MECP2 mutations in Serbian Rett syndrome patients.

BACKGROUND: Rett syndrome is a severe neurodevelopmental X-linked dominant disorder affecting 1/15,000 girls worldwide. Eight years ago, the MECP2 gene was associated with the devastating clinical features observed in Rett syndrome patients. OBJECTIVES: To investigate the spectrum and the frequency of MECP2 mutations in Serbian Rett syndrome patients. PATIENTS AND METHODS: We screened the MECP2 coding region by conventional mutational screening (single-strand conformation polymorphism/sequencing) in 24 patients of Serbian origin and in their 41 unaffected family members. In search for gene dosage alterations in seemingly mutation-negative girls, we developed a new, specific quantitative PCR method. RESULTS: Nineteen patients (79%) carried MECP2 mutations, five of which were novel (one nonsense mutation, one duplication and three deletions). Fourteen previously described disease-causing sequence changes and one polymorphism were also detected. Detailed case reports are given for the carriers of the novel mutations. Large MECP2 rearrangements cause Rett syndrome in a significant number of girls without 'classic' mutations in this gene. Therefore, we developed a specific quantitative PCR method, covering MECP2 exons 3 and 4, which previously has not been used for screening. No dosage alterations of the two exons were found in the four tested mutation-negative girls. CONCLUSIONS: This is the first genetic study of Rett syndrome in Serbian patients describing the MECP2 mutational and phenotypic spectrum in this population. Detailed clinical descriptions of this ethnically homogeneous patient population add to our knowledge of genotype/phenotype correlations in this severe condition.

Neurovascular stalk of the superficial sural flap: human fetus anatomical study.

BACKGROUND: The neurovascular stalk of the superficial sural flap, which is the most frequently used, is composed of the lesser saphenous vein, the sural nerve, and the median superficial sural artery. However, it has many variations. This is very important for its application in the reconstruction of soft-tissue defects. The aim of the authors' research was to detect the presence of the superficial sural blood vessels and to investigate their relationships with the sural nerve and the lesser saphenous vein. METHODS: The study group consisted of 42 fetal lower extremities. Fetuses were fixed in 10% formalin and their blood vessels were injected with Micropaque solution (barium sulfate). RESULTS: The median superficial sural artery was detected in 83.3 percent of the cases, whereas the sural nerve and lesser saphenous vein were detected in all cases. The median superficial sural artery was located lateral to the medial cutaneous sural nerve and sural nerve, whereas the lesser saphenous vein was located medially. CONCLUSIONS: All three superficial sural arteries (medial, median, and lateral) were detected in fetuses with different gestational ages. The median superficial sural artery was the most frequently detected one and had the constant relationship with the other elements of the neurovascular stalk of the superficial sural arteries.

[Diffuse and binodular goiter in a patient with tuberous sclerosis]. / Difuzna i binodozna struma kod bolesnice sa tuberoznom sklerozom.

Tuberous sclerosis complex is hereditary multisystemic, neurocutaneous disorder characterized by hamartomatous lesions mainly involving the brain, skin and viscera. Thyroid gland is rarely affected in these patients. Case history of a patient with tuberous sclerosis complex and diffuse and binodular goiter was presented. No epileptic seizures occurred despite multiple intracranial calcifications, cortical and subependimal tubers, revealed by neuroimaging. Borderline intellectual functioning was determined. Ultrasonographic findings of the heart, abdomen and pelvis were normal. Bone radiographs revealed multiple cystic lesions of the metacarpal, metatarsal and terminal phalanges. Clinical, endocrine and metabolic evaluations were performed because of anxiety, mild weight loss, transient problems of swallowing and breathing complaints, developing when she was 39 years old. Palpable diffuse and asymmetric thyroid enlargement was found. Laboratory tests of thyroid function, ultrasonographic examination and scintiscanning of the thyroid gland and patohistological analysis of fine-needle aspiration biopsy revealed the euthyroid, inactive, diffuse and binodular goiter. Unusual association of goiter and tuberous sclerosis was discussed.

[Miraculous recoveries in the frescoes of the Decani Monastery]. / Cudesna isceljenja u zivopisu manastira decani.

