Salt shield: intracellular salts provide cellular protection against ionizing radiation in the halophilic archaeon, Halobacterium salinarum NRC-1.

The halophilic archaeon Halobacterium salinarum NRC-1 was used as a model system to investigate cellular damage induced by exposure to high doses of ionizing radiation (IR). Oxidative damages are the main lesions from IR and result from free radicals production via radiolysis of water. This is the first study to quantify DNA base modification in a prokaryote, revealing a direct relationship between yield of DNA lesions and IR dose. Most importantly, our data demonstrate the significance of DNA radiation damage other than strand breaks on cell survival. We also report the first in vivo evidence of reactive oxygen species scavenging by intracellular halides in H. salinarum NRC-1, resulting in increased protection against nucleotide modification and carbonylation of protein residues. Bromide ions, which are highly reactive with hydroxyl radicals, provided the greatest protection to cellular macromolecules. Modified DNA bases were repaired in 2 h post irradiation, indicating effective DNA repair systems. In addition, measurements of H. salinarum NRC-1 cell interior revealed a high Mn/Fe ratio similar to that of Deinococcus radiodurans and other radiation-resistant microorganisms, which has been shown to provide a measure of protection for proteins against oxidative damage. The work presented here supports previous studies showing that radiation resistance is the product of mechanisms for cellular protection and detoxification, as well as for the repair of oxidative damage to cellular macromolecules. The finding that not only Mn/Fe but also the presence of halides can decrease the oxidative damage to DNA and proteins emphasizes the significance of the intracellular milieu in determining microbial radiation resistance.

Adhesion of erythrocytes to endothelial cells after acute exercise: differences in red blood cells from juvenile and adult rats.

Erythrocytes (RBC) from untrained male Wistar rats and rat glomerular endothelial cells (EC) were used to investigate the effects of acute exercise (speed: 20 m/min, slope: 0, duration: 1 hour) on RBC membrane protein oxidation and adhesion to cultured EC. Experimental animals were divided into juvenile (age 10 weeks) and adult (age 30 weeks) groups for these studies. Immediately following exercise, juvenile rat RBC membrane protein oxidation was significantly enhanced. Adult rat RBC showed significantly higher basal protein oxidation than juvenile RBC; but the level of adult rat RBC membrane protein oxidation was unaffected by exercise. Prior to exercise, adult rat RBC showed significantly higher adhesion to EC than RBC of juvenile rat. There was no difference in plasma fibronectin or fibrinogen levels following exercise. Only juvenile rat RBC showed a significant decrease in sialic acid residue content following exercise. These experiments show that there are changes in RBC-EC interactions following exercise that are influenced by animal age.

The effect of interferon alpha administration on acute attacks of familial Mediterranean fever: A double-blind, placebo-controlled trial.

BACKGROUND: About a quarter of familial Mediterranean fever (FMF) patients are partially or totally resistant to colchicine. A previous observation reported that acute attacks may be shortened by administration of interferon alpha (IFN). OBJECTIVE: We designed a double-blind, placebo-controlled trial to test our initial observations of a beneficial response with IFN in FMF attacks. METHODS: We treated 34 acute abdominal attacks with IFN 5 million IU or placebo sc in the early phase of the attack. Leucocytes, thrombocytes, the erythrocyte sedimentation rate, fibrinogen, C-reactive protein (CRP), serum amyloid A protein (SAA), haptoglobin, transferrin, IL-1beta and TNF-alpha were measured at hours 0, 6, 12, 24 and 48. RESULTS: The median time to recovery in those treated with IFN and placebo was not significantly different, while the leucocytosis and high levels of fibrinogen were significantly more prolonged in placebo-treated patients. CRP and SAA were extremely elevated and peaked at 24h, remaining less marked in the IFN-treated patients but the difference was not statistically significant. Observations regarding the other parameters were unremarkable. CONCLUSIONS: Although there were some clues indicating a depressed inflammatory response with IFN, we could not demonstrate a definitive effect of this agent in this double-blind trial. The drug may suppress the acute inflammation of FMF only if administered at the earliest phase. CRP and SAA may be more sensitive indicators of an attack than ESR or fibrinogen.

Significance of heat shock protein-27 expression in patients with renal cell carcinoma.

