Informal in-hospital care in a rehabilitation setting in Greece: an estimation of the nursing staff required for substituting this care.

PURPOSE: To explore: (a) the type and frequency of care-giving activities provided by family members in the Rehabilitation Setting (RS), (b) opportunities for family members to receive training in care-giving activities, (c) to what extent caregivers feel free to ask the nursing staff for help and (d) to estimate the number of nursing staff required to substitute this care and thus to estimate the money saved by the RS due to the in-hospital informal care. METHOD: A convenience sample of 80 family members was selected. A questionnaire was developed to investigate several aspects of informal in-hospital care. Data was analysed using SPSS for Windows (Release 10.1). RESULTS: Cultural reasons and nursing staff shortage led 78.8% (n = 63) of the sample to provide informal in-hospital care. Oral and facial care (67.5%), help with getting dressed (62.5%), help with feeding (61.25%, n = 49), making patients' beds (57.5%, n = 46) and assistance with transferring patients from one hospital department to another (56.25%, n = 45) was provided on a daily basis by the subjects. 48.75%, (n = 39) changed sheets 1 - 2 times per week, while assistance with transfers from bed to wheel-chair and vice-versa (43.75%, n = 35) was provided 3 - 4 times per week. The estimated total time spent per week by the subjects on care-giving activities was 34,034 minutes that corresponds to a total of 75.6 working days or 15.12 working weeks. In order to substitute this care, the RS would need to hire 17 more assistant nurses, entailing a cost of from Euro 14,450 to Euro 20,060 per month. CONCLUSIONS: Informal in-hospital care is provided by Greek families in the RS. Nursing care staff shortage combined with cultural factors are the main reasons for this phenomenon. However, it saves the RS and the Greek State money and policy makers should be looking for ways to overcome the nursing shortage.

Blockade of Kupffer cells by gadolinium chloride reduces lipid peroxidation and protects liver from ischemia/reperfusion injury.

BACKGROUND/AIMS: The implication of lipid peroxidation in the inhibitory effect of GdCl3 (gadolinium chloride) on Kupffer cells activation has not been extensively investigated. The aim of this study was to examine the effect of GdCl3 inhibition of Kupffer cells activation on lipid peroxidation after severe total hepatic ischemia/reperfusion. METHODOLOGY: Male Wistar rats (n = 40) were randomly divided into a sham-operation group, a control ischemia/reperfusion group, and two ischemia/reperfusion groups pretreated with GdCl3 (10 mg and 20 mg/kg bw intravenously, 48 and 24 h prior to operation). Following 60 min of total hepatic ischemia and 120 min of reperfusion, the rats were sacrificed, and liver samples were taken for determination of malondialdehyde and light microscopy examination. Blood samples were also taken for assay of aspartate and alanine transaminase. Additional animals (n = 60) were followed up for a 7-day survival rate determination. RESULTS: Ischemia/reperfusion decreased the survival rate to 13.3%, increased (p < 0.001) the levels of aspartate and alanine transaminase in serum to 2387 +/- 75 and 2157 +/- 87 IU/L, respectively, and increased (p < 0.001) malondialdehyde levels in liver to 1.609 +/- 0.096 nmoles/g compared with 1.164 +/- 0.060 in the sham operation group. Pretreatment with GdCl3 increased the survival rate to 60%, and decreased (p < 0.001) the levels of aspartate transaminase in serum to 1549 +/- 66 and 1496 +/- 55 IU/L, the levels of alanine transaminase in serum to 1302 +/- 48 and 1305 +/- 63 IU/L, and the levels of malondialdehyde in liver to 1.132 +/- 0.034 and 1.149 +/- 0.57 nmoles/g for the lower and the higher doses of GdCl3, respectively. Histological examination showed protection of liver parenchyma in the animals treated with GdCl3. CONCLUSIONS: Experimental data suggest that GdCl3 inhibition of Kupffer cells activation protects liver from ischemia/reperfusion injury by a mechanism that reduces lipid peroxidation.

