Differential Effects of Ethanol on Serum GABAergic 3α,5α/3α,5β Neuroactive Steroids in Mice,Rats, Cynomolgus Monkeys,and Humans

Background: Acute ethanol administration increases plasma and brain levels of progesterone and deoxycorticosterone‐derived neuroactive steroids (3α,5α)‐3‐hydroxypregnan‐20‐one (3α,5α‐THP) and (3α,5α)‐3,21‐dihydroxypregnan‐20‐one (3α,5α‐THDOC) in rats. However, little is known about ethanol effects on GABAergic neuroactive steroids in mice, nonhuman primates, or humans. We investigated the effects of ethanol on plasma levels of 3α,5α‐ and 3α,5β‐reduced GABAergic neuroactive steroids derived from progesterone, deoxycorticosterone, dehydroepiandrosterone, and testosterone using gas chromatography‐mass spectrometry. Methods: Serum levels of GABAergic neuroactive steroids and pregnenolone were measured in male rats, C57BL/6J and DBA/2J mice, cynomolgus monkeys, and humans following ethanol administration. Rats and mice were injected with ethanol (0.8 to 2.0 g/kg), cynomolgus monkeys received ethanol (1.5 g/kg) intragastrically, and healthy men consumed a beverage containing 0.8 g/kg ethanol. Steroids were measured after 60 minutes in all species and also after 120 minutes in monkeys and humans. Results: Ethanol administration to rats increased levels of 3α,5α‐THP, 3α,5α‐THDOC, and pregnenolone at the doses of 1.5 g/kg (+228, +134, and +860%, respectively, p < 0.001) and 2.0 g/kg (+399, +174, and +1125%, respectively, p < 0.001), but not at the dose of 0.8 g/kg. Ethanol did not alter levels of the other neuroactive steroids. In contrast, C57BL/6J mice exhibited a 27% decrease in serum 3α,5α‐THP levels (p < 0.01), while DBA/2J mice showed no significant effect of ethanol, although both mouse strains exhibited substantial increases in precursor steroids. Ethanol did not alter any of the neuroactive steroids in cynomolgus monkeys at doses comparable to those studied in rats. Finally, no effect of ethanol (0.8 g/kg) was observed in men. Conclusions: These studies show clear species differences among rats, mice, and cynomolgus monkeys in the effects of ethanol administration on circulating neuroactive steroids. Rats are unique in their pronounced elevation of GABAergic neuroactive steroids, while this effect was not observed in mice or cynomolgus monkeys at comparable ethanol doses.     

Free Light Chains of Immunoglobulins in Serum from Patients with Leukaemias and Multiple Myeloma

The serum concentration of free kappa and lambda light chains of immunoglobulins was measured in 114 patients with myelo- and lymphoproliferative disorders including multiple myeloma. Increased concentrations of a single light chain type, suggesting monoclonal origin, were found with high frequency in B-cell diseases only. Thus 6 out of 9 patients with chronic lymphatic leukaemia and 24 of 28 patients with multiple myeloma had increased concentrations of a single chain type. The highest values reported in chronic lymphatic leukaemia were approximately 10 and in multiple myeloma 1000 times normal mean. Cytostatic treatment of chronic lymphatic leukaemia was followed by a decrease in the light chain levels. The levels were, however, not correlated to the number of circulating lymphocytes, the lymphatic infiltration of tissue or clinical activity. Increased concentrations of both chain types, suggesting a polyclonal origin, were found in both of 2 patients with acute monocytic leukaemia, 6 of 7 with acute myelomonocytic leukaemia, 2 of 23 with acute myeloid leukaemia and 1 of 7 with acute lymphoblastic leukaemia. The highest levels of light chains in these groups were 5 times normal mean. All patients with myeloproliferative disorders revealed normal values of both light chain types.     

Two cases of γD Myeloma‡

Summary: The clinical and immunological features of 2 cases of γD myeloma are compared with other reported cases of γD myeloma and other types of multiple myeloma. Both cases showed extra-medullary plasmacytoma which has recently been described as a frequent finding in γD myeloma but which is unusual in other types of multiple myeloma. Bence Jones proteinuria was present in both cases. This is in line with the higher incidence in γD myeloma compared with other types of multiple myeloma. Immunoelectrophoresis showed one case with the more typical L type light chain, the other with the less common K type. Serum levels of the γD paraprotein were low in both instances, reflecting the higher catabolic rate of γD immunoglobulin.     