OBJECTIVES: To investigate heat shock protein (HSP)-27 expression in patients with renal cell carcinoma (RCC) and examine its biologic significance. HSPs were first defined as proteins induced by heat shock and other environmental and pathophysiologic stresses. They are implicated in protein-protein interactions and are thought to play an important role in cancer. The expression of HSP-27 has been demonstrated in some human tumors. METHODS: The expression of HSP-27 was studied in tumor and normal parenchyma tissue specimens from 76 patients with RCC by immunohistochemistry. The findings were correlated with clinical stage, lymph node metastasis, histologic grade, and survival. RESULTS: Of the 76 RCC tissue specimens studied, the presence of HSP-27 was demonstrated in 73 (96%). The expression was low in 10 patients (14%), intermediate in 38 (50%), and high in 25 (33%). HSP-27 expression was greater in RCC tissue compared with adjacent noncancerous renal tissue (P <0.001). An inverse relationship was found between tumor stage and HSP-27 expression (r = -0.281, P = 0.016). However, no statistically significant difference was observed in progression-free survival with respect to HSP-27 expression. No relationship was found between HSP-27 expression and tumor grade, lymph node metastasis, distant metastasis, or cause-specific survival. CONCLUSIONS: Our data suggest that HSP-27 expression is not a powerful and statistically significant prognostic indicator for disease-free survival for patients with RCC.

Nitric oxide as an activity marker in multiple sclerosis.

Nitric oxide (NO) molecules have one of the most important roles in the pathogenesis of multiple sclerosis (MS). It has been stated that a continuous and high concentration of NO metabolites in CSF and in the serum of MS patients in relapse may cause toxic damage to myelin and oligodendroglia. The aim of this study was to investigate whether NO is a marker of disease activity and is correlated with other disease activity markers such as active lesions on brain magnetic resonance imaging (MRI) and increased immunoglobulin G (IgG) index. Cerebrospinal fluid (CSF) and peripheral serum (PS) samples were taken from patients with definite MS (n = 24) during relapse and remission and from control subjects (n = 18). The Griess reaction was used to measure the NO metabolites, nitrite and nitrate in CSF and PS. Cranial MRI was carried out with triple dose (0,3 mmol/kg) gadolinium and the IgG index was determined. Nitrite and nitrate concentrations (NNCs) of CSF were 11.16 +/- 8.60 micromol/ml in relapse and 6.72 +/- 3.50 micromol/ml in remission, whereas in PS they were 12.89 +/- 7.62 micromol/ml during relapse and 12.35 +/- 6.62 micromol/ml during remission. In control subjects NNCs in CSF and PS were 7.42 +/- 2.81 micromol/ml and 4.37 +/- 1.63 micromol/ml respectively. NNCs in CSF during relapse period were significantly higher than those of both remission phase and control subjects (p = 0.000). Although serum NNCs did not differ in relapse and remission, they were still higher than normal controls. Validity analysis revealed that NNC measurement in CSF was 71 % specific and 66 % sensitive to disease activity. The most important result was the significant correlation of increased NNCs with the existence of active lesion in cranial MRI and an increase in IgG index (p < 0.05).In conclusion, these results add background data to assist in further outlining the possible role of NO in the pathogenesis of MS. Together with the other markers it may be used as an activity marker in relapses of MS.

The relationship between neutrophils and incisional wound healing.

The systemic administration of granulocyte-macrophage colony-stimulating factor (GM-CSF) is used clinically to increase circulating neutrophils, but its wound healing effects after intraperitoneal treatment have not been studied yet. We planned to investigate the effect of neutrophils on wound healing under cyclophosphamide and GM-CSF treatment. Forty rats were divided into three groups: control group (group I, n = 12) receiving saline, group II (n = 14) receiving cyclophosphamide and group III (n = 14) receiving GM- CSF. The rats in all groups underwent incisional wounding and were euthanized after 7 days. Blood neutrophil counts and functions, tensile strengths and the hydroxyproline level of skin were determined, and a histopathological evaluation of healing was made. Neutrophil counts and phagocytosis significantly increased in group III and decreased in group II. Although the skin hydroxyproline level did not differ, there was a difference in tensile strength of the wounded skin between group II and group III. The wound score in group II was lower than that in groups III and I. As a result we suggest that systemically given GM-CSF - by increasing the neutrophil count and neutrophil phagocytosis index - can enhance the tensile strength of surgical incisions.

Significance of tissue laminin P(1) elastase and fibronectin levels in transitional cell carcinoma of the bladder.

