On the cytochemical demonstration of basic proteins in the cell nucleus, including the nucleolus.

A combined Feulgen-alkaline fast green method is described for simultaneous demonstration of DNA and basic proteins in the cell nucleus. The method is based on preserving both types of substances in the tissue section and releasing in them reactive groups for the 2 kinds of staining. These conditions are best provided, as proved by staining tests on tissue hydrolysates, if formalin-containing mixtures (SERRA's or LILLIE's fluids) are employed for fixation, and cold 5 N HCl is used for FEULGEN hydrolysis. In this way, a good cytological picture is also achieved. Nuclear euchromatin stains with this method red, while heterochromatin, pycnotic nuclei and sperm heads exhibit a deep violet to blue-violet colour. Prominent nucleoli of metabolically active cells display a distinct blue-green staining thus manifesting their high content of basic proteins. Acetylation test reveals that these proteins are of lysine-rich type. The known negative reaction of the nucleoli with the routine alkaline fast green method according to ALFERT and GESCHWIND must be attributed to an extraction of the nucleolar basic proteins with the hot TCA used in this method. Certain analogy in the cytochemical behaviour between the nucleolous and the chromatin under various conditions of hydrolysis leads to the suggestion that the nucleolar basic proteins demonstrated should be in the form of a ribonucleoprotein complex, probably of the pre-ribosomal material of the nucleolus.

Histochemical demonstration of the effect of partial hepatectomy on the rat liver enzyme activity of 3 alpha-, 17beta-hydroxisteroid dehydrogenase and glucose-6-phosphate dehydrogenase.

The dynamics of the partial hepatectomy effect on the activity of 3alpha-HSD, 17beta-HSD and G-6-PD in rat liver has been followed by means of histochemical methods. In all stages of the investigation the 3alpha-HSD and G-6-PD did not show any deviation from the normal histologic findings in the sham-operated control animals. The reaction for 17beta-HSD decreased abruptly and more or less approximately evenly in the liver cells of the hepatectomized animals as early as in the 24th h after operation. That went on until the 48th h. During the 72nd, 96th and 120th h a certain increase of the 17beta-HSD activity was established. On the 8th day the intensity of the histochemical reaction was identic with that in the liver cells of control animals. The histochemical changes of the 17beta-HSD enzyme activity coincide fully in time, strength and direction with the rate of liver regeneration. During the first 2 days after partial hepatectomy, during the rapid mitotic division and loss of the highly specialized function of the hepatic cells, the activity of the 17beta-HSD enzyme greatly decreases, probably in connection with the involvement of cell proteins in the synthesis of new structures. When the regeneration rate diminishes, about the 5th to 8th day, the liver cells gradually return to a quiet ("differentiated") state, regenerate their enzyme systems (17beta-HSD) and renew the normal function of metabolizing steroid hormones.

Psychophysiological comorbidity and computerized biofeedback.

Our previous research on basic and applied psychophysiology comprised the quantum model for subcellular brain processes, the psychosomatic health risk assessment and management, as well as the electrodermal biofeedback psychometrics and therapy. Based on this experience, in the present paper the stress-related psychophysiological disorders in patients dependent on hemodialysis and insulin are considered. The most frequent comorbid psychological disorders appeared to be anxiety and depression, and they are generally under-diagnosed and under-treated. It is concluded that the quality of life of these patients needs to be considerably enhanced. The application of biofeedback, as a complementary cost-effective and non-invasive psychophysiological tool is recommended. In terms of ESAO vocabulary, this technique could be viewed as some kind of artificial ANS/CNS support. Simultaneously, the concept of biocompatibility may be viewed in a larger perspective as "psychobiocompatibility".

The need for regular screening of postpartum depression.

AIM: The aim of this study was to examine the level of depression during the first week of the postpartum period in a sample of women hospitalized for childbirth at the University Obstetric Clinic in Skopje and correlate it with some demographic characteristics. METHOD AND SAMPLE: The sample comprised 150 randomly selected women with a mean age of 29.23 years (SD±5.11). As a psychometric test the Beck Depression Inventory was used. It is a multiple choice, self-report inventory for measuring the severity of depression. For statistic evaluation (descriptive analysis and correlations) software Statistica 7 was used. RESULTS: The random study showed that 68% of postpartum women have minimal, 25% mild and 5% moderate depression. Fortunately only 2% manifested severe depression. Our findings showed that depression is negatively correlated with the level of education (r=-0.17), as well as with age (r=-0.15). The comparison of the level of depression with a group of chronic dialyzed patients showed that PPD is not so serious a problem. However, this study shows that PPD is under diagnosed. CONCLUSION: As a common mental health problem PPD is not systematically screened in our country. The study showed the presence of minimal (68%) and mild (25%) depression in the examined women. The age of the woman is important for PPD; younger women are more susceptible to depressive reactions. The level of education is negatively correlated with depression. It was pointed out that PPD must be recognized, treated and followed for a long period because it is a risk not only to the mental health of the mother but also for the development of child.

