Eric Diamond

Saliva Analysis in the Clinical Setting: Revisiting an Underused Diagnostic Tool

Background One purpose of this study was to compare various biochemical and immunological parameters in blood and saliva that are routinely evaluated only in the blood for general medical requirements. Another purpose was to concomitantly compare these and other oral/salivary parameters differentially in whole, parotid, and submandibular and sublingual saliva to examine the source of those parameters and their specific concentrations. Methods Twelve healthy individuals (6 women, 6 men) were examined in the blood-saliva comparison study, and 30 healthy individuals (15 women, 15 men) were studied in the intersalivary comparison study. Results On the basis of the results we obtained, we suggest a classification scheme using a whole saliva compositional profile as a diagnostic tool in the evaluation of systemic and/or local pathologies. This system may be used to analyze various components of saliva beyond those analyzed in this study, thereby increasing the clinician/s ability to locate and assess specific pathologies. We also suggest that consideration be given to the use of compositional saliva analysis in the diagnosis of general medical conditions in which there is a high correlation between the salivary and blood concentrations of relevant components. Conclusion We think that saliva analysis is a useful, worthwhile diagnostic tool because saliva collection is noninvasive, easy, and inexpensive and may be performed by the patient with no need for the involvement of medical personnel, if so desired.

Diagnostic coronary angiography induces a systemic inflammatory response in patients with stable angina.

BACKGROUND Systemic markers of inflammation increase after percutaneous coronary intervention (PCI). The rise in inflammatory markers after PCI is frequently attributed to the inflammatory stimulus associated with coronary artery injury during balloon inflation and coronary stent implantation. The aim of this study was the determine whether diagnostic coronary angiography performed in patients with stable angina triggers a systemic inflammatory response. METHODS We prospectively studied patients with chronic stable angina undergoing either coronary angiography (n = 13) or coronary angiography followed by PCI (n = 13). Peripheral blood samples were obtained before and 24 hours, 48 hours, and 4 weeks after the procedure and analyzed for C-reactive protein (CRP), interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNF-alpha). Patients with periprocedural myocardial necrosis were excluded. RESULTS There was a significant increase in CRP levels at 24 and 48 hours in both the coronary angiography (P <.05) and PCI (P <.01) groups. IL-6 levels peaked at 24 hours in both the coronary angiography (median, 2.5-9.5 pg/mL; P =.01) and PCI (median, 3.0-8.2 pg/mL; P <.005) groups. At 4 weeks, both CRP and IL-6 returned to baseline levels. TNF-alpha levels were unchanged with either coronary angiography or PCI. The magnitude of the rise of CRP and IL-6 levels was not significantly different between the groups. There was a fair correlation between baseline and peak postprocedural levels of CRP (r = 0.67, P =.008) and IL-6 (r = 0.48, P =.016). CONCLUSION Uncomplicated diagnostic coronary angiography triggers a systemic inflammatory response in patients with stable angina. The contribution of coronary angiography should be considered in interpreting the significance of the systemic inflammatory response observed after PCI.

Tissue-specific p53 expression in the nervous system

P53 is a transcription factor that has been found to be expressed in association with cell proliferation and apoptosis. Previously, bacterial chloramphenicol acetyl transferase (CAT) enzymatic expression was predominantly found in the testes of p53 promoter driven-CAT transgenic mice. In the current study, we extended this study to survey p53 expression across both the central and peripheral nervous systems of the same strain of transgenic mice as well as their parental strain. High levels of p53 promoter driven-CAT activity was observed in the cerebellum, hippocampus, hypothalamus, pons, thalamus and upper cerebral spine. Furthermore, we consistently found unexpectedly high levels of p53 promoter-driven CAT expression in the eyes. These observations were reinforced by p53 protein analysis using a p53 pan ELISA assay. Immunohistochemical studies confirmed and further defined p53 expression in several regions of the nervous system. Significantly, p53 promoter-driven CAT expression was visualized in the Ammon horn of the hippocampus, in the Purkinje cells of the cerebellum and in the cornea as well as in the retina of the eye. Furthermore, strong p53 protein expression was found in the cornea of the parental mouse strain. p53 ELISA demonstrated a profile of p53 protein concentration, which correlate well with the high p53 promoter-driven CAT activities observed in the cerebellum, hindbrain, hypothalamus, thalamus, hippocampus, whole eyes as well as with the low CAT activities observed in the cortex and spinal cord. In both of these assays considerable p53 promoter activity and p53 protein levels were found in post-mitotic non-dividing cells.

Plasma endothelin-1 levels in patients with complex regional pain syndrome

The clinical characteristics of complex regional pain syndrome (CRPS) – spontaneous and stimulus‐evoked pain, autonomic abnormalities, motor dysfunction, and trophic changes in the affected limb—are well known. However, its pathogenesis is unclear, and the diagnosis is often delayed, in part due to lack of objective laboratory tests. Endothelin‐1 (ET‐1) is a potent vasoconstrictor that has recently been shown to produce pain, allodynia, edema, and muscle weakness, as well as to exert a direct excitatory effect on nociceptive afferents. Furthermore, new evidence indicates that ET‐1 is involved in various cancer‐ and non‐cancer‐related painful conditions. The aim of the present explorative study was to determine the ET‐1 plasma levels in patients with CRPS in an attempt to identify a ‘laboratory marker’ for CRPS and to search for evidence suggesting that ET‐1 may be involved in the pathogenesis of CRPS. ET‐1 plasma levels were determined in 20 severely affected CRPS patients, in eight patients with non‐CRPS chronic painful conditions, and in 10 healthy volunteers. The results showed that there were no significant differences in ET‐1 plasma levels between the three groups. We conclude that the plasma level of ET‐1 cannot be regarded as a ‘marker’ for CRPS. Yet, the possibility that ET‐1 is involved in the pathophysiology of CRPS has not been excluded and deserves further investigation.

Parenteral selenium supplementation in extremely low birth weight infants: inadequate dosage but no correlation with hypothyroidism.

OBJECTIVE:Selenium is an essential trace element, known to be important in thyroid metabolism. We speculated that parenteral selenium supplementation is inadequate in preterm infants and may contribute to the development of hypothyroidism.STUDY DESIGN:Serum selenium and thyroid function were evaluated on day 10 of life in extremely low birth weight infants. Selenium intake provided by parenteral nutrition was prospectively evaluated.RESULTS:Selenium intake was close to the recommended 2 μg/kg per day. Serum selenium values were 0.54 ± 0.13 μM (mean ± SD, n = 29). Selenium serum levels were low in 26 of 29 infants. In infants with subnormal serum selenium levels, free T4 was transiently low in 10 of 26 infants but was normal in 16 of 26 infants. No significant correlation was found between serum selenium levels and hypothyroidism.CONCLUSION: Current selenium supplementation guidelines may be inadequate in extremely low birth weight infants. However, selenium deficiency does not seem to play a major role in neonatal hypothyroidism.

Intestinal and Systemic Effects of Oral Insulin Supplementation in Rats After Weaning

Oral insulin has intestinal trophic effects in suckling animals. In mice, lower glucose and lipid levels may be seen when oral insulin is given after the weaning period. The purpose of the present study is to examine local and systemic effects of oral insulin supplementation in rats in the postweaning period. Immediately after weaning, Sprague–Dawley rats received either drinking water (controls) or oral insulin in their drinking water (1 U/ml) for either 1 week or 6 weeks. Intestinal mucosal parameters (bowel and mucosal weight, mucosal DNA and protein) and histological changes were examined in all study groups. Glucose levels were monitored weekly, and at the end of the study, blood levels of glucose, lipids, and insulin were measured in the fasting state. After 1 week of insulin supplementation, mucosal weight in duodenum and jejunum as well as jejunal DNA content were significantly higher in insulin-supplemented rats compared to controls. Duodenal circumference and villus height in jejunum were significantly higher in insulin-supplemented rats compared to controls on both day 7 and day 42. Total cholesterol levels were significantly lower in the study group compared with the controls. We conclude that oral insulin supplementation exerts intestinal trophic effects, as well as systemic effects in the postweaning period in rats.

The Prevalence of Low Selenium Levels in Newly Diagnosed Pediatric Cancer Patients

Low selenium (Se) levels have been found in assoiciation with high incidences of various types of adult cancer. Much less is known about this issue among pediatric cancer patients. Forty-two pediatric patients with a variety of newly diagnosed malignancies were divided into two groups, 20 with localized disease (LD) and 22 with widespread disease (WSD). Analysis of serum collected before the commencement of treatment showed that half the patients had low Se serum levels, lower and more common in WSD than in LD. There was no significant difference in the prevalence of low albumin levels among patients with low Se levels, and most of the newly diagnosed children did not suffer from malnutrition. It was concluded that Se deficiency is common among newly diagnosed pediatric cancer patients, Se levels are lower in WSD than LD, and low Se levels are more prevalent in WSD patients than in LD patients.

Effect of zinc-deficient diet of varying duration on intestinal disaccharidase activity in the rat.

To determine whether zinc has a specific role on weight gain and intestinal disaccharidase activity, 42 male Sprague-Dawley rats were assigned to one of seven groups (n =6 each). These were a baseline control group (O) that was killed to analyze initial intestinal disaccharidase (sucrase and maltase) activity, a second group (A) fed a zinc-deficient diet for 1 week, a third group (B) pair-fed control for A, a fourth group (C) fed a zinc-deficient diet for 2 weeks, a fifth group (D) pair-fed control for C, a sixth group (E) fed a zinc-deficient diet for 3 weeks, and a seventh group (F) pair-fed control for E. All experimental groups received distilled deionized drinking water, whereas control groups received zinc-enriched (25 μg of zinc/ml) distilled deionized water. Water was given ad libitum. After killing, the mucosa of the proximal half of the small intestine was analyzed for protein and disaccharidase activity, and liver, kidney, and heart were analyzed for zinc concentration. Protein content and disaccharidase activity of the jejunal mucosa in the experiment and control groups did not differ significantly. However, animals on the zinc-deficient diet demonstrated mildly depressed growth rates that were proportional to the duration of the experiment, and significantly lower zinc concentration in the kidney in the experimental groups. The data indicate that administration of a zinc-deficient diet for up to 3 weeks did not result in significant changes in intestinal mucosa protein content or in disaccharidase activity.

Serum levels of bile salt-stimulated lipase and breast feeding.

OBJECTIVES Bile salt-stimulated lipase (BSSL) is present in the sera of healthy humans, may affect lipoprotein structure and composition, and reduce atherogenicity of oxidized LDL-cholesterol. Our aims were to examine serum levels of BSSL in breast- and formula-fed infants, and explore the influence of BSSL on serum lipid profile and oxidative status. METHODS Infants (2-8 weeks old) were prospectively enrolled. Blood was drawn for serum levels of BSSL, total antioxidant status (TAS), and lipid profile. RESULTS Serum levels of BSSL were similar in breast-fed (0.28 +/- 0.15 microg/l, n = 18) and formula-fed (0.31 +/- 0.09 microg/l, n = 15) infants, and were much lower than reported levels for adults. In breast-fed infants only, BSSL levels were correlated with LDL-cholesterol serum levels (r = -0.53, p = 0.04). Total cholesterol (119.2 +/- 34.3 mg/dl vs 97 +/- 27.2, and p = 0.05) and LDL-cholesterol serum levels (50.5 +/- 26.1 mg/dl vs 33.3 +/- 20.3, p = 0.05), were elevated in breast-fed compared with formula-fed infants, but TAS was similar in both groups (1.02 +/- 0.18 mmol/l and 0.98 +/- 0.12 mmol/l, respectively). CONCLUSIONS Lack of difference in BSSL serum levels between formula- and breast-feeding, and lower BSSL levels in infants compared to adults, suggest that human milk does not contribute to BSSL serum levels.

Effects of Hypothyroidism on Jejunal Mucosal Function: Study by In Situ Luminal Perfusion in Rats

ABSTRACT: To assess the effects of hypothyroidism (HT) on small-intestinal function, HT was induced in rats (120–150 g) by methimazole in drinking water. After 6 wk of methimazole, intestinal absorption studies were performed in HT and in control (C) rats by in situ luminal perfusion of a 20-cm proximal jejunal loop with a bicarbonate buffer containing sodium, glucose or fructose, glycine or lysine, and phenol red as a nonabsorbable marker for determination of water fluxes. Mucosa from the perfused segment was taken for assay of disaccharidases and ATPases and for light and electron microscopy. Compared with C rats, HT rats had significantly lower jejunal transport rates of water (2.54 ± 0.36 versus 5.02 ± 0.7 μL/min/μg mucosal protein, p < 0.03), sodium (37.1 ± 10.3 versus 102.7 ± 18.6 μmol/min/μg protein, p < 0.05), and glucose (1.49 ± 0.28 versus 5.17 ± 0.82 μmol/min/μg protein, p < 0.02). A reduction in glycine transport was also observed but did not attain statistical significance (p = 0.058). Fructose and lysine transport was unchanged. Mucosal sucrase and Iactase activities were similar in both groups, but Na,K-ATPase was significantly lower in HT rats (1.17 ± 0.3 versus 4.03 ±1.5 μmol inorganic phosphate/h/mg protein; p < 0.05), with a diminution of ouabain binding sites by 41.5%. Light microscopy of jejunal mucosa from HT rats did not differ from that from C rats; electron microscopy showed mild mitochondrial swelling in HT enterocytes. A group of HT rats were treated with L-thyroxine during 4 wk; these rats had absorption rates, mucosal enzyme activities, ouabain binding, and mucosal morphology not different from C rats. We conclude that HT in the rat can depress jejunal mucosal Na,K-ATPase activity and reduce Na,K-ATPase-dependent transport without structural changes other than mild enterocyte mitochondrial swelling. A diminished jejunal functional capacity might add a nutritional component to the growth retardation that is observed in HT.