OBJECTIVE: Elastase is a serine protease which hydrolyses connective tissue components. Laminin and fibronectin also play an important role in progression and invasion of cancer. The purpose of this study is to investigate the relation between tissue elastase, laminin P(1) and fibronectin levels and tumor characteristics, and analyze the potential of these as prognostic factors in transitional cell carcinoma (TCC) of the bladder. METHODS: Thirty-four patients with TCC of the bladder and 11 controls were included in this study. Elastase and fibronectin levels in tissue homogenates were determined using an enzyme immunoassay and laminin P(1) by radioimmunoassay. Mean follow-up was 43 months. RESULTS: The mean elastase level in bladder carcinoma tissue was 120+/-11.42 ng/homogenate protein, while normal tissue level was 12.36+/-2.70 (p<0.01). The carcinoma and normal tissue mean laminin P(1) levels were 7.02+/-0.37 U and 0.65+/-0.10 U/mg homogenate protein, respectively (p<0.01). The mean fibronectin level was 19.97+/-1.45 ng/mg homogenate protein in the carcinoma tissue and 2.16+/-0.40 in normal tissue (p<0.01). There was no correlation between tumor stage, grade, size, multiplicity and elastase, laminin P(1) and fibronectin levels. CONCLUSION: These results provide evidence that tissue elastase, laminin P(1) and fibronectin levels increase in TCC of the human bladder. Further studies including serum and urine levels should be performed in order to analyze their value as tumor markers in a larger group of patients.

Serum ferritin as a clinical marker for renal cell carcinoma: influence of tumor size and volume.

OBJECTIVES: There is no established tumor marker for renal cell carcinoma (RCC). Ferritin is shown to be expressed by the tumor, and proposed as a tumor marker. The aim of this study is to assess the relation between ferritin levels and tumor volume, size and prognosis in RCC. METHODS: We studied ferritin levels in serum from peripheral and renal veins of 52 patients with RCC who underwent surgery. Ferritin levels were measured by an enzyme immunoassay method. Tumor volume and the largest tumor diameter were calculated from the pathologic specimens. RESULTS: The mean serum ferritin level from the renal vein (RVF) was statistically higher than the ferritin level from the peripheral vein (PVF) (p = 0.028). Although mean RVF level increased with increasing stage, it was not significant. While there was a correlation with tumor size, volume and RVF, PVF was in correlation with disease status. PVF in patients with metastatic and/or locally advanced disease was significantly higher than the patients with localized disease (p = 0.023). The initial RVF and PVF levels were predictive of survival (p = 0.028 and p = 0.034, respectively). CONCLUSIONS: Higher levels in the renal vein, its positive correlation with tumor size and volume suggest that ferritin is expressed by RCC. Initial peripheral serum values of ferritin can be indicative of disease status and also be a prognosticator of survival.

The role of neutrophils in the formation of peritoneal adhesions.

The most common cause of intraperitoneal adhesions which may result in infertility and intestinal obstruction is previous abdominal surgery. Surgical trauma of the peritoneum in the absence of infection elicits a rapid and transient influx of polymorphonuclear leukocytes (PMN) into the peritoneal cavity. The role of neutrophils in intraperitoneal adhesion formation has not been studied. We aimed to study the effects of PMN counts and PMN functions on peritoneal adhesion formation. Forty peritoneal adhesion-induced rats were randomly divided into three groups; group I, receiving saline; group II, receiving cyclophosphamide; and group III, receiving granulocyte-macrophage colony-stimulating factor (GM-CSF). In all groups, peritoneal lavage was performed to determine PMN counts the day after adhesion induction. Blood neutrophil counts and neutrophil functions were also determined. Adhesions were evaluated blindly 14 days after the operation. Adhesion tissue samples were microscopically evaluated. Tissue hydroxyproline and collagen concentrations were measured. The neutrophil counts and phagocytosis significantly increased in group III and neutrophil counts decreased in group II (P < 0.05). The score of adhesion formation in group II was significantly less than that in groups I and III (P < 0.05). Hydroxyproline concentrations of adhesion tissue were significantly decreased in group II when compared with group III (P < 0.05). The present study shows that neutropenia lowers the degree of postoperative adhesion formation. It is concluded that PMN may have a role to play in modulating post-operative adhesion formation.

Effects of granulocyte-macrophage colony-stimulating factor on incisional wound healing in an experimental diabetic rat model.

