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ABSTRACTWhen production of bispecific antibodies requires the co-expression and assembly of three or four polypeptide chains, low expression of one chain can significantly limit assembly and yield. κλ bodies, fully human bispecific antibodies with native IgG structure, are composed of a common heavy chain and two different light chains, one kappa and one lambda. No engineering is applied to force pairing of the chains, thus both monospecific and bispecific antibodies are secreted in the supernatant. In this context, stoichiometric expression of the two light chains allows for maximal assembly of the bispecific antibody. In this study, we selected a κλ body with suboptimal characteristics due to low kappa chain expression. Codon optimization to increase expression of the kappa chain did not improve bispecific yield. Surprisingly, progressive introduction of non-optimal codons into the sequence of the lambda chain resulted in lowering its expression for an optimal tuning of the relative distribution of monospecific and bispecific antibodies. This codon de-optimization led to doubling of the κλ body yield. These results indicate that assembly of different proteins into a recombinant complex is an interconnected process and that reducing the expression of one polypeptide can actually increase the overall yield.

Background The antiretroviral regimens including tenofovir and a ritonavir-boosted protease inhibitor (r/PI) have been associated with a reduced bone mineral density (BMD), increased bone turnover markers and renal tubular dysfunction.Methods An observational, prospective study was performed including HIV-1-infected, virologically suppressed patients treated with tenofovir/emtricitabine plus an r/PI for at least 12 months who switched to raltegravir plus nevirapine. The primary end point was changes after 48 weeks in estimated glomerular filtration rate (eGFR), prevalence of tubular dysfunction, BMD and concentration of two serum markers of bone turnover: collagen type-1 cross-linked C-telopeptide (CTX) and bone-specific alkaline phosphatase (BAP).Results A total of 46 patients were enrolled: 78% were male, 96% were Caucasian, the mean age was 45 years and the mean CD4+T-lymphocyte count was 681 cells/mm3. A renal impairment was present in 72% of patients and was the main reason for the switch. After 48 weeks, prevalence of proximal tubular dysfunction decreased significantly (-72%; P<0.001), whereas the mean value of eGFR did not change significantly. At the same time, after 48 weeks a significant increase in both lumbar spine and total hip BMD, T-score and Z-score was reported (+11.5% in lumbar spine T-score; P<0.001), and there was a significant reduction in both CTX and BAP mean serum concentrations (-15% and -13%, respectively; P<0.001). Two (4.3%) patients had virological failure due to suboptimal adherence and one (2.2%) subject discontinued treatment due to a skin rash.Conclusions Switching virologically suppressed patients from tenofovir/emtricitabine plus one r/PI to raltegravir plus nevirapine after 48 weeks significantly improved proximal tubular function, increased BMD and reduced serum markers of bone turnover.

Evidence has recently emerged that butyrophilins, which are members of the extended B7 family of co-stimulatory molecules, have diverse functions in the immune system. We found that the human and mouse genes encoding butyrophilin-2A2 (BTN2A2) are regulated by the class II trans-activator and regulatory factor X, two transcription factors dedicated to major histocompatibility complex class II expression, suggesting a role in T cell immunity. To address this, we generated Btn2a2-deficient mice. Btn2a2−/− mice exhibited enhanced effector CD4+ and CD8+ T cell responses, impaired CD4+ regulatory T cell induction, potentiated antitumor responses, and exacerbated experimental autoimmune encephalomyelitis. Altered immune responses were attributed to Btn2a2 deficiency in antigen-presenting cells rather than T cells or nonhematopoietic cells. These results provide the first genetic evidence that BTN2A2 is a co-inhibitory molecule that modulates T cell–mediated immunity.

