2013 European guideline on the management of lymphogranuloma venereum.

WHAT IS NEW IN THIS UPDATED GUIDELINE?: This is the update version of the 2010 European guideline on the management of lymphogranuloma venereum (LGV). New issues are: EPIDEMIOLOGY: Based on clonal relatedness of prevalent LGV strains there is evidence that the LGV epidemic among men who have sex with men (MSM) in the Western world prevailed already in the United States in the 1980s and was introduced into Europe by the end of the last century. AETIOLOGY AND TRANSMISSION: A new LGV variant causing severe proctitis was unveiled and designated L2c. The L2b LGV variant causing the vast majority of infections among MSM is now also found among a few heterosexual women. MANAGEMENT: Apart from HIV and STI screening, Hepatitis C Virus (HCV) testing should be offered to all LGV patients. To exclude reinfections, STI screening during a follow-up visit 3 months after an LGV diagnosis should be offered.

Aberrant in vivo T helper type 2 cell response and impaired eosinophil recruitment in CC chemokine receptor 8 knockout mice.

Chemokine receptors transduce signals important for the function and trafficking of leukocytes. Recently, it has been shown that CC chemokine receptor (CCR)8 is selectively expressed by Th2 subsets, but its functional relevance is unclear. To address the biological role of CCR8, we generated CCR8 deficient (-/-) mice. Here we report defective T helper type 2 (Th2) immune responses in vivo in CCR8(-/)- mice in models of Schistosoma mansoni soluble egg antigen (SEA)-induced granuloma formation as well as ovalbumin (OVA)- and cockroach antigen (CRA)-induced allergic airway inflammation. In these mice, the response to SEA, OVA, and CRA showed impaired Th2 cytokine production that was associated with aberrant type 2 inflammation displaying a 50 to 80% reduction in eosinophils. In contrast, a prototypical Th1 immune response, elicited by Mycobacteria bovis purified protein derivative (PPD) was unaffected by CCR8 deficiency. Mechanistic analyses indicated that Th2 cells developed normally and that the reduction in eosinophil recruitment was likely due to systemic reduction in interleukin 5. These results indicate an important role for CCR8 in Th2 functional responses in vivo.

CD69-triggered ERK activation and functions are negatively regulated by CD94 / NKG2-A inhibitory receptor.

CD69 represents a functional triggering molecule on activated NK and T cells, capable of inducing cytotoxic activity and costimulating cytokine production. It belongs to the C-lectin type superfamily, and its gene maps in the NK gene complex, close to other genes coding for NK receptors. CD94 / NKG2-A complex is the inhibitory receptor for the non classical MHC class I molecule HLA-E on human NK cells. To investigate CD69-initiated signal transduction pathways, and to evaluate CD94 / NKG2-A interference on CD69 triggering ability, we have generated transfectants expressing both receptors in the RBL cell line. Here we report that CD69 engagement leads to the activation of extracellular signal-regulated kinase (ERK) enzymes belonging to the MAPK family, and that this event is required for CD69-mediated cell degranulation. Moreover, we show that the co-engagement of CD94 / NKG2-A inhibitory receptor effectively suppresses both CD69-triggered cell degranulation in RBL transfectants, through the inhibition of ERK activation, and CD69-induced cytotoxicity in human NK cells. Thus, here we provide new information on the molecular mechanisms initiated by CD69 activation receptor, and show that CD69-initiated signaling pathways and functional activity are negatively regulated by CD94 / NKG2-A inhibitory complex.

CD94/NKG2-A inhibitory complex blocks CD16-triggered Syk and extracellular regulated kinase activation, leading to cytotoxic function of human NK cells.

The CD94/NKG2-A complex is the inhibitory receptor for the nonclassical MHC class I molecule HLA-E on human NK cells. Here we studied the molecular mechanisms underlying the inhibitory activity of CD94/NKG2-A on NK cell functions by analyzing its interference on CD16-initiated signaling pathways involved in the control of cytolytic activity. Both tyrosine phosphorylation and activation of Syk kinase together with tyrosine phosphorylation of CD16 receptor zeta subunit are markedly inhibited by the coengagement of CD94/NKG2-A complex. As a downstream consequence, CD94/NKG2-A cross-linking impairs the CD16-induced activation of extracellular regulated kinases (ERKs), a pathway involved in NK cytotoxic function. The block of ERK activation is exerted at an early, PTK-dependent stage in the events leading to p21ras activation, as the CD16-induced tyrosine phosphorylation of Shc adaptor protein and the formation of Shc/Grb-2 complex are abrogated by CD94/NKG2-A simultaneous engagement. Our observations indicate that CD94/NKG2-A inhibits the CD16-triggered activation of two signaling pathways involved in the cytotoxic activity of NK cells. They thus provide molecular evidence to explain the inhibitory function of CD94/NKG2-A receptor on NK effector functions.

The chemokine receptor CCR8 is preferentially expressed in Th2 but not Th1 cells.

In this paper we report on the cloning and characterization of mouse CCR8. Like its human homologue, it is predominantly expressed in the thymus. In the periphery, murine CCR8 mRNA was found most abundantly expressed in activated Th2-polarized cells and in NK1.1+ CD4+ T cells. Human CCR8 is also preferentially expressed in human Th2-polarized cells and clones. This pattern of expression suggests that CCR8 is part of a Th2-specific gene expression program. The CCR8 ligands I-309 and its mouse homologue T cell activation gene 3 (TCA-3) are potent chemoattractants for Th2-polarized cells. Taken together, these observations strongly suggest that CCR8 plays a role in the control of Th2 responses, and may represent a potential target for treatment of allergic diseases.

[A cohort study of art glass workers in the Empoli area]. / Studio di coorte dei lavoratori del vetro artistico nel territorio empolese.

The investigation aimed at studying cause-specific mortality of art glass workers employed in 17 industrial facilities in Tuscany, Italy. A cohort of 3390 workers employed for at least 1 year was obtained from company payrolls. Follow-up was between the year each factory started operations, mostly in the mid-fifties, and the end of 1993. The cause specific expected mortality was computed relative to Tuscany rates, specified for gender, 5-year age groups and calendar year. Separate analyses were carried out for the job titles of makers, batch mixers and grinders. For males, 3180 individuals, the observed mortality for cancer causes was above the expected for the lung [standardized mortality ratio (SMR) 123, 10 observed (Obs)], larynx (SMR 166, 10 Obs), stomach (SMR 105, 30 Obs) and brain (SMR 150, 7 Obs). For non-cancer causes observed mortality was above expected for hypertensive diseases (SMR 178, 10 Obs) and diseases of the genitourinary system (SMR 169, 11 Obs). Increases for the above listed causes were shown also among makers. Mortality for larynx and lung cancer increased with time since first exposure and significantly increased SMRs were observed for 21 or more years since first exposure: this pattern was still present with smoking adjustment. The results showed consistently increased mortality for lung and larynx cancer in the overall cohort and among makers. Stomach cancer, brain cancer, hypertensive diseases and diseases of the genitourinary system were also increased in the overall cohort and among makers.

Isolation and chromosomal localization of GPR31, a human gene encoding a putative G protein-coupled receptor.

The screening of a human genomic library with a chemokine receptor-like probe allowed us to obtain a putative member of the G protein-coupled receptor gene (GPCR) family, designated GPR31. Its deduced amino acid sequence encodes a polypeptide of 319 amino acids that shares 25-33% homology with members of the chemokine, purino, and somatostatin receptor gene families. Amino acid sequence comparison reveals that the best match in the protein databases is with the human orphan GPCR called HM74 (33% identity). Southern genomic analysis of the GPR31 gene shows a hybridization pattern consistent with that of a single-copy gene. Using fluorescence in situ hybridization, we have determined the chromosomal and regional localization of the GPR31 gene at 6q27. The GPR31 mRNA is expressed at low levels by several human cell lines of different cellular origins. The phylogenetic analysis suggests that the GPR31 receptor may represent a member of a new GPCR subfamily.

[Radiotherapy of the breast: changes in the volume of the lung covered in the treatment fields and resulting changes in dose distribution]. / Radioterapia della mammella: variazioni del volume del polmone compreso nei campi del trattamento e conseguenti modificazioni della distribuzione della dose.

In breast cancer adjuvant therapy, respiratory movements continuously modify the irradiated volumes and the anatomical shape of this body region. Fifteen patients were submitted to 3 Computed Tomography (CT) sequences for treatment planning: the first one without any indications to the patient (the standard sequence) and the second and the third one with spontaneous stopped inspiration and expiration, respectively; the patient was always in the same position. The treatment was planned on standard CT images and then applied to the other sequences, maintaining all parameters unvaried, including isocenter position and treatment time. The lung volumes within the fields (and those included in the 95%, 100%, 105% isodoses referred to the prescribed dose) were evaluated with dose/volume histograms. The average irradiated lung was 69 cm3 (DS 28) in standard sequences, 136 cm3 (DS 67) in inspiration and 41 cm3 (DS 25) in expiration. The pulmonary volume within the above isodoses exhibited similar changes. In other words, the lung volume actually irradiated during the whole treatment is smaller than the one which can be calculated on standard CT sequences and it corresponds to expiration volume. The remaining part falls into a wide "twilight zone" relative to dose. Therefore, the true risk of lung toxicity can be similarly lower than the calculable one on standard CT images. Thus, the complication risk (based on dose/volume histograms and normal tissue control probability parameters) could be assessed in new prospective studies, introducing a corrective factor for the irradiated lung volume, because the latter is smaller than that shown by standard CT.

Molecular cloning of TER1, a chemokine receptor-like gene expressed by lymphoid tissues.

Several chemokine receptors have been cloned and shown to belong to a superfamily of seven transmembrane, G protein-coupled receptors. We report here the molecular cloning of TER1, a novel human chemokine receptor-like gene. The amino acid sequence deduced from the TER1 cDNA shows 43, 40, 40, and 39% identity to CCR4, CCR5, CCR1, and CCR2B beta chemokine receptors, respectively. By the use of fluorescent in situ hybridization, we have mapped the TER1 gene to chromosome 3p21, clustered with other chemokine receptor genes. By Northern blot analysis, TER1 mRNA is found to be expressed in the thymus, spleen, and at barely detectable levels in peripheral blood lymphocytes. Moreover, TER1 message in abundant in the NK cell line NK3.3 and in the T cell line MOLT-4. The restricted TER1 expression in cells and tissues of the lymphoid lineage suggests that this receptor may play a role in regulating immune functions.

[Volume of the irradiated small intestine during pelvic radiotherapy. Comparison of various calculation methods]. / Il volume dell'intestino tenue irradiato durante la radioterapia pelvica. Confronto tra diverse metodiche di calcolo.

In pelvic irradiation, the small bowel portion included in the planning treatment volume is one of the major factors of acute enteropathy. Three different methods are used to calculate the bowel volume: Gallagher's grid method and two systems based on specific algorithms using CT data. We compared the results of these different methods in a series of nine patients submitted to treatment volume planning simulation for pelvic irradiation, after oral barium administration. The small bowel volumes were calculated with the grid method on orthogonal radiographs. About one hour later, the patients were submitted to CT for radiotherapy planning. The small bowel regions to be irradiated were drawn manually on all CT slices on a Varian Cadplan 2.62 console. Two different algorithms were used to calculate the small bowel volumes: one of them based on polyhedral and the other on cylindric approximation. The average volumes, the variance and the determination coefficient with linear and polynomial regression were in substantial statistical agreement in the three series; the correlation index between the grid and the CT methods ranged 0.84-0.87. Therefore, the authors believe that enteric side-effects can be correlated with the irradiated small bowel volume, independent of the calculation method.

[Assessment of the tumor bed dose in the conservative treatment of breast carcinoma]. / Valutazione della dose al letto tumorale nel trattamento conservativo del carcinoma mammario.

We investigated the variations in the total dose given to primary tumor sites in breast irradiation after conservative surgery. Fifty patients consecutively submitted to CT for radiotherapy treatment planning were entered into this study. Treatment was planned with Varian Cadplan 2.61 for both whole breast irradiation (2 Gy/fraction, up to 50 Gy) and boost (2 Gy/fraction, up to 10 Gy) according to the ICRU 50 report. The doses were calculated localizing the tumor site with preoperative mammography or breast US, surgery description, the possible presence of clips and treatment planning CT. The doses to tumor volumes ranged 46.5 to 53.4 Gy [average: 50.7, standard deviation (SD): 1.47] with the tangential fields. The relative biological effects (RBE) ranged 55.2 to 64.9 Gy (average: 61.05, SD: 2.06). The total doses to tumor beds ranged 57.8 to 65.2 Gy (average: 61.6, SD: 1.63) and the relative RBE from 67.8 Gy to 79.4 Gy (average: 74.3, SD: 2.30). In conclusion, in our opinion, the assessment of radiotherapy efficacy in breast irradiation should be related also to tumor bed dose and not only to the prescribed dose. Indeed, its wide range (and, consequently, the marked differences seen in RBE) might be misleading, especially when the relationship between local relapse and boost usefulness is considered.

[Stage 1 glottic laryngeal carcinoma: observations on 76 cases treated with transcutaneous radiotherapy]. / Il carcinoma della laringe--piano glottico--nel primo stadio: osservazioni su 76 casi trattati con radioterapia transcutanea.

In this paper the authors retrospectively review various parameters which influenced local control and survival in a series of 76 patients with Tis or T1 glottic carcinoma. All the patients (89% smokers) received radical irradiation with two oblique anterior isocentric fields (45-315 degrees), with a Cobalt unit, 5 x 5 or 6 x 6, using compensator wedges; 2 Gy were given per fraction, up to 60-66 Gy at the minimum reference isodose. The mean total dose to ICRU reference point was 68 Gy (range: 63-72 Gy). All patients achieved complete remission. Four patients only had a local recurrence and none had regional relapses. Few late side-effects were observed and among them only two grade I and one grade II, the latter requiring tracheotomy. Eight patients died of cardiovascular diseases and nine of secondary cancer. Two patients are alive with a secondary malignancy. The five-year overall survival rate is 77.5%, while NED survival is 93%; no differences in survival rates were found according to T stage, grading, field size, treatment time and total dose delivered. With reference to the current literature, the authors conclude that: a) radiotherapy permits very good local control; b) lymph nodes should not be irradiated; c) very high doses do not improve local control, but only increase late side-effects; d) the role of biologic dose inhomogeneity and overall treatment time remain to be investigated.

Food allergy and infantile autism.

The etiopathogenesis of infantile autism is still unknown. Recently some authors have suggested that food peptides might be able to determine toxic effects at the level of the central nervous system by interacting with neurotransmitters. In fact a worsening of neurological symptoms has been reported in autistic patients after the consumption of milk and wheat. The aim of the present study has been to verify the efficacy of a cow's milk free diet (or other foods which gave a positive result after a skin test) in 36 autistic patients. We also looked for immunological signs of food allergy in autistic patients on a free choice diet. We noticed a marked improvement in the behavioural symptoms of patients after a period of 8 weeks on an elimination diet and we found high levels of IgA antigen specific antibodies for casein, lactalbumin and beta-lactoglobulin and IgG and IgM for casein. The levels of these antibodies were significantly higher than those of a control group which consisted of 20 healthy children. Our results lead us to hypothesise a relationship between food allergy and infantile autism as has already been suggested for other disturbances of the central nervous system.

Molecular cloning and expression of cDNA encoding the rat UDP-N-acetylglucosamine:alpha-6-D-mannoside beta-1,2-N-acetylglucosaminyltransferase II.

UDP-N-acetyl-D-glucosamine:alpha-6-D-mannoside beta-1,2-N-acetylglucosaminyltransferase II (EC 2.4.1.143) (GnT II) is a Golgi resident enzyme that catalyzes an essential step in the biosynthetic pathway leading from high mannose to complex N-linked oligosaccharides. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of the enzyme purified from rat liver revealed a polypeptide of 42 kDa. Amino acid sequences were obtained from the N terminus and a tryptic peptide. Overlapping cDNA clones coding for the full-length rat GnT II were obtained. The complete nucleotide sequence revealed a 1326-base pair open reading frame that codes for a polypeptide of 442 amino acids, including a presumptive N-terminal membrane-anchoring domain. The region of cDNA coding for the C-terminal 389 amino acids of rat GnT II was linked in frame to a cDNA segment encoding the cleavable signal sequence of the human interleukin-2 receptor and transiently expressed in COS-7 cells. A 77-fold enhancement of GnT II activity over a control carrying the GnT II cDNA out-of-frame was detected in the culture medium at 72 h after transfection. 1H-NMR spectroscopy confirmed that the oligosaccharide synthesized in vitro by the recombinant enzyme was the product of GnT II activity. These data verify the identity of the cloned GnT II cDNA and demonstrate that the C-terminal region of the protein includes the catalytic domain.

[Conservative treatment of stage I-II breast carcinoma. Side-effects and esthetic results]. / Trattamento conservativo del carcinoma mammario negli stadi I-II. Effetti collaterali e risultato estetico.

A good cosmetic result is one of the main goals in conservative breast cancer treatment (quadrantectomy plus radiotherapy and, sometimes, adjuvant chemotherapy). In a retrospective study on 302 patients treated at Cremona Hospital Radiotherapy Department, a group of 68 patients was selected because of its homogeneity: all these patients had been operated by the same surgeon and irradiated according to the same technical rules. Acute reactions, late side-effects and cosmetic results are analyzed in this group. Acute postoperative complications were seen in 15% of the cases and late or persistent changes in 54%. Radiotherapy caused acute reactions in 73.5% of the cases and late side-effects in 47%. An acceptable cosmetic result was obtained in 95.5% of the patients: an excellent result in 31%, a good result in 41% and a sufficient one in 23.5%. The dose to whole breast, the dose to the quadrant and the kind of boost have no influence on the cosmetic results, while concomitant chemotherapy (p = 0.028) and high inhomogeneity in dose distribution (p = 0.04) caused a worse outcoming. The authors believe that better results may be obtained by improving treatment planning (to reduce inhomogeneity) and by optimizing chemotherapy and radiotherapy combination.

Food allergy in cystic fibrosis.

Food allergy was investigated in 20 children with cystic fibrosis (CF), who still suffered from diarrhea and failed to thrive, in spite of adequate diet and enzyme treatment (group A). The study also included two age-matched control groups, comprising 10 CF children without intestinal symptoms and/or failure to thrive (group B), and 20 healthy children (group C). Skin tests were positive and total IgE higher than the mean + 2SD respectively in 14/20 and 11/20 patients of group A, in 3/10 and 2/10 patients in group B and in none in group C. The specific IgE were present in 6/14 children in group A whose skin tests were positive and in none in group B. There was no significant difference between group A and group B (p > 0.05). The levels of specific antibodies IgG, IgA and IgM were overall higher than the mean + 2SD of the normal in 18/20 in group A, in 6/10 in group B and in none in group C. The measurement by ELISA of specific antibodies for cow milk and egg proteins showed a statistically significant difference for casein, beta-lactoglobulin and ovalbumin between the IgG (p < 0.05) and IgA (p < 0.001) levels in group A and the other groups (B and C). Symptoms improved in 90% of CF patients (group A) when the implicated foods were eliminated from the diet and in 78% the oral provocation test resulted positive. The occurrence of food allergy must be considered in CF patients who do not improve with the conventional treatment. In these patients immunological investigations, in particular the measurement of IgG, IgA and IgM specific antibodies, are useful for diagnosis and in selecting an appropriate diet leading to an improvement in nutritional status.

[Hemicrania and food allergy in children]. / Emicrania e allergia alimentare nel bambino.

Ninety-two children affected by migraine were studied, of which 49 had positive skin tests to one or more foods. Forty of those who tested positive (87%) improved after following an elimination diet for 4-6 weeks and were positive for at least one challenge test for the same types of foods which resulted in positive skin tests. Thirty-one children were cured following the elimination diet and 9 improved. At two years, despite the reintroduction of the suspected food or foods 6 to 12 months after the start of the elimination diet, these results remained unchanged.