medlennykh potentialov gladkikh myshts zheudochno-kishechnogo trakta i migriruiushchii mioélektricheskii kompleks.

In dogs with electrodes implanted in the smooth muscle wall of the stomach, small intestine and colon, i.v. administration of bombesin (50, 100, 200 and 300 ng/kg) during the 2nd phase of the migrating myoelectric complex (MMC) increased the dose-dependent frequency of slow potential in the small intestine and stomach leaving unchanged the frequency of slow potentials in the colon. Single doses of 50 or 200 ng/kg bombesin during the 2nd phase of the MMC caused a short-term inhibition of the gastric and intestinal spike activity followed by its increase, the active period of the MMC being prolonged.

Participation of cholinergic mechanisms in the realization of the effect of bombesin on the migrating myoelectrical complex.

The experiments were carried out on dogs with chronically implanted bipolar ball-shaped silver electrodes in the antral part of the stomach and duodenum. The effect of bombesin (injected intravenously in a dose of 200 ng/kg) on the migrating myoelectrical complex (MMC) was examined after pretreatment with hexamethonium (10 mg/kg) or atropin (100 micrograms/kg), or alpha-antagonists (phentolamine, 2 mg/kg) and beta-antagonists (propranolol, 2 mg/kg) of the adrenergic receptors. Introduced during the second MMC phase, bombesin caused inhibition of the spike activity, accompanied by an increase of the slow potential frequency. Prolonged intensified spike activity occurred 5-10 min later. Early brief appearance of spike potentials was observed after bombesin in the presence of hexamethonium, which was not manifested on the background of atropin. After hexamethonium or atropin, however, the phase of the late activation of the spike activity did not occur and there was no change in the effect of bombesin on the frequency of the slow potential. The alpha- and beta-antagonists of the adrenergic receptors do not change the effect of bombesin on the electrical activity of the stomach and the intestines.

Development of the second contraction component after restoration with isoprenaline of pre-inhibited contractile activity of isolated myocardium.

Transmembrane action potentials and contractile activity were registered under isometric conditions during in vitro experiments on the papillary muscle of guinea-pig myocardium. The blocking agents of the slow transmembrane inward current: D 600 (10(-6) to 5 X 10(-6) M) and Mn2+ (5 X 10(-3)M), induce strong inhibition of the contraction amplitude. Applied on this background, isoprenaline (5 X 10(-7) to 10(-6)M) completely restores the contraction amplitude. The action potential is characterized by a high-amplitude plateau, followed by fast repolarization. The muscle was stimulated with suprathreshold rectangular impulses, 1 ms duration and 1-60 s interstimulus intervals. Directly proportional dependence was observed between the duration of the action potentials and the interstimulus intervals. A second component appears in the relaxation phase of the contraction upon stimulation with 10 s interstimulus interval, being particularly pronounced for 40-60 s intervals. As a consequence, the contraction acquires a marked two-component configuration. The data obtained are interpreted in the light of the hypothesis about the existence of two pools providing calcium for the contraction, assuming the participation of intracellular stores for the first component and transmembrane calcium for the second one. A scheme of the possible pathways of the Ca-ions, through which the contractile process in mammalian muscles takes place, is proposed.

Changes in the conductivity and in the functional refractory period of the atrioventricular node during hypoxia.

The experiments were carried out on preparations from rabbit right atrium, containing the sinoatrial node, the interatrial barrier, the atrioventricular node and the His-bundle. Surface electrodes were used to record the atrial and His-electrograms. The conduction time (AH-interval) was measured and the functional refractory period of the node was determined during normoxia (95% O2, 5% CO2) and hypoxia (30% O2, 65% N2 and 5% CO2) for 45 min. Hypoxia causes a considerable delay in conduction. This delay is greater for the shorter coupling-intervals and at the higher frequencies of the imposed rhythm. The shifting of the conduction times towards the higher values during hypoxia is not parallel compared with hypoxia. In the case of hypoxia the refractory curve of the A-V node is situated higher than the curve during normoxia, owing to the greater duration of the H1H2 intervals. The functional refractory period is increased and shifted toward the longer coupling-intervals. The results obtained show that during hypoxia the functional refractory properties of the atrioventricular node are impaired, which, combined with the delayed conduction through it, creates conditions for serious disturbances in the function of the atrioventricular conduction system.

