Broad clonal heterogeneity of antigen-specific CD4+ T-cells localizing at the site of disease during tuberculosis.

The repertoire of CD4+ T-lymphocytes was investigated in six patients affected by tuberculosis, who had a negative PPD skin test at diagnosis. Polyclonal CD4+ T-cell lines from the peripheral blood failed to proliferate to PPD and to the 16- or 38-kDa proteins of Mycobacterium tuberculosis, while CD4+ T-cell lines from the site of disease responded to PPD, and to the 16- and 38-kDa proteins, and derived epitopes in vitro. The repertoire of CD4+ T-cells accumulating at the site of disease was found to be widely heterogeneous as demonstrated by the finding that at least seven different peptides from the 16- and 38-kDa proteins were recognized by every patient. These results indicate that CD4+ T-cells localized at the site of disease in tuberculosis recognize a vast array of M. tuberculosis epitopes.     

Efficacy of Hilotherapy face mask in improving the trend of edema after orthognathic surgery: a 3D analysis of the face using a facial scan app for iPhone

Purpose

Cryotherapy after orthognathic surgery is essential for the control of facial edema. The aim of our study is to evaluate the efficacy of Hilotherapy face mask in reducing facial edema after orthognathic surgery, studying facial surfaces with an innovative, fast, economical 3D facial scan system based on an iPhone app.

Methods

Eighty-four patients with Class III were included: 35 patients treated with Hilotherm after orthognathic surgery (Group 1), 32 patients with ice packs (Group 2), 7 patients who refused cryotherapy (not 1 - not Group 2). Their facial scans performed immediately after surgery (T0), at 24 (T1), 48 (T2) and 72 h (T3) after surgery, were acquired in specific software, and the discrepancies between them were studied in an accurate 3D volumetric method.

Results

We measured a significantly better edema trend in Group 1 in the tragus–nasal wing line and in the tragus–labial commissure line at T1, and also in the tragus–menton line at T2 and T3.

Conclusions

In conclusion, Hilotherapy represents a more comfortable and more effective cryotherapy system in controlling the trend of facial edema after orthognathic surgery. The method we used for the facial scans is accurate, cheap, smart, and fast. As demonstrated by the 3D volumetric study of the face, the regions of the middle third of the face are those in which the difference is most noticeable.

     

Laboratory diagnosis of boutonneuse fever by enzyme-linked immunosorbent assay

Little is known about the use of the enzyme-linked immunosorbent assay (ELISA) in the diagnosis of boutonneuse fever, and the reports which do exist have concerned the use of a strain of African origin and a peroxidase-labelled conjugate. We have used, as antigen, a Sicilian strain of Rickettsia conorii recently isolated from a patient with boutonneuse fever and anti-human immunoglobulins G and M labelled with alkaline phosphatase as conjugate. 432 (84.6%) of 500 sera from patients with boutonneuse fever, 4.4% of 384 sera from patients with various other pathologies, and 2.9% of 204 sera from blood donors gave positive reactions. The ELISA was highly sensitive and specific in the diagnosis of boutonneuse fever, and seems to be the test of choice for sero-epidemiological investigations, and when a large number of specimens must be examined. The test is more sensitive than the indirect immunofluorescence test, at least in the early stage of the disease (44.9% vs 21.3% positive respectively), and a long time after an acute attack.     

N-valproyl-L-tryptophan for CNS-targeting: synthesis, characterization and efficacy in vitro studies of a new potential antiepileptic drug

A new aminoacidic derivative of valproic acid (VPA) has been synthesized and characterized by analytical and spectral data. The rationale for the preparation of such potential antiepileptic agent is based on the observation that chemical combination of the anticonvulsant pharmacophore, VPA with essential aminoacids could afford more effective and less toxic actives. The synthesis, characterization, physico-chemical parameters functional for crossing Blood Brain Barrier of N-valproyl-L-tryptophan (4) are reported. The Log D (pH7.4) (0.3) indicates that (4) is adequate to cross biological membranes. Its chemical and enzymatic stability were assessed. The experiments indicate high stability of compound (4) at pH conditions of physiological fluids. Moreover, both in plasma and in cerebral enzymatic environments compound (4) doesn't undergo cleavage after 24 h. The anticonvulsant activity of the new compound was assessed against epileptic burst discharges evoked in vitro in rat hippocampal slices (Seizure like events - SLEs) and compared with that of the widely used VPA. Compound (4), even at the lower tested concentration, when compared to VPA, showed an improved protective effect against hippocampal seizures. The collected data suggest that compound (4) could be considered a very valuable candidate for subsequent in vivo evaluation as new potential antiepileptic drug.     

