Ethanol-induced apoptosis in human HL-60 cells.

The mechanisms responsible for aberrant immune function associated with chronic ethanol use remain obscure, but a decrease in monocyte numbers is often reported for individuals who chronically abuse ethanol. We investigated, using human HL-60 promyelocytic cell line, the possibility that ethanol induces apoptosis which contributes to decreased monocyte numbers. Characteristic features of apoptosis were observed 4 days after ethanol treatment, as documented by increased DNA fragmentation; enhanced expression of phosphatidylserine, an early marker of apoptosis; and the appearance of a hypodiploid apoptotic cell population identified by flow cytometry analysis of the cell cycle. Treatment with the protein kinase C inhibitor, GF 109203X, potentiated ethanol-induced apoptosis. Direct induction of human HL-60 cell apoptosis by ethanol and potentiation of ethanol-induced apoptosis by inhibiting protein kinase C provides a partial explanation for the cytotoxic effects of ethanol on hematopoietic progenitor cells and establishes a link between inhibiting protein kinase C activity and ethanol-induced apoptosis.

Molecular variants of epidermal growth factor in malignant astrocytoma.

The present study has measured EGF levels in primary brain tumor tissues. EGF levels were measured by specific radioimmunoassay (RIA) and further analyzed by reversed-phase high performance liquid chromatography (RP-HPLC) followed by RIA and radioreceptor binding. The levels of EGF-like immunoreactivity (EGF-LI) in astrocytoma-IV tumors were fourfold greater than those in normal brain tissues. In astrocytoma-II and astrocytoma-III tumors, however, levels of EGF-LI were not different from those in normal brain. HPLC analysis of extracts from normal brain tissue and astrocytoma-II showed one peak of EGF-LI that coeluted with standard human EGF (retention time 22 min). Interestingly, EGF-LI in extracts of astrocytoma-IV tumors eluted in two distinct peaks with retention times of 24 and 26 min (Astro-A and Astro-B, respectively). Materials in both Astro-A and Astro-B peaks reduced the specific binding of [125I]hEGF to EGF receptors in human placental membranes. These studies demonstrate elevated levels of EGF-LI in malignant astrocytoma, but not in benign tumors. Furthermore, two different EGF-like molecules that are different from native EGF are present in malignant astrocytoma.

Hexamethylene bisacetamide-induced differentiation of Friend virus-transformed murine erythroleukemia cells is associated with parallel changes in casein kinase II and guanine nucleotide exchange factor activities.

In mammalian cells, the guanine nucleotide exchange factor (GEF, eIF-2B) plays a major role in the regulation of initiation of protein synthesis. It catalyzes the exchange of eukaryotic chain initiation factor (eIF)-2-bound GDP for GTP and facilitates the recycling of eIF-2 during polypeptide chain initiation. We used the Friend virus-transformed murine erythroleukemia (MEL) cell system to elucidate the translational regulatory processes that occur during growth and hexamethylene bisacetamide (HMBA)-induced cell differentiation. GEF activity is increased during growth and decreased during MEL cell differentiation, and this parallels the overall changes in protein synthesis during this period. Inhibition of GEF activity in induced cells may occur indirectly by phosphorylation of the alpha-subunit of eIF-2. However, the decrease in GEF activity in induced cells cannot be reversed by increasing the concentration of eIF-2-GDP added as a substrate in the GEF assay. This is diagnostic for the presence of eIF-2 alpha(P)-GDP in cell lysates and suggests that regulation of GEF activity may occur by one or more mechanisms other than eIF-2(alpha) phosphorylation. We have previously shown that the activity of GEF may be influenced directly by phosphorylation with casein kinase II (CK-II) of the 82-kD subunit of the factor. CK-II activity parallels the changes in GEF activity and the rate of protein synthesis during growth and differentiation of MEL cells. Addition of 1mM spermidine, a stimulator of CK-II but not of purified GEF, in induced MEL cell extracts enhances both CK-II and GEF activities approximately 48 and 32%, respectively. The results presented suggest that the inhibition of protein synthesis during MEL cell differentiation may be linked to the decreased CK-II and GEF activities.