Cotinine concentrations in amniotic fluid and urine of smoking, passive smoking and non-smoking pregnant women at term and in the urine of their neonates on 1st day of life.

Cotinine was measured in the amniotic fluid and urine of 31 pregnant women and in the urine of their offspring. Amniotic fluid cotinine was 8 times higher in active and 2.5 times higher in passive smokers than in non-smokers. In general, amniotic fluid cotinine was considerably higher than urinary cotinine both in active and in passive smokers. Estimation of cotinine both in amniotic fluid immediately before delivery and in urine of the newborn on the 1st day of life aids in assessing the degree of prenatal exposure to tobacco smoke.

Effects of steroid sex hormones on chick embryo gonads in organ culture, with special reference to hormonal control of gonadal sex differentiation.

At the initial stages of sex differentiation (7.5 and 8.5 days of incubation), chick embryo gonads were treated directly with testosterone or estradiol-17 beta in organ cultures. Chemically-defined media containing cholesterol as a steroid precursor were used. The differentiation of gonads in the 10 to 12-day controls, cultured in media containing no hormones, was close to that of gonads of equivalent age in ovo. Testosterone added to the medium exerted an inhibitory effect on the cortex of the female gonad and a masculinizing one on its medulla. The results of estradiol treatment confirmed the known feminizing effect of that hormone on the male gonad, the meiotic prophase in the genetically male germ cells being initiated in the induced cortex. These data may be interpreted in favour of a bihormonal theory of gonadal sex differentiation in birds, where the predominantly-synthesized male or female hormone in the gonad determines the male or female pattern of development of the corresponding gonad.

Meiosis-inducing and meiosis-preventing effects of sex steroid hormones on hamster fetal ovaries in organ culture.

Day 11 to day 15 p.c. female gonads were cultured for 6-8 days in chemically-defined media. In day 11 and day 12 p.c. ovaries grown in a non-hormonal medium, the germ cells were unable to enter meiosis; they were retained at a stage of oogonia or more frequently at a preleptotene stage. Ovaries of the same ages cultured in an estradiol-containing medium showed germ cells progressing through meiotic prophase in a way close to that in ovaries of equivalent age in vivo. That was the case of the germ cells in day 13 to day 15 p.c. ovaries maintained in a non-hormonal medium. In a testosterone-containing medium, the germ cells in day 13 and day 14 p.c. ovaries were prevented from entering meiosis; by contrast, those in day 15 p.c. ovaries underwent meiotic prophase normally. These results indicated that each of both hormones was able to exert its corresponding (meiosis-inducing or meiosis-preventing) effect before a definite critical time of ovarian development. The possibility is suggested that the germ cell differentiation in the female and male gonads in vivo would also depend on estrogens or androgens precociously synthesized in the gonads or supplied from other organs via the fetal blood.

[Asymmetry in artificially deformed skulls from a Protobulgarian mass grave of the 9th century]. / Die Asymmetrie künstlich deformierter Schädel aus einem protobulgarischen Massengrab vom 9. Jh

A series of 18 skulls with pronounced asymmetry, picked up out of a total of 52 artificially deformed and accordingly restored skulls are studied. It is a matter of material discovered in a circular tomb near by the village of Devnja, dating back to the end of the 9th century. To obtain metrical determination of the asymmetry, the authors performed measurements of chords, resp. arches between the anthropological points sphenion and asterion, as well as of the distance from the latter and from the porion on either side up to the glabella, inion, bregma and lambda. Besides the 18 sizes directly recorded, 18 chords constructed on the craniograms (three per skull) mainly between the above mentioned points were also utilized. For each dimension the absolute differences between left and right, the mean value of the differences and the asymmetry index after Duncker were calculated. Using the method outlined, an accurate and quantitative determination was made of the asymmetry both in the occipito-parietal and temporo-clinoidal parts of the skull (compensatory and persisting asymmetry). It is demonstrated that the discrepancies between some of the size values established in individual skulls, and the skull asymmetry, recorded scopically with other measurements, are due to displacements of the anthropological points (sphenion, asterion, inion), produced by differences in growth rate and size of bones. The arrested development of bones and the resulting depressions along their surfaces under the effect of the artificial deformation dressing are of utmost importance for the processes developing within the deformed skulls.

