Lower serum zinc in major depression is a sensitive marker of treatment resistance and of the immune/inflammatory response in that illness.

The aims of the present study were to examine i) serum zinc (Zn) and copper (Cu) in treatment resistant depression (TRD); ii) the effects of subchronic antidepressant therapy on these trace elements; and iii) the relationships between serum Zn and Cu and immune/inflammatory markers. Serum Zn was significantly lower in TRD than in normal controls. There was a significant inverse correlation between baseline serum Zn and staging of depression based on severity of prior treatment resistance. There were no significant effects of antidepressive treatment on serum Zn, whereas serum Cu was significantly reduced. There were highly significant correlations between serum Zn and the CD4+/CD8+ T-cell ratio (negative), and total serum protein, serum albumin, and transferrin (all positive). The results suggest that lower serum Zn is a marker of TRD and of the immune/inflammatory response in depression. It is suggested that treatment resistance may bear a relationship with the immune/inflammatory alterations in major depression.

Lower serum L-tryptophan availability in depression as a marker of a more generalized disorder in protein metabolism.

Recently, it has been reported that major and melancholic depression are accompanied by a lower availability of total L-tryptophan (L-TRP) to the brain and by significant changes in electrophoretically separated protein fractions, such as albumin and alpha 2-globulin. The aim of this study was to examine the relationships between serum L-TRP availability and total serum protein, albumin, and alpha 2-globulin in 42 depressed and 24 normal subjects. In depressed and normal subjects, alone and together, there were significant and positive correlations between serum L-TRP and total serum protein or albumin concentrations. In the depressed subjects, but not in normal controls, there were significant inverse relationships between the L-TRP/competing amino acid ratio and the alpha 2-globulin fraction. Serum L-TRP and albumin were significantly lower in melancholic subjects than in normal and minor depressed subjects. Depressed subjects had a significantly lower L-TRP/competing amino acid ratio and significantly higher serum alpha 2-globulin than normal controls. Total serum protein was significantly lower in major depressed subjects than in normal controls. The results suggest that lower L-TRP availability to the brain in depression is related to lower serum albumin and to increased alpha 2-globulin fraction, which are both hallmarks of the acute phase response in depression. the results further corroborate the hypothesis that lowered L-TRP availability in depression is related to the acute phase response in that illness.

A simple and economical strategy for downstream processing of specific antibodies to human transferrin from egg yolk.

A rapid and economical procedure for extraction of antibodies from egg yolk is described. Laying hens were immunized with human transferrin and extracts of egg-yolk were purified with a procedure based on affinity chromatography. The resulting purified antibodies were evaluated in a nephelometric system for the assay of transferrin in human sera. The results agreed closely with those obtained with a commercially available anti-human transferrin serum from rabbits.

Effects of psychological stress on serum immunoglobulin, complement and acute phase protein concentrations in normal volunteers.

The aim of this study was to examine the effects of academic examination stress on serum immunoglobulins (Igs), i.e. IgA, IgG, IgM, complement factors, i.e. C3c and C4, and acute phase proteins, i.e. alpha 1-acid glycoprotein (alpha 1-S), haptoglobin (Hp) and alpha 2-macroglobulin (alpha 2-M). Thirty-seven university students participated in this study. Serum was sampled a few weeks before and after as well as one day before a difficult academic examination. On the same occasions, students completed the Perceived Stress Scale (PSS). Students were divided into two groups, i.e. those with high- and low-stress perception as defined by changes in the PSS score. Academic examination stress induced significant increases in serum IgA, IgG, IgM, and alpha 2-M in students with high-stress perception, but not in these with low-stress perception. The stress-induced changes in serum IgA, C3c, and alpha 1-S concentrations were significantly higher in students with high-stress perception than in those with a low-stress perception. The stress-induced changes in serum IgA, IgM, C3c, C4, alpha 1-S, Hp and alpha 2-M were normalized a few weeks after the stress condition, whereas IgG showed a trend toward normalization. There were significant positive relationships between the stress-induced changes in the PSS and serum IgA, IgG, IgM and alpha 2-M. These findings suggest that psychological stress is accompanied by an altered secretion of serum Igs, complement factors and some acute phase proteins.

Simultaneous use of Plasmodium falciparum crude antigen and red blood cell control antigen in the enzyme-linked immunosorbent assay for malaria.

An enzyme-linked immunosorbent assay for malaria, based upon duplicate testing of serum samples on both a crude Plasmodium falciparum antigen and a red blood cell control antigen, is evaluated. Results were analyzed using the Student's t-test for identification of positive serum samples (t greater than or equal to 2.92, P less than or equal to 0.05) and for calculation of the mean difference in absorbance values (delta ABS) obtained between the P. falciparum wells and the control wells. Cross-evaluation with the IFA test for P. falciparum antibodies gave 89.6% concording positive or negative results. Among discrepant sera 8.35% were ELISA+/IFA- and 2.05% ELISA-/IFA+. In addition, delta ABS values in ELISA were highly correlated to titers obtained in immunofluorescence (r = 0.80, P less than 0.001). The results confirm the high degree of species-specificity of the ELISA using P. falciparum crude antigen. The necessity of the simultaneous use of red blood cell control antigen with a crude plasmodial antigen is demonstrated by comparing the presented results with those obtained on the P. falciparum antigen only.

Detection of antibodies to Plasmodium vivax by indirect immunofluorescence: influence of the geographic origin of antigens and serum samples.

The results of a double-blind serological study of 15 sera sampled in a residual focus of vivax malaria transmission in Algeria, and of 7 sera from patients with slide-proven P. vivax infections acquired in India, are analyzed. The reactivity of each of these serum samples was tested by indirect immunofluorescence using 6 different batches of antigen, including 3 batches of P. vivax antigen prepared with isolates from Zaire (Africa), India and the Solomon Islands, respectively. The geometric mean of reciprocal titers (GMRT) calculated on the 7 sera from proven vivax infections fell from 289.8 using the homologous antigen from the same geographic origin (India) to 48.7 using a homologous (vivax) antigen originating from a different continent (Africa). Among the 15 samples from Algeria, the percentage of seropositives decreased from 100% using the homologous P. vivax antigen originating from the same continent (Africa) to 53.3% using a homologous antigen from India. Two aspects are included in the discussion: in seroepidemiological studies, sensitivity could be improved by the use of a homologous antigen from the same geographic origin; in detection of clinical cases of malaria and species identification based on serology, our results stress the need for caution in interpreting serological titers and for taking into account the geographic origin of the isolates used as antigen.

Inflammatory markers in younger vs elderly normal volunteers and in patients with Alzheimer's disease.

BACKGROUND: Recently it has been reported that activation of the inflammatory response system (IRS) may play a role in the aging process and in the pathogenesis of the degenerative changes associated with Alzheimer's disease (SDAT). The aims of the present study were to examine the peripheral IRS in normal aging and in SDAT patients. METHODS: Serum zinc (Zn), total serum protein (TSP), albumin (Alb), SP electrophoresis, and serum interleukin-6 (IL-6) and the stimulated production of tumor necrosis factor-alpha (TNFalpha) were determined in younger versus elderly healthy subjects and in SDAT patients vs. age-matched, healthy volunteers. RESULTS: Serum Zn and Alb were significantly lower in elderly than in younger healthy volunteers and were significantly and inversely correlated with age. The production of TNFalpha was significantly higher in elderly than in younger healthy volunteers and was significantly and positively correlated with age. In SDAT patients, no significant changes in serum Zn or TNFalpha production could be found. Serum Alb was significantly lower and serum IL-6 and the alpha1 and alpha2 globulin fractions significantly higher in SDAT patients than in controls. CONCLUSIONS: Activation of the IRS appears to accompany the normal aging process, i.e. lower serum Zn and Alb and increased TNFalpha production, as well as SDAT, i.e. lower serum Alb and increased serum IL-6 and alpha1 and alpha2 globulin fractions. The findings suggest that not all indicators of IRS activation in SDAT are related to those of the normal ageing process.

Lower serum zinc in major depression in relation to changes in serum acute phase proteins.

There is now some evidence that major depression is accompanied by activation of the inflammatory response system (IRS). Other signs of IRS activation, which have been reported in major depression are lowered serum zinc (Zn) and serum albumin (Alb) concentrations. In serum, Zn is closely bound to Alb. The aims of the present study were to replicate previous findings that major depression is accompanied by lowered serum Zn and Alb and to examine whether the decrease in serum Zn may be explained by that in serum Alb. The above variables were determined in 48 major depressed patients and in 15 age-sex-matched healthy volunteers. Serum Zn and Alb were significantly lower in major depressed patients than in normal volunteers. In healthy volunteers and major depressed patients, there were significant and positive correlations between serum Zn and Alb. We found that 53.8% of the variance in serum Zn could be explained by the combined effects of serum Alb and diagnostic classification. The results suggest that lower serum Zn in depression is in part explained by lowered serum Alb and by another depression-related mechanism. It is suggested that lower serum Zn in depression may be secondary to sequestration of metallothionein in the liver, which may be related to increased production of interleukin-6.

Alterations in iron metabolism and the erythron in major depression: further evidence for a chronic inflammatory process.

There is now some evidence that major depression is accompanied by biochemical and immune changes pointing to the presence of a chronic inflammatory response. The latter condition is reportedly characterized by changes in iron (Fe) metabolism and the erythron, such as decreased serum Fe and transferrin (Tf) and increased serum ferritin, lower number of red blood cells (RBC), lower hematocrit (Htc) and hemoglobin (Hb). The aim of the present study was to examine Fe metabolism and the erythron in 38 major depressed subjects versus 15 normal volunteers, as well as the effects of antidepressant treatments on these variables. Major depressed subjects had significantly lower serum Fe and Tf, a lower number of RBC, lower Htc and Hb, and a significantly increased number of reticulocytes than normal controls. Serum ferritin was significantly higher in major depressed patients with melancholia than in those with simple major depression and normal controls. Mean corpuscular volume (MCV), MC Hb (MCH), MC Hb concentration (MCHC) and RBC distribution width (RDW) were not significantly different between major depressed subjects and normal controls. Treatment with antidepressants during 5 weeks had no significant effect on the alterations in number of RBC and reticulocytes, Htc, Hb, Fe and Tf. There were significant relationships between the above Fe and erythron variables and established immune-inflammatory markers of major depression, e.g., lowered serum albumin and zinc and the increased electrophoretically-separated alpha 1-globulin fraction. The results suggest that the disorders in Fe metabolism and the erythron during major depression may be induced by the immune-inflammatory response in that illness.

Lower serum vitamin E concentrations in major depression. Another marker of lowered antioxidant defenses in that illness.

OBJECTIVE: Major depression is associated with defective antioxidant defenses. Vitamin E is the major fat soluble antioxidant in the body. The aim of the present study is to examine serum vitamin E concentrations in major depressed patients versus normal volunteers. METHOD: Serum vitamin E concentrations were measured in 26 healthy volunteers and 42 major depressed patients by means of HPLC. Since vitamin E is a fat soluble vitamin, and serum vitamin E concentrations are strongly related to these of low-density-lipoprotein cholesterol (LDL-C) and triglycerides, we have adjusted the results for possible differences in these lipids. The numbers of peripheral blood leukocytes were measured. RESULTS: Patients with major depression had significantly lower serum vitamin E concentrations than healthy controls. The area under the ROC (receiver operating characteristics) curve was 83%. There were significant and negative correlations between serum vitamin E and number of total leukocytes and neutrophils. CONCLUSIONS: Major depression is accompanied by significantly lower serum vitamin E concentrations, suggesting lower antioxidant defenses against lipid peroxidation. The results could, in part, explain previous findings, which suggest increased lipid peroxidation in major depression.

Lower total serum protein, albumin, and beta- and gamma-globulin in major and treatment-resistant depression: effects of antidepressant treatments.

Strong evidence has recently been reported that major depression is accompanied by an acute phase response (APR), characterized by elevated levels of positive acute phase proteins (APPs) and decreased levels of negative APPs. The APR is also reflected in lowered total serum protein (TSP) and specific changes in the major electrophoretically separated protein fractions. The present study examined pretreatment and posttreatment serum TSP and the concentrations and percentages of the major electrophoretically separated serum protein fractions in 37 major depressed subjects, of whom 29 had treatment-resistant depression (TRD), and in 29 normal controls. We found that TSP and the percentage and concentration of serum albumin (Alb) and gamma-globulin fraction were significantly lower in major depression and TRD than in normal controls. Serum beta-globulin concentrations were significantly lower in major depressed and TRD subjects than in normal controls. The percentages of the alpha 1- and alpha 2-globulin fractions were significantly higher in major depressed subjects than in normal controls. There were no significant effects of subchronic treatment with antidepressants on TSP, the percentage or concentration of the major electrophoretically separated protein fractions, i.e. alpha 1-, alpha 2- and beta-globulin. There was a significant increase in percentage of the gamma-globulin fraction after subchronic treatment with antidepressants. The results support the hypothesis that major depression and TRD are accompanied by a chronic APR.

Influence of academic examination stress on hematological measurements in subjectively healthy volunteers.

Some recent reports showed that a brief exposure to a mental stressor during 3-20 min may induce hematological changes in humans. The aim of the present study was to examine the effects of academic examination stress on erythron variables, such as the number of red blood cells (RBC), hemoglobin (Hb), hematocrit (Ht), mean corpuscular volume (MCV), mean cell Hb (MCH), mean cell Hb concentration (MCHC), RBC distribution width (RDW), and serum iron and transferrin (Tf). The above variables were determined in 41 students in three conditions, i.e. the stress condition (the day before a difficult oral exam) and two baseline conditions, i.e. a few weeks earlier and later. At the same occasions, subjects completed the Perceived Stress Scale (PSS), the state version of the State-Trait Anxiety Inventory (STAI) and the Profile of Mood States (POMS). Academic examination stress significantly increased Ht, Hb, MCV, MCH and MCHC and significantly decreased RDW. There were significant relationships between the stress-induced changes in the PSS, STAI and POMS scores and those in Ht, Hb, MCV and MCH (allpositive) and RDW (negative). It is concluded that academic examination stress induces significant hematological changes indicative of an increased number of large RBC and increased hemoglobinisation, which cannot be explained by shifts of fluid out of the intravascular space, concentrating non-diffusible blood constituents.

The conversion of a standard gas chromatograph to a fused silica capillary system: applications in human toxicology and tropical medicine.

A simple and inexpensive modification of the injection port and detector configuration of a standard gas chromatograph, equipped with a nitrogen-phosphorus detector, permits the installation of a fused silica capillary column. Different ways of connecting the column to the injector and its impact on the interchangeability of the columns are discussed. The application of the system to toxicological determinations of clinical and forensic interest has been investigated, using a direct injection technique. The method allows the quantitative determination of different alkaloids and drugs in human biological samples in the low nanogram range, with reliable reproducibility and linear response.

Application of the combined use of fused silica capillary columns and NPD for the toxicological determination of codeine and ethylmorphine in a human overdose case.

The applicability of capillary gas chromatography to the toxicological analysis of extracts of human tissue samples was investigated for codeine and ethylmorphine in a case of drug overdose. After column extraction, the samples were injected onto a fused silica capillary column, using a direct injection technique. The nitrogen-phosphorus detector provided excellent stability and sensitivity without the need for extensive clean-up procedures.

Combined use of fused silica capillary columns and nitrogen-phosphorus detection for the toxicological analysis of illicit heroin samples.

Capillary GC is applied to the toxicological analysis of illicit heroin samples. The combined use of a permanently deactivated fused silica capillary column, and a nitrogen-phosphorus detector (NPD), allows the separation and determination of nanogram amounts of underivatised opiates, using a direct injection technique.

Evaluation of the REMEDi drug profiling system.

One hundred and sixteen samples for toxicological analysis (78 urines and 38 stomach contents) were analysed both with the Bio-Rad REMEDi Drug Profiling System and with a set of immunoassays and standard chromatographic procedures (HPLC, GC/NPD and GC/MS). As a screening method, REMEDi performed only moderately well, compared with immunoassays. Amphetamines and tricyclic antidepressants were detected by REMEDi in 80% and 68%, respectively, of the positive immunoassay results. Positive immunoassay screening tests were more often confirmed by the conventional chromatographic methods (80%) than by REMEDi (35%). The highest confirmation score for REMEDi was observed in the amphetamine group (9 of 12), while low scores were obtained for barbiturates (1 of 7) and benzodiazepines (13 of 75). However, in these samples REMEDi detected 53 additional compounds, which were neither reactive in the immunoassay screening tests, nor detected by the standard chromatographic procedures. These results clearly demonstrate that REMEDi can be a useful complementary technique in the clinical toxicology laboratory, since it expands the range of drugs covered by the immunoassays and provides a rapid, preliminary report in emergency situations.