Inhibition of rabbit reticulocyte guanine nucleotide exchange factor activity by heparin and its reversal by polyamines.

We have previously demonstrated that phosphorylation of the 82-kDa subunit of the guanine nucleotide exchange factor, eIF-2B, by casein kinases I and II stimulates eIF-2B activity. In the present report, we show that heparin binds to eIF-2B (KD approximately 0.3 microM) and inhibits the GDP/GTP exchange activity of eIF-2B. The effect is dose dependent, with half-maximal inhibition (IC50) achieved at 0.03 microM heparin. Although spermidine alone does not stimulate or inhibit guanine nucleotide exchange, it reverses the inhibitory effect of heparin up to 90 percent (IC50 = 0.22 mM). ATP and NADPH which interact with the 65 and 55-kDa subunits also partially relieve heparin inhibition of nucleotide exchange, and the effects of ATP and NADPH are additive. By using a gel overlay technique, we demonstrate that 125I-heparin binds to the 82, 65 and 55-kDa subunits of eIF-2B.

Circulating immune complexes in intracranial neoplasms.

The levels of circulating immune complexes (CIC) were assayed in the sera of 109 patients with intracranial space occupying lesions. The CIC levels were significantly increased in all the brain tumours. After treatment, the CIC levels were still significantly increased when compared to the controls but showed no change when compared to their respective pre-operative values. Further, no change was observed in the CIC levels between the malignant and benign tumour case. Moreover, in brain tumours, 90% of the CIC precipitate consisted of IgG. However, the CIC levels fail to prognosticate the process of the disease in these patients.

Translational control of eukaryotic gene expression. Role of the guanine nucleotide exchange factor and chain initiation factor-2.

In mammalian cells, the guanine nucleotide exchange factor (GEF or eIF-2B) is a key regulator of polypeptide chain initiation. The exchange of GDP bound to chain initiation factor 2 (eIF-2) for GTP by GEF is a rate limiting step in protein synthesis. The multisubunit characteristics of GEF suggest that this protein is composed of several distinct structural and functional domains, and is regulated by allosteric means and by phosphorylation. The activity of GEF may be regulated indirectly by the phosphorylation state of the smallest subunit of eIF-2 (alpha-subunit). On the other hand, phosphorylation of the largest subunit of GEF (82-kD subunit) by casein kinase (CK) I or II stimulates GDP/GTP exchange. GEF contains NADPH which is required for structural integrity of the protein. Upon stimulation of cells by insulin and growth factors, allosteric activation of GEF by sugar phosphates and other effector molecules may also play an important role in the regulation of polypeptide chain initiation. In this article, recent information about structure-function relationship of eIF-2 and GEF in nucleotide exchange and the regulatory mechanisms that influence the rate of polypeptide chain initiation under various physiological and pathological conditions are presented.

Serum adenosine deaminase activity in brain tumours.

Adenosine deaminase (ADA) activity in serum was estimated in 86 patients with intracranial tumours and 40 healthy volunteers. Although high ADA concentrations in biological fluids and tumour tissues were observed in several neoplastic conditions, there was no significant difference in the ADA in sera of brain tumour patients when compared to the control values. Therefore, cell-mediated immunity probably does not play a significant role in brain tumours.

Urinary excretion of pseudouridine in patients with brain tumours.

Elevated urinary levels of modified nucleosides, especially pseudouridine (psi), have been observed in patients with a variety of malignant neoplasms. In this report, the urinary psi levels were estimated by high performance liquid chromatography in 93 patients with brain tumours and in 40 age and sex matched controls. The psi levels were found to be excreted in equal amounts in the control and the tumour groups. There was no significant difference in psi when patients with malignant diseases were compared with those with benign tumours. Following therapy, the psi levels remained unchanged when compared to the controls and their preoperative values. It is concluded that urinary psi concentration is not a useful marker for brain tumours.

