Search

The COVID-19 pandemic highlighted lateral flow immunoassay (LFIA) strips as the most known point-of-care (POC) devices enabling rapid and easy detection of relevant biomarkers by nonspecialists. However, these diagnostic tests are usually associated with the qualitative detection of the biomarker of interest. Alternatively, electrochemical-based diagnostics, especially known for diabetes care, enable quantitative determination of biomarkers. From an analytical point perspective, the combination of the two approaches might represent a step forward for the POC world: in fact, electrochemical transduction is attractive to be integrated into LFIA strips due to its simplicity, high sensitivity, fast signal generation, and cost effectiveness. In this work, a LFIA strip has been combined with an electrochemical transduction, yielding an electrochemical LFIA (eLFIA). As a proof-of-concept method, the detection of prostate-specific antigen has been carried out by combining a printed-electrochemical strip with the traditional LFIA tests. The electrochemical detection has been based on the measurement of Au ions produced from the dissolution of the gold nanoparticles previously captured on the test line. The analytical performances obtained at LFIA and eLFIA were compared, highlighting how the use of differential pulse voltammetry allowed for a lower detection limit (2.5-fold), respectively, 0.38 and 0.15 ng/mL, but increasing the time of analysis. Although the correlation between the two architectures confirmed the satisfactory agreement of outputs, this technical note has been thought to provide the reader a fair statement with regard to the strength and drawbacks about combining the two (apparently) competitor devices in a diagnostics field, namely, LFIA and electrochemical strips.

To enhance pollinator loyalty, which is vital for plant reproduction, flowers have evolved rewards like nectar or excess pollen. Besides nutrients, nectars contain a plethora of secondary metabolites (SMs). Although the primary function of SMs is to protect plants from herbivores, growing evidence suggests that some of them may have adaptive functions in plant-pollinator interactions. GABA, which is prevalent in the nectar of many plants, interferes with learning and memory in restrained bees. Crucially, it is yet to be tested whether GABA affects pollinator foraging behaviour. The study aims to test whether free-flying bumble bees (Bombus terrestris) show a preference for artificial flowers dispensing GABA and whether GABA impacts bees’ learning of visual floral cues and promotes flower fidelity. We used three concentrations of GABA throughout the experiments, including two within the natural range and a third above it, to investigate any dose-dependent effects on foraging activity in flight arenas. The results showed that GABA did not affect bumble bee foraging preference at any concentration tested. Bees successfully learned to visit rewarding flowers regardless of the concentration of GABA, and no concentration affected the speed of learning. After learning, they tended to stay loyal to these flowers even when they became unrewarding. However, bumble bees that experienced GABA at the natural doses, but not at the unnatural high dose, remained faithful significantly longer to unrewarding flowers. The study provides insights into the use of SMs by plants to influence pollinator behaviour, with potential applications in agriculture.

Background: Widespread vaccination and emergence of less aggressive SARS-CoV2 variants may have blunted the unfavourable outcomes of COVID-19 in nursing home (NH) residents. We analysed the course of COVID-19 epidemic in NHs of Florence, Italy, during the “Omicron era” and investigated the independent effect of SARS-CoV2 infection on death and hospitalization risk. Methods: Weekly SARS-CoV2 infection rates between November 2021 and March 2022 were calculated. Detailed clinical data were collected in a sample of NHs. Results: Among 2044 residents, 667 SARS-CoV2 cases were confirmed. SARS-CoV2 incidence sharply increased during the Omicron era. Mortality rates did not differ between SARS-CoV2-positive (6.9%) and SARS-CoV2-negative residents (7.3%, p = 0.71). Chronic obstructive pulmonary disease and poor functional status, but not SARS-CoV2 infection independently predicted death and hospitalization. Conclusions: Despite that SARS-CoV2 incidence increased during the Omicron era, SARS-CoV2 infection was not a significant predictor of hospitalization and death in the NH setting.

Background: Nursing home (NH) residents have been dramatically affected by COVID-19, with extremely high rates of hospitalization and mortality. Aims: To describe the features and impact of an assistance model involving an intermediate care mobile medical specialist team (GIROT, Gruppo Intervento Rapido Ospedale Territorio) aimed at delivering “hospital-at-nursing home” care to NH residents with COVID-19 in Florence, Italy. Methods: The GIROT activity was set-up during the first wave of the pandemic (W1, March–April 2020) and became a structured healthcare model during the second (W2, October 2020–January 2021). The activity involved (1) infection transmission control among NHs residents and staff, (2) comprehensive geriatric assessment including prognostication and geriatric syndromes management, (3) on-site diagnostic assessment and protocol-based treatment of COVID-19, (4) supply of nursing personnel to understaffed NHs. To estimate the impact of the GIROT intervention, we reported hospitalization and infection lethality rates recorded in SARS-CoV-2-positive NH residents during W1 and W2. Results: The GIROT activity involved 21 NHs (1159 residents) and 43 NHs (2448 residents) during W1 and W2, respectively. The percentage of infected residents was higher in W2 than in W1 (64.5% vs. 38.8%), while both hospitalization and lethality rates significantly decreased in W2 compared to W1 (10.1% vs 58.2% and 23.4% vs 31.1%, respectively). Discussion: Potentiating on-site care in the NHs paralleled a decrease of hospital admissions with no increase of lethality. Conclusions: An innovative “hospital-at-nursing home” patient-centred care model based on comprehensive geriatric assessment may provide a valuable contribution in fighting COVID-19 in NH residents.

