Spectroscopic trends for the determination of illicit drugs in oral fluid

The present work aims to review all of the articles published so far, focusedon the determination of drugs of abuse in oral fluid. This fluid provides a simpler,faster, and more controllable sampling in comparison with the other biological fluids,such as blood or urine. Actually, the main goal of the researchers is to lower the limitof detection (LOD) to detect quantities of drugs smaller than the cut-off limitsestablished by law for drug controls. Advances in Raman, infrared (IR), and nuclearmagnetic resonance (NMR) spectroscopy applications are discussed. SurfaceenhancedRaman spectroscopy (SERS) has been shown as the most sensitive techniquefor the detection of illicit drugs in oral fluid. The use of IR spectroscopy fordetermining drugs of abuse in oral fluid is growing, although the LODs obtained untilnow do not yet satisfy the necessities in the forensic field. Finally, NMR spectroscopyhas seldom been used to determine drugs in oral fluid. Another future trend seems tobe related with the use of portable instrumentation, which would allow us to performin-situ analysis. This last application seems to be particularly promising to performroadside drug tests and to identify overdose drugs in patients in emergency conditions

Analysis of street cocaine samples in nasal fluid by Raman spectroscopy

The principal objective of this work was to demonstrate the capability of Raman spectroscopy to detectsmall amounts of cocaine in nasal fluid, and to identify the main drug and the most widely used cuttingagents. Initially, standard samples were analysed and sampling conditions were studied by comparingdifferent swabs used for the sample collection. Once the most appropriate swab was selected, whichpermitted a relatively simple detection of the standard cocaine hydrochloride, qualitative analyses of realsamples were carried out. Three street cocaine samples were analysed, and the presence of cuttingsubstances was highlighted by the appearance of different bands not corresponding to the ones of thestandard cocaine. To identify the substances present in each sample, the spectra of the street cocainesamples were collected and compared with a digital library created on purpose with the spectra of themost common cutting agents. In this case, correlation coefficients permitted to recognize the most importantsubstances presumably present in the samples, and gave an estimation of the purity of the cocaine.However, when nasal fluid was present, its strong signal could overlap or interfere with thesmaller signal of the cutting substances, hindering their identification

Isolation of a 25-kDa protein binding to a curved DNA upstream the origin of the L strand replication in the rat mitochondrial genome

The presence of a curved DNA sequence in the gene for the NADH-dehydrogenase subunit 2 of rat mitochondrial genome, upstream from the origin of the light strand replication have been demonstrated through theoretical analysis and experimental approaches. Gel retardation assays showed that this structure makes a complex with a protein component extracted from the mitochondrial matrix. The isolation and purification of this protein is reported. With a Sepharose CL-6B and magnetic DNA affinity chromatography a polypeptide was purified to homogeneity having 25-kDa mass as shown by gel electrophoresis. To functionally characterize this protein, its capability to bind to other sequences of the homologous or heterologous DNA and to specific riboprobes was also investigated. A role for this protein as a trans-acting agent required for the expression of the mammalian mitochondrial genome is suggested

Ultraviolet resonance Raman spectroscopy for the detection of cocaine in oral fluid

Detecting and quantifying cocaine in oral fluid is of significant importance for practical forensics. Up to date, mainly destructive methods or biochemical tests have been used, while spectroscopic methods were only applied to pretreated samples. In this work, the possibility of using resonance Raman spectroscopy to detect cocaine in oral fluid without pretreating samples was tested. It was found that ultraviolet resonance Raman spectroscopy with 239-nm excitation allows for the detection of cocaine in oral fluid at 10 mu g/mL level. Further method development will be needed for reaching the practically useful levels of cocaine detection. (C) 2017 Elsevier B.V. All rights reserved

Antibody levels after BNT162b2 vaccine booster and SARS-CoV-2 Omicron infection

In the present study, immunogenicity data in 61 vaccinated healthcare workers (HCWs) either infection naïve (naïve HCWs) or with infection of Delta and/or Omicron COVID-19 (experienced HCWs) were evaluated up to 270 days after the second dose of BNT162b2 vaccine and up to 90 days after a booster dose. A decrease in antibody levels at 270 days following administration of the second dose (p = 0.0335) was observed, although values did not fall below the positivity threshold (33.8 BAU/ml). After booster vaccination, antibody levels increased after 30 days (p = 0.0486), with much higher values than after first and second vaccination. Antibody levels then decreased at 60 and 90 days after the booster dose. A comparison between mean antibody levels of naïve and experienced HCWs revealed higher values in experienced HCWs, resulting from both natural and vaccination-induced immunity. A total of 14.7% of HCWs contracted the Omicron virus variant after the vaccine booster, although none showed severe symptoms. These results support that a booster dose results in a marked increase in antibody response that subsequently decreases over time

Complex chromosome rearrangements related 15q14 microdeletion plays a relevant role in phenotype expression and delineates a novel recurrent syndrome

Complex chromosome rearrangements are constitutional structural rearrangements involving three or more chromosomes or having more than two breakpoints. These are rarely seen in the general population but their frequency should be much higher due to balanced states with no phenotypic presentation. These abnormalities preferentially occur de novo during spermatogenesis and are transmitted in families through oogenesis

Diagnostic utility of FGF-23 in mineral bone disorder during chronic kidney disease

Our data confirm that intact fibroblast growth factor 23 (iFGF-23) concentration is increased in patients with chronic kidney disease (CKD) and that it increases with disease progression (stages I-V). Therefore, iFGF-23 could be considered an early biomarker in the course of chronic kidney disease-mineral bone disorder (CKD-MBD), which has several aspects that make it potentially useful in clinical practice. The availability of an automated method for iFGF-23 assay may represent an added value in the management of the patient with CKD-MBD already from the early stages of the disease, before the increase of the routinely used laboratory parameters, 1-84 parathyroid hormone (PTH) and 25-OH-vitamin D (25-OH-vitD), which occur in more advanced stages of the disease

Gold nanorods as SERS substrate for the ultratrace detection of cocaine in non-pretreated oral fluid samples

This work shows the capability of surface-enhanced Raman spectroscopy (SERS) to detect ultratraces of cocaine(COC) in oral fluid (OF). It proposes a new solid substrate made of gold nanorods (Au NRs) to perform sensitiveanalyses of this complex matrix without any sample pretreatment. The Au NRs were synthetized optimizing thesynthesis conditions, and then characterized and tested as SERS substrate. The presented results demonstratedthat the SERS methodology was able to detect COC in OF with a limit of detection (LOD) as low as 10 ng/mL.This value was five orders of magnitude smaller than the one obtained with Raman spectroscopy (RS) and in theorder of the cut-off value (8 ng/mL) established for confirmatory test of COC in OF. Besides, a multivariateanalysis (OPLS-DA) applied on the samples analysed by SERS evidenced that it was possible to discriminateamong various COC concentrations. These are quite positive results because even the 1 ng/mL COC concentration sample could be identified as different from the OF sample. In addition, the absence of samplepretreatment and the ease of the proposed method, allowed performing rapid and non-destructive analyses,making the method suitable for in situ forensic analyses

Prevalence of mutations in BRCA and MMR genes in patients affected with hereditary endometrial cancer

Endometrial cancer (EC) is the fifth most common cancer in women from developed countries, accounting for 4.8% of new cases and 2.1% of deaths. The genetic basis for the familial risk of endometrial cancer has not been completely defined. Mostly, hereditary EC is part of two syndromes as Lynch syndrome (LS) and Hereditary Breast and Ovarian Cancer syndrome (HBOC). LS is the prototypical hereditary cancer syndrome in EC and accounts for 2-6% of all endometrial cancers. This disease is caused by autosomal dominant mutations in DNA mismatch repair (MMR) genes. Patients carrying a germline mutation in one of the MMR genes have a cumulative lifetime risk to develop EC of 20-70%. HBOC is an autosomal dominantly inherited disease, which mostly predisposes to breast and ovarian cancers, but it can be also associated with other malignancies. HBOC results from germline mutations in BRCA1/2 genes. The aim of this study was to determine the mutational status of a cohort of 40 EC patients, 19 belonging to families with LS and 21 to HBOC. Mutation analysis of MLH1, MSH2, BRCA1 and BRCA2 genes showed pathogenic variants in 17/40 (42.5%) patients. Out of 19 patients belonging to LS families, 8 (42.1%) showed a pathogenic variant. Out of 21 patients belonging to HBOC families, 9 (42.8%) showed a pathogenic variant. 1/21 (4.8%) patient report 1 variant of unknown significance (UV), c.599 C > T (p.T200I), in BRCA2. Moreover, in 1/21 (4.8%) patient we identified a novel missense variant in BRCA2, c.9541A > T (p.Met3181Leu). Mutational analysis was extended to family members, both healthy and cancer affected, of mutated patients; all the tested relatives affected with cancer displayed the pathogenic variant. Our data suggest that patients with hereditary EC have a high percentage of mutations in the LS and HBOC main susceptibility genes; therefore, the surveillance for EC, already indicated in LS patients, should also be recommended for patients with HBOC