Sperm DNA damage in potentially fertile homozygous beta-thalassaemia patients with iron overload

BACKGROUND: To test the hypothesis that human sperm DNA could sustain iron-induced oxidative damage and reduce its fertilizing ability, we studied patients with homozygous beta-thalassaemia major (HbTh) as a model of iron overload. METHODS: Sperm from six thalassaemic patients and five age-matched controls were assessed by the sperm chromatin structure assay (SCSA) and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labelling (TUNEL) assay. Semen parameters, endocrine markers of testicular function, iron profiles and the presence of organ dysfunction were also determined. RESULTS: All patients with HbTh were iron overloaded (median ferritin: 2251 microg/l) and had evidence of spontaneous spermatogenesis. Thalassaemic patients had more sperm DNA damage than the controls (P < 0.01). The sperm DNA damage by SCSA and TUNEL were positively correlated (P < 0.05). Sperm motility and TUNEL results were negatively correlated (P < 0.05), while the age of onset of chelation and sperm DNA damage were positively associated with both SCSA (R(2) = 0.80, P = 0.016) and TUNEL data (R(2) = 0.67, P < 0.044). No other biochemical or clinical data were associated with sperm DNA damage. CONCLUSIONS: The increase in sperm DNA damage and the negative correlation between sperm motility and DNA damage suggest that iron overload in HbTh predisposes sperm to oxidative injury. This finding has important implications in assisted reproductive procedures such as ICSI where there is increased risk of transmitting defective DNA to the offspring.     

Serum urate concentrations in homozygous sickle cell disease

Serum and urinary urate concentrations were studied in 44 patients with homozygous sickle cell (SS) disease, and in 27 controls with normal haemoglobin. Hyperuricaemia (>0·39 mmol/l (6·5 mg/100 ml)) occurred in 41% of SS patients and inversely correlated with renal urate clearance but not with indices of bone marrow turnover. Higher serum urate concentrations occurred in patients with proteinuria, probably due to associated tubular damage. Higher serum urate concentrations and lower urate clearance occurred in males compared to females.     

Serum immunoreactive trypsin in beta-thalassaemia major.

To assess the exocrine pancreatic function in beta-thalassemia major with iron overload, serum immunoreactive trypsin (IRT) was measured in 38 patients with this condition. In 23 (60%) patients' IRT was abnormal: it was subnormal in 16 patients and supranormal in seven. Whereas subnormal IRT concentrations were more frequent in patients of more than 12 years old, supranormal IRT concentrations were more frequent in younger patients. These data provide the first antemortem evidence of exocrine pancreatic damage in this condition. They also suggest that this acinar cell damage is initially associated with a rise in IRT which is followed by a fall in the serum concentration of this enzyme.     

Serum amylase levels in homozygous sickle cell patients.

Serum amylase was evaluated in a group of 26 steady-state homozygous sickle cell (SS) patients and 11 age-matched HbAA controls. Half (50%) of the SS patients had values above normal while only two of the control group had slightly elevated values. The SS patients had a mean +/- SD value of 301.46 +/- 119.40 iu/L while the control group had 274.36 +/- 89.70 iu/L. The difference between these two values is statistically significant (P less than .05). Males in both groups had significantly higher mean serum amylase values. The values in the SS group did not show an association with age. Higher levels in SS patients suggest a predisposition to chronic pancreatitis.     

Elution of antispectrin antibodies from red cells in homozygous beta-thalassaemia.

It is well-established that autoantibodies against red cell membrane antigens, such as spectrin are present in plasma of humans and animals. It is shown here that antispectrin IgG antibodies can be eluted from the red cells of patients with beta-thalassaemia intermedia. It is suggested that these antibodies could play a part in the increased rate of destruction of red cells in homozygous beta-thalassemia, either by reacting with spectrin which is abnormally exposed on the red cell surface or by a cross-reaction with a different membrane component. No rise was found in serum antispectrin levels in beta-thalassaemia; anti-red cell membrane IgG levels were slightly raised, but there was considerable overlap between the levels in patients and in normals. No autoantibody could be found in eluates from cells of normal subjects or patients with sickle cell anaemia.     

Serum kynurenine and neopterin concentrations in patients with cardiomyopathy.

Serum kynurenine and neopterin concentrations were determined in patients with hypertrophic (n = 22) and dilated (n = 23) cardiomyopathy. Significantly increased kynurenine and neopterin levels have been observed in cases of dilated cardiomyopathy. In these patients we found a positive correlation between serum kynurenine and neopterin levels (RS = 0.52, P less than 0.001). There existed several associations between the laboratory parameters neopterin and kynurenine and clinical findings, such as left ventricle (LV) systolic volume and LV circumferential fiber shortening velocity in patients with dilated cardiomyopathy (all P less than 0.05). In vitro, interferon-gamma induces the formation of neopterin in human monocytes/macrophages. Also the degradation of tryptophan via the kynurenine pathway is induced by interferon-gamma. Our data show that increased kynurenine as well as neopterin accumulation in serum are associated with severity of dilated cardiomyopathy. Thus, the data point to a role of immune activation in the pathogenesis of the disease.     

Raised IgE levels in beta-thalassaemia: correlation with splenectomy and hepatitis B virus infection.

IgE values obtained in 117 beta-thalassaemia patients were significantly higher than in age matched normal subjects. In 31 patients (26.5%) IgE levels were above 2 s.d. of normal values for age, but the frequency of IgE with reaginic activity was lower in patients (5.1%) than in controls (11.9%). The highest values were observed in splenectomized patients who were also positive for one or more serological markers of hepatitis B virus infection. The increase of IgE levels was directly correlated with the number of years after splenectomy, and patients with biopsy proven chronic liver disease had higher IgE levels than those without evidence of liver damage. On the other hand, IgE levels were not correlated with the number of transfusions, age, IgG, IgA, IgM levels or T cell subsets and mitogen responsiveness. These results show that beta-thalassaemia patients develop elevated IgE levels to which splenectomy and hepatitis B virus infection contribute in a synergistic manner.     

ITP患者脾切除前后外周血Th亚群细胞因子的变化及意义

目的: 探讨辅助性T淋巴细胞(T helper lymphocyte,Th)亚群细胞因子在原发性免疫性血小板减少症(ITP)患者脾切除术前后的表达及意义。方法: 采用QuantiGene Plex (QGP)方法检测22例ITP患者腹腔镜脾切除术前后外周血Th1(IL-2和IFN-γ)、Th2(IL-4、IL-5、IL-6和IL-10)、Th3(TGF-β₁)和Th17(IL-17)细胞因子mRNA表达水平的变化。30例健康体检者作为对照组。结果: (1)术前组IL-2表达水平较对照组明显降低(P<0.05),IL-17表达水平较对照组明显升高(P<0.05),其它细胞因子表达水平在术前组和对照组之间无显著差异(P>0.05)。术后组TGF-β₁表达较术前组和正常对照组均明显升高(P<0.05);术后组IL-2 表达较术前有所升高(P<0.05),但仍低于对照组(P<0.05)。其它细胞因子表达水平在术前组和术后组之间无显著性差异(P>0.05)。(2)术前IL-2与IFN-γ之间成正相关(r=0.647,P<0.01),术前其它细胞因子之间均无明显相关性(P>0.05)。术后3对细胞因子之间成正相关,分别是IL-2与IFN-γ(r=0.787, P<0.01),IL-17与IL-2(r=0.554,P<0.01),IL-17与IFN-γ(r=0.461,P<0.05);术后其它细胞因子之间均无明显相关性(P>0.05)。 结论: ITP患者存在Th亚群细胞因子失衡,脾切除术可改善ITP患者部分Th亚群细胞因子的失衡。     

Immunologic studies before and after splenectomy in a patient with the Wiskott-Aldrich syndrome

Sequential studies of cellular and humoral immunity were conducted in an infant with the Wiskott-Aldrich syndrome prior to and after a splenectomy for uncontrollable hemorrhage. All measures of cellular immunity showed gradual improvement during the 8-month period after surgery. Serum isohemagglutinins, diphtheria and tetanus antibodies, and the percentage of immunoglobulin-bearing B cells did not change significantly from presplenectomy values. The serum IgE concentration declined from a high of 10,800 IU/ml at 1 month post-splenectomy to a low of 860 IU/ml at 5 months after surgery and the IgG concentration gradually decreased from a high of 1880 mg/dl presplenectomy to a low of 620 mg/dl 8 months later. The platelet count ranged from 64,000 to 206,000/mm³ for the first 6 months after splenectomy. It decreased precipitously 6.5 months after the operation; at the same time there was a marked rise in platelet-bound IgG antibody (PB-IgG). The PB-IgG declined rapidly following vincristine therapy and, after another rise, declined more gradually following steroid therapy.     

Serum beta-lipoprotein and other specific protein concentrations in patients with immunocytoma.

Serum beta-lipoprotein and other specific protein concentrations were measured in 56 patients suffering from multiple myelomatosis, "benign" paraproteinaemia or Waldenström''s macroglobulinaemia and in 56 control subjects. The mean level of B-lipoprotein in untreated patients with multiple myelomatosis and macroglobulinaemia was significantly lower than that of the controls. Patients who responded to chemotherapy showed a rapid return to normal of the beta-lipoprotein concentration, while the level remained unchanged in most of those who did not.     

Serum concentrations of nitrite in patients with HIV-1 infection.

AIMS: To measure circulating concentrations of nitrite in patients with HIV-1 infection. METHODS: Nitrite concentrations were measured using the Griess reaction adapted to microtitre plates in the serum of 10 asymptomatic HIV-1 positive patients, 33 patients with AIDS with cerebral disorders, 17 patients with AIDS with pulmonary involvement, and in eight patients with AIDS with other disorders. Nitrite concentrations were also measured in bronchoalveolar lavage (BAL) fluid and cerebrospinal fluid (CSF) of patients with AIDS with pulmonary involvement and cerebral disorders, respectively. RESULTS: Increased serum concentrations of nitrite were observed in patients with pulmonary involvement, and in particular in serum and in BAL samples of patients with interstitial pneumonia (36.2 (26.2) mumol/l and 0.3 (0.4) mumol/l, respectively). Increased serum concentrations of nitrite were also noted in patients with retinitis caused by infection with cytomegalovirus. Serum nitrite concentrations were also raised in patients with cerebral toxoplasmosis, whereas normal serum concentrations were found in patients with HIV-1 encephalopathy and cryptococcal meningitis. Nitrite concentrations in CSF were not raised in patients with cerebral disorders. CONCLUSIONS: These results suggest that production of nitrite in patients with AIDS with concomitant opportunistic infections may be part of the host defense against opportunistic organisms.     

Serum concentrations of cytokines in patients with active tuberculosis (TB) and after treatment

During TB cytokines play a role in host defence. To determine the cytokine pattern during various disease stages of TB, serum levels of IL-12, interferon-gamma (IFN-γ), IL-4, IL-6 and IL-10 were measured in 81 patients with active TB, 15 patients during therapy and 26 patients after anti-tuberculous therapy as well as in 16 persons who had been in close contact with smear-positive TB and in 17 healthy controls. IFN-γ was elevated during active TB when compared with healthy controls, declining during and after treatment. IL-12 (p40 and p70) serum levels were not significantly higher in patients with active TB compared with any of the other groups. IL-4 levels were low in all groups. IL-6 and IL-10 serum levels were elevated in patients with active TB and during treatment. In patients with active TB serum levels of IFN-γ and IL-6 were higher in patients with fever, anorexia and malaise. IL-12 levels were higher in patients with a positive smear. Cytokine levels did not correlate with localization of TB (pulmonary versus extrapulmonary), or skin test positivity. Cytokines directing a Th1 response (IL-12) or a Th2 response (IL-4) were not elevated in sera of this large group of patients with pulmonary and extrapulmonary TB. In patients with active TB, cytokines that were elevated in serum were IFN-γ, IL-6 and IL-10.     

Serum procalcitonin concentrations in transplant patients with acute rejection and bacterial infections.

Procalcitonin (PCT) represents a new marker of systemic inflammatory reactions of the body to infections. PCT is selectively induced by severe bacterial infections leading to sepsis or multiorgan dysfunction syndrome. The aim of our study was to test PCT as a postoperative infection marker in heart and kidney transplant patients compared with healthy subjects and patients with localized lung-inflammatory processes without a manifest systemic response. PCT concentrations were measured by an immunoluminometric assay (ILMA) in a total of 419 serum samples. Normal serum levels were in the range of 0.08-0.6 ng/ml. Operative trauma associated with heart (not kidney) transplantation induced a transient increase in PCT levels to 7-10 ng/ml with a decline to normal levels within 2-3 days in most patients. Severe bacterial infections dramatically augmented serum PCT concentrations reaching values of 46-297 ng/ml in the most critical periods. Good response to antibiotic therapy was associated with a decline in serum PCT concentrations. Acute rejection or cytomegalovirus infections did not significantly increase the serum PCT levels. Localized pulmonary infections showed either no, or only a limited increase, in the serum PCT levels (max. 7 ng/ml). We conclude from our data that PCT can be used as a sensitive marker to differentiate systemic bacterial infections from other complications in organ transplantation.     

Serum nickel concentrations in hemodialysis patients with environmental exposure

Nickel was analyzed by electrothermal atomic absorption spectrophotometry in serum specimens from 22 healthy hospital workers and 30 patients with end-stage renal disease treated by extracorporeal hemodialysis, who resided in Sudbury, Ontario, Canada, a city with extensive nickel mines and smelters. Samples of tap water from Sudbury contained 109 +/- 46 micrograms Ni per L (P less than 0.01 vs 0.4 +/- 0.2 micrograms Ni per L in corresponding water samples from Hartford, Connecticut). Serum nickel concentrations averaged 0.6 +/- 0.3 micrograms Ni per L in Sudbury hospital workers (P less than 0.05 vs 0.2 +/- 0.2 micrograms Ni per L in corresponding serums from 43 healthy hospital workers in Hartford). In serums collected post-treatment from Sudbury hemodialysis patients, nickel concentrations averaged 8.5 +/- 2.8 micrograms Ni per L, (i.e., 14-times the corresponding mean in Sudbury hospital workers, P less than 0.01), but were not significantly higher than the nickel concentrations in serums from 42 Hartford hemodialysis patients (7.2 +/- 2.2 micrograms Ni per L). This study confirms the presence of hypernickelemia in hemodialysis patients, but does not suggest that hemodialysis patients have significantly increased risk of nickel toxicity in Sudbury, compared to Hartford, despite the high nickel concentrations in Sudbury tap water. This favorable outcome attests to the efficient deionization of water used to prepare hemodialysis solutions in Sudbury.     

Serum C-reactive protein and neopterin concentrations in patients with viral or bacterial infection.

Serum C-reactive protein and neopterin concentrations were measured in samples taken at an early stage in different types of infection to see whether the combination of markers could contribute to the diagnosis of infection and help distinguish between bacterial and viral infections, tuberculosis, and infections due to "other" pathogens. Both markers were significantly raised in all categories of infection compared with controls, and there were significant differences between the means of both markers when comparing several of the categories of infection. Only C-reactive protein concentrations in bacteraemic patients, however, were both sensitive and specific at distinguishing the type of infection. The additional use of neopterin estimation played only a minor part in increasing the specificity of diagnosis in tuberculosis and in viral infections. On the basis of this study it was not considered worth the time and expense of performing neopterin assays in addition to C-reactive protein estimations to differentiate viral from bacterial infection.     

Male and female salivary testosterone concentrations before and after sexual activity.

Salivary testosterone concentrations were measured in male and female members of four heterosexual couples on a total of 11 evenings before and after sexual intercourse and 11 evenings on which there was no intercourse. Testosterone increased across the evening when there was intercourse and decreased when there was none. The pattern was the same for males and females. Early evening measured did not differ on the two kinds of days, suggesting that sexual activity affects testosterone more than initial testosterone affects sexual activity.