Role of germinal center and CD39highCD73+ B cells in the age-related tonsillar involution.

BACKGROUND: The tonsils operate as a protection ring of mucosa at the gates of the upper aero-digestive tract. They show similarities with lymph nodes and participate as inductive organs of systemic and mucosal immunity. Based on the reduction of their size since puberty, they are thought to experience involution in adulthood. In this context, we have used tonsillar mononuclear cells (TMC) isolated from patients at different stages of life, to study the effect of ageing and the concomitant persistent inflammation on these immune cells. RESULTS: We found an age-dependent reduction in the proportion of germinal center B cell population (BGC) and its T cell counterpart (T follicular helper germinal center cells, TfhGC). Also, we demonstrated an increment in the percentage of local memory B cells and mantle zone T follicular helper cells (mTfh). Furthermore, younger tonsils rendered higher proportion of proliferative immune cells within the freshly isolated TMC fraction than those from older ones. We demonstrated the accumulation of a B cell subset (CD20+CD39highCD73+ cells) metabolically adapted to catabolize adenosine triphosphate (ATP) as patients get older. To finish, tonsillar B cells from patients at different ages did not show differences in their proliferative response to stimulation ex vivo, in bulk TMC cultures. CONCLUSIONS: This paper sheds light on the changing aspects of the immune cellular landscape, over the course of time and constant exposure, at the entrance of the respiratory and digestive systems. Our findings support the notion that there is a re-modelling of the immune functionality of the excised tonsils over time. They are indicative of a transition from an effector type of immune response, typically oriented to reduce pathogen burden early in life, to the development of an immunosuppressive microenvironment at later stages, when tissue damage control gets critical provided the time passed under immune attack. Noteworthy, when isolated from such histologic microenvironment, older tonsillar B cells seem to level their proliferation capacity with the younger ones. Understanding these features will not only contribute to comprehend the differences in susceptibility to pathogens among children and adults but would also impact on vaccine developments intended to target these relevant mucosal sites.

Biomarkers of exposure in urine of active smokers, non-smokers, and vapers.

The exposure to smoking related products has been evaluated through urine illness risk marker determination through the analysis of urine samples of smokers and vapers. Biomarkers and their metabolites such as N-acetyl-S-(2-cyanoethyl)-L-cysteine (CEMA), N-acetyl-S-(3,4-dihydroxybutyl)-L-cysteine (DHBMA), N-acetyl-S-[1-(hydroxymethyl)-2-propen-1-yl)-L-cysteine (MHBMA), N-acetyl-S-(3-hydroxypropyl)-L-cysteine (3HPMA), 2R-N-acetyl-S-(4-hydroxybutan-2-yl)-L-cysteine (HMPMA), and N-acetyl-S-(3-carboxy-2-propyl)-L-cysteine (CMEMA) together with nicotine and cotinine were identified and quantified by LC-HRMS and LC-MS/MS, and data found normalized to the creatinine level. One hundred two urine samples were collected from smokers, non-smokers, and vapers, spanning an age range from 16 to 79 years. Results obtained showed that CEMA was only detected in urine samples from smokers and MHBMA was in the same order of magnitude in all the urine samples analyzed. HMPMA was found in the urine of vapers at the same order of concentration as in non-smokers. 3HPMA in vapers was lower than in the urine of smokers, presenting an intermediate situation between smokers and non-smokers. On the other hand, DHBMA in vapers can reach similar values to those found for smokers, while CMEMA shows concentrations in the urine of vapers higher than in the case of non-smokers and traditional smokers, requiring new research to link this metabolite to the use of electronic cigarettes and possible alternative metabolomic routes. In general, this study seems to verify that traditional smoking practice constitutes a major source of carcinogenic chemicals compared with substitutive practices, although those practices are not free of potential harm.

Green near-infrared determination of copper and mancozeb in pesticide formulations.

A green analytical procedure has been successfully developed for the simultaneous determination of copper and mancozeb in phytosanitary products. The method is based on different direct measurements of diffuse reflectance near-infrared (DR-NIR) spectra. Accuracy of the method has been evaluated by comparison of the obtained copper and mancozeb concentrations with those provided by reference methodologies based on titrimetric procedures. The average relative prediction error was 0.7 and 1.6 % for copper and mancozeb, respectively. The evaluation of the greenness of the DR-NIR procedure provided 100 points, which is the maximum value in the Green Certificate ranking, because of the absence of consumed reagents and waste generation and energy consumption lower than 0.1 kWh.

The chemical memory of smoking tobacco.

The concentration in urine of N-acetyl-hydroxy-propyl-cisteine (3HPMA), an acrolein metabolite, has been employed as a marker of the risk of illness of smokers and the relative concentration of creatinine has been evaluated to verify the effect of moving from the practice of burning tobacco to nicotine vaping. From the results concerning the urine samples of 38 subjects, collected from 2021 to 2023 and analyzed by LC-MS/MS, corresponding to 5 active smokers, 13 previously heavy smokers who replaced traditional tobacco by vaping, and 20 non-smokers, a dramatic reduction was found in 3HPMA/creatinine in urine. 3HPMA varied from values of 2150-3100 µg gcreatinine-1 to levels of 225-625 µg gcreatinine-1 found for non-smokers, with the time decay described by the equation y = 0.3661x2 - 94.359x + 6246.4 (R2: 0.757), providing a time of approximately 10 years for tobacco memory after the cessation of the consumption of burned tobacco.

Fast determination of fish mineral profile. Application to Vietnamese panga fish.

A methodology, based on inductively coupled plasma optical emission spectrometry after microwave-assisted digestion with HNO3/H2O2, was developed to determine the mineral profile of panga fish samples imported from Vietnam. A total of 42 essential and toxic elements were studied in seven samples taken from the local market. Preliminary studies were focused on selecting the best wavelength and the required dilution of samples in order to provide the highest sensitivity to maximize the number of analytes to be determined without spectral or matrix interferences. Adequate accuracy was assured by the analysis of certified reference material TORT-2. Mercury was also determined by a direct method based on atomic absorption spectrometry. Results obtained indicated a low mineral profile, fourteen elements were found at quantitatively levels, Na (6000 µg g(-1)) K (1800 µg g(-1)) Mg (173 µg g(-1)), Ca (80 µg g(-1)), Zn (2.44 µg g(-1)), Fe (1.6 µg g(-1)), Al (1.1 µg g(-1)), Sr (0.4 µg g(-1)) and B, Ba, Hg, Mn, V (under 0.1 µg g(-1)). Additionally data were compared with those previously reported in literature and an estimation of daily intake was calculated and compared with recommended or tolerable guidelines values. Levels of As, Cd, Pb and Hg were far below the established values by the European Community.

A green methodology for the determination of cocaine in camouflaged samples.

A methodology based on the ultrasound-assisted extraction with ethanol and the dry film attenuated total reflectance infrared spectroscopy (DF-ATR-FTIR) measurement of extracts has been developed for a fast evaluation of non-conventional ("exotic") solid-sized cocaine samples. The method provides quantitative results in less than three minutes with a limit of detection in the solid sample of 1.6 µg g-1 of cocaine with a variation coefficient lower than 7%. Results found for seized samples of different natures were compared with those obtained by a reference gas chromatography method and the greenness of the whole proposed procedure was evaluated and compared using the analytical eco-scale, green analytical procedure index (GAPI), and analytical greenness metric (AGREE). The green evaluation of the proposed methodology provided green scores by considering different evaluation criteria.

Ecofriendly in-line process monitoring: a case study. Anthracene photodegradation in the presence of refuse-derived soluble bio-organics.

Photodegradation of anthracene has been studied in aqueous solutions containing soluble bio-organic substances isolated from urban refuse. To perform a preliminary rapid feasibility study of this process while reducing the amount of analytical effort and reagents, an experimental set-up was developed comprising a Teflon coil surrounding a UV-lamp and coupled with an in-line spectrofluorimeter. In this fashion only few millilitres of solution are needed to study the degradation process. Furthermore, the in-line spectroscopic approach enables monitoring of the process without consumption of reagents. Additional studies by liquid chromatography and use of toxicity tests clearly indicated that the apparent inhibition effect of bio-organic compounds on anthracene degradation is not relevant. The results imply that urban refuse may be used as an auxiliary in the recovery of polycyclic aromatic hydrocarbons from contaminated soil by washing, without deleterious effects on the photodegradation of anthracene and other aromatic pollutants.

Protein determination in serum and whole blood by attenuated total reflectance infrared spectroscopy.

Attenuated total reflectance mid-infrared spectra of serum and blood samples were obtained from 4,000 to 600 cm(-1). Models for the determination of albumin, immunoglobulin, total globulin, and albumin/globulin coefficients were established for serum samples, using reference data obtained by capillary electrophoresis. Based on the use of the amide bands I and II regions, the relative root mean square error of prediction (RRMSEP) was 4.9, 14.9, 4.5, and 7.1% for albumin, immunoglobulin, total globulin, and albumin/globulin coefficients, respectively, determined in an independent validation set of 120 samples using 200 samples for calibration. Additionally, the use of Kennard-Stone method for the selection of a representative calibration subset of samples provided comparable results using only 60 samples. For whole blood analysis, hemoglobin was determined in 40 validation samples using models built from 40 calibration independent samples with RRMSEP of 8.3, 5.5, and 4.9% with models built from direct spectra in the first case and from sample spectra recorded after lysis by sodium dodecyl sulfate and freezing, respectively, for the last two ones. The developed methodologies offer green alternatives for patient diagnosis in a few minutes, minimizing the use of reagents and residues and being adaptable for its use as a point-of-care method.

Date-rape evidence through fast determination of γ-butyrolactone in adulterated beverages.

An infrared spectroscopy (IR) based methodology has been developed to determine γ-butyrolactone (GBL) in adulterated beverages. The proposed method permits the direct screening of GBL in beverages and involves a minimum sample treatment requiring less than 2 min for quantitative determination of GBL. Sensitivity of IR method was improved by using liquid-liquid extraction (LLE) providing detection limits of 0.023 mg g-1. Accuracy of the proposed methodology was evaluated through the analysis of soft beverages and alcoholic cocktails spiked with GBL at concentration levels ranging from 0.075 to 10 mg g-1 providing recovery values from 91 to 100%. GBL was determined in twelve blind-spiked beverages, including from mineral water to wine and cocktails. Results obtained were statistically comparable to those provided by a liquid chromatography (LC) reference methodology and consistent with the spiked values. Therefore, the use of LLE-FTIR allowed a simple, sensitive and quantitative determination of GBL in soft beverages and alcoholic cocktails, thus evidencing its use for sex submission intention.

The use of near-infrared spectrometry in the olive oil industry.

The enormous possibilities offered by near-infrared (NIR) spectroscopy for the (on/in/at-line) quality control process of olive fruits, pastes, and oils are summarized throughout this paper. Special attention has been paid to the combination of NIR and chemometric treatments for the on-line analysis of olive fruits and also for the quality parameters evaluation on olive oils and pastes which can enhance the production of a high quality olive oil and the selection of olive fruit with superior properties. The implementation of NIR sensors in olive mills with successful results has also been reviewed and the commercial olive fruit and oil analyzers highlighted.

Inductively coupled plasma optical emission spectroscopy determination of trace element composition of argan oil.

A methodology based on inductively coupled plasma optical emission spectroscopy (ICP-OES) after microwave assisted acid digestion has been developed to determine the trace element content of Moroccan argan oil. Limit of detection values equal or lower than few mg/kg were obtained for all elements under study. To assure the accuracy of the whole procedure, recovery studies were carried out on argan oil samples spiked at different concentration levels from 10 to 200 µg/L. Quantitative average recovery values were obtained for all elements evaluated, demonstrating the suitability of this methodology for the determination of trace elements in argan oil samples. Aluminum, calcium, chromium, iron, potassium, lithium, magnesium, sodium, vanadium and zinc were quantitatively determined in Moroccan argan oils being found that their concentration is different of that found in other edible oils thus offering a way for authentication and for the evaluation of possible adulterations.

A clinical case of necrotizing sialometaplasia and conjunctival hemorrhage.

The first case reported in the literature of a rare disease called necrotizing scialometaplasia (NS), dates back to 1973 when Abrams et al. described the main histological features of this disease. In this article we describe the rare clinical case of a young woman came to our observation for a double ulcer in the middle portion of the hard palate, aching, that histological examination showed compatible with a diagnosis of NS and preceded his appearance a haemorrhagic conjunctival suffusion left. We have provided a complete description of all the investigations in which the patient underwent and its treatment. We have also outlined the major etiological hypotheses of SN, histological features that point to a correct diagnosis, clinical features and prognostic and finally we reflected on the rare and interesting overlap in clinical manifestations palatal and conjunctival those trying to find a possible explanation.

Determination of total Sb, Se, Te, and Bi and evaluation of their inorganic species in garlic by hydride-generation-atomic-fluorescence spectrometry.

A sensitive and simple analytical method has been developed for determination of Sb(III), Sb(V), Se(IV), Se(VI), Te(IV), Te(VI), and Bi(III) in garlic samples by using hydride-generation-atomic-fluorescence spectrometry (HG-AFS). The method is based on a single extraction of the inorganic species by sonication at room temperature with 1 mol L(-1) H2SO4 and washing of the solid phase with 0.1% (w/v) EDTA, followed by measurement of the corresponding hydrides generated under two different experimental conditions directly and after a pre-reduction step. The limit of detection of the method was 0.7 ng g(-1) for Sb(III), 1.0 ng g(-1) for Sb(V), 1.3 ng g(-1) for Se(IV), 1.0 ng g(-1) for Se(VI), 1.1 ng g(-1) for Te(IV), 0.5 ng g(-1) for Te(VI), and 0.9 ng g(-1) for Bi(III), in all cases expressed in terms of sample dry weight.

Partial least squares modelization of energy dispersive X-ray fluorescence.

As a proof of concept, a green methodology has been developed for the energy dispersive X-ray fluorescence (ED-XRF) determination of calcium, potassium, iron, magnesium, aluminum, chromium, strontium, phosphorus and nickel in the peel of untreated kaki fruit (Diospyros kaki. L) samples. ED-XRF spectra of fifty-six kakis purchased in the local area of LLombay (Valencia) were obtained directly from samples without any previous treatment and without sample damage just after cleaning the fruit with distilled water. Inductively Couple Plasma Optical Emission Spectrometry (ICP-OES) was used as a reference method to determine the mineral elements after microwave assisted acid digestion. XRF spectra and concentration values obtained by ICP-OES were processed using partial least squares (PLS) data treatment to build the corresponding chemometric models for prediction of mineral profile of samples. PLS-ED-XRF permits a direct and accurate determination of Ca and K in kaki peel. For Al, Fe, Mg, Ni and Sr screening semiquantitative results were obtained. Concentrations obtained directly by the internal calibration of instrument, using GeoChem Trace model, were also compared with data predicted by chemometric models being found that PLS models must replace the calibration of the instrument for thus kind of analysis.

On-line gel permeation chromatography-attenuated total reflectance-Fourier transform infrared determination of lecithin and soybean oil in dietary supplements.

Gel permeation chromatography (GPC) with attenuated total reflectance-Fourier transform infrared (ATR-FTIR) spectrometry detection has been proposed for the simultaneous determination of lecithin and soybean oil in dietary supplements. The method involves the extraction of analytes with dichloromethane in an ultrasound water bath and the injection of 2 ml of centrifuged and filtered extracts into the system integrated by two Envirogel GPC columns (19 mm x150 mm, 19 mm x 300 mm) coupled on-line. Dichloromethane was used as mobile phase. A method has been developed to select the most appropriated wavenumber to be used for the determination of each considered compound from the calculation of a factor which maximizes the analyte signal minimizing the interferent contributions, being selected the detection wavenumbers of 1034 and 1138 cm(-1) for lecithin and soybean oil, respectively in the first order derivative ATR-FTIR spectra. The method provides limits of detection of 2 and 4 mg ml(-1) for lecithin and soybean oil and repeatability values, measured as relative standard deviation, of 2.5% and 3.4% being extended the linear range till 100 mg ml(-1) for lecithin and up to 50 mg ml(-1) for soybean oil. Accurate results were found for 10 synthetic samples and 7 commercial dietary supplement preparations.

Determination of glycolic acid in cosmetics by online liquid chromatography-Fourier transform infrared spectrometry.

An isocratic online liquid chromatography Fourier transform infrared procedure has been developed for the determination of glycolic acid in cosmetics. The method involves the ultrasound-assisted extraction of glycolic acid from the samples with an acetonitrile:phosphate buffer (25 mM, pH 2.7) (3:97 v/v). The extracts were centrifuged and filtered before their injection into the chromatography system, which was equipped with a C18 column and used a flow rate of 150 microL min(-1). FTIR spectra were acquired using a time-resolved rapid scan mode. To calculate the chromatograms, the spectral area was integrated between 1288 and 1215 cm(-1), with baseline correction established between 1319 and 1150 cm(-1), after correcting for the eluent spectral background. Peak area values of the extracted sample chromatograms were interpolated from an external calibration curve. The method provided a limit of detection of 0.034 mg mL(-1) and a relative standard deviation of 6% for five measurements at the 0.174 mg mL(-1) concentration level. Recovery values obtained by spiking 400 mg of three commercially available samples with amounts of glycolic acid from 3.7 to 9.8 mg ranged between 99.6 and 101%. The results obtained for the commercial samples agree well with their declared concentrations. An attempt to directly determine glycolic acid by attenuated total reflectance measurements using partial least squares calibration showed that results were strongly influenced by compounds coextracted from the matrix.

Trace analysis by ion mobility spectrometry: From conventional to smart sample preconcentration methods. A review.

Ion mobility spectrometry (IMS) is a rapid and high sensitive technique widely used in security and forensic areas. However, a lack of selectivity is usually observed in the analysis of complex samples due to the scarce resolution of the technique. The literature concerning the use of conventional and novel smart materials in the pretreatment and preconcentration of samples previous to IMS determinations has been critically reviewed. The most relevant strategies to enhance selectivity and sensitivity of IMS determinations have been widely discussed, based in the use of smart materials, as immunosorbents, aptamers, molecularly imprinted polymers (MIPs), ionic liquids (ILs) and nanomaterial. The observed trend is focused on the development of IMS analytical methods in combination of selective sample treatments in order to achieve quick, reliable, sensitive, and selective methods for the analysis of complex samples such as biological fluids, food, or environmental samples.

Ion mobility spectrometry and high resolution mass-spectrometry as methodologies for rapid identification of the last generation of new psychoactive substances.

A new drug trafficking trend has been observed in the last years by the introduction in the black market of new psychoactive substances (NPS) in order to difficult competent authority controls. In this study, ion mobility spectrometry (IMS) and high-resolution mass-spectrometry (HRMS) were proposed as vanguard and rearguard methodologies for the rapid identification of the last generation of NPS in seizures. The combined use of IMS and HRMS has been evaluated through the analysis of 24 NPS seized from 2016 to 2018 in Valencia (Spain) to demonstrate the utility of this approach. The characteristic reduced mobility (K0) values for seized NPS were determined and mass-mobility relationships were proposed and evaluated for the main NPS families: amphetamine and cathinone derivatives, and synthetic cannabinoids. IMS did not allow a unequivocal identification by itself; so, HRMS analysis was employed as rearguard confirmation methodology for the right identification of NPS. Thus, the combined use of IMS and HRMS can be considered as promising alternative for the rapid screening and identification of NPS in seizures.

Fast authentication of tea tree oil through spectroscopy.

Two new procedures, based on infrared spectroscopy in the near infrared (NIR) and mid infrared (MIR), have been developed for the authentication of tea tree oil (TTO) commercial samples. Infrared measurements were made on untreated samples by transmission NIR and attenuated total reflectance (ATR) followed by partial least square discriminant analysis (PLS-DA). These methods offer a fast and low cost alternative to enantiomeric two-dimensional gas chromatography coupled to mass detection usually employed to discriminate between authentic and non-authentic samples. In these studies, a set of 267 samples, including authentic and non-authentic labelled tea tree oil samples, were used to build the models based on the wavenumber range, data pre-processing and latent variables number selection. Infrared methods can be discriminant for authentic and non-authentic TTO samples with a 98% certainty for both ATR and NIR methodologies, employing 92 and 142 external samples respectively. Developed PLS-DA infrared based methodologies and the reference methodology have been evaluated and compared from a Green Analytical point of view.

Radiation Safety Aspects of Iodine-131 metaiodobenzylguanidine (131I mIBG) Therapy Program Startup.

As a medical center without a pre-existing radiopharmaceutical therapy program, it was a daunting endeavor to implement a I metaiodobenzylguanidine (mIBG) high-dose treatment regimen. It took several years of planning with hospital administration, vendors, and Texas Department of Health Radiological Control regulators to establish a viable program. Effective communication between physicians, nursing, nuclear medicine, environmental services, maintenance, and other support staff is essential and paramount for the successful execution and continued sustainability of the mIBG therapy program. Besides providing an effective treatment for patients, an additional goal for the program is to keep radiation exposure As Low As Reasonably Achievable (ALARA) for staff and patient caregivers. As such, start-up presented many training, logistical, and radiation safety challenges. The location of the isolation room and shielding specifications were designed to keep radiation exposure to public access areas to less than 2 microsieverts per hour. Before the first patient was treated the policies and procedures for training, radiation safety, product quality control, and infusion process needed to be developed, tested, and approved by various committees. Furthermore, a similar process was required for developing room set-up, post therapy cleanup, and waste storage procedures. Throughout the maturation process of the program, the departments involved have found that our safety culture has continually improved by the re-enforcement of knowledge and lessons learned, as both the ancillary and treatment staff grew more confident in each other's ability during more patient treatments are performed. This article describes the process and lessons learned during the time leading up to the startup and early years of the mIBG therapy program.