Contemporary genetic structure and postglacial demographic history of the black scorpionfish, Scorpaena porcus, in the Mediterranean and the Black Seas.

Understanding the distribution of genetic diversity in the light of past demographic events linked with climatic shifts will help to forecast evolutionary trajectories of ecosystems within the current context of climate change. In this study, mitochondrial sequences and microsatellite loci were analysed using traditional population genetic approaches together with Bayesian dating and the more recent approximate Bayesian computation scenario testing. The genetic structure and demographic history of a commercial fish, the black scorpionfish, Scorpaena porcus, was investigated throughout the Mediterranean and Black Seas. The results suggest that the species recently underwent population expansions, in both seas, likely concomitant with the warming period following the Last Glacial Maximum, 20 000 years ago. A weak contemporaneous genetic differentiation was identified between the Black Sea and the Mediterranean Sea. However, the genetic diversity was similar for populations of the two seas, suggesting a high number of colonizers entered the Black Sea during the interglacial period and/or the presence of a refugial population in the Black Sea during the glacial period. Finally, within seas, an east/west genetic differentiation in the Adriatic seems to prevail, whereas the Black Sea does not show any structured spatial genetic pattern of its population. Overall, these results suggest that the Black Sea is not that isolated from the Mediterranean, and both seas revealed similar evolutionary patterns related to climate change and changes in sea level.

Biomonitoring of trace metals in the Black Sea (Romania) using mussels Mytilus galloprovincialis.

In a preliminary biomonitoring study, accumulated trace metal concentrations (Cd, Cu, Zn, Hg, Fe, Mn) have been measured in the mussel Mytilus galloprovincialis collected in the Black Sea, at 100 km from the Danube Delta in February 2001 and February 2002. Mussels were collected from four sites. In 2001 metal concentrations were determined in the whole soft body whereas, in 2002, the distribution of metals was evaluated in three different organs (gills, visceral mass and remaining tissues). The results obtained in 2002 confirmed those of 2001. For a given site, concentrations were always higher in the gills and visceral mass of mussels than in the remaining tissues. Principal component analysis allowed separating stations as a function of the metal concentrations in the organs. It is concluded that the mussels M. galloprovincialis are suitable biomonitors to assess changes in metal pollution in this coastal area of the Black Sea.

Mixed tumor: mucinous papillary cystadenocarcinoma associated with arrhenoblastoma.

A case of a 41-year old woman with an enlargement of the inferior part of the abdomen is presented. An echographic examination established the diagnosis of a right ovarian cyst. During the operation, a 14-cm (5 1/2") tumor, presenting a large ruptured cyst and solid mass, was found. From the cystic activity, a viscous fluid was oozing. On the left ovary, a 4 cm (1 1/2") cyst was also found. Within this case report, we underline the unusual association of a mucinous papillary cystadenocarcinoma with an arrhenoblastoma in the right ovary together with the presence of an cystadenoma in the left ovary.

HBsAg positive secondary autoimmune active chronic hepatitis. Immunogenetic aspects.

Observations were made in a group of 120 patients with active chronic hepatitis (ACH), using demonstration of HBsAg presence, structural study of the peripheral blood mononuclear cell population by means of scanning electron microscopy (SEM) and typing of HLA-A, B and C antigens. From the whole group 22 patients were proved to have an autoimmune form of ACH secondary to infection with hepatitis B virus (HBV). The authors consider that the diagnostic value of the ratio between B and T lymphocytes (studied by SEM) is relative but the demonstration of an increased percentage of monocytomacrophage cells and the presence of "killing phenomenon" clearly reflect a more severe hepatic inflammation. A higher incidence of the HLA-Bw35-Cw4 was observed particularly in the HBsAg positive forms of ACH and in the HBsAg positive secondary autoimmune forms. The authors discuss the utility of these observations when the opportunity of corticotherapy has to be established.

Electrophoretogram of lysates deriving from mononuclear cells. Preliminary observation.

Electrophoresis of mononuclear lysates was applied for the first time in the study of the lymphocyte protein content as an additional method beside the rosette test and scanning electron microscopy. The specification of T and B lymphocytes in the presence of discordant results sometimes obtained by the latter methods is facilitated by the additional data supplied by the study of their endoplasmic protein contents. The electfophorogram contributes to a better knowledge of the immunologic status of patients with autoimmune and lymphoproliferative diseases as well as to a more precise determination of the effects following the administration of immunomodulating treatment.