Frequency of δ+27-thalassaemia in Sardinians

Summary To determine the incidence of δ⁺27 thalassaemia in Northern Sardinia we examined blood samples from 750 Sardinian schoolboys by PCR-based molecular analysis. The incidence of δ⁺27 mutation was 1.2% in this study, i.e. twice as high as previously described on the basis of phenotypical studies; the frequency of the β-thalassaemia is 10.5% and their interaction has been calculated at 0.0003. The majority of δ⁺27 carriers are characterized by a HbA₂ level lower than 1.9% and the mean HbA₂ level is significantly lower than in normal subjects. All compound heterozygotes for δ⁺27 and β-thalassaemia show a silent β-thalassaemic phenotype related to normalization of their HbA₂ levels. This study suggests that δ⁺27 thalassaemia should be borne in mind in counselling at-risk couples in which one member has the typical high HbA₂ β-thal trait while the other shows normal or borderline HbA₂ level. In these subjects, PCR-based ECO O 109 I digestion of the δ globin gene allows rapid detection of the δ⁺27 mutation.     

Peripheral T cell receptors αβ and γδ in patients with Hodgkin's lymphoma

Antigen recognition by T cells is determined by an antigen specific T cell receptor (TCR). Two heterodimeric TCR structures associated with CD3 have been defined: TCR αβ and TCR γδ. TCR αβ and its function are well described but the role of TCR γδ in normal and lymphoproliferative disorders is not well established. In newly diagnosed or relapsed/refractory Hodgkin's disease (HD), a disease associated with defective T cell functions and increased sIL-2R, We determined levels of seven TCR αβ variable regions [βV5(a), βV5(b), βV6(a), βV12(a), αβV(a), αV2(a)] and TCR γδ by using monoclonal antibodies (MCA). TCR γδ levels did not show any difference, but several variable regions of the TCR αβ differed when groups are compared with each other and the control group.     

Hemoglobinopathies in the dogon country: Presence of βs, βc,and δa'genes

The population of the Dogon, located in Mali, is divided in an endogamic Noble class and two endogamic servant castes (Tanners and Blacksmiths). We find that the polymorphic frequencies of β^c, β^s, and, unexpectedly, a mutation of the δ-chain (δ^A'), are geographically (valley vs. plateau) as well as social status dependent. © 1994 Wiley-Liss, Inc.     

Genetic interactions in thalassemia intermedia: Analysis of β-Mutations, α-Genotype, γ-Promoters,and β-LCR hypersensitive sites 2 and 4 in Italian patients

In order to verity the genetic factors influencing the clinical expression of β-thalassemla we have studied 292 Kalian patients, 165 with thalassemia intermedia and 127 with thalassemia major. The β-globin gene mutations were defined in all cases. The number of α-globin genes and the integrity of specific control regions of the β-globin cluster—γ promoters and β-Locus Control Region (β-LCR)—were studied in selected cases. Homozygosity for mild mutations (group I) accounts for 24% of the intermedia patients and it is not represented among major patients. Forty-four percent of intermedia patients had combinations of mild/severe (group II) mutations and 32% had homozygosity or double heterozygosity for severe mutations (group III). Seventy-six percent of patients with thalassemia major were classified in group III end 24% in group II. Deletion type —α^3.7 thalassemia, assessed in a part of the cases, was found in 5% of thalassemia major and 19.5% of Intermedia patients in groups II and III. Structural analysis of γ promoters and β-LCR HS2 and HS4 regions, carried out in order to look for alterations associated with Hb F increase, did not reveal new mutations. Only rare polymorphic changes were observed at the HS2 and HS4 level. The ? 158 γ C T change was found with an increased incidence in intermedia patients in groups II and III. A subset of 10 β-thalassemia heterozygotes with mild intermedia phenotype resulted from coinheritance of a triplicated α-locus. We have been unable to find a molecular basis for the benign clinical course in approximately 20% of patients with thalassemia intermedia. Other genetic or acquired factors must be hypothesized which ameliorate the clinical condition.     