[Asymmetry in the structure of the middle part of the facial skull]. / Die Asymmetrie im Bau des Mittelteils des Gesichtsschädels.

The results of study of the middle third of the facial region, performed on a total of 412 skulls of male individuals, lead to the following conclusions: 1. In the investigated upper section, the zygomatic bone inclusive, downwards to its lower section along the alveolar process of the maxillary bone, a complete and approximative symmetry displayed by the width values is more frequently met with, whilst asymmetry is rather rarely. In the same direction, from above downwards, the number of skulls with differences in favour of the right side decrease, whereas the number in favour of the left side increase, with the rarely observed asymmetry in the lowermost section usually involving the leftside. 2. The height of the upper jaw-bone region near the sagittal line shows more frequently symmetry (complete and approximative), and rather rarely asymmetry, whilst insofar as the zygomatic bone is concerned the opposite is observed; therefore, in the direction from medial to lateral-wards the incidence of asymmetry increases, whereas that of the symmetry decreases. In the same direction the number of skulls with differences in favour of the right side increases, while the number in favour of the left side decreases. 3. The coincidence of absolute differences recorded in 69%, respectively 61.6%, between right and left side of closely positioned dimensions (20 and 31, resp. 20 and 42) proves that the data about these dimensions are apt to determine symmetry and asymmetry manifestations with a sufficient reliability. 4. The individual values of the pairs of breadth dimensions of one and the same region of the studies craniofacial portion show a higher degree of correlation than those of different regions which is due to the differences between the latter in terms of incidence of symmetry and asymmetry in their build up. 5. The width of the middle third of the median craniofacial portion and the lateral distance of the skull Zygomaxillare--Porion do not correlate, i.e. there is no parallelism between the fluctuations of their individual values. 6. In most of the cases the bones of the external nose are symmetrical, and hence its asymmetry is attributed to the soft parts as cartilage and skin.

[Relationship between the retromolar space and the mandibular measurements in Bulgarians]. / Zusammenhang zwischen dem retromolaren Raum und den Massen des Unterkieferknochens beim Bulgaren.

We examined 412 mandibles from adult males. Using the classic anthropometric methods according to Martin-Saller, we took 17 measurements: length, width, height and angle of the mandible. The data obtained were subjected to variance and correlation analyses. We observed poor correlations for the length of the dental arch, the height of the ascending ramus, the condylar width and the width of the chin; there were medium correlations for the following measurements: mental plane, median plane and angular width in the region of 46, 47, 36, 37 and the thickness of the alveolar portion. The correlations for all the other measurements were high; in some cases, they approximated the normal value.

Adaptation of the ammoniacal silver reaction to cytochemical demonstration of myelin basic protein.

A modification of Black and Ansley's ammoniacal silver reaction (ASR) for histones is proposed for visualizing myelin basic protien (MBP) in the nervous system. The reaction is performed on histological sections of tissues fixed in neutralized formalin-alcohol and delipidized in the course of the routine paraffin embedding. The deparaffinized sections are again treated with formalin in order to make the "unmasked" by the delipidization basic groups of MBP reactive to ammoniacal silver. After treatment with this reagent MBP of the myelin sheaths of the nerve fibres is impregnated brownish-black. Deparaffinized sections subjected to an extraction of MBP with hydrochloric acid exhibit a negative reaction at the level of the myelin sheaths the same reaction being preserved at the level of the nuclear histones. The reaction is positive in paper spots of nervous tissue extracts obtained with the same acid. These assays indicate the specificity of the modified ASR. The method can be used for studies on the processes of myelination and demylination in normal histogenesis and in pathology of the nervous tissue.

Symbolic trepanations of skulls from the Middle Ages (IXth-Xth century) in Bulgaria.