Serbian medieval medicine can be said to have the same characteristics as western medicine of those times in terms of practical medical science, whereas the ancillary medical branches were under Byzantine influence. Besides the compositions of Christ's miracles and paraboles are underlying part of the fresco/painting, the scenes of Christ's miraculous recoveries have an important place in the christian iconography of the New Testament events. One of the most impressive cycles of Christ's miracles in the Serbian and Byzantine medieval art was painted in c. 1340 year in the church dedicated to the Ascension of Lord Christ at Dechani Monastery and constructed between 1327 and 1335. A considerable number of 22 frescoes of different artistic value and technique displays the miraculous recovery of patients afflicted by various chronic ailments: paralytics (razlabljeni), lepers, and handicapped (blind, lame and deaf). One scene presents the bleeding (krvotociva) women. Scenes of recovery from nervous and mental diseases are especially impressive. Patients with atrophic arm, these with hemiplegy, epilepsy, an somnambulist boy, lunatics and possessed are there painted. The magnificent compositions of the miraculous recovery of a patient with ascites (debela, vodena bolest) and these affected by disorders of mainly neuromuscular and locomotor systems are performed by talented masters. The King Stefan Urosh Dechanski Third (1322-1331) founded the hospital in the Dechani Monastery, as a purely medical institution for providing treatment. This hospital was organized and modelled on the hospital in the Pantocrator Monastery in Constantinople. His biographer Gregory Camblak described that this hospital took in patients suffering from epilepsy (svestenoga neduga radi stradalcem) and serious nervous disorders, lepers, cripples, paralytics and those with lung diseases. Regardless of differences in the artistic level, most of these frescoes are impressively natural and realistic. This supports the ascertain that masters painted even some of patients from Monastery Hospital. The scenes of Christ's miraculous recoveries in the Dechani Monastery are undoubtelly significant for both the cultural history and the Serbian medieval medicine. This considerable cycle of the monumental painting and frescoes with medical matter, constitutes the basis for research of the Serbian medicine of the Middle-Ages.

[Antiphagocytic activity of sera treated with organic solvents (chloroform, ether)]. / Antifagocytárna aktivita sér opracovaných organickými rozpúst'adlami (chloroform, éter).

The opsonization of C. albicans by means of sera treated with chloroform and ethyl ether resulted in significantly decreased values of PMN leucocyte phagocytic activity. In contrast to ethyl ether treated sera (decrease of phagocytic activity about 45%), chloroform processed sera possessed increased antiphagocytic activity (decrease of phagocytic activity about 63%). Significant changes were observed in the candidacidal capacity to C. albicans (decrease about 50%) and in the capacity of NBT (INT) reduction measured spectrophotometrically (decrease about 66% in chloroform treated sera and about 42% in ethyl ether treated sera). (Fig. 1, Ref. 22.)

[Inflammation, immunity and surgery]. / Zápal, imunita a chirurgia.

The activation of endothelial and phagocytic cells, with concomitant formation of a range of adhesive molecules and inflammatory mediators, are integral parts of the host response to injurious agents (trauma, infection, altered antigens, toxins, chemicals etc.). The excessive mediator responses are associated with increased morbidity and lethality. Cytokines, soluble mediators secreted by cells, play an integral role in the metabolic and immune responses to the injurious agents. Widespread tissue damage, when associated with fulminant sepsis may induce massive release of cytokines (TNF, IL-1, IL-6), triggering certain steps of reactions involving multiple organs and culminating in the systemic inflammatory response syndrome, sepsis syndrome or multiple organ failure syndrome. (Fig. 2, Ref. 21.)

[Antiphagocytic activity of human amniotic fluid]. / Antifagocytárna aktivita l'udskej plodovej vody.

The preincubation of PMN leukocytes in amniotic fluid obtained at the end of the gestation period resulted in decreased values of phagocytic activity (about 37%), phagocytic index (about 21.5%) and mainly of the candidacidal capacity to C. albicans (about 72%). The level of lysozyme (11.8 +/- 7.2 mg.l-1) and the level of alpha 1-antitrypsin (0.745 +/- 0.53 g.l-1) were also determined in amniotic fluid. The level of alpha 2-macroglobulin and the proteolytic activity at acidic and mildly alkaline pH could not be established by the methods applied. (Fig. 1, Ref. 23.)