Influence of stenopril on the isometric myogram and the action potential of isolated myocardium of warm-blooded animals.

The effect of the original Bulgarian drug Stenopril (1-phenyl-2/1-phenylpropyl-/3/-amino/propane hydrochloride) on the papillary muscle of guinea-pig right ventricle was studied in vitro. The isometric myogram and the transmembrane cellular activity were recorded. At low Stenopril concentrations (1-5 x 10(-6)M) three effects were observed: weak positive inotropic effect, phasic response (initial increase and subsequent decrease of the amplitude of the contractions) and weak negative inotropic effect. Concentrations of 10(-5) and 10(-4)M inhibit the contractile activity of the myocardium. The changes in the action potentials at the low concentrations are small. At the higher concentrations the Stenopril-negative inotropy is accompanied by lowering of the plateau and extension of the potential at the expense of the late repolarization, as well as decrease of the repolarization velocity. In a number of experiments the excitability decreased until the appearance of local responses. Isoprenaline (10(-6)M) completely restores the Stenopril-inhibited electrical and contractile activity of the drug. It is assumed that Stenopril influences the inward calcium current, similar to the calcium antagonists, and that it also inactivates the Na-permeability of the membrane.

Effects of some calcium antagonists on the excitation-contraction coupling in the myocardium under steady-state and nonsteady-state conditions of operation.

The specific features of excitation-contraction coupling in guinea-pig myocardium were studied by applying two different calcium antagonists--Mn2+ and methoxyverapamil (D 600). The muscle preparations were placed under two different conditions of operation: upon stimulation with equal interstimulus intervals (0.5, 1, 5 and 10 s)--steady state and with presentation of the same test-stimuli with the same intervals after a priming high-frequency series--nonsteady-state. The well-known coupling takes place under steady-state conditions of operation under the effect of Mn2+ (5 mM) and D 600 (1-5 x 10(-6) M). However, under nonsteady-state conditions of operation, in spite of the presence of antagonists of the slow inward current, potentiated contractions are observed. It is assumed that under nonsteady-state regime conditions are created for activation of the intracellular calcium stores and that the two calcium antagonists act on different sites in the electro-mechanical coupling in the myocardium.

Changes of certain cardio-vascular parameters and the sympathetic nervous activity upon afferent vagal stimulation in cats.

Afferent stimulation of the cervical vagus in cats for 5 s upon bilateral vagotomy reduces systemic blood pressure by 15--50 percent. The maximum of this depressor reaction occurs 10--15 s after the beginning of the stimulation, its latency being 2--3 s. The decrease in the blood pressure is preceded by inhibition of the electrical activity of some preganglionic (n. splanchnicus) and postganglionic (n. cardiacus and n. renalis) sympathetic nerves with 100--200 ms latency. During the maximum pressure decrease activity appears in the above-mentioned nerves, afterwards the pressure begins to rise until its normalization. In addition to the decrease in systemic blood pressure changes are also observed in other cardio-vascular parameters, namely, decrease in the pressure in the left ventricle, decrease in the contractile force of the heart (dp/dt max) and of the heart rate.

On the mechanism of the depressor reaction upon afferent vagal stimulation in cats. II. Role of the beta-adrenergic structures for the development of the depressor reaction.

Electrical stimulation of the central section of the cervical vagus in cats with bilateral vagotomy results in lowering of the arterial blood pressure. A study is made of the effect of some beta-adrenotropic drugs on the depressor reaction. Propranolol (3 mg/kg i.v.) causes considerable blocking of the depressor reaction in all experiments. On the other hand, Isoprenaline ( a. 10 mug/kh/min) results in a considerable increase of this reaction. The administration of Propranolol (3 mg/kg) and Phentolamine (2 mg/kg) causes almost complete blocking of the reaction. A conclusion is made on the basis of these data that both alpha- and beta-adrenergic structures take part in the realization and development of the depressor reaction upon afferent vagal stimulation.