Serum γ-glutamyl transpeptidase activity in liver disease

Serum gamma-glutamyl transpeptidase (GGT) activity correlates closely with the activities of alkaline phosphatase (ALP) and 5'-nucleotidase (5NT) in various forms of liver disease. Maximum elevations of all three enzyme activities are observed in diseases which particularly affect the biliary tract. Compared with the other two enzymes GGT is generally increased to a greater extent and is thus the most sensitive indicator of biliary-tract disease, while estimations of serum GGT are more reproducible than those of 5NT. However, a group of patients who had been treated with phenytoin and barbiturates were found to have elevated serum GGT activities without any other evidence of liver disease. The apparent effect of certain drugs on serum GGT activity indicates the need for caution in interpreting the results of this test.     

Serum Concentrations of Immunoglobulins and Free Light Chains before and after Vascular Events in Essential Hypertension

Serum concentrations of immunoglobulins (Ig) and free light chains (LC) of Ig have been determined at regular intervals during five years in patients with essential hypertension and correlated to development of vascular complications. In 6 cardiac event patients serum IgG increased by 11.9% (p<0.05) and LC by 19.8% (p<0.05). IgG levels correlated positively with LC levels (r= 0.59, p<0.001) and changes in IgG correlated positively with changes in LC (r=0.46, p<0.01). The post-event increments in IgG persisted up to 21 months. No significant changes in serum concentrations of Ig A, IgM, albumin and creatinine were found. In 10 cerebral event patients and in matched groups of 15 uncomplicated hypertensive patients, no significant changes in the above parameters could be demonstrated. It is concluded that heart tissue damage in essential hypertension induces a long-lasting stimulation of B-lymphocytes, and it is suggested that the high Ig levels found in as yet uncomplicated hypertensive patients may reflect an ongoing damage of the vascular bed.     

Fidelity of G protein β-subunit association by the G protein γ-subunit-like domains of RGS6, RGS7, and RGS11

Several regulators of G protein signaling (RGS) proteins contain a G protein gamma-subunit-like (GGL) domain, which, as we have shown, binds to Gbeta5 subunits. Here, we extend our original findings by describing another GGL-domain-containing RGS, human RGS6. When RGS6 is coexpressed with different Gbeta subunits, only RGS6 and Gbeta5 interact. The expression of mRNA for RGS6 and Gbeta5 in human tissues overlaps. Predictions of alpha-helical and coiled-coil character within GGL domains, coupled with measurements of Gbeta binding by GGL domain mutants, support the contention that Ggamma-like regions within RGS proteins interact with Gbeta5 subunits in a fashion comparable to conventional Gbeta/Ggamma pairings. Mutation of the highly conserved Phe-61 residue of Ggamma2 to tryptophan, the residue present in all GGL domains, increases the stability of the Gbeta5/Ggamma2 heterodimer, highlighting the importance of this residue to GGL/Gbeta5 association.     

Serum Neopterin and β2-Microglobulin Concentrations in Monoclonal Gammopathies

ABSTRACT Serum neopterin and β₂-microglobulin concentrations were investigated in 46 patients with multiple myeloma and in 28 patients with asymptomatic monoclonal gammopathy followed for long periods (median 9.6 years) and showing an absence of evolution. Seventy-two per cent of the patients with multiple myeloma showed β₂-microglobulin concentrations higher than 3 mg/1 with a mean of 6.84 mg/1, whereas all the patients with asymptomatic monoclonal gammopathies had concentrations lower than 3 mg/1 with a mean of 1.64 mg/1. Concerning serum neopterin concentrations, 91% of the patients with multiple myeloma had values with in pathological limits (>8 nmol/1) with a mean of 34 nmol/1, whereas all but one of the patients with asymptomatic monoclonal gammopathy had normal values with a mean of 5.19 nmol/1. The differences thus observed in these two groups of patients are highly significant (p<0.001). Serum neopterin concentration, unrelated to renal insufficiency, seems to be useful in the differentiation of malignant or benign asymptomatic monoclonal gammopathies.