Normal blood cell values in groups of population from various geographic areas of Romania.

The aim of this work was to determine the normal values of erythro-, leuko-, and thrombocytes in several groups of population situated in various geographic areas (hills, plain, sea-side), considering that such data could be useful for the delimitation between the normal conditions and the near-normal ones. Sixteen hematologic parameters have been investigated in 2,053 healthy subjects, insisting particularly on those most frequently required in the clinic, i.e., erythrocytes/microliter, hemoglobin (g%), hematocrit (%), leukocytes/microliter, and thrombocytes/microliter. No significant differences related to the geographic areas were recorded. The normal values of the hematologic parameters are presented in separate tables.

Study of the relationship between the granulocyte LDH, alkaline phosphatase and Zn at the level of the leukocyte in patients with chronic myeloid leukemia.

Mixed leukocyte suspensions were prepared from heparinized blood collected from healthy subjects and from patients with chronic myeloid leukemia (CML). In all the suspensions determinations were made for: zinc, by atomic absorption; granulocyte alkaline phosphatase (GAP), using the method with p-nitrophenylphosphatase; granulocyte LDH, by means of the enzymatic autoanalyser LKD 8,600. In the patients with CML, the values of zinc and of granulocyte alkaline phosphatase activity were very low while the granulocyte LDH values were higher than normal. The chromatogram of the granulocyte LDH isoenzymes on DEAE-Sephadex A50 minicolumn (0.5 X 12 cm) showed an "alpha type abnormality" revealed by the increased activity of the isoenzymes with high electrophoretic mobility LDH2 and LDH1 specific for tissues with intense oxidative phosphorylation. In the normal subjects the chromatogram of the leukocyte LDH isoenzymes showed a type M (skeletal muscle) prevalence denoting intense anaerobic glycolysis. Therefore the low zinc concentrations (0.55 micrograms mg N2 as compared with the normal 1.24 micrograms mg N2) in these patients cause the decrease of GAP activity by the lack of zinc in the active center of the enzyme and the decrease of cellular permeability thus allowing the extracellular release of granulocyte LDH.

The metabolic behaviour of the cellular populations in the pleural or peritoneal effusions from patients with malignant diseases.

Complex morphologic, biochemical and cyto-enzymochemical investigations have been performed in the pleural or peritoneal effusions which occurred in 80 patients with various neoplastic diseases (pleural, pulmonary, hepatic, genital or breast cancers). As shown by the morphologic and biochemical tests, the proportion of fluids with a malignant or benignant character varied widely in the different forms of neoplasias. In the same effusions, the authors have studied the metabolic behaviour of acid phosphatase activity, the glycogen and nucleolar RNA amounts in the lymphocytes, in the malignant cells and in the mesothelial ones. High values of these cyto-enzymochemical components were found in the lymphocytes and the malignant cells from fluids with a positive malignant character, although similar changes were also observed in many cells of the effusions with morphologically negative reactions, and in a lesser proportion in those from patients with adenocirrhosis. It is concluded that such investigations could contribute to a better understanding of the metabolic behaviour of the cells from neoplastic effusions thus increasing the possibility of detecting the malignant character of the cells in the pleural or peritoneal effusions with a morphologically negative character.

Eosinophilia in the pleural and peritoneal effusions.

Of 248 patients with various benign or malignant chronic diseases who presented pleural or peritoneal effusions, the authors have selected 16 cases in which the proportion of eosinophils in these fluids exceeded 5% of the total cell count. High values of eosinophilia have been observed mostly in the serous membrane fluids from patients with tuberculosis and various neoplasms. In the authors' opinion, this suggests the participation of eosinophils in the immune processes occurring in the course of these pathologic conditions, in which the lymphocytes are also involved.

The metabolic behaviour of the lymphocytes from serous effusions in various benign internal diseases.

The functional status of the lymphocytes from pleural or peritoneal effusions occurring in 148 patients with various internal diseases such as tuberculosis, liver cirrhosis, heart failure and pneumonias was studied by laboratory investigations including morphologic examination, cytoenzymatic, cytochemical and biochemical tests for determining the scores of acid phosphatase activity and of endolymphocytic glycogen, as well as the presence of nucleolar RNA. These values were found increased in the fluids from tb patients and in some cases of liver cirrhosis, heart failure and metapneumonic pleurisies. The variations of these parameters are assumed to provide information on the metabolic behaviour of the lymphocytes present in these fluids and on their participation to the immune, inflammatory processes which occur in the course of some of the diseases investigated.