Symbolic trepanations (ST) of skulls (IXth-Xth century) found in north-eastern Bulgaria near the West coast of the Black Sea were Studied. It was established that trepanations were performed on subjects well over the age of thirty and cover 41% from the total material which is the highest frequency documented so far. Tracks of the ST are localized most often on and round the sagittal suture, bregma and lambda. Their number ranges from 1 to 9 per cranium with a diameter of 3-4 to 20-28 mm and a depth of 1 to 6 mm. They are usually round and encircled with a prominent ridge. In this article we try to explain the reasons for the symbolic trepanations as well as the link between the traditions of the ancient Turkish nations and Chinese medicine.

Interaction of the anti-tumour drug cis-Pt(en)Cl2 with Cytidilyl-3'--5'-guanosine.

Chemical shifts occurring in 1H nuclear magnetic resonance and Sephadex G 25 chromatography are used to follow the reaction of the dinucleotide C3'p5'G (cytidilyl-3'phosphate5'-guanosine) in aqueous solution and to separate its products. Binding of the platinum occurs in first place at the cytosine, then at the guanine base. Two major complexes are produced, Pt:CpG and (Pt:CpG)2, which account for respectively for an internal and an external cross-linking effect.

Structural investigations by extended X-ray absorption fine structure spectroscopy of the iron center of mitochondrial aconitase in higher plant cells.

We have obtained iron K-edge extended x-ray absorption fine structure spectra of the plant mitochondrial aconitase in its active state, in the presence (aconitase (+)) and absence (aconitase (-)) of the substrate citrate. Analysis of the data indicates that oxygens are present in the first coordination shell, at an average Fe-O distance of 1.96/1.98 A (aconitase (+)/aconitase(-)). Part of these oxygens is provided by the citrate, which binds at 1.99 A from the iron in aconitase (+). The second shell (sulfur) contribution is split and is consistent with Fe-S distances of 2.30/2.29 and 2.56/2.59 A, and the third shell (iron) is consistent with an Fe-Fe distance of 2.83/2.84 A. Both Fe-S and Fe-Fe distances are longer than similar distances found in most Fe-S centers. A strong scattering at approximately 5 A has been identified as originating from an iron atom which is near to, but not part of, the Fe-S cluster. These data indicate that active plant mitochondrial aconitase contains a novel type of iron center.

[Passive smoking and positive intracutaneous allergen tests in children with respiratory diseases]. / Passives Rauchen und positive intracutane Allergenteste bei Kindern mit respiratorischen Erkrankungen.

In children with respiratory disease (bronchitis, pneumonia and bronchial asthma) skin hypersensitivity towards tobacco leaf crude extract has been tested and related to urinary cotinine level, the latter being most informative in assessing exposure to cigarette smoke. Besides routine intracutaneous tests a mixture of seven main brands of Bulgarian tobaccos (Burley, Virginia, Oriental etc.) has been applied as allergen. Urinary cotinine has been estimated by the method of BARLOW et al. [1]. Skin sensitization towards tobacco was positive in 2 (8%) of the children hospitalized for pneumonia and bronchitis and in 8 (47%) of children with bronchial asthma, followed up as outpatients. Cotinine concentrations were 66 mumol.l-1 and 107 mumol.l-1, respectively compared to 34 mumol.l-1 and 37 mumol.l-1 in healthy controls. Results outline the great importance of passive exposure to environmental tobacco smoke, an all the year round domestic pollutant in the home of smoking parents.

Ultrastructure of developing interstitial cells in chick embryonic gonad in relation to their genesis and steroidogenic function.

Electron microscopic studies on the development of interstitial (steroidogenic) cells in embryonic chick gonads were carried out in a chronological sequence from the time of their appearance to the end of incubation, in a comparative aspect in both female and male sexes, and through a comparison with light microscopic findings. An asynchrony in the development of these cells in both sexes is established. In the ovaries (left and right) individual steroidogenic cells are detected for the first time towards the 7th embryonic day, grouping themselves towards the 9th...10th embryonic day in nests, interstitial organoids. In the testis this takes place towards the 10th and 14th...15th days, respectively, the interstitial organoids forming as incomplete muffs around the seminiferous cords and later on (towards the 16th embryonic day) also as nests in stroma. On the basis of observations that the precursers of the steroidogenic cells differentiate within the sex cords (first proliferation) and separate from them, falling into the stroma, the authors adopt the concept of the mesothelial (epithelial) origin of those cells. Particular protrusions (multivesicular structures) of the cytoplasm of the interstitial cell are described whose role is believed to be connected with the mechanism of the steroid secretion of the gonads. The results are discussed in relation to some biochemical and experimental data on the morphogenetic function of the embryonic sex hormones.