The exact nature of poor wound healing in diabetes is uncertain. Neutrophils play a critical role in the host defense mechanism, and it is suggested that impaired neutrophil functions cause healing difficulties with or without infections in diabetic patients. Granulocyte-macrophage colony-stimulating factor (GM-CSF) is used clinically when given systematically to increase the circulating neutrophils, but its wound-healing effects have not been systematically studied. This study was undertaken to examine the effects of GM-CSF on incisional wound healing in an experimental diabetic rat model. Forty rats were randomly divided into three groups, group I receiving saline as control, diabetes-induced group II receiving saline and diabetes-induced group III receiving GM-CSF. The anesthetized rats in all groups were wounded 21 days after diabetes induction by streptozotocin. Blood neutrophil counts and neutrophil fractions were also determined three days after wounding. Tensile strengths of wounded skin and the hydroxyproline (hyp) level of the wound were determined and wound healing processes were evaluated by light and electron microscopy, fourteen days after wounding. Neutrophil counts and phagocytosis were significantly increased in group III and neutrophil counts decreased in group II (p < 0.05). Although the hydroxyproline level of wound tissue significantly decreased in group II as compared with group III (p < 0.05), there was no differences of tensile strength between group II and III (p < 0.05). Wound score in group II was less than that in groups I and III (p < 0.05). It is concluded that PMN may have a role in modulating wound healing. GM-CSF may be useful for creating better wound healing healing. GM-CSF may be useful for creating better wound healing in risky patients such as diabetics.

Ischemia/reperfusion injury in flow-through venous flaps.

The effects of ischemia/reperfusion (I/R) injury in flow-through venous flaps were evaluated in rabbits. The rates of flap survival and the levels of lipid peroxidation, protein oxidation, and sulfhydryl groups were compared between flow-through venous flaps, conventional flaps after an I/R period (experimental groups), and flow-through venous flaps without being subjected to I/R injury (control groups) in 20 animals. On the seventh day after the onset of reperfusion, 3 of 10 flow-through venous flaps (30%) and 6 of 10 arteriovenous flaps (60%) survived in the experimental groups. Flow-through venous flaps showed a decreased survival rate compared with control and conventional flaps (p < 0.05). Tissue lipid peroxide levels were found to be higher in venous flaps during reperfusion after secondary ischemia (p < 0.05). Tissue protein oxidation and total sulfhydryl groups levels did not show any difference among groups. This study suggests that more free radical damage occurs in flow-through venous flaps during I/R injury.

Significance of serum laminin P1 values in patients with transitional cell carcinoma of the bladder.

OBJECTIVES: The goal of this study was to investigate the relation between serum laminin P1 expression and the grade, stage, size, and multiplicity of the tumors to elucidate the potential role of laminin as a diagnostic and prognostic factor in transitional cell carcinoma (TCC) of the bladder. METHODS: Serum concentration of laminin P1 was measured by radioimmunoassay in 38 patients with TCC of the bladder and compared with the serum laminin P1 values obtained from 34 healthy control subjects. Patients were grouped according to their tumor stage, grade, size, and multiplicity to compare the serum laminin P1 values. RESULTS: The mean serum laminin P1 level in the patient group was statistically higher than the controls (P = 1.3 x 10(-8); with the destruction of basement membrane (stages pT1 or greater), a significant increase in the serum value was observed (P = 0.00023). Laminin level was found to be positively related to the tumor size and number; however, no correlation was observed with the grade. In tumors invading the lamina propria and beyond, the sensitivity of laminin P1 was 78.9% and specificity was 97.1%. The positive and negative predictive values of laminin P1 were 93.7% and 89.5%, respectively. The patients whose serum laminin P1 levels were above the upper limit of normal (1.6863 U/mL) were found to have a higher recurrence rate (72%) than the patients who had lower serum values in the control cystoscopies performed 3 months after the first admission (P < 0.01). CONCLUSIONS: Serum laminin P1 seems to be a valuable adjunctive marker to predict tumor invasion and recurrence. This warrants further research.

Effect of extracorporeal shockwave lithotripsy on plasma and urine endothelin concentrations.

Since the first reports of extracorporeal shockwave lithotripsy (SWL), there have been increasing numbers of articles in the literature documenting renal blood flow impairment and blood pressure elevation as complications. However, little is known about the pathophysiology and prevention of these complications. In this prospective study, the influence of high-energy shockwaves on plasma and urine endothelin concentrations was investigated in 20 patients with renal stones. The patients were randomly assigned to receive a calcium channel blocker, 10 mg of nitrendipine (Bypress; Bayer) (N = 10) 2 hours before SWL or no medication (control group; N = 10). Blood samples were taken just before and 1 minute after application of 3000 shocks. Urine samples were collected by ureteral catheters. The plasma endothelin-1 concentrations were significantly elevated after SWL in the control group (P = 0.003). On the other hand, nitrendipine significantly reduced plasma endothelin concentrations after SWL (P = 0.003). No significant change was observed in urine samples and blood pressure measurements. These results suggest that endothelin release after SWL may be a cause for lithotripsy-induced hemodynamic changes. Medical prevention with calcium channel blockers warrants further investigation.