The value of some cytoenzymochemical investigations of the leukocytes and platelets in estimating the effects of occupational exposure to benzene, vinyl chloride and carbon disulphide.

Cytoenzymochemical investigations were performed on the leukocytes and platelets from 2,782 workers occupationally exposed to benzene, vinyl chloride or carbon disulphide, in view of detecting the eventual occurrence of cytologic metabolic disorders induced by these chemical substances. The cytomorphologic aspects and the cytoenzymochemical parameters studied, such as the leukocyte alkaline phosphatase, endolymphocytic glycogen, lymphocytic nucleolar RNA amounts (revealed by the lymphocyte nucleologram), as well as the platelet counts have shown some abnormal values, whose severity was related to the time of exposure to toxic chemicals. The results obtained demonstrate the utility of such tests for an early detection of some metabolic disorders of the endolymphocytic proteins, carbohydrates or lipids, in view of preventing the occurrence of more severe pathologic consequences.

Alkaline phosphatase activity improved by the addition of zinc in granulocytes from leukemic subjects (in vitro study).

Granulocyte alkaline phosphatase (GAP) was extracted from the leukocyte suspensions of 15 subjects with chronic myeloid leukemia (CML), determining the Zn content by atomic absorption spectrophotometry. The purified enzymatic preparation was dialyzed against a ZnCl2 solution (.01 mg%) in TRIS-HCl 0.1 M, pH 7.5, for 48 hours. After dialysis Zn concentration and GAP activity reached normal values. This suggests that the GAP molecule isolated from CML subjects is not altered as it is able to receive at most 2 Zn moles responsible for the enzymatic function.

Value of some leukocyte cytoenzymochemical investigations in detecting the noxious effects of the vinyl chloride monomer or polymer.

A group of 100 workers exposed to vinyl chloride (VC) (mean duration of exposure-13.2 years) were subjected to a complex medical examination for the detection of eventual alterations of the biologic parameters, induced by occupational exposure. Leukocyte cytoenzymochemical determinations (lymphocytic RNA--Plenert's method modified by Micu and glycogen--Hotchkiss McManus method), which might give some information on two important metabolic processes--of proteins and of carbohydrates--were performed in peripheral blood smears. Lymphocytic nucleologram and the relative and absolute nucleolar area showed an increase of endolymphocytic RNA, due to the high percentage of binucleolated cells and of spherical macronucleoli. The increase of the endolymphocytic glycogen score in parallel with the duration exposure was in correlation with high titers of serum gamma GT.

The relationship zinc--alkaline phosphatase activity in normal and pathologic granulocytes.

To study the relationship granulocyte alkaline phosphatase (AP)--zinc (Zn) in mixed leukocyte suspensions, the enzyme from 51 mixed leukocyte suspensions was separated and purified. In the pure extract, determinations were carried out for enzymatic activity--using the autoanalyzer LKB 8600--and for Zn content--using atomic absorption spectrophotometry. From the data obtained it resulted that, in agreement with the results in the literature Zn concentration in the mixed leukocyte suspensions from chronic myeloid leukemia, (CML) patients is about half that in the normal controls (0.65 +/- 0.15 as compared with 1.24 +/- 0.15 micrograms/mg total nitrogen). Granulocyte AP separated from normal leukocyte suspensions was found to bind about 95% of the total Zn content, i.e., 1.21 +/- 0.15 micrograms/mg total nitrogen and its enzymatic activity was about 16 U/g total nitrogen. In CML patients the enzyme bound a mean Zn concentration of 0.55 +/- 0.15 micrograms/mg total nitrogen and its mean activity was 2.25 U/g total nitrogen, i.e., about 86% of the normal values found in the controls. The enzymatic activity decreased to zero after only 20 min dialysis with 8-hydroxyquinoleine-5-sulfonic acid (8HC5S) as compared with the 60-80 min in the controls. It can be concluded that the leukemic leukocyte is unable to form the metal-enzyme complex owing to the deficit of the Zn involved in the enzymatic activity.