Plasma atrial natriuretic peptide in essential hypertension after treatment with terazocin.

The aim of this study was to evaluate the medium term effects of the selective alpha1-adrenenergic- blocker terazocin on atrial natriuretic peptide (ANP) levels in patients with moderate essential hypertension. The drug was given orally for 30 days. The daily dose was 1 mg for the first 7 days, 2 mg for the next 7 days and 5 mg for the remaining period of this clinical trial. At the end of this clinical trial, plasma ANP levels increased by 16.40% despite the drop in blood pressure while left atrial and ventricular diameters remained unchanged. These findings indicate that the increase of ANP plasma levels is not the result of a mechanical load on the left cardiac chambers but the result of a pharmacological action. These observations also indicate that terazocin exerts part of its antihypertensive action by increasing ANP plasma levels.

The pathogenetic influence of I-parathyroid hormone on slipped capital femoral epiphysis. Towards a new etiologic approach?

The displacement of the femoral head along the upper femoral physis that occurs during adolescence or slipped capital femoral epiphysis (SCFE) is not a very common traumatic entity. Ever since Muller1 first described it in 1888, its symptoms, clinical manifestations, diagnosis, treatment and complications have been thoroughly described and studied. Nevertheless little progress has been accomplished as far as its etiology is concerned. In order to assess the potential pathologic influence of any parathyroid hormone (PTH) disturbances on the development of SCFE, we conducted a prospective clinical study with 14 patients, 7 boys and 7 girls (16 hips), suffering from SCFE (Group A). Another 5 patients who had been treated for SCFE a few years before the study, were used as a control group (Group B). We measured the level of I-PTH along with serum calcium (Ca) and phosphorus (P) levels. Furthermore, we checked all the necessary anthropometric characteristics of the patients (i.e., age, height, weight and sexual maturation). Each patient of Group A was categorized from grade I to grade V according to the progress of the slipping. The results showed an increased incidence of serum PTH level abnormalities (both decrease and increase) in Group A while Group B patients had normal results. The detected I-PTH serum level abnormalities were not in any pattern related to the Ca and P serum levels. We believe that a temporary parathyroid hormone disorder during the early years of adolescence may play a potentially significant role (along with other etiologic factors) in the development of SCFE.

Severe total hepatic ischemia and reperfusion: relationship between very high alpha-tocopherol uptake and lipid peroxidation.

Reperfusion injury of the liver occurs in liver transplantation and in major hepatectomies. It triggers a severe oxidative stress that leads to increased lipid peroxidation. In our study we examined the effect of parenteral supranutritional administration of alpha-tocopherol, a vitamin that plays a key role in the endogenous antioxidant system, to rats subjected to severe ischemia/reperfusion (I/R) injury of the liver. alpha-Tocopherol was administered to the animals at doses of 30 and 300 mg/kg bw, whereas total hepatic ischemia was induced for 60 min followed by 120 min reperfusion. Tissue and blood samples were collected for malonyldialdehyde (MDA) and serum alpha-tocopherol assay, respectively. In the sham operation group, mean MDA level in liver was 1.14 nmole/g wet tissue in the control subgroup, and 1.01 or 0.74 nmole/g wet tissue in the subgroups given 30 or 300 mg/kg alpha-tocopherol. In the I/R group, mean MDA level was 1.57 nmole/g wet tissue in the control subgroup, and 0.97 and 0.77 nmole/g wet tissue in the subgroups given 30 or 300 mg/kg alpha-tocopherol. Mean levels of alpha-tocopherol in serum (mumole/l) were 10.20 and 1.80 in the control subgroups, 25.28 and 11.25 in the subgroups treated with 30 and 300 mg/kg bw of alpha-tocopherol, and 31.00 and 13.02 in the subgroups treated with 30 and 300 mg/kg bw of alpha-tocopherol, within the sham-operation and I/R groups, respectively. A significant decrease of MDA accompanied by a significant increase of serum alpha-tocopherol was documented in the alpha-tocopherol-treated rats within both groups. Ischemia/reperfusion triggered a significant increase of the MDA level in the liver of the rats not treated with alpha-tocopherol as compares with the treated animals.

Transforming and insulin-like growth factors in the aqueous humour of patients with exfoliation syndrome.

BACKGROUND: Growth factors circulating with the aqueous may play an important role in the pathogenesis of exfoliation syndrome (XFS), which is characterized by excessive synthesis and accumulation of abnormal extracellular material. METHODS: We investigated the concentration of three ubiquitous growth factors (TGF-beta1, TGF-beta2 and IGF-1) in the aqueous humour of 50 patients with XFS (27 from Erlangen, 23 from Thessaloniki) and 54 age-matched controls (27 from Erlangen, 27 from Thessaloniki). This study was performed in two centres, independently of each other, using different assay systems. RESULTS: In the aqueous humour samples collected in Erlangen, both the levels of total TGF-beta1 (P<0.001) and mature TGF-beta1 (P<0.05) were significantly increased in XFS patients compared with controls. Specifically, for total TGF-beta1 patients with XFS exhibited higher a mean value (90.5 +/- 37.4 pg/ml) than controls (30.2 +/- 8.3 pg/ml). The mean level of mature TGF-beta1 was also higher in XFS (14.2 +/- 2.8 pg/ml) than in controls (4.9 +/- 5.5 pg/ml). No difference was found between XFS and controls in the levels of total or mature TGF beta2 in the aqueous or in the level of these two growth factors in the serum. In aqueous humour samples collected in Thessaloniki a significant difference between XFS and controls was also observed for mature TGF-beta1 (XFS 17.06 +/- 11.02 pg/ml vs controls 9.01 +/- 5.69 pg/ml; P=0.006). No difference was observed in TGF-beta2 concentration or IGF-1 concentration in the aqueous. No correlation could be established between protein concentration and the levels of the three growth factors measured. A significant correlation was found between age and protein concentration in XFS, but not in the controls. CONCLUSION: Since TGF-beta1 induces the synthesis and accumulation of extracellular matrix, it is hypothesized that TGF-beta1 plays an important role in the pathogenesis of XFS. Our data suggest that the increased levels of TGF-beta1 are most likely due to enhanced local synthesis.

Phosphorylation of laminin-1 by protein kinase C.

Several extracellular proteins have been reported to be phosphorylated. Previous studies of our laboratory indicated that laminin-1 can be phosphorylated by protein kinase A (PKA). Moreover, it has been reported that protein kinase C (PKC), although known to be intracellular, can phosphorylate extracellular proteins in the case of cellular damage and/or platelet activation. In the present study we examined the possibility of laminin-1 serving as a substrate of PKC. Amino acid analysis revealed that laminin-1 is phosphorylated by this enzyme on serine residues. Self assembly, heparin binding, and cell attachment on the phosphorylated molecule were then studied. Phosphorylated laminin-1 showed an increased and more rapid self assembly than the non-phosphorylated molecule. Heparin binding and cell attachment experiments indicated enhanced heparin and cell binding capacity of the phosphorylated molecule in comparison to the non- phosphorylated control. These results indicate that laminin-1 can be phosphorylated by protein kinase C. Furthermore, phosphorylation by protein kinase C seems to alter several properties of the molecule, though, the in vivo significance of this phenomenon remains to be studied.

Effect of treatment with GnRH agonists on new markers of bone metabolism.

To assess the utility of new markers in monitoring bone turnover during treatment with GnRH agonists, alkaline phosphatase (total and bone specific) and urinary N-telopeptides were measured. 16 women undergoing treatment with GnRH agonists for endometriosis or leiomyomas were studied before and 3 months after the onset of treatment. N-telopeptide levels increased significantly (44% of baseline, p < 0.05). Bone specific alkaline phosphatase (BALP), measured with a new ELISA assay, was more elevated (40% of baseline, p = 0.001) than total ALP (15% of baseline, p < 0.001). In conclusion, in estrogen deficiency states, urinary N-telopeptide measurements provide a quantitative measure of bone resoption. In the assessment of bone formation, BALP determination is move sensitive than total ALP and this may be clinically useful.

Post traumatic paraplegics living in Athens: the impact of pressure sores and UTIs on everyday life activities.

This paper contains the findings of a scientific research which was done on post-traumatic paraplegics who live in the area of Athens. Greece, and measured the impact of pressure sores and UTIs on their everyday life activities. The target population was 127 individuals out of which 98 were observed and interviewed. A semi-structured questionnaire which consisted of two parts used; the first part of the questionnaire included general questions whereas the second contained the Sarno Functional Life Scale (SFLS) which is a tool measuring the level of independence of disabled people. Sixty-two per cent of the population was male. The mean age of the sample population at the time they had the accident which caused the paraplegia was 31.5, while the median was 28.5. The majority of the subjects comes from the lower socioeconomic class. Traffic accidents accounted for the 55% of the occurrences of paraplegia, falls for 37%, surgical complication for 4% and athletic activities for the remaining 4%. Forty-nine percent of the sample had been hospitalised in a rehabilitation centre. However, the multiple regressions used did not reveal any statistically significant relation between the hospitalisation in a rehabilitation centre and the paraplegic's scores in any of the Sarno Functional Life Scale's (SFLS) variables. Instead, the multiple regressions employed yielded paraplegics' scores in the SFLS's everyday life activities that were negatively related to age (P = 0.004) and pressure sores (P = 0.021). The paraplegics' scores on the SFLS's indoors activities are positively related to the years since injury (P = 0.048) and health education on daily fluid consumption (P = 0.003). The scores of the subjects on the SFLS's outdoor activities are positively related to the years of education (P = 0.008), the years since injury (P = 0.011), while are negatively related to pressure sores (P = 0.034) and UTIs (0.044). The subjects' scores on the SFLS variables regarding social relations had a negative relation to sex, (female) (P = 0.0001), age (P = 0.001) and pressure sores (P = 0.019), while they have a positive relation with the years since injury (P = 0.024).

Interleukin-4 suppresses antifungal activity of human mononuclear phagocytes against Candida albicans in association with decreased uptake of blastoconidia.

Pathogenesis of invasive candidiasis may involve regulatory activities of Th2 immunity on phagocytic host defenses. The effects of interleukin (IL)-4 on antifungal capacity of human mononuclear phagocytes against Candida albicans were studied. Incubation of adherent mononuclear leukocytes from healthy donors with IL-4 (1-5 ng ml(-1)) at 37 degrees C for 2-4 days suppressed uptake of C. albicans blastoconidia in the presence of human serum (P < or = 0.01), and anti-IL-4 inhibited its suppressive effect. The effect of IL-4 was protein synthesis-dependent. Interferon-gamma (0.25-25 ng ml(-1)), granulocyte-macrophage colony-stimulating factor (CSF, 20 ng ml(-1)), macrophage-CSF (15 ng ml(-1)) but not IL-10 (100 ng ml(-1)) somewhat counteracted the suppressive effect of IL-4. In contrast, mannose receptor-mediated uptake of blastoconidia in the absence of serum was increased by IL-4. Killing of conidia was decreased after incubation of morphonuclear leukocytes with IL-4 for 2 days (P < 0.05). While superoxide anion production in response to phorbol myristate acetate was decreased by IL-4 (P < 0.05), it was not altered in response to blastoconidia and pseudohyphae. Morphonuclear leukocyte-induced pseudohyphal damage also remained unaltered. These findings suggest that IL-4 plays its detrimental role in invasive candidiasis by predominantly suppressing uptake and killing of blastoconidia by morphonuclear leukocytes. Anti-IL-4, IFN-gamma, GM-CSF and M-CSF appear to counteract suppression of morphonuclear leukocyte phagocytic activity suggesting new approaches to the management of disseminated candidiasis.

Hepatic tissue microcirculation, oxygenation and energy charge in ischemia-reperfusion subjected cirrhotic rat liver.

BACKGROUND/AIMS: The hemodynamic disturbances in the cirrhotic liver following severe variceal bleeding and subsequent restoration by blood transfusion is an ischemia/reperfusion injury event which represents the clinical situation of liver dysfunction. Therefore, the aim of this study was to evaluate the microcirculation, oxygenation and energy charge of the cirrhotic rat liver after ischemia/reperfusion. METHODOLOGY: In eight carbon tetrachloride-induced cirrhotic rats and an equal number of controls subjected to 30 minutes of ischemia and 60 minutes of reperfusion by hepatoduodenal ligament clamping, the following parameters were assessed: hepatic microcirculation by laser-Doppler fluxmetry, hepatic tissue oxygenation by a Clark-type electrode, hepatic energy charge by tissue sampling and adenine-nucleotides determination by means of high-performance liquid chromatography. RESULTS: At baseline, liver microcirculation was found to be significantly decreased in the cirrhotics versus controls groups. Ischemia led to a reduction in both groups, while reperfusion improved microcirculation, but not to the baseline level. Oxygenation was reduced during ischemia and restored after reperfusion in both groups. Hepatic energy charge was reduced in the cirrhotics versus controls at baseline, and significantly decreased during ischemia in both groups. At reperfusion, a further reduction was found in the cirrhotic group, while in the control group it was restored to baseline. CONCLUSION: Hepatic microcirculation, oxygenation and energy charge are subjected to different degrees of diminution after ischemia/reperfusion in the cirrhotic rat liver.

IL-10 exerts suppressive and enhancing effects on antifungal activity of mononuclear phagocytes against Aspergillus fumigatus.

Invasive aspergillosis has emerged as a frequent and serious infection in immunocompromised patients. We studied the effects of human IL-10 on antifungal activity of monocytes (MNCs) from healthy adults against Aspergillus fumigatus after incubation with IL-10 at 37 degrees C for 2 to 4 days. IL-10 (2-20 ng/ml)-pretreated MNCs exhibited approximately 40% suppression of superoxide anion (02-) production in response to PMA and FMLP (p < or = 0.003), and anti-IL-10 containing supernatant neutralized the IL-10 effect. IL-10 (20 ng/ml)-pretreated MNCs exhibited decreased damage of Aspergillus hyphae after 2 to 4 days (55-98% decrease, p < or = 0.04). The MNC phagocytic activity against conidia, as assessed by microscopy (percentage of phagocytosing MNCs and number of intracellular conidia per MNC) as well as by colony counting (colonies grown from intracellular conidia), was enhanced by 127% (p = 0.006), 14% (p = 0.03), and 23% (p = 0.009), respectively. MNC capacity to inhibit intracellular germination was marginally enhanced (p = 0.04) and intracellular conidiocidal activity was unaffected by IL-10. IL-4 (5 ng/ml) did not change the up-regulatory IL-10 effect on phagocytosis. IFN-gamma (25 ng/ml) and granulocyte-macrophage CSF (20 ng/ml), but not macrophage CSF (15 ng/ml), appeared to counteract suppressive IL-10 effects. Thus, IL-10 suppresses oxidative burst and antifungal activity of MNCs against Aspergillus hyphae, while increasing their phagocytic activity. These findings further elucidate a potential role of IL-10 in the pathogenesis of invasive aspergillosis, which may lead to new treatment strategies.

Pyridoxine deficiency: new approaches in immunosuppression and chemotherapy.

Pyridoxine deficiency leads to impairment of immune responses. It appears that the basic derangement is the decreased rate of production of one-carbon units necessary for the synthesis of nucleic acids. The key factor is a pyridoxine enzyme, serine hydroxymethyltransferase. This enzyme is very low in resting lymphocytes but increases significantly under the influence of antigenic or mitogenic stimuli, thus supplying the increased demand for nucleic acid synthesis during an immune response. Serine hydroxymethyltransferase activity is depressed by deoxypyridoxine, a potent antagonist of pyridoxal phosphate, and also by known immunosuppressive or antiproliferative agents. The combination of these agents is additive. Our results lead us to suggest the following medical applications: (a) combination of deoxypyridoxine with immunosuppressive or chemotherapeutic drugs may be effective in cases of immunosuppressive therapy or organ transplantation, (b) the development of special agents directed against the serine hydroxymethyltransferase apoprotein may prove to be a valuable medical tool, since this enzyme presents an excellent target for chemotherapy, (c) lymphocytes of individual patients could be used to design tailor-made specific immunosuppressive or chemotherapeutic treatment, and (d) the serine hydroxymethyltransferase activity of lymphocyte culture presents an excellent indicator for the evaluation of potency of immunosuppressive, chemotherapeutic or genotoxic compounds in a simple and rapid test.

Haemoglobin O Arab in the Pomak population of Thrace.

A 21 year old woman from Thrace with clinical and laboratory findings of mild hemolysis, was found to be homozygous for haemoglobin O Arab. Blood samples from 15 members of her family and from 42 inhabitants of her village were examined for the presence of Hb O Arab. A high incidence of this variant allele was also detected among the patient's family members as well as in other inhabitants of the village (p = 0.274 +/- 0.049). The possibility that Pomaks, a culturally unique tribe with controversial ethnic origin, may represent one of the original pools for the geographic distribution of the gene in the broader area of the Mediterranean basin is discussed.

Lipid peroxidation after acute renal ischemia and reperfusion in rats: the effect of trimetazidine.

Lipid peroxidation is a critical pathway of reactive oxygen species inducing tissue injury in postischemic acute renal failure. In order to evaluate the effect of renal ischemia reperfusion on kidneys, renal tissue malondialdehyde (MDA, nmol/g wet weight) concentration was measured in 29 male Wistar rats subjected to a midline abdominal incision and 60 min occlusion of the left renal artery. A right nephrectomy was performed at the beginning of the ischemic period. The animals were separated in four groups. Groups 1 (n = 7) and 3 (n = 7) underwent 60 min of ischemia and 15 min of reperfusion, respectively. Groups 2 (n = 8) and 4 (n = 7) were subjected to the same procedure but, in addition, they received 2.5 mg/kg TMZ into the tail vein 2 h prior to the left renal artery occlusion. A significant elevation of MDA after 60 min of ischemia (1.43 vs. 2.1, p < 0.001), which was augmented after 15 min of reperfusion (1.4 vs. 3.72, p < 0.001) was observed. Furthermore, there was a significant reduction of renal tissue MDA in ischemic rats treated with TMZ (group 3) (2.1 vs. 1.52, p < 0.001). The maximum reduction of renal tissue MDA was observed in ischemic-reperfused rats (group 4) that had received TMZ (3.72 vs. 1.36, p < 0.001). It is suggested that lipid peroxidation is a critical event in postischemic acute renal failure, and TMZ is a useful protective agent of renal damage from oxygen free radicals.

Possible role of transferrin in exfoliation syndrome.

Protein concentration was measured in 30 exfoliation syndrome samples and 22 age matched controls. Exfoliation samples contained significantly more protein than controls. We also analyzed by SDS gel electrophoresis 32 aqueous samples, 12 with exfoliation syndrome and 20 age matched controls. A novel finding in all samples was a double band with 77 kDa and 78 kDa. These bands probably represent transferrin isoforms, with different carbohydrate side chains. Exfoliation samples contained a lower amount of the 77 kDa band in comparison to the amount of the 78 kDa band. The lower relative concentration of this transferrin isoform in the exfoliation syndrome samples may be indicative of its possible involvement in the disorder. A different oligosaccharide side chain in combination with a relatively high protein concentration may lead to sedimentation of this molecule. On the other hand, laminin, a glycoprotein detected previously in exfoliation material, contains similar oligosaccharide side chains with transferrin. Thus the oligosaccharide side chains of both proteins may be influenced by the same factors.

In vivo binding of the radioiodinated peptide YIGSR on B16 melanoma cells.

It has been reported that metastatic melanoma cell lines selectively bind in vitro with the synthetic laminin pentapeptide tyrosyl-isoleucyl-glycyl-seryl-arginine (YIGSR). The aim of this study was to investigate whether the same peptide can bind on melanoma cells in vivo as well. Iodine-125-labeled YIGSR was administered to B16 melanoma-bearing animals. Microscopic autoradiography of tumor and organ sections taken 24 h after peptide administration showed that the peptide did accumulate on the surface of certain tumor cells. The peptide binding cells were frequent in metastatic sites and tumors grown for 24 days and rare in tumors grown for 10 days. A similarly radiolabeled control pentapeptide (peptide DRLKY) did not bind to any tumor cell. It is suggested that the YIGSR binding tumor cells may represent a distinct melanoma cell population with a high metastatic potential.

Detection of a high molecular weight heparin binding acidic growth factor in human placenta.

A heparin binding fibroblast growth factor with a molecular weight of approximately 45 kDa has been detected in human placenta. The protein was extracted from placenta and purified by a combination of ammonium sulfate precipitation, DE-52 anion exchange chromatography, G100 gel exclusion and heparin sepharose affinity chromatography. The molecule was shown to be monomeric after treatment with beta-mercaptoethanol, and had an acidic isoelectric point. The properties of the polypeptide described in this paper (45 kDa, Pi 4-5, monomeric, heparin binding fibroblast growth factor), suggest that it may represent an as yet undescribed growth factor.

Rat liver endoplasmic reticulum protein kinases.

1. Rat liver microsomal membranes were studied for the presence of protein kinases. Microsomal proteins solubilized with Triton X-100 were analyzed by means of ion exchange chromatography. 2. Protein kinase activity was detected in the column fractions using specific assays for cAMP-dependent protein kinase, cGMP-dependent protein kinase, protein kinase C, Ca2+/calmodulin-dependent protein kinase and casein kinases. 3. Fractions with protein kinase activity were further analyzed by SDS-polyacrylamide gel electrophoresis. 4. The results indicate that cAMP-dependent protein kinase type I and II, casein kinases I and II, protein kinase C proenzymes I and II and Ca2+/calmodulin kinase II are associated with the membranes of endoplasmic reticulum (ER).

Effect of combination of deoxypyridoxine with known anti-proliferative or immunosuppressive agents on lymphocyte serine hydroxymethyltransferase.

Measurements of serine hydroxymethyltransferase (SHMT) in resting lymphocyte cultures showed that the level of activity of this enzyme is very low. Under the influence of mitogenic stimuli serine hydroxymethyltransferase activity is induced 5-20-fold. Addition in the cultures of 4-deoxypyridoxine (dB6), a potent antagonist of vitamin B6 coenzymes, concurrently with the mitogen, inhibits the induction of SHMT. Separate addition in the cultures of four anti-proliferative (AP) and immunosuppressive (IMS) agents, namely actinomycin, cytarabine, asparaginase and cyclosporine, led to the following observations. (1) The AP and IMS agents produce a decrease in the mitogen-induced activity of SHMT. The higher the concentration of the AP/IMS compound, the greater the decrease of enzymatic activity. (2) When a AP/IMS agent is combined with dB6 its effect on SHMT is considerably greater. (3) Ineffective concentrations of AP/IMS agents become effective when combined with dB6. (4) The observed changes in SHMT activity are not, as one would expect, the same in the case of all four drugs. (5) The combination makes it possible to use much smaller doses of these agents with much better results, at least as far as the decrease of enzymic activity is concerned. This is very promising for clinical use of AP agents in cancer chemotherapy and IMS agents in transplantation especially of the heart and lungs, because combining these compounds with dB6 will make possible to use smaller doses over a longer period of time with greater effectiveness.(ABSTRACT TRUNCATED AT 250 WORDS)