Effect of ejaculation on prostate-specific antigen levels in normal men.

While several manipulations such as digital rectal examination and cystoscopy have been shown to increase prostate-specific antigen (PSA) levels, lack of ejaculation for a period of time has been implicated as a cause of decreased serum PSA concentrations. However, the effect of ejaculation on the serum PSA concentration is not clear. This work determined the effect of ejaculation on serum PSA levels in normal men. Blood samples of 19 healthy subjects were taken before and 5 consecutive days after ejaculation. One subject was withdrawn from the study because of ejaculation during sleep, since another ejaculation was prohibited during the study. The mean PSA levels of the men before and 5 consecutive days after ejaculation were 0.79, 0.72, 0.63, 1.01, 0.71, and 0.74 ng/ml--not statistically significantly different. We conclude that ejaculation does not seem to affect the serum PSA concentration.

Ferritin: a tumor marker expressed by renal cell carcinoma.

Renal cell carcinoma (RCC) has been shown to secrete several hormones and biologically active substances that influence the host metabolism or induce paraneoplastic syndromes. Observation of anemia in 20% of patients with RCC and the spontaneous recovery of anemia following nephrectomy drew attention to the body iron metabolism. Ferritin was previously proposed as a tumor marker for RCC. In order to determine whether RCC cells actually produce ferritin, we studied ferritin levels in serum from peripheral and renal veins as well as from the tumor tissue and the healthy parenchyma from radical nephrectomy specimens of 22 patients with RCC. Ferritin levels both in sera and cytosols were measured by an enzyme immunoassay method. The mean serum ferritin level from the renal vein was 419.9 +/- 72.4 ng/ml, and it was 157.3 +/- 18.3 ng/ml from the peripheral vein (p < 0.05). Renal vein ferritin correlated with stage and had a significant impact on prognosis (p < 0.05). The mean cytosolic ferritin level of the cancer tissue was 705.6 +/- 56.9 ng/mg cytosol protein, whereas in the normal parenchyma it was 95.9 +/- 10.1 ng/mg cytosol protein. This was also highly significant (p = 1.15 x 10(-13)), suggesting that RCC cells probably express ferritin. As currently there exists no reliable tumor marker for RCC, the value of ferritin as a marker should be investigated further before drawing any clinical conclusions.

The effect of extracorporeal electromagnetic shock waves on renal proximal tubular function.

In order to investigate the effect of electromagnetic shock waves on proximal tubular renal function, we measured the urine levels of the lysosomal enzyme N-acetyl-beta-glucosaminidase (NAG) prior to, 1 hour, 1 day, and 1 week after extracorporeal electromagnetic lithotripsy (ESWL) in 50 patients with urinary stone disease. Mean NAG level which was 5.18 + 0.09 U/l prior to ESWL increased to 5.29 + 0.09 U/l in the first hour and to 10.7 + 0.19 U/l on the first day after ESWL (p < 0.001). The level returned to pre-ESWL values one week after ESWL (5.27 + 0.09 U/l). These findings show that extracorporeal electromagnetic shock waves cause significant damage to the proximal tubular epithelium of the kidney; however, these changes return to normal in the first week.

The effect of extracorporeal shock wave lithotripsy on pancreatic enzymes.

In order to assess the effect of electromagnetic shock waves on pancreatic tissue, we studied serum and urinary amylase and serum lipase levels prior to, 1 hour, 1 day and 1 week after extracorporeal shock wave lithotripsy (ESWL) in 150 patients with upper urinary tract stones. A control group consisted of 22 patients with lower ureteric stones who underwent ESWL. All three parameters increased after ESWL (p < 0.001); however, this increase was within normal laboratory limits. There was no difference between the right and left renal units. In the control group there was no change of pancreatic enzymes after ESWL. High energy shock waves may cause some elevations of pancreatic enzymes, probably due to altered cell membrane permeability, however, ESWL with an electromagnetic shock wave emitter system does not cause significant injury to the pancreatic tissue.