Introduction Since highly sensitive and specific serologic tests have been available (e.g., AGA [antigluten antibody] and more recently EMA [antiendomysium antibody]), it has been possible to identify a high prevalence [...]

The design, preparation and evaluation of molecularly imprinted polymers for roxarsone (4-hydroxy-3-nitrophenylarsonic acid), an organo-arsenic swine and poultry feed additive, using bi-substituted ureas and squaramide receptors as the functional monomers, are demonstrated. Pre-polymerisation studies of the template–monomer complexation performed by 1H NMR experiments show that squaramide-based monomers provide association equilibrium constant values higher than urea-based monomers. Equilibrium rebinding experiments in methanol show that two squaramide-based materials have good molecular recognition properties towards roxarsone, with high affinity (Keq= 16.85 × 103L mol−1and 14.65 × 103L mol−1, respectively), high imprinting factors (4.73 and 3.64 respectively) and good selectivity towards two roxarsone-related compounds, acetarsone (3-acetamido-4-hydroxyphenylarsonic acid) and nitarsone (4-nitrophenylarsonic acid). Polymer MIP-SQ2 was successfully used to setup an experimental protocol for the direct solid phase extraction of roxarsone from surface water samples. The method gives clean HPLC traces, with recoveries between 91% and 95% at concentration levels of 5.0, 10, and 25 mg L−1. Sample preconcentration with good recoveries between 87% and 97%, are shown, confirming that it is possible to employ the developed materials to measure roxarsone down to 1 μg L−1in water samples.

Stable and efficient conjugates between antibodies and gold nanoparticles (GNP-Ab) are sought to develop highly sensitive and robust biosensors with applications in medicine, toxicology, food safety controls, and targeted drug delivery. Several strategies have been proposed for directing the antibody attachment to GNPs thus preserving antibody activity, including covalently coupling the antibody to a polymer grafted on GNP surface and exploiting the high affinity of bioreceptors as mediators for the binding. Both approaches also allow for shielding GNPs with a protective layer that guarantees the robustness of the conjugate. Notwithstanding, antibodies freely adsorb to GNP with high binding efficiency. The nonspecific adsorption is far more simple, fast, and inexpensive than any mediated coupling. Therefore, it is preferred for most applications, although it is considered to produce GNP-Ab with a limited activity. In this work, we compared three strategies for producing GNP-Ab, such as (i) covalent coupling mediated by a chemical layer, (ii) affinity-based binding mediated by a biomolecular layer composed of Staphylococcal protein A, and (iii) direct attachment via adsorption. The so-prepared GNP-Ab were employed as probes in a colorimetric lateral flow immunoassay (LFIA) for measuring salivary cortisol as a model biosensor that relies on the use of active GNP-Ab conjugates. Unexpectedly, the biosensors fabricated using the three probes were completely comparable in terms of their ability to measure salivary cortisol. Furthermore, we observed that the sensitivity of the LFIA primarily depended on the amount of the antibody bound to GNPs rather than on the method by which it was bound. The probes prepared using both the direct adsorption approach and mediated coupling via the biochemical mediator enabled development of point-of-care devices for the fast, sensitive, and reliable measurement of human salivary cortisol.

The development of reliable molecularly imprinted sorbent assays is ensured by in-depth knowledge of the binding between the tracer, conventionally based on a template analogue conjugated to an enzyme, and the imprinted polymer used as a recognition element. To this end, the binding properties of a cortisol-3-(O-carboxymethyl)oxime-horseradish peroxidase conjugate to cortisol-imprinted microparticles previously adsorbed at the bottom of microplates were assessed. The effect of different blocking agents as well as of different percentages of Tween 20 in the working buffer were investigated in order to minimise the non-specific binding of the enzyme tracer to the adsorbed imprinted microparticles. The capability of the enzyme tracer to bind the imprinted solid phase and to compete with free cortisol was assessed by measuring the apparent equilibrium dissociation constant, KD(39.7 ± 13.5 pmol L−1), and the apparent binding site concentration, Bmax(21.7 ± 4.3 nmol L−1), whereas the IC50value for the cortisol competition curve was found to be 5.32 ± 1.15 ng mL−1. Moreover, binding selectivity measured for several cortisol-related steroids confirmed the experimental results previously published for cortisol-imprinted polymers. A competitive assay for the determination of cortisol in human saliva was developed with a limit of detection of 1.02 ng mL−1, providing quantitative results comparable to those of a commercial ELISA.

We compared 3 major cross-match (XM) tests to identify dog erythrocyte antigen (DEA) 7 blood incompatibilities in dogs as a result of anti–DEA 7 antibodies: gel (GEL), standard tube (TUBE) agglutination, and immunochromatography strips (STRIP). Blood samples from 42 dogs were typed for DEA 7; 2 tested DEA 7–positive (DEA 7+). The 40 DEA 7–negative (DEA 7–) plasma samples were cross-matched against the 2 DEA 7+ and 3 DEA 7– red blood cell (RBC) samples by GEL to identify samples with anti–DEA 7 antibodies. Twenty DEA 7– plasma samples without and with anti–DEA 7 antibodies were cross-matched with samples of the 2 DEA 7+ RBCs in a double-blind fashion using the TUBE and STRIP XM methods. GEL results were used as the reference method for comparison. To determine relationships between results, 2 × 2 tables were used. Cohen kappa coefficient (κ) was calculated between results of GEL and the other 2 methods. With GEL, 21 of 40 XM tests were positive and 19 of 40 negative for anti–DEA 7 antibodies. The same results were obtained by TUBE, whereas only 1 of 40 XM tests was positive by STRIP. There was a statistically significant relationship between results of GEL and TUBE (p< 0.000) with perfect agreement (κ = 1.000), but not between GEL and STRIP results (p= 1.000) in which agreement was equivalent to chance (κ = 0.0453). The GEL and TUBE XM tests, but not STRIP, are useful methods for identification of DEA 7 incompatibilities caused by anti–DEA 7 antibodies.

Improvement in morbidity of pancreatoduodenectomy (PD) largely depends on the reduction in the incidence of clinically relevant (CR) postoperative pancreatic fistula (POPF). After internal validation of the clinical risk score (CRS) of POPF, and identification of other predictive factors for POPF, robotic (RPD), and open (OPD) PDs were stratified into risk categories and matched by propensity scores. The primary endpoint of this study was incidence of CR-POPF. Secondary endpoints were 90-day morbidity and mortality, and sample size calculation for randomized controlled trials (RCT). No patient undergoing RPD was classified at negligible risk for POPF, and no CR-POPF occurred in 7 RPD at low risk. The matching process identified 48 and 11 pairs at intermediate and high risk for POPF, respectively. In the intermediate-risk group, RPD was associated with higher rates of CR-POPF (31.3% vs 12.5%) (p= 0.0026), with equivalent incidence of grade C POPF. In the high-risk group, CR-POPF occurred frequently, but in similar percentages, after either procedures. Starting from an unadjusted point estimate of the effect size of 1.71 (0.91–3.21), the pair-matched odds ratio for CR-POPF after RPD was 2.80 (1.01–7.78) for the intermediate-risk group, and 0.20 (0.01–4.17) for the high-risk group. Overall morbidity and mortality were equivalent in matched study groups. Sample size calculation for a non-inferiority RCT demonstrated that a total of 31,669 PDs would be required to randomize 682 patients at intermediate risk and 1852 patients at high risk. In patients at intermediate risk, RPD is associated with higher rates of CR-POPF. Incidence of grade C POPF is similar in RPD and OPD, making overall morbidity and mortality also equivalent. A RCT, with risk stratification for POPF, would require an enormous number of patients. Implementation of an international registry could be the next step in the assessment of RPD.

In humans, blood groups are associated with varying prevalence of infections. The aim of this study was to determine if associations exist between the feline AB blood group system and haemoplasma infection. Data from two studies were combined. In the first study, DNA samples from 131 haemoplasma‐infected and 132 haemoplasma‐uninfected UK cats underwent pyrosequencing to determine their blood genotype as AA, Abor bb. In the second study, blood samples from 160 Italian cats of known blood phenotype A, B or AB underwent PCR testing for feline haemoplasma species DNA. Haemoplasma infection was demonstrated in cats of all phenotypes and genotypes. A significantly higher number of Abgenotype cats tested positive for overall haemoplasma infection status (p= 0.04) and for Mycoplasma haemofelisinfection (p= 0.03). Haemoplasma‐infected Italian cats were few, possibly increasing the chance of type II error, and the presence of purebred cats in the sample population may have had a confounding effect. Feline haemoplasmas do not appear to preferentially use either blood type A or B antigens as attachment sites for erythrocyte colonisation. Further investigations in a larger number of haemoplasma‐infected cats of known blood phenotype are warranted to explain the association between genotype Aband haemoplasma infection.

The reported prevalence of naturally occurring anti–dog erythrocyte antigen (DEA) 7 antibodies in DEA 7–negative dogs is as high as 50%. Characterization of these antibodies may better define their importance in canine transfusion medicine. We determined in vitro activity, specificity, and titer of anti–DEA 7 antibodies in DEA 7–negative dogs. Plasma samples from 317 DEA 7–negative dogs were cross-matched with DEA 7–positive red blood cells (RBCs) using gel column technology. Agglutination occurred with DEA 7–positive RBCs but not with DEA 7–negative RBCs in 73 samples (23%), which were hence classified as containing anti–DEA 7 antibodies. These samples were evaluated for hemolytic and agglutinating activity, strength of agglutination, and antibody specificity and titers. All samples showed agglutination but none showed hemolysis. Gel agglutination was graded as 1+ for 20 samples (27%), 2+ for 49 samples (67%), 3+ for 4 samples (6%); no samples were graded 4+. The agglutination titer was <1:2 for 51 samples (73%), 1:2 for 13 samples (19%), 1:4 for 4 samples (5%), and 1:8 for 2 samples (3%). Of 16 samples treated with 2-mercaptoethanol, 11 samples (69%) contained only IgM, 4 samples (25%) exhibited only IgG activity, and 1 sample (6%) had both IgG and IgM activity. Low titers of warm, weakly agglutinating, mostly naturally occurring IgM anti–DEA 7 antibodies were found in 23% of DEA 7–negative dogs. The presence of naturally occurring anti–DEA 7 antibodies suggests that cross-matching of canine blood recipients is advisable, even at first transfusion, to minimize delayed transfusion reactions.

Current selection methods for neurosurgical residents fail to include objective measurements of bimanual psychomotor performance. Advancements in computer-based simulation provide opportunities to assess cognitive and psychomotor skills in surgically naive populations during complex simulated neurosurgical tasks in risk-free environments. This pilot study was designed to answer 3 questions: (1) What are the differences in bimanual psychomotor performance among neurosurgical residency applicants using NeuroTouch? (2) Are there exceptionally skilled medical students in the applicant cohort? and (3) Is there an influence of previous surgical exposure on surgical performance?

The article offers the author's insights on the impact of the political situation in Crimea, Ukraine and the involvement of Russia in the country on the foreign economic relations of China and Ukraine.

Galgos (Spanish Greyhounds), in common with other sighthounds, have higher hematocrits, hemoglobin concentrations, and red blood cell counts than other breeds. In addition to these hematological characteristics, the physical characteristics of these dogs (medium to large dogs with an easily accessible jugular vein and a good temperament) make galgos ideal blood donors. However, to date, there are only published reports concerning dog erythrocyte antigen (DEA) 1 in this breed. Information on DEAs 4 and 7 would be useful when recruiting blood donors to donation programs, as DEA 1 and 7–negative and DEA 4–positive dogs can be considered universal donors. Ethylenediamine tetra-acetic acid–anticoagulated jugular blood samples were collected from 205 galgos. Dogs were aged between 1 and 10 years, 102 were female (49.8%) and 103 male (50.2%), and all were living in South Madrid, Spain. All 205 blood samples were tested for DEA 1 by card agglutination, and 150 of these samples were tested for DEA 4 and DEA 7 by gel column agglutination using polyclonal anti-DEA antibodies. Of the 205 galgos blood samples typed, 112 out of 205 (54.6%) were positive for DEA 1. Of the 150 blood samples tested, all (150/150, 100%) were positive for DEA 4, and 12 out of 150 (8%) samples tested positive for DEA 7. Of these samples, 70 out of 150 (46.7%) were positive only for DEA 4. There was no relationship between blood types and sex. In addition to the hematological characteristics previously reported and the physical characteristics of these dogs, the relative prevalence of blood types DEA 1, 4, and 7 make galgos good candidates for blood donation in blood donor programs.

Objective and Design.Legionellacontrol still remains a critical issue in healthcare settings where the preferred approach to health risk assessment and management is to develop a water safety plan. We report the experience of a university hospital, where a water safety plan has been applied since 2002, and the results obtained with the application of different methods for disinfecting hot water distribution systems in order to provide guidance for the management of water risk.Interventions.The disinfection procedures included continuous chlorination with chlorine dioxide (0.4–0.6 mg/L in recirculation loops) reinforced by endpoint filtration in critical areas and a water treatment based on monochloramine (2-3 mg/L). Real-time polymerase chain reaction and a new immunoseparation and adenosine triphosphate bioluminescence analysis were applied in environmental monitoring.Results.After 9 years, the integrated disinfection-filtration strategy significantly reduced positive sites by 55% and the mean count by 78% (P< .05); however, the high costs and the occurrence of a chlorine-tolerant clone belonging to Legionella pneumophilaST269 prompted us to test a new disinfectant. The shift to monochloramine allowed us to eliminate planktonic Legionellaand did not require additional endpoint filtration; however, nontuberculous mycobacteria were isolated more frequently as long as the monochloramine concentration was 2 mg/L; their cultivability was never regained by increasing the concentration up to 3 mg/L.Conclusions.Any disinfection method needs to be adjusted/fine-tuned in individual hospitals in order to maintain satisfactory results over time, and only a locally adapted evidence-based approach allows assessment of the efficacy and disadvantages of the control measures.

Objective.Our aim was to evaluate the impact of aerators on water microbiological contamination in at-risk hospital departments, with a view to quantifying the possible risk of patient exposure to waterborne microorganisms.Design.We analyzed the microbiological and chemical-physical characteristics of hot and cold water in some critical hospital departments.Setting.Two hospitals in northern Italy.Methods.We took 304 water samples over a 1-year period, at 3-month intervals, from taps used by healthcare personnel for handwashing, surgical washing, and the washing of medical equipment. We analyzed heterotrophic plate counts (HPCs) at 36°C and 22°C, nonfastidious gram-negative bacteria (GNB-NE), and Legionella pneumophila.Results.The percentages of positivity and mean values of HPCs at 22°C, HPCs at 36°C, and GNB-NE loads were significantly higher at outlet points than in the plumbing system. In particular, GNB-NE positivity was higher at outlet points than in the plumbing system in both the cold water (31.58% vs 6.58% of samples were positive) and hot water (21.05% vs 3.95%) supplies. Our results also revealed contamination by L. pneumophilaboth in the plumbing system and at outlet points, with percentages of positive samples varying according to the serogroup examined (serogroups 1 and 2-14). The mean concentrations displayed statistically significant (P< .001) differences between the outlet points (27,382.89 ± 42,245.33 colony-forming units [cfu]/L) and the plumbing system (19,461.84 ± 29,982.11 cfu/L).Conclusions.These results reveal a high level of contamination of aerators by various species of gram-negative opportunists that are potentially very dangerous for immunocompromised patients and, therefore, the need to improve the management of these devices.

We applied a direct competitive immunoassay for measuring corticosterone and glucocorticoid metabolites in faeces (FGMs) as a non-invasive tool for monitoring the stress response of African penguins (Spheniscus demersus) held in captivity in a zoological facility. The developed assay, validated in-house, proved to be rapid (the test could be completed in 90 minutes), sensitive (LOD for corticosterone 0.2 μg l−1, dynamic range 0.75–75 μg l−1) and broad-selective, as it cross-reacted with the major corticosteroids, thus allowing the detection of excreted FGMs resulting from a biological stressor. Matrix interference, due to components of faecal samples, was overcome by diluting sample extracts (1 + 4 or 1 + 9, depending on the sample). The assay enabled us to investigate the response to stress in five animals – three adult males and two adult females – over a period of 30 hours, and to identify the peak of FGM production as being 7–10 hours after the stressful event.

OBJECTIVE: To assess the occurrence of adverse events in a cohort of patients with polymyalgia rheumatica (PMR), treated with low-dose glucocorticoids (GC). METHODS: This was a retrospective study by review of medical records. RESULTS: We identified 222 patients who had a mean duration of followup of 60 ± 22 months and a mean duration of GC therapy of 46 ± 22 months. We found that 95 patients (43%) had at least 1 adverse event after a mean duration of GC therapy of 31 ± 22 months and a mean cumulative dose of 3.4 ± 2.4 g. In particular, 55 developed osteoporosis, 31 had fragility fractures; 27 developed arterial hypertension; 11 diabetes mellitus; 9 acute myocardial infarction; 3 stroke; and 2 peripheral arterial disease. Univariate analysis showed that the duration of GC treatment was significantly associated with osteoporosis (p < 0.0001), fragility fractures (p < 0.0001), arterial hypertension (p < 0.005), and acute myocardial infarction (p < 0.05). Cumulative GC dose was significantly associated with osteoporosis (p < 0.0001), fragility fractures (p < 0.0001), and arterial hypertension (p < 0.01). The adverse events occurred more frequently after 2 years of treatment. Multivariate analysis showed that GC duration was significantly associated with osteoporosis (adjusted OR 1.02, 95% CI 1.02–1.05) and arterial hypertension (adjusted OR 1.03, 95% CI 1.01–1.06); GC cumulative dose was significantly associated with fragility fractures (adjusted OR 1.4, 95% CI 1.03–1.8). CONCLUSION: Longterm, low-dose GC treatment of PMR is associated with serious adverse events such as osteoporosis, fractures, and arterial hypertension; these adverse events occur mostly after 2 years of treatment.

Plasma samples from human cord blood, and fetuses, newborns, and adults of different mammalians species were analyzed by gel-filtration chromatography, to ascertain whether gamma-glutamyltransferase (GGT) fractions reflect liver maturation. Human cord blood plasma showed higher b-, m-, and s-GGT fraction as compared to adult women. In rat and mouse fetuses and in newborns, b-GGT was the most abundant fraction. As in adult humans, in adult rats, mice, rabbits, sheep, and mini pigs, f-GGT was the most abundant fraction. GGT fractions are a common feature of all mammalian species tested. Their pattern changes seem to reflect liver postnatal maturation, function.

The paper analyses the problem related to dispersion and substantial unconsciousness of the knowledge necessary for actions and decisions to coordinate within the economy at best. By the approach of the Austrian school of economics insights on various features of economic activity are exposed, which show the opportunity to realise a knowledge economy within the framework of a new free market experiment; as a consequence, governments must focus on how to improve the diffusion of existing knowledge instead of achieving their preferred industrial structure. Developments of information and communication technologies in emerging economies, especially the African ones, are here exposed as the perfect example of how some infrastructural deficiencies can be overcome by information and communication technologies; the current African renaissance therefore offers a real chance to implement knowledge economy prescriptions.

Thyroid surgery during the last century was characterized by a development of Kocher’s concepts: through a relentless work of surgeons from all over the world thyroidectomy reached a standard of quality in terms of overall results which was unimaginable in the first half of the XX century.The flattering data collected in the literature until the 90’s were all concordant in assuming that there would be little space for a real improvement in the quality standard of thyroid surgery. The introduction of laparoscopic surgery, though, changed very quickly the attitude of surgeons towards their operative behavior and countless new mini-invasive techniques were soon proposed for almost any field of surgery. In 1994, Gagner published the first series of laparoscopic adrenalectomies. Soon after, parathyroid adenomas seemed to offer an ideal field of application of these new surgical concepts. The first report of an endoscopic parathyroidectomy was in 1996. One year later other videoscopic procedures were described whose results seemed quite encouraging so as to push surgeons to try the same access and the same technique also for operations on thyroid. During the following decade several endoscopic or video-assisted approaches were proposed for the removal of thyroid gland. This paper aims to evaluate the results of minimally invasive thyroid and parathyroid surgery through an extensive review of the literature, in particular as far as minimally invasive video-assisted thyroidectomy is concerned.

Background:The incidence and costs of acute myocardial infarction (AMI) in Europe represent a substantial problem due to population aging. Methods:Between 2001 and 2005, Italian hospitalization records were examined to evaluate hospital admissions and costs of AMI in adults aged ≥45 and in elderly people ≥65 or ≥75. Hospital costs were calculated on the basis of Diagnosis Related Groups (DRGs). Results:75,586 men and 43,164 women were hospitalized because of AMI in 2005, showing respectively increases of 17.2% and 29.2% across five years. In the youngest age group (45–64), 29,925 hospitalizations in men and 6443 in women due to AMI were registered during 2005. In the subgroup of patients aged 65–74, 21,621 men and 10,145 women were hospitalized for AMI; in the oldest group (≥75) 24,040 and 26,576 hospitalizations were recorded. The increasing rates across the five examined years were 8.3% and 22.0% in the first age group, 14.3% and 17.4% in people aged 65–74, and 31.8% and 36.3% in the oldest subgroup, respectively in men and women. Among AMI patients aged ≥75, the number of women was always higher than that of men. Overall hospitalization costs due to AMI in Italy were 305 million Euros in 2001 and 370 million in 2005, with an average cost of 3115 Euros per patient in the latter year. Conclusion:Our findings confirm AMI as a leading health problem and a leading cause of health care costs.

CONTEXT: Evaluation of the degree of neoplastic infiltration beyond the thyroid capsule remains a unique parameter that can be evaluated by histopathological examination to label a papillary thyroid carcinoma (PTC) of 20 mm or less in size as a pT1 or pT3 tumor. OBJECTIVE: We correlated the BRAF V600E mutation with both clinical-pathological features and the degree of neoplastic infiltration to redefine the reliability of the actual system of risk stratification in a large selected group of PTCs smaller than 20 mm. DESIGN: The presence of BRAF mutations was examined in 1060 PTCs less than 20 mm divided into four degrees of neoplastic infiltration: 1) totally encapsulated; 2) not encapsulated without thyroid capsule invasion; 3) thyroid capsule invasion; and 4) extrathyroidal extension. RESULTS: The overall frequency of the BRAF V600E mutation was 44.6%. In both univariate and multivariate analyses, BRAF mutations showed a strong association with PTC variants (classical and tall cell), tumor size (11–20 mm), multifocality, absence of tumor capsule, extrathyroidal extension, lymph node metastasis, higher American Joint Commission on Cancer stage, and younger patient age. In PTCs staged as pT1 with thyroid capsule invasion, the frequency of BRAF mutations was significantly higher than in pT1 tumors that did not invade the thyroid capsule (67.3 vs. 31.8%, respectively; P < 0.0001). No statistically significant difference in BRAF alterations was found between pT1 tumors with thyroid capsule invasion and pT3 tumors (67.3 and 67.5%, respectively). CONCLUSION: We suggest that evaluation of BRAF status would be useful even in pT1 tumors to improve risk stratification and patient management, although follow-up data are necessary to confirm our speculations.

A cross‐linked methacrylate molecularly imprinted polymer (poly‐4‐vinylpyridine‐co‐trimethylolpropane‐trimethacrylate) selective for bisphenol A (BPA) was synthesized, using a fluorinated BPA derivative (4,4′‐(hexafluoroisopropylidene)‐diphenol) as a mimic template, and applied to the analysis of real‐world samples of process and potable waters. The molecularly imprinted polymer also showed a high affinity and selectivity for 17‐β‐estradiol and ethynylestradiol. A method to analyze BPA, 17‐β‐estradiol, and ethynylestradiol at ultratrace levels was thus developed from a screening procedure to monitor endocrine‐disrupting chemicals in water samples. The method consists of the BPA‐selective cleanup by molecularly imprinted SPE using cartridges packed with the polymer developed, its recovery by stir bar sorptive extraction after ad hoc derivatization to obtain the corresponding BPA‐acetate, and its analysis by GC‐time window‐SIM‐MS after online thermal desorption. The method showed good linearity in the working range (R2=0.9969), high repeatability (RSD% <10.1), recoveries always above 90%, and very low LOD (10 pg/L) and LOQ (1 ng/L) and can easily be extended to the determination of 17‐β‐estradiol and ethynylestradiol ultratraces. The method's effectiveness was evaluated by analyzing the real‐world water samples; it enabled preconcentration and detection of BPA at ultratrace levels.

Mycotoxins are toxic compounds produced by various moulds, which can affect a variety of crops. Due to their high toxicity and wide diffusion, mycotoxins constitute a severe risk for human health, therefore maximum tolerance levels in food and feed products have been set up all over the world and analytical controls are mandatory for many commodities. Despite validated analytical methods for assessing mycotoxin contamination already existing, a number of papers describing new methods of extraction, identification or measurement appear daily in literature. Nevertheless, the extraction and determination of such contaminants in food and feed is a topic of constant and increasing interest, as also attested by the number of related patents which have been applied for in the last few years. Scientific papers dealing with mycotoxin occurrence, potential risk and determination have recently been reviewed. Nevertheless, the objective of this review is to focus on patent activity rather than on the scientific breakthroughs on this subject. Therefore, the most recent patents regarding the whole analytical protocol to measure mycotoxins in food, starting from 2006 to date are presented and discussed. The possibility of a technology transfer for the various innovations presented is also discussed as are future developments in the field.

Serum γ-glutamyltransferase (GGT) is thought to derive from the liver, but its values predict morbidity and mortality for several diseases, such as cardiac infarction, stroke, diabetes, renal failure and cancer. We assessed total GGT and its fractions in the culture supernatants of human cell lines (melanoma, prostate cancer, bronchial epithelium) by gel filtration chromatography. We also compared the GGT elution profile in plasma and the corresponding very-low-density lipoprotein (VLDL) fraction. All the cell lines tested released soluble GGT whose activity increased in parallel with the cell growth. Released GGT presented a molecular weight of 2000 kDa, identical to the b-GGT fraction of human plasma and corresponding to that of VLDL. But ultracentrifugation studies showed that b-GGT had a higher density than VLDL. The b-GGT present in human plasma can be produced by tissues other than the liver, thus explaining the increase of serum GGT observed in diseases of other organs.

A method for molecularly imprinted SPE of banned Sudan azo‐dyes from food samples was investigated. The molecularly imprinted polymer was obtained by suspension polymerization using 1‐(4‐chlorophenyl)azonaphthalen‐2‐ol as the mimic template. The molecular recognition properties of imprinted beads were evaluated for use as a SPE sorbent, in order to develop a selective extraction protocol for the Sudan class of dyes. The optimized extraction protocol resulted in a reliable molecularly imprinted SPE (MISPE) method suitable for HPLC analysis. It was selective for the main analyte, Sudan I, and the related azo‐dyes Sudan II, III, IV, Sudan Red B, and Sudan Red 7B, while the permitted azo‐dyes Allura Red AC, Neococcin, and Sunset Yellow FCF were not extracted. The method was tested for Sudan I, II, III, and IV in five different food samples (hot chilli pepper, hot chilli tomato sauce, sausage, tomato sauce, and hard boiled egg yolk) at three concentration levels (15, 100, and 300 μg/g). It demonstrated itself to be insensitive to the presence of different complex matrices, precise, accurate, and with good recovery rates (85–101%). The LOD and LOQ were satisfactory for most analytical determinations.

Molecular imprinted polymers are man-made materials with predetermined selectivity towards one or more analytes. They have been used as selective stationary phases in several analytical separative techniques. This review discusses the possible uses of imprinted polymers, with a particular emphasis on their use as high pressure and electrochromatographic stationary phases for resolution of optical enantiomers, materials for fast screening of libraries and selective solid-phase extraction for sample clean-up and preconcentration.

Oral absorption of the antihyperglycaemic agent metformin hydrochloride (MF-HCl) is confined to the upper part of the intestine, therefore rational controlled-release formulations of this drug should ensure a complete release during transit from stomach to jejunum. The aim of this study was the preparation of a system able to sustain release of high MF-HCl doses in compliance with the above requirement. Matrices (6 mm diameter; 50 mg weight) comprising varying drug-Precirol ATO 5 ratios were prepared by compression. The matrix containing 70% drug was coated on one face with Eudragit L100–55. Drug release to simulated gastric (SGF), jejunal (SJF) and ileal (SIF) fluids in sequence was studied using a modified USP rotating basket method. Release depended on drug load whereas it was independent of dissolution medium pH and hydrodynamics. Release kinetics were of √t type and were determined by drug diffusion in aqueous pores created in the matrix by drug dissolution. An equation correlating rate-determining factors was developed, whereby the release pattern could be optimized. The half-coated matrix started release in SGF and completed it in SJF. The half-coated matrix, synchronizing drug release and matrix transit across the small intestine, may improve drug bioavailability and reduce side effects.

Oral absorption of the antihyperglycaemic agent metformin hydrochloride (MF‐HCl) is confined to the upper part of the intestine, therefore rational controlled‐release formulations of this drug should ensure a complete release during transit from stomach to jejunum. The aim of this study was the preparation of a system able to sustain release of high MF‐HCl doses in compliance with the above requirement. Matrices (6 mm diameter; 50 mg weight) comprising varying drug‐Precirol ATO 5 ratios were prepared by compression. The matrix containing 70% drug was coated on one face with Eudragit L100–55. Drug release to simulated gastric (SGF), jejunal (SJF) and ileal (SIF) fluids in sequence was studied using a modified USP rotating basket method. Release depended on drug load whereas it was independent of dissolution medium pH and hydrodynamics. Release kinetics were of √t type and were determined by drug diffusion in aqueous pores created in the matrix by drug dissolution. An equation correlating rate‐determining factors was developed, whereby the release pattern could be optimized. The half‐coated matrix started release in SGF and completed it in SJF. The half‐coated matrix, synchronizing drug release and matrix transit across the small intestine, may improve drug bioavailability and reduce side effects.

A molecularly imprinted polymer which recognises the mycotoxin ochratoxin A was prepared using the mimic N-(4-chloro-1-hydroxy-2-naphthoylamido)-(L) -phenylalanine as a template. The polymer was obtained by dissolving the template, methacrylic acid and ethylendimethacrylate in chloroform and polymerising the mixture by thermal treatment at 60 °C. The monolith obtained was crushed, sieved to 30–90 μm and extensively washed till the template could no longer be found in the washing solution. The binding properties towards the template, ochratoxin A and several related molecules were measured by eluting with acetonitrile and chloroform a HPLC column packed with the imprinted polymer. The experimental results show that the polymer recognises not only the template well, but also the ochratoxin A. The specific molecular recognition effect is due to hydrogen bond interactions but in order to assure the full recognition effect adjunctive steric factors are necessary. The magnitude of these interactions can be controlled by the use of limited amounts of acetic acid in the mobile phase. From the measurement of the relative selectivity it was found that only the simultaneous presence of the carboxyl, the phenolic hydroxyl and certain peculiar substructures such as the chlorine atom assures the whole recognition of the template. [ABSTRACT FROM AUTHOR]

The binding and selectivity features of an immobilised anti-progesterone antiserum were studied by the use of four different enzyme tracers: progesterone 11α-hemisuccinate–horseradish peroxidase (P-11α-HS–HRP), progesterone 11α-carboxymethyl ether–horseradish peroxidase (P-11α-CME–HRP), progesterone 11β-carboxymethyl ether–horseradish peroxidase (P-11β-CME–HRP) and progesterone 3-(O-carboxymethyl)oxime–horseradish peroxidase (P-3-CMO–HRP). The antiserum–tracer affinities generally showed a remarkable reduction in respect to the affinity of the analyte because of the steric hindrance of the enzyme and, among the four tracers, the higher affinity value was evaluated for the P-11α-HS–HRP (homologous to the immunogen molecule). The concentration of antibody binding sites interacting with the tracers showed the presence of different classes of antibodies able to react with variable affinity with tracers and analyte, as confirmed by the cross-reactivity values measured towards different progesterone derivatives. The assays performed with the tracers showed that an increase of sensitivity can be obtained using enzyme tracers provided with heterologous structure features with respect to the immunogen molecule.

We measured growth hormone (GH), insulin-like growth factor-I (IGF-I), and both total and glycosylated prolactin (PRL) levels in 131 blood samples obtained by cordocentesis in normal and abnormal fetuses from 19 to 40 weeks of gestation. In normal fetuses, IGF-I and PRL levels showed a positive correlation and GH a negative correlation with gestational age. A negative relation between GH and IGF-I levels was observed, while PRL did not show any correlation with both GH and IGF-I concentrations. IGF-I increased from 5.6±3 (at 19–22 weeks) to 10.7±5 nmol/l at term; GH decreased from 31±10 to 7.7±4 μg/l and PRL increased from 16±18 to 139±76 μg/l. Glycosylated PRL accounted for about 15% of total PRL, a value similar to that found in normal adults. In 27 fetuses of 27–37 weeks with intra-uterine growth retardation, GH and PRL levels were higher and IGF-I levels lower than in normal fetuses matched for week of gestation. In 8 anencephalic fetuses of 19–26 weeks of gestation, both GH and IGF-I levels were lower, and PRL levels were higher than in matched controls. Altogether these data support the views that a) both GH and PRL secretion are under the hypothalamic control during fetal development, b) the serum GH decrease from midgestation to the end of pregnancy is mediated by the negative feed-back mechanism of increasing IGF-I levels and c) IGF-I production is mainly regulated by fuel supply and only partially by GH.