Background: The association between abacavir use and increased risk of myocardial infarction has been heavily debated, but cohort studies and randomized trials have provided conflicting results. Aim of our study is to compare the effect of abacavir and tenofovir on the inflammation and endothelial activation markers. Methods: We performed an observational study of HIV-infected naïve patients starting tenofovir/emtricitabine (group A) or abacavir/lamivudine (group B) plus efavirenz. In the present analysis, we measured serum levels of high-sensitivity C-reactive protein (hsCRP), interleukin-6 (IL-6), tumor necrosis factor- (TNF-), soluble vascular adhesion molecule-1 (VCAM-1), soluble intercellular adhesion molecule-1 (ICAM-1), E-selectin and P-selectin at baseline and during a 48-week follow-up. Results: As a whole, 118 patients (93 males; mean age ± SD of 42.8 ± 10.1 years) were enrolled: 61 in group A and 57 in group B. In group A at weeks 24 and 48 the mean concentrations of IL-6, TNF-, ICAM-1, VCAM-1, E-selectin and Pselectin decreased significantly in comparison with respective baseline values. In group B at week 24 a significant increase in mean values of these markers was reported in comparison with group A, but after 48 weeks they significantly decreased in group B too and no significant differences between groups A and B were found. Conclusion: In our study, naïve patients starting tenofovir/emtricitabine or abacavir/lamivudine plus efavirenz showed after 48 weeks a significant and comparable decrease in serum concentrations of IL-6, TNF-, ICAM-1, VCAM-1, Eselectin and P-selectin, while the mean level of hs-CRP did not change significantly in any group.

Myocardial fatty foci were found in 50% (24 of 48) of adult patients with tuberous sclerosis complex (TSC) but were not found in any control subjects, enabling us to confirm these lesions were a highly specific indicator for TSC; their presence was associated with TSC1and TSC2gene mutations, brain or multiorgan involvement, and degree of multiorgan involvement.

The present study investigated the effect of parity on plasma insulin level around parturition in Saanen goats. On d –14, –7, 0, 3, 7, 10, and 14 from parturition, plasma glucose, NEFA, free AA, cortisol, and insulin concentrations were analyzed in 10 primiparous and 10 multiparous goats. At parturition, BW of primiparous goats was about 75% of that of multiparous ones (P< 0.001) and then their milk production was lower than that of multiparous ones (P< 0.001). At parturition, glucose increased (P< 0.01) in both primiparous and multiparous goats and then decreased (P< 0.01) on d 3 of lactation, remaining higher (P< 0.01) in primiparous than multiparous goats until the end of the study period. In both groups, free AA decreased (P< 0.01) at parturition, returning to prepartum levels (P< 0.01) on d 3 of lactation without difference between groups. Only in multiparous goats, plasma NEFA increased at parturition (P< 0.01), returning to prepartum levels on d 14 (P< 0.01). Changes in glucose and AA could have been caused by cortisol, which increased (P< 0.01) at parturition in both primiparous and multiparous goats, returning to prepartum levels (P< 0.01) on d 7 of lactation, without difference between the parity groups. In multiparous goats, insulin decreased soon after parturition (P< 0.05), remaining at low levels until the end of the study period, whereas in primiparous goats, insulin did not vary until d 14 of lactation, when it decreased (P< 0.05) also in this group. Therefore, between d 3 and 14 of lactation, insulin was higher in primiparous than multiparous goats (P< 0.05). Only in primiparous goats, at kidding, insulin was negatively correlated to BW (P< 0.01), and after parturition it was negatively correlated with milk yield (P< 0.05) and plasma NEFA (P< 0.05). We hypothesize that higher insulin levels in primiparous Saanen goats, which are still immature at their first breeding season, acted to limit both the mobilization of bodily reserves and the capture of nutrients by the lactating mammary gland, thus providing nutrients for their own physical and physiological development.

Increasing evidence suggests that neutrophils may participate in the regulation of adaptive immune responses, and can reach draining lymph nodes and cross-prime naive T cells. The aim of this study was to identify the mechanism(s) involved in the migration of neutrophils to the draining lymph nodes. We demonstrate that a subpopulation of human and mouse neutrophils express CCR7. CCR7 is rapidly expressed at the membrane upon stimulation. In vitro, stimulated human neutrophils migrate in response to the CCR7 ligands CCL19 and CCL21. In vivo, injection of complete Freund adjuvant induces a rapid recruitment of neutrophils to the lymph nodes in wild-type mice but not in Ccr7−/− mice. Moreover, intradermally injected interleukin-17–and granulocyte-macrophage colony-stimulating factor–stimulated neutrophils from wild-type mice, but not from Ccr7−/− mice, migrate to the draining lymph nodes. These results identify CCR7 as a chemokine receptor involved in the migration of neutrophils to the lymph nodes.

Introduction: Pancreatic cancer is one of the most aggressive gastrointestinal cancer with less than 10% long-term survivors. The apoptotic pathway deregulation is a postulated mechanism of carcinogenesis of this tumour. The present study investigated the prognostic role of MUC2 and MUC5 apomucin expression in a series of surgically resected pancreatic cancer patients. Material and Methods: All patients affected by pancreatic ductal adenocarcinoma and treated with surgical resection from 1988 to 2003 were considered for the study. MUC2 and MUC5 expression were evaluated by immunohistochemical staining. Tumour specimens of 59 resected patients were included in the study. Results: By univariate analysis, survival was influenced by MUC2 expression but not by MUC5 expression. The MUC2 overexpression was associated with better prognosis (P=0.003). By a multivariate Cox regression analysis, MUC2 overexpression maintained the prognostic statistical value. In particular, patients with high MUC2 staining showed a longer survival. Moreover the present study does report the absence of a prognostic role of MUC5 expression in this type of cancer. Conclusions: the study demoinstrated the prognostic relevance of MUC2 expression in pancreatic cancer and underlined its potential role as target gene in the field of therapy research.

The article focuses on the Reform Böhm system clarinets from Germany and France. This clarinet was developed by German clarinetist Oskar Oehler, a founding member of the Berlin Philharmonic Orchestra, in Berlin, Germany in 1887. The model offers major improvements in the clarinet during the 19th century and offers solutions for problematic technical issues, intonation and volume. A brief history of the Reform Böhm system based on the studies of Ernst Schmidt and Louis Kolbe is also provided.

Chemokines are important mediators of the immune response that are responsible for the trafficking of immune cells between lymphoid organs and migration towards sites of inflammation. Using phage display selection and a functional screening approach, we have isolated a panel of single-chain fragment variable (scFv) capable of neutralizing the activity of the human chemokine CXCL10 (hCXCL10). One of the isolated scFv was weakly cross-reactive against another human chemokine CXCL9, but was unable to block its biological activity. We diversified the complementarity determining region 3 (CDR3) of the light chain  variable domain (VL) of this scFv and combined phage display with high throughput antibody array screening to identify variants capable of neutralizing both chemokines. Using this approach it is therefore possible to engineer pan-specific antibodies that could prove very useful to antagonize redundant signaling pathways such as the chemokine signaling network.

Across the animal kingdom, multivalency discriminates antibodies from all other immunoglobulin superfamily members. The evolutionary forces conserving multivalency above other structural hallmarks of antibodies remain, however, incompletely defined.

Methylenetetrahydrofolate reductase (MTHFR) plays a central role in the metabolism of folate, which provides a methyl donor for DNA methylation and deoxynucleoside synthesis. We performed a case-control study to explore the relationship between two common MTHFR polymorphisms (C677T and A1298C), their combination and interaction with environmental exposures, on gastric adenocarcinoma susceptibility and progression in an Italian population. One hundred and two cases and 254 hospital controls, matched by age and gender, were enrolled. Individuals carrying the MTHFR 677T allele showed an increased risk of gastric cancer (odds ratio (OR) 1.62, 95% confidence interval (CI) 0.98-2.67), particularly among ever smokers (OR 2.10, 95% CI 1.07-5.33) and, among 677 TT individuals, those with a low intake of fruit and vegetables (OR 2.18, 95% CI 1.05-4.54). The strongest effect, however, was noted for the MTHFR 677 TT genotype among the diffuse gastric cancer histotype (OR 2.92, 95% CI 1.12-7.60). No association was detected for the effect of MTHFR A1298C polymorphism. Survival analysis did not show any association between each polymorphism on the overall survival, although when the analysis was restricted to the first year of follow-up after the surgical intervention an improved survival was noted among MTHFR 677 CC subjects compared with the T allele carriers (p value for log-rank test 0.02). In conclusion, MTHFR 677 (any T genotype) appears to modulate an individual's susceptibility to gastric cancer, particularly when combined with cigarette smoking and among those with a low intake of fruit and vegetables. Our results also suggest that an aberrant DNA methylation pattern, through impaired folate metabolism, might play a key role in gastric carcinogenesis. A possible survival effect of the MTHFR C677T genotype in gastric cancer patients deserves further investigations with larger sample sizes.

Polydisperse Ficoll mixtures have been used to explore glomerular sieving. Ficoll appears to be neither absorbed nor secreted by the renal tubule, and so urinary Ficoll concentrations reflect only the glomerular filtration barrier. The literature is contradictory regarding Ficoll's behavior as an idealized spherical solute. Further definition of Ficoll transport will inform interpretation of in vivo results. Flat-sheet membranes comprising a uniform array of slit pores measuring 8 nm by 45 μm were perfused with FITC-labeled Ficoll 70 and BSA. Ficoll and BSA concentrations were quantified by gel-permeation chromatography and Bradford assay, respectively. BSA and Ficoll molecules with diameters equal to approximately half of the slit pore width displayed hindered transport in agreement with modeled rigid sphere transport through slit-shaped pores. Ficoll molecules larger than ∼0.65 slit width displayed transport rates in excess of predictions. Ficoll molecules with Stokes-Einstein diameters greater than the pore dimension were observed in permeate samples. We present data for Ficoll filtration through a novel array of well-defined pores, which illustrate that Ficoll is well modeled as an ideal sphere in one size domain, but the model breaks down as molecular diameter approaches pore size. These data inform the present debate regarding glomerular filtration and affect conclusions drawn from the use of Ficoll as a tracer molecule. The apparent hyperpermeability of Ficoll through slit-shaped pores suggests that further modeling incorporating deformation of the molecule is necessary when using Ficoll solutions to characterize membranes.

The article features clarinetist Luigi Amodio. He had to make a living by playing freelance in Italy and Switzerland in various groups. Chamber music was performed by Amodio in German concert halls with success. Critics praised all of his performances in Italy and abroad, particularly for his peculiar tonal qualities and interpretative finesse. A short contract of a few months with the Rome Radio Orchestra was accepted by Amodio in the summer of 1936.

Hemangiomas are common benign tumors; they frequently occur in the liver but very rarely in the gallbladder, with only seven cases reported in the scientific literature to date. We here report an additional patient, a 49-year-old white woman presenting with an echogenic lesion of the gallbladder that was incidentally discovered. Cholecystectomy was performed after computed tomography had revealed a gallbladder neoplasm; pathological examination showed the mass to be a cavernous hemangioma. No postoperative complications occurred and the patient is alive and free of recurrence five years after the operation. We present a review of the literature on this topic with special emphasis on the diagnostic and therapeutic challenges these lesions may entail. Hemangiomas of the gallbladder may have extremely variable presentations (from non-specific abdominal pain to acute syndromes resembling cholangitis or choledocholithiasis) and can mimic different lesions (liver tumors, sarcoma). Only surgical exploration can provide a correct diagnosis. Excision is indicated as these lesions may grow to huge sizes, compress adjacent structures or bleed.Conclusions Gallbladder hemangiomas are uncommon benign tumors. A preoperative diagnosis is difficult to make. Surgical excision is mandatory both in reaching a final diagnosis and in preventing bleeding or compression of vital structures.

The article focuses on the "Concerto in B flat major, Op. 101," composed by Saverio Mercadante for clarinet and orchestra. It states that Italian composer Mercandante was born in Altamura, Italy and used the clarinet in his compositions in a serious way. It mentions that his two clarinet concertos belong to the first decades of the 19th century and the B flat major concerto is the most frequently performed. An analysis of the concerto is presented.

Outer membrane protein 3;A (OmpA) is a class of bacterial cell wall protein that is immunogenic without adjuvant. As specific immune responses are initiated in the lymph nodes (LN), we analyzed the effect of the OmpA from Klebsiella pneumoniae (KpOmpA) on chemokine/chemokine receptor expression by APC and on cell migration to the LN. Upon contact with KpOmpA, human immature DC andmacrophages acquire CCR7 expression and responsiveness to CCL21. In parallel, CCR1 and CCR5 expression is down-regulated and CXCL8, CCL2, CCL3 and CCL5 production is up-regulated. Mice injected subcutaneously with KpOmpA present a transient inflammatory reaction at the site of injection accompanied by an enlargement of the draining LN with a higher proportion of DC and macrophages. Lastly, when exposed to KpOmpA prior injection, DC but not macrophages migrate to the draining LN. In conclusion, KpOmpA confers a migratory phenotype to DC and triggers their migration to the regional LN. This property contributes to explain how innate cells initiate adaptive immune response upon recognition of conserved bacterial components and also why OmpA is immunogenic in the absence of adjuvant.

Human monocytes differentiate into dendritic cells (DCs) or macrophages according to the nature of environmental signals. Monocytes stimulated with granulocyte-macrophage colony-stimulating factor (GM-CSF) plus interleukin 4 (IL-4) yield DCs. We tested here whether interferon-γ (IFN-γ), a potent activator of macrophages, may modulate monocyte differentiation. Addition of IFN-γ to IL-4 plus GM-CSF–stimulated monocytes switches their differentiation from DCs to CD14−CD64+macrophages. IFN-γ increases macrophage colony-stimulating factor (M-CSF) and IL-6 production by IL-4 plus GM-CSF–stimulated monocytes by acting at the transcriptional level and acts together with IL-4 to up-regulate M-CSF but not IL-6 production. IFN-γ also increases M-CSF receptor internalization. Results from neutralizing experiments show that both M-CSF and IL-6 are involved in the ability of IFN-γ to skew monocyte differentiation from DCs to macrophages. Finally, this effect of IFN-γ is limited to early stages of differentiation. When added to immature DCs, IFN-γ up-regulates IL-6 but not M-CSF production and does not convert them to macrophages, even in the presence of exogenous M-CSF. In conclusion, IFN-γ shifts monocyte differentiation to macrophages rather than DCs through autocrine M-CSF and IL-6 production. These data show that IFN-γ controls the differentiation of antigen-presenting cells and thereby reveals a new mechanism by which IFN-γ orchestrates the outcome of specific immune responses.

Aims We describe the fourth reported case of granulomatous gastritis associated with gastric adenocarcinoma, with a review of the literature and considerations about the prognostic implications of this association.Results A 48-year-old woman who had been suffering from gastritis for ten years was admitted to our institute for increasing left epigastric pain associated with vomiting. After an endoscopic biopsy had revealed an ulcerated signet ring cell carcinoma, the patient was submitted to subtotal gastrectomy with regional lymph node dissection. Pathological examination of the resected specimen revealed a superficial signet ring cell carcinoma (early cancer) associated with multiple granulomas. The granulomas, which were observed within the mucosa and the submucosa at the periphery of the carcinoma, were composed of CD68-positive, CD15-negative epithelioid and giant cells of the Langhans type, confirming their true histiocytic nature, and were also extensively found within the dissected lymph nodes. Since no ocular, skin, pulmonary or other gastrointestinal lesions were found and the granulomas were negative for acid-fast and fungal stain, a diagnosis of granulomatous gastritis was made.Conclusions To the best of our knowledge this is the fourth example of gastric adenocarcinoma and granulomatous gastritis. These cases suggest an association between granulomatous gastritis and early gastric cancer.

CD86 is a costimulatory molecule constitutively expressed by human antigen presenting cells which interacts with CD28 and CTLA-4 expressed by T cells. We have recently reported the identification of an alternatively spliced CD86 mRNA variant (CD86ΔTM) characterized by the deletion of exon 6 which encodes for the transmembrane domain. We report here the identification of an alternatively spliced variant (called CD86ΔEC) expressed by nonstimulated human monocytes and characterized by the deletion of exons 4 and 5 which encode for the extracellular V-like and C-like domains, respectively. The activation of monocytes by IFNγ (i) induces the preferential expression of the transcript encoding for the membrane form and (ii) increases the expression of CD86 and of the accessory molecules CD40, CD49d and CD54. These results suggest that resting human monocytes may constitutively express different forms of CD86 which can then influence the activation of T cells.

Using the sequence of cosmids derived from chromosome 19p12, we have identified a gene encoding a novel protein, BSMAP (brain-specific membrane-anchored protein) and cloned cDNA encoding the full-length open reading frame. Northern blot analysis revealed that BSMAP mRNA is preferentially expressed at a high level in the brain. BSMAP has a putative transmembrane domain and is predicted to be a type-I membrane glycoprotein. Genomic sequence analysis revealed that the gene encoding BSMAP consists of eight exons spanning approximately 8 kb and lies 6 kb away from the gene encoding CLF-1 in a reverse orientation. Although no candidate genetic disorders were found to map either to this precise region of chromosome 19 or to the syntenic region of the mouse genome, the highly specific expression of BSMAP mRNA suggests a role for the protein in CNS function.

Following T cell antigen receptor (TCR) engagement, the protein tyrosine kinase (PTK) ZAP-70 is rapidly phosphorylated on several tyrosine residues, presumably by two mechanisms: an autophosphorylation and a trans-phosphorylation by the Src-family PTK Lck. These events have been implicated in both positive and negative regulation of ZAP-70 activity and in coupling this PTK to downstream signaling pathways in T cells. We show here that Tyr315 and Tyr319 in the interdomain B of ZAP-70 are autophosphorylated in vitro and become phosphorylated in vivo upon TCR triggering. Moreover, by mutational analysis, we demonstrate that phosphorylation of Tyr319 is required for the positive regulation of ZAP-70 function. Indeed, overexpression in Jurkat cells and in a murine T cell hybridoma of a ZAP-70 mutant in which Tyr319 was replaced by phenylalanine (ZAP-70-Y319F) dramatically impaired anti-TCR-induced activation of the nuclear factor of activated T cells and interleukin-2 production, respectively. Surprisingly, an analogous mutation of Tyr315 had little or no effect. The inhibitory effect of ZAP-70-Y319F correlated with a substantial loss of its activation-induced tyrosine phosphorylation and up-regulation of catalytic activity, as well as with a decreased in vivo capacity to phosphorylate known ZAP-70 substrates, such as SLP-76 and LAT. Collectively, our data reveal the pivotal role of Tyr319 phosphorylation in the positive regulation of ZAP-70 and in TCR-mediated signaling.

CTLA-4, expressed by activated T cells, transduces an inhibitory signal. We show here that PCR amplification of the coding sequence of CTLA-4 in nonstimulated human T lymphocytes results in the amplification of two transcripts of 650 and 550 bp. Sequencing shows that the larger form codes for membrane CTLA-4 and the 550-bp transcript is a spliced variant in which exon 2 coding for the transmembrane region is deleted. This spliced cDNA has been named CTLA-4delTM. The splicing induces a frame shift which results in the addition of 22 extra amino acids before a translational termination. Activation of T cells with phorbol 12-myristate 13-acetate plus ionomycin or anti-CD3 plus anti-CD28 monoclonal antibodies induces a suppression of CTLA-4delTM mRNA expression associated with a preferential expression of the membrane CTLA-4 mRNA, showing that CTLA-4delTM mRNA expression is restricted to nonactivated T cells. A soluble immunoreactive form of CTLA-4 was detected in the serum of 14 / 64 healthy subjects. These results suggest that nonstimulated T cells may constitutively produce a soluble form of CTLA-4 which may have an important role in the regulation of immune homeostasis.

We have evaluated the expression and the involvement of membrane-associated TNF-α (mTNF-α) in human NK cell-mediated cytotoxicity. Results from FCM analysis show that peripheral blood NK cells constitutively express mTNF-α. In contrast, mTNF-α expression is undetectable on resting T cells, B cells and monocytes. Western blotting analysis confirmed that freshly purified NK cells express the 17-kDa soluble form (sTNF-α) and the 26-kDa transmembrane form of TNF-α. Stimulation with IL-2, IL-15 and IL-18 up-regulates TNF-α mRNA, sTNF-α and mTNF-α expression in NK cells. The role of mTNF-α in the cytotoxic activity of resting NK cells has been evaluated in in vitro cytotoxic assays using freshly purified NK cells fixed with paraformaldehyde as effector cells (in order to avoid the participation of cytotoxic soluble mediators such as perforin, granzymes or sTNF-α) and the TNF-α-sensitive Fas ligand- and TRAIL-resistant cell line KYM-1-D4 as target cell. Results show that fixed NK cells kill the KYM-1-D4 cells and that neutralizing anti-TNF-α antibodies partly prevent this effect. In contrast to the other types of peripheral blood mononuclear cells NK cells from adult blood constitutively express functional mTNF-α in the absence of prior contact with target cells or activation. These data demonstrate a novel mechanism of cell-mediated cytotoxicity by non-acitvated human peripheral blood NK cells.

Aims and background Surgical resection offers the only potential cure for pancreatic carcinoma. Although the overall prognosis remains a dismal, several recent series have reported an encouraging increase in 5-year survival after resection, ex-ceding 20%. As the reasons for this improvement are not clearly understood, numerous clinico-pathological parameters (demographic, intraoperