Inappropriateness in laboratory medicine: An elephant in the room?

Appropriateness of diagnostic testing can be conventionally described as prescription of the right test, using the right method, at the right time, to the right patient, with the right costs and for producing the right outcome. There is ongoing debate about the real burden of inappropriateness in laboratory diagnostics. The media coverage of this issue has also recently led to either over- or under-emphasizing the clinical, organizational and economic consequences. This is quite problematic, inasmuch as some reliable data are available in the current scientific literature, showing that inappropriateness of laboratory testing can be as high as 70%. This is especially evident for, though not limited to, cancer biomarkers testing, in which the practice of avoidable tests ordering is dramatically magnified. The reasons beyond inappropriateness are many and multifaceted, entailing wrong habits, resistance to changes, poor culture, insufficient education and healthcare inefficiencies. There are many unfavorable consequences attributable to avoidable testing, including unjustified incremental costs, derangement of laboratory efficiency and potential patient safety issues. The tentative solutions to this important problem necessitate that policymakers, local hospital administrators, laboratory professionals, clinicians, patients' associations and diagnostic companies join the efforts and embark in the same landmark effort for disseminating a better culture of appropriateness

A rapid spectroscopic method to detect the fraudulent treatment of tuna fish with carbon monoxide

Carbon monoxide (CO) can be used to treat fresh meat and fish in order to retain its 'fresh' red colour appearance for a longer period of time. In fact, upon aging, myoglobin is oxidized to met-myoglobin with the concomitant blue-shift and broadening of the Soret maximum, which brings about a change in the colour of the fish, revealing that it is no longer fresh. The use of carbon monoxide, which reacts with the oxy-myoglobin to form a fairly stable cherry red carboxy-myoglobin complex may mask spoilage, because the CO-complex can be stable beyond the microbiological shelf life of the meat. The presence of CO in tuna fish has been investigated by optical spectroscopy as the formation of the CO adduct can be easily detected by the combined analysis of electronic absorption spectra in their normal and second derivative modes, monitoring the intense Soret band at 420 nm. The presence of met- and oxy-myoglobin can obscure the presence of small amounts of the CO adduct; however, it can be revealed by chemically reducing the met- and oxy-forms to the deoxy-form in an anaerobic environment. This spectroscopic method provides a qualitatively rapid laboratory screening procedure for food control to unmask the presence of CO in frozen or fresh fish. (c) 2006 Elsevier Ltd. All rights reserved

Monocyte to lymphocyte blood ratio in tuberculosis and HIV patients: Comparative analysis, preliminary data

Recent data confirmed the hypothesis suggested by historical studies that the ratio of peripheral blood monocytes to lymphocytes (M/L) is associated with the risk of tuberculosis (TB) disease. We retrospectively analyzed the electronic health records of tuberculosis and HIV-positive patients who had followed day-care programs at the AIDS Center of the University of Palermo, Italy. 261 patients were recruited and divided into 6 groups as follows: healthy control group (HCG: 47 pts), latent HIV negative infected TB group (LIG, 43 pts), active HIV negative tuberculosis (TAG: 61 pts), treated tuberculosis HIV negative (TTG: 44 pts), HIV drug-naive patients tested TST and QFT-IT-negative with negative chest x-Ray (HIVnG: 44 pts), and HIV-tuberculosis coinfection (HIVTB-G: 22 pts). For each group, absolute lymphocyte (L), monocyte (M) and M/L ratio by peripheral blood was calculated. The mean value of monocytes in the TAG group was significant, the highest (0.70\uc2\ub10.37 1x103/\uce\ubcl) in comparison to HGC (0.70>0.44, p-value <0.05), HIVnG (0.70>0.40, p-value <0.05) and HIVTB-G (0.70>0.45, p-value<0.05). Monocyte to lymphocyte blood RATIO showed a significant difference between groups (p-value <0.001). In particular, the mean score of M/L ratio was higher in the TAG group compared to the HGC (0.49>0.27, p-value<0.05), LIG (0.49>0.29, p-value<0.05), TTG (0.49>0.32) and HIVTB-G groups (0.49>0.27, p-value<0.05). Our data confirm a significant difference in monocyte to lymphocyte blood ratio in tuberculosis disease. These data may be useful for monitoring and revising implementation plans for the different phases of tuberculosis disease (latent Mycobacterium tuberculosis (MTB) infection versus TB active disease). Regarding HIV samples, the small sample size is somewhat offset by the need, fully satisfied in our sample, to enlist specific patients such as co-infected HIV/TBC who voluntarily submit to clinical trials in our geographical area

Procalcitonin and community-acquired pneumonia (CAP) in children

The role of procalcitonin (PCT) as a biomarker for sepsis in adults is well documented, while its role in infections affecting neonatal children remains controversial. Among these infections, Community-Acquired pneumonia (CAP) has been studied extensively, because it's the second cause of death in children in developing countries, and one of the most frequent causes of hospitalization in industrialized countries. The PubMed database and the Cochrane Library were used to search for the following keywords: CAP, procalcitonin, children. Thirteen articles were studied to determine the role of PCT in CAP management, specifically its usefulness for distinguishing pneumococcal infections from viral and unknown infections, for predicting severity and the correct antibiotic treatment. This paper focuses on the studies performed to identify the best inflammatory biomarker for CAP management. Although there is an increase in studies confirming the usefulness of PCT in CAP management in children, further studies are needed to have better understanding of its role for pediatric CAP management

Somatostatin: A Novel Substrate and a Modulator of Insulin-Degrading Enzyme Activity

Insulin-degrading enzyme (IDE) is an interesting pharmacological target for Alzheimer's disease (AD), since it hydrolyzes beta-amyloid, producing non-neurotoxic fragments. It has also been shown that the somatostatin level reduction is a pathological feature of AD and that it regulates the neprilysin activity toward beta-amyloid. In this work, we report for the first time that IDE is able to hydrolyze somatostatin [k(cat) (s(-1)) = 0.38 (+/-0.05); K-m (M) = 7.5 (+/-0.9) x 10(-6)] at the Phe6-Phe7 amino acid bond. On the other hand, somatostatin modulates IDE activity, enhancing the enzymatic cleavage of a novel fluorogenic beta-amyloid through a decrease of the K-m toward this substrate, which corresponds to the 10-25 amino acid sequence of the A beta(1-40). Circular dichroism spectroscopy and surface plasmon resonance imaging experiments show that somatostatin binding to IDE brings about a concentration-dependent structural change of the secondary and tertiary structure(s) of the enzyme, revealing two possible binding sites. The higher affinity binding site disappears upon inactivation of IDE by ethylenediaminetetra acetic acid, which chelates, the catalytic Zn2+ ion. As a whole, these features suggest that the modulatory effect is due to an allosteric mechanism: somatostatin binding to the active site of one IDE subunit (where somatostatin is cleaved) induces an enhancement of IDE proteolytic activity toward fluorogenic beta-amyloid by another subunit. Therefore, this investigation on IDE-somatostatin interaction contributes to a more exhaustive knowledge about the functional and structural aspects of IDE and its pathophysiological implications in the amyloid deposition and somatostatin homeostasis in the brain. (C) 2008 Elsevier Ltd. All rights reserved

Ferrous Campylobacter jejuni truncated hemoglobin P displays an extremely high reactivity for cyanide - A comparative study

Campylobacter jejuni hosts two hemoglobins (Hbs). The Camplylobacter jejuni single-domain Hb (called Cgb) is homologous to the globin domain of flavohemoglobin, and it has been proposed to protect the bacterium against nitrosative stress. The second Hb is called Ctb (hereafter Cj-trHbP), belongs to truncated Hb group III, and has been hypothesized to be involved in O 2 chemistry. Here, the kinetics and thermodynamics of cyanide binding to ferric and ferrous Cj-trHbP [Cj-trHbP(III) and Cj-trHbP(II), respectively] are reported and analyzed in parallel with those of related heme proteins, with particular reference to those from Mycobacterium tuberculosis. The affinity of cyanide for Cj-trHbP(II) is higher than that reported for any known (in)vertebrate globin by more than three orders of magnitude (K = 1.2 × 10-6 m). This can be fully attributed to the highest (ever observed for a ferrous Hb) cyanide-binding association rate constant (kon = 3.3 × 103 m-1·s-1), even though the binding process displays a rate-limiting step (kmax = 9.1 s -1). Cj-trHbP(III) shows a very high affinity for cyanide (L = 5.8 × 10-9 m); however, cyanide association kinetics are independent of cyanide concentration, displaying a rate-limiting step (l max = 2.0 × 10-3 s-1). Values of the first-order rate constant for cyanide dissociation from Cj-trHbP(II)-cyanide and Cj-trHbP(III)-cyanide (koff =5.0 × 10-3 s -1 and loff ≥ 1 × 10-4 s-1, respectively) are similar to those reported for (in)vertebrate globins. The very high affinity of cyanide for Cj-trHbP(II), reminiscent of that of horseradish peroxidase(II), suggests that this globin may participate in cyanide detoxification. © 2008 The Authors

The Met80Ala and Tyr67His/Met80Ala mutants of human cytochrome cshed light on the reciprocal role of Met80 and Tyr67 in regulating ligand access into the heme pocket.

The spectroscopic and functional properties of the single Met80Ala and double Tyr67His/Met80Ala mutants of human cytochrome c have been investigated in their ferric and ferrous forms, and in the presence of different ligands, in order to clarify the reciprocal effect of these two residues in regulating the access of exogenous molecules into the heme pocket. In the ferric state, both mutants display an aquo high spin and a low spin species. The latter corresponds to an OH- ligand in Met80Ala but to a His in the double mutant. The existence of these two species is also reflected in the functional behavior of the mutants. The observation that (i) a significant peroxidase activity is present in the Met80Ala mutants, (ii) the substitution of the Tyr67 by His leads to only a slight increase of the peroxidase activity in the Tyr67His/Met80Ala double mutant with respect to wild type, while the Tyr67His mutant behaves as wild type, as previously reported, suggests that the peroxidase activity of cytochrome c is linked to an overall conformational change of the heme pocket and not only to the disappearance of the Fe-Met80 bond. Therefore, in human cytochrome c there is an interplay between the two residues at positions 67 and 80 that affects the conformation of the distal side of the heme pocket, and thus the sixth coordination of the hem

Averaging lifetimes for B hadron species

The measurement of the lifetimes of the individual B species are of great interest. Many of these measurements are well below the 10 $\%$ level of precision. However, in order to reach the precision necessary to test the current theoretical predictions, the results from different experiments need to be averaged. Therefore, the relevant systematic uncertainties of each measurement need to be well defined in order to understand the correlations between the results from different experiments. \par In this paper we discuss the dominant sources of systematic errors which lead to correlations between the different measurements. We point out problems connected with the conventional approach of combining lifetime data and discuss methods which overcome these problems