Urinary excretion of 6-hydroxymethylpterin in brain tumours.

The urinary 6-hydroxymethylpterin(Pt-6-CH2OH) excretion was determined in 87 patients with brain tumours and in 50 control patients. The Pt-6-CH2OH levels were significantly elevated in all patients with tumours. No difference was observed when malignant tumours were compared with benign neoplasms. Following therapy, the Pt-6-CH2OH levels were partially reduced when compared with control patients and their pre-operative values.

Elevation of serum ceruloplasmin levels in brain tumours.

Serum ceruloplasmin levels have been estimated in 80 patients with various intracranial space occupying lesions and in 30 controls. The ceruloplasmin levels were significantly increased in all brain tumours except in meningiomas. After therapy, the ceruloplasmin levels were still significantly increased when compared to controls and their respective preoperative values. However, the rise in levels of ceruloplasmin in malignant tumours compared to benign was statistically not significant. It is concluded that ceruloplasmin may have a role to play as an acute phase reactant protein in brain tumours.

Evaluation of serum beta 2-microglobulin in oral cancer.

Beta 2-microglobulin (beta 2-m) low molecular mass protein was measured in serum of normal subjects and in patients with oral cancer. There was a significant increase in beta 2-m levels in oral cancer patients. The possible role of beta 2-m as a biochemical parameter and its significance in oral cancer is discussed.

Serum immunoglobulins in brain tumours.

Immunoglobulin level in the sera of 62 patients with intracranial space occupying lesions was assayed using the radial immunodiffusion method. Serum IgM levels showed a highly significant increase in all types of brain tumour when compared to controls. Serum IgG levels were also increased in benign as well as malignant cases. Serum IgA levels were high only in benign cases. Hence, increased serum Ig levels may be of prognostic value in these cases.

Clinical utility of serum fibrinogen degradation products (FDP) in the diagnostic and prognostic evaluation of oral cancer.

Serum Fibrin Degradation Products (FDP) were determined in 50 oral cancer patients and 50 normal individuals prior to any kind of treatment. The mean FDP values were significantly increased with the advancement of stage of oral cancer patients in comparison to the normal individuals.

A case of pre-leukaemia masquerading as haemolytic anaemia due to multi-enzymopathy of the red cell.

A 19 year old woman presented as a case of haemolytic anaemia due to multi-enzyme deficiency of the erythrocyte. After a transient improvement with folic acid therapy, she developed acute myeloblastic leukaemia. This is the second reported case of a myelodysplastic syndrome presenting with a haemolytic picture and subsequently developing an acute myeloblastic leukaemia.

Diagnostic significance of estimation of serum apolipoprotein A along with alpha-fetoprotein in alcoholic cirrhosis and hepatocellular carcinoma patients.

The serum apolipoprotein A (Apo A) and alpha-fetoprotein (AFP) were evaluated in histologically verified 30 cases of alcoholic cirrhosis and 18 cases of hepatocellular carcinoma (HCC). The latter were also divided into subgroups depending on the presence or absence of associated cirrhosis. Serum Apo A levels were found to be significantly decreased in cirrhotics (p less than 0.001) compared to controls and non-cirrhotic HCC patients. In 22 cases of alcoholic cirrhosis (AFP less than 10 ng/ml) and 12 cases of HCC (AFP greater than 600 ng/ml), the AFP levels itself were diagnostic, but in the remaining cases, AFP levels (100-600 ng/ml) were not able to differentiate between cirrhosis and malignancy. In this later group of patients with low pathological range of AFP, serum Apo A levels found to be significantly decreased in alcoholic cirrhotic patients (p less than 0.001) compared to HCC patients. Thus, estimation of Apo A levels may be helpful to interpret the AFP values at lower pathological range due to suspected liver pathology.