Vaginal leukocyte characteristics in urogenital trichomoniasis.

Morphological and functional characteristics of vaginal exudate leukocytes were examined in 47 patients with urogenital trichomoniasis. Electron microscopic morphology, viability, phagocytosis of Candida albicans blastospores and ability to undergo respiratory burst in the iodonitrotetrazolium reductase test were evaluated in these cells. Vaginal inflammatory leukocytes were almost exclusively polymorphonuclear neutrophils, and their concentration was positively correlated (r = 0.58; p less than 0.001) with the number of trichomonads in the exudate. Median leukocyte viability reached 39% and both phagocytic and tetrazolium reductase activities of these cells were significantly reduced in comparison with those of circulating polymorphonuclear leukocytes. Patients with a clinical picture of severe mucosal inflammation had significantly higher vaginal exudate leukocyte concentrations and viability than those without inflammatory signs. The possible role of vaginal leukocytes in the pathogenesis of urogenital trichomoniasis is discussed.

[Trichomonas in girls during the period of hormonal inactivity]. / Trichomoniáza dievcat v období hormonálneho pokoja.

Trichomoniasis occurs also in childhood in the period of hormonal inactivity, though its rate is not so high as in women of childbearing age and full sexual activity. In our series of children in the period of sexual inactivity, established by functional cytology, trichomoniasis was diagnosed in 3.8% of the total number of 1,562 children. Preparations of imidazol proved to be fully effective in the treatment of trichomoniasis in girls. In diagnosis the semisolid Szenes's culture medium in Valent's modification was used to advantage. Its standard use provides a basic precondition for good recovery of the protozoon. Trichomoniasis as such does not present so serious a problem as do the consequences of the unrecognized and untreated disease when the proliferated secondary flora potentiates the severity of the infection. This condition may lead to salpingitis and peritonitis causing sterility, and it may also spread into the uropoietic system.

Proteolytic activity in Tritrichomonas mobilensis.

Cell extracts of an entero-invasive protozoon of squirrel monkeys, Tritrichomonas mobilensis, contained relatively high proteolytic activity, measured on hide powder azure (HPA). Multiple proteinase forms, optimally active at pH 5-7, were detected by electrophoretic analysis in gelatin-containing polyacrylamide gels. Three major proteinase bands of apparent low molecular weights, Mr 18, 23 and 30 kDa, were seen on gels. Inhibition-activation studies suggest that only cysteine proteinases were involved in HPAase and gelatinolytic activities of T. mobilensis cell extracts.

Quantification of proliferative and suppressive responses of human T lymphocytes following ConA stimulation.

The mitogenic response of human T lymphocytes to graded doses of concanavalin A (ConA) has been measured by means of an MTT tetrazolium dye metabolic assay. Three groups of healthy subjects, representing children, younger adults and elderly persons, were investigated. It was shown that a typical bell-shaped course of the ConA dose-response curve is the result of a proliferative response to suboptimal concentrations of ConA and a toxic action of ConA at supraoptimal concentrations. The ascending part of the response curve reflects in its shape the regulatory interaction of responding cells. A decrease in suppressive functions is accompanied by a shift of this part of the curve to lower concentrations of ConA. By means of a mathematical model derived from enzyme kinetics, an attempt was made to quantify the suppressive functions from the course of the individual dose-response curve. It was found that after suitable data processing, suppression-related shape changes can be assigned to a single parameter. The value of this parameter as a diagnostic tool was tested in a study of the age dependence of human T lymphocyte responses to ConA. While the proliferative response decreased with age, the suppressive functions exhibited their maximum effect in the group of adults. Thus it could be demonstrated that ConA induced proliferative and suppressive responses are due to two different pathways which can be independently extracted from the dose-response curve.

Family study of natural killer cell activity in C1q-deficient patients with systemic lupus erythematosus-like syndrome: association between impaired natural killer cell function and C1q deficiency.

Impaired natural killer (NK) cell activity has been found in patients with systemic lupus erythematosus (SLE)-like syndrome. The mechanism by which NK cell function is impaired in SLE patients is not quite clear. We report here a family study of NK cell activity in C1q-deficient patients with SLE-like syndrome. In both SLE-active and SLE-inactive stages of the disease, NK cell function was significantly impaired when compared with the healthy controls (10.6 +/- 2.3% and 16.9 +/- 4.8% to 34.7 +/- 9.6%, p less than 0.025). On the other hand, differences in NK cell cytotoxicity between SLE-active and SLE-inactive members of the family were not statistically relevant (p less than 0.1). Further, we found no correlation between NK cell activity and clinical or laboratory values, except for a positive correlation between function of NK cells and C1q and CH50 values, respectively (rs = 0.93, 0.01 less than p less than 0.02). To our knowledge, this is the first report on a notable association between impaired NK cell activity and C1q deficiency. The type of inheritance of C1q deficiency in this family is also discussed.

[The development of cytotoxic T lymphocytes in HLA-D/DR identity]. / Vznik cytotoxických lymfocytov T pri identite HLA-D/DR.

The development of cytotoxic T-lymphocytes is controlled by genes of the HLA complex and occurs, in the majority of cases, when the responding and stimulating subjects differ both in HLA-A, -B and HLA-D/DR antigens. We examined a married couple for kidney transplantation and found that they differed in HLA-A, -B antigens and were identical in HLA-DR antigens. The negative results of the MLC test confirmed the serological findings. Nevertheless, cytotoxic T-lymphocytes developed despite HLA-D/DR identity. The presented case is one of the few published exceptions to the rule governing the development of cytotoxic T-lymphocytes.

HLA-D/DR identity results in a positive CML reaction.

Investigating HLA-A, HLA-B, HLA-C, and HLA-DR antigens, MLR, CML, and PLT reactivity in two unrelated persons it was found that despite their HLA-D/DR identity cytotoxic T lymphocytes (CTL) could be induced in the CML assay. The HLA-DP antigens proved to provide the necessary proliferative impetus for the generation of CTL.

A review: HLA and diseases--current status.

Since the discovery of polymorphism of the HLA complex the relationship between HLA antigens and various diseases has been followed. A number of HLA disease associations have been found. A review of the most important ones is given in this article. Explanation of the relations: "HLA and diseases" remains obscure till now what however does not hinder to take profit from them in practical medicine at diagnosis, differential diagnosis, therapy, prevention and also in genetic advisory service.

HLA-DP antigens provide the proliferative impetus for the generation of cytotoxic T lymphocytes.

Investigating HLA-A, -B, -C, -DR and -Dw antigens and MLR, CML, and PLT reactivity in two unrelated persons, it was found that, despite their HLA-D/DR identity, cytotoxic T lymphocytes (CTL) could be induced in the CML assay. The HLA-DP antigens proved to provide the proliferative impetus for the generation of CTL.

Specific parasitism of purified vaginal epithelial cells by Trichomonas vaginalis.

Human vaginal epithelial cells (VECs) from vaginal swabs obtained from normal women or from patients with trichomoniasis were purified, and VEC parasitism by Trichomonas vaginalis was examined. Trichomonads bound equally well to live or dead VECs, and up to 20% of VECs were parasitized. Trichomonal cytadherence of human VECs was time, temperature, and pH dependent. Saturation binding levels of live trichomonads to VECs gave approximately 2 organisms adherent to parasitized VEC. No differences in cytadherence levels were detected by different isolates to VECs from the same patient compared with adherence to VECs from normal individuals. Trypsinized, live T. vaginalis organisms failed to recognize VECs. A ligand assay identified four adhesin candidates, and only organisms without a prominent immunogen on the surface (negative phenotype) cytadhered to VECs and synthesized the adhesins, confirming the results of a recently published report by us on adherence to HeLa cell monolayers (J. F. Alderete and G. E. Garza, Infect. Immun. 56:28-33, 1988). These data show the ability of T. vaginalis to parasitize human vaginal epithelial cells in a specific receptor-ligand manner.