A Novel Oral Fluid Assay (LC-QTOF-MS) for the Detection of Fentanyl and Clandestine Opioids in Oral Fluid After Reported Heroin Overdose

Introduction

The adulteration of heroin with non-pharmaceutical fentanyl and other high-potency opioids is one of the factors contributing to striking increases in overdose deaths. To fully understand the magnitude of this problem, accurate detection methods for fentanyl and other novel opioid adulterant exposures are urgently required. The objective of this work was to compare the detection of fentanyl in oral fluid and urine specimens using liquid chromatography quadrupole time-of-flight mass spectrometry (LC-QTOF-MS) in a population of heroin users presenting to the Emergency Department after overdose.

Methods

This was a prospective observational study of adult Emergency Department patients who presented after a reported heroin overdose requiring naloxone administration. Participants provided paired oral fluid and urine specimens, which were prepared, extracted, and analyzed using a dual LC-QTOF-MS workflow for the identification of traditional and emerging drugs of abuse. Analytical instrumentation included SCIEX TripleTOF® 5600+ and Waters Xevo® G2-S QTOF systems.

Results

Thirty participants (N = 30) were enrolled during the study period. Twenty-nine participants had fentanyl detected in their urine, while 27 had fentanyl identified in their oral fluid (overall agreement 93.3%, positive percent agreement 93.1%). Cohen’s Kappa (k) was calculated and demonstrated moderately, significant agreement (k = 0.47; p value 0.002) in fentanyl detection between oral fluid and urine using this LC-QTOF-MS methodology. Additional novel opioids and metabolites, including norfentanyl, acetylfentanyl, and U-47700, were detected during this study.

Conclusion

In this study of individuals presenting to the ED after reported heroin overdose, a strikingly high proportion had a detectable fentanyl exposure. Using LC-QTOF-MS, the agreement between paired oral fluid and urine testing for fentanyl detection indicates a role for oral fluid testing in surveillance for nonpharmaceutical fentanyl. Additionally, the use of LC-QTOF-MS allowed for the detection of other clandestine opioids (acetylfentanyl and U-47700) in oral fluid.

     

Long-term pertussis-specific immune responses to a combined diphtheria,tetanus, tricomponent acellular pertussis and hepatitis B vaccine in pre-term infants

Immunoglobulin G (IgG) antibody titres against pertussis antigens, Bordetella pertussis-specific proliferation and cytokine production by peripheral blood mononuclear cells (PBMCs) were evaluated at the age of 5-6 years in 13 children who had been pre-term infants with a gestational age (GA) of < or = 31 weeks, 10 who had been pre-term infants with a GA of 32-37 weeks, and 15 who had been term infants with a GA of 38-42 weeks. All of the infants had been immunised with a combined diphtheria, tetanus, tricomponent acellular pertussis and hepatitis B vaccine (DTaP-HBV) at 3, 5, and 11 months of post-natal age. Our results show that the long-term immune responses induced by primary pertussis vaccination in the pre-term infants (especially those with a GA of < or = 31 weeks) were qualitatively and quantitatively lower than those observed in the term infants. In order to maintain an adequate pertussis-specific immune response, in term children a booster at 5-6 years of age may be suggested, whereas in pre-term an earlier booster should be useful.     

Antimicrobial Susceptibility of Respiratory Isolates of Enterobacteriaceae and Staphylococcus aureus in Italy: Incidence and Trends Over the Period 1997–1999

The antibiotic susceptibility of members of the family Enterobacteriaceae and of Staphylococcus aureus strains isolated from the respiratory tract was assessed over the period 1997–1999 as part of the Italian Epidemiological Observatory survey sponsored by the SmithKline Foundation. A standardised method was used to determine the MICs of 22 antibiotics against isolates of Klebsiella pneumoniae (n=870), Escherichia coli (n=684), Enterobacter cloacae (n=342), Enterobacter aerogenes (n=187) and Serratia marcescens (n=135) as well as the MICs of 11 antibiotics against isolates of Staphylococcus aureus (n=1,606). Overall, the susceptibility rate of Enterobacteriaceae isolates was ≥90% to 5 agents (meropenem, imipenem, amikacin, cefepime and gentamicin); 89–80% to 2 agents (ciprofloxacin and tobramycin); and <80% to 11 agents (cefotaxime, piperacillin-tazobactam, trimethoprim-sulfamethoxazole, cefetamet, ceftriaxone, ceftazidime, aztreonam, ticarcillin-clavulanate, tetracycline, piperacillin, cefuroxime, chloramphenicol, ticarcillin, amoxicillin-clavulanate and amoxicillin). During the 3-year monitoring period, antibiotic susceptibility increased in Klebsiella pneumoniae against amoxicillin-clavulanate, in Escherichia coli against third-generation cephalosporins and aztreonam, in Enterobacter aerogenes against amoxicillin and piperacillin-tazobactam and in Serratia marcescens against most of the antibiotics. In contrast, Enterobacter cloacae showed a tendency to develop resistance to cefetamet, amikacin and ciprofloxacin. Of the total number of Staphylococcus aureus strains, 38% were methicillin resistant. Nearly 80% of the methicillin-resistant strains displayed a multiresistance pattern (additional resistance to 2 or more non-beta-lactam antibiotics). Rates of susceptibility of particular species (Klebsiella pneumoniae, Escherichia coli, Staphylococcus aureus) were compared using strains from different geographical areas of Italy (northern, central and southern) and from different nosocomial areas (outpatients, intensive care unit [ICU] inpatients, non-ICU inpatients). Susceptibility of Klebsiella pneumoniae to several antibiotics was lower in southern Italy, whereas the incidence of methicillin-resistant strains was higher in northern and central Italy. The susceptibility of Escherichia coli was similar in all three areas. No significant differences in susceptibility of Klebsiella pneumoniae or Escherichia coli were found between strains from inpatients and outpatients or from inpatients admitted to ICU and non-ICU units. The incidence of methicillin-resistant Staphylococcus aureus was higher in ICU inpatients (52%) than in non-ICU inpatients (38%) and lower in outpatients (19%) than in inpatients. Electronic Publication     

Sequestration of T lymphocytes to body fluids in tuberculosis: reversal of anergy following chemotherapy.

The specificity of CD4 T lymphocytes was investigated in 6 patients affected by tuberculosis who had negative tuberculin purified protein derivative (PPD) skin tests at diagnosis. Polyclonal CD4 T cell lines from the peripheral blood failed to proliferate to PPD and to the 16- or 38-kDa proteins of Mycobacterium tuberculosis, while CD4 cell lines from the disease site responded to PPD and to the 16- and 38-kDa proteins and derived epitopes in vitro. Four months after chemotherapy, the patients became responsive to PPD. The proliferative response to PPD and to the 16- or 38-kDa proteins and their derived peptides decreased in CD4 T cell lines from the disease site and increased in lines from the peripheral blood. These results indicate that CD4 T cells recognizing a vast array of M. tuberculosis epitopes are compartmentalized at the site of disease in anergic patients but appear in peripheral blood after chemotherapy.     

Cryptomarket drug acquisition leading to furanyl fentanyl overdose

Forensic Toxicology -     

Change of Th0 to Th1 cell-cytokine profile following tuberculosis chemotherapy

T cells mediate protection against tuberculosis, but little is known about their role during chemotherapy of patients with active disease. Here we examined the cytokine profile of CD4 T cells before and after four months of chemotherapy in six initial skin test anergic cases. Purified protein derivative (PPD) and 16-kDa antigen-reactive CD4 T-cell clones prior to therapy resided mostly in disease-associated body fluids and were of the Th0 (interferon (IFN)-γ + interleukin (IL)-4) secreting profile. In contrast, the majority of postchemotherapy CD4 T-cell clones originated from blood and were of the IFN-γ secreting Th1 type. However, the recognition of several peptides derived from the 16-kDa antigen was not significantly different between the Th1 and Th0 clones. We conclude that chemotherapy shifts CD4 T cells from the affected body fluids to the blood circulation, accompanied by a change from Th0 to Th1 cytokine profile.