Determination of leucocyte subsets in human saliva by flow cytometry.

OBJECTIVE: Validation of a flow cytometry-based method for the determination of major leucocyte subsets [polymorphonuclear (PMN) cells, monocytes, T cells and B cells] in paraffin-stimulated whole human saliva. DESIGN: Salivary leucocyte subsets were determined by four-colour flow cytometry in eight healthy volunteers on three consecutive days. Comparison of leucocyte subsets between saliva and whole blood was also performed. Day-to-day variability and intraclass correlation coefficients (ICC) were determined as indicators of assay reliability. RESULTS: It was observed that PMN cells were the predominant cells in the saliva. Percentages of mononuclear cells ranged from 0.3% to 7.2%, with monocytes composing the highest percentage, followed by T cells and B cells. Regardless of high intra-individual day-to-day variability, proportions of leucocyte subsets did not significantly change over three measurements, and high ICCs were calculated for T cells and monocytes. CONCLUSION: Flow cytometry can be used as non-invasive and reproducible method for the analysis of leucocyte subsets in human saliva. Further investigation of pathological and other conditions that have the potential to influence salivary leucocyte subsets is warranted.

The association between brain-derived neurotrophic factor Val66Met variants and psychotic symptoms in posttraumatic stress disorder.

OBJECTIVES: Psychotic symptoms frequently occur in veterans with combat-related posttraumatic stress disorder (PTSD). Brain-derived neurotrophic factor (BDNF) plays a major role in neurodevelopment, neuro-regeneration, neurotransmission, learning, regulation of mood and stress responses. The Met allele of the functional polymorphism, BDNF Val66Met, is associated with psychotic disorders. This study intended to assess whether the Met allele is overrepresented in unrelated Caucasian male veterans with psychotic PTSD compared to veteran controls. METHODS: The BDNF Val66Met variants were genotyped in 576 veterans: 206 veterans without PTSD and 370 veterans with PTSD subdivided into groups with or without psychotic features. RESULTS: Veterans with psychotic PTSD were more frequently carriers of one or two Met alleles of the BDNF Val66Met polymorphism than veterans with PTSD without psychotic features and veterans without PTSD. CONCLUSIONS: The study shows that veterans with psychotic PTSD carried more Met alleles of the BDNF Val66Met than non-psychotic veterans with PTSD or veterans without PTSD. The results might add further support to the hypothesis that psychotic PTSD is a more severe subtype of PTSD.

Exaggerated platelet reactivity to physiological agonists in war veterans with posttraumatic stress disorder.

An association between traumatic stress and cardiovascular disease (CVD) is supported by various epidemiological studies. Platelet activation and binding of activated platelets to leukocytes contributes to the pathophysiology of CVD. Evidence of hyperactive sympathetic nervous system, altered expression of platelet α(2)-adrenoreceptors (α(2)AR), and altered platelet adenylate cyclase activity in patients with posttraumatic stress disorder (PTSD) suggest that platelet reactivity in PTSD may be altered as well. We tested whether platelet reactivity to increasing doses of adenosine-diphosphate (ADP), epinephrine (EPI), or their combination differs between war veterans with PTSD (n=15) and healthy controls (n=12). For this purpose, citrated whole blood was incubated with increasing concentrations of ADP (0.1, 1, 10 µM), EPI alone (10 nM, 100 nM, 1000 nM), or EPI (10 nM, 100 nM, 1000 nM) in combination with 0.1 µM ADP. A subset of samples was also incubated with 10 µM yohimbine (YOH), α(2)AR antagonist, to distinguish receptor-specific effects. Platelet CD62P expression and formation of platelet-leukocyte aggregates (PLA) [platelet-monocyte (P-Mo), -lymphocyte (P-Ly), and -neutrophil (P-Ne) aggregates] were measured using three-color flow cytometry. Platelet reactivity was higher in war veterans with PTSD when compared to controls, as determined by greater CD62P expression and formation of PLA in response to ADP alone or in combination with EPI. Platelet reactivity also correlated with the severity of PTSD symptoms. Preliminary experiments with YOH indicate that stress-associated EPI elevations may contribute to platelet activation through a α(2)AR-dependent mechanism. The enhanced platelet reactivity observed in our study may be the underlying mechanism contributing to the development of CVD in PTSD patients.

Homocysteine and serum lipids concentration in male war veterans with posttraumatic stress disorder.

The evidence of increased cardiovascular disease (CVD) risk in posttraumatic stress disorder (PTSD) is accumulating. The present study aimed to determine whether chronic, combat-related PTSD is associated with serum lipid and homocysteine concentrations that could indicate higher CVD risk. The authors tested 66 war veterans with PTSD, 33 war veterans without PTSD, and 42 healthy volunteers for serum concentrations of homocysteine, total cholesterol, high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C) and triglycerides. All the subjects were men and the analyses were adjusted for age, body mass index and smoking. Potential influences of depression, anxiety, and psychotic symptoms on the outcome measures were checked by introducing the scores from the Hamilton Depression Rating Scale (HAM-D-17), the Hamilton Anxiety Scale (HAMA), and the Positive and Negative Syndrome Scale (PANSS) into the overall statistical model. No differences in total cholesterol, LDL-C, HDL-C and triglycerides were found between the groups. Non-smoking PTSD war veterans had higher homocysteine concentrations (mean=10.4 micromol/L, SD=1.7) when compared to non-smoking war veterans without PTSD (mean=8.2 micromol/L, SD=4.0, P=0.014) and both smoking (mean=8.7 micromol/L, SD=2.3, P=0.008) and non-smoking healthy volunteers (mean=8.8 micromol/L, SD=2.2, P=0.021). The results of our cross-sectional study are possibly confounded by many factors, especially behavioral and life-style related which are difficult to control comprehensively and might have influenced serum lipids and homocysteine concentration in a complex manner. An increase in the homocysteine concentration observed in the non-smoking PTSD patients needs further investigation with a carefully designed prospective study to confirm associated, possibly enhanced CVD risk.

Psychiatric heredity and posttraumatic stress disorder: survey study of war veterans.

AIM: To explore the prevalence of psychiatric heredity (family history of psychiatric illness, alcohol dependence disorder, and suicidality) and its association with the diagnosis of stress-related disorders in Croatian war veterans established during psychiatric examination. METHODS: The study included 415 war veterans who were psychiatrically assessed and diagnosed by the same psychiatrist during an expert examination conducted for the purposes of compensation seeking. Data were collected by a structured diagnostic procedure. RESULTS: There was no significant correlation between psychiatric heredity of psychiatric illness, alcohol dependence, or suicidality and diagnosis of posttraumatic stress disorder (PTSD) or PTSD with psychiatric comorbidity. Diagnoses of psychosis or psychosis with comorbidity significantly correlated with psychiatric heredity (Phi=0.111; P=0.023). There was a statistically significant correlation between maternal psychiatric illness and the patients' diagnoses of partial PTSD or partial PTSD with comorbidity (Phi=0.104; P=0.035) and psychosis or psychosis with comorbidity (Phi=0.113; P=0.022); paternal psychiatric illness and the patients' diagnoses of psychosis or psychosis with comorbidity (Phi=0.130; P=0.008), alcohol dependence or alcohol dependence with comorbidity (Phi=0.166; P=0.001); psychiatric illness in the primary family with the patients' psychosis or psychosis with comorbidity (Phi=0.115; P=0.019); alcohol dependence in the primary family with the patients' personality disorder or personality disorder with comorbidity (Phi=0.099; P=0.044); and suicidality in the primary family and a diagnosis of personality disorder or personality disorder with comorbidity (Phi=0.128; P=0.009). CONCLUSION: The study confirmed that parental and familial positive history of psychiatric disorders puts the individual at higher risk for developing psychiatric illness or alcohol or drug dependence disorder. Psychiatric heredity might not be necessary for the individual who was exposed to severe combat-related events to develop symptoms of PTSD.

Platelet serotonin in combat related posttraumatic stress disorder with psychotic symptoms.

BACKGROUND: Combat-related posttraumatic stress disorder (PTSD) is severe form of PTSD, frequently associated with psychotic symptoms. Platelet serotonin (5-hydroxytryptamine, 5-HT) was used as a peripheral 5-HT marker to identify particular symptoms in PTSD. METHODS: Platelet 5-HT was determined fluorimetrically in 67 war veterans with combat related PTSD, 36 combat exposed veterans who did not develop PTSD, 35 veterans with PTSD complicated with psychotic features. PTSD diagnosis of current and chronic PTSD, and clinical symptoms of PTSD and psychoses were assessed according to DSM-IV criteria, using the Clinician Administrated PTSD Scale, and Positive and Negative Syndrome Scale (PANSS). RESULTS: Platelet 5-HT concentration was significantly higher in veterans with psychotic PTSD than in veterans with non-psychotic PTSD, veterans without PTSD, or in control subjects. Platelet 5-HT was significantly positively correlated with the positive symptoms in PANSS subscale, and with the symptoms of delusions within PANSS positive subscale. LIMITATIONS: The results were obtained on peripheral 5-HT marker, i.e. platelet 5-HT concentration. CONCLUSIONS: Since the delusions are the core psychotic symptoms occurring in our psychotic PTSD patients, the result of the increased platelet 5-HT concentration, associated with delusions, indicate that platelet 5-HT might be used as a trait marker of psychotic symptoms in PTSD, but not as a state marker for PTSD.

Epidemiological study of suicide in the Republic of Croatia -- comparison of war and post-war periods and areas directly and indirectly affected by war.

We examined the differences in the suicide characteristics between areas directly and indirectly affected by war activities and in war and post-war periods according to the following variables: suicide rate, sex, age and method of suicide. Analysis was done on 5349 suicides committed in the period 1993-1998 (war and post-war years). The suicide rates in the Republic of Croatia oscillated in the pre-war, war and post-war periods (1985-2000) but without significant differences. In the areas directly affected by war, the suicide rate was significantly lower than in other areas during the study period 1993-1998 (chi-square = 10.3245; P = 0.0017). The number of suicides in both sexes declined in the areas directly affected by war-more in men than in women; the difference between sexes was statistically significant (chi-square = 3.6697; P = 0.055). Middle- and old-aged people were the population with high suicide risk in both areas (t = 1.76; P = 0.078). There were significant differences in the methods of suicides between war and non-war areas (chi-square = 108.8473; P = 0.001). Firearms or explosive devices were the methods used more significantly for suicides in the areas directly affected by war than in other areas, whereas hanging was more frequently used in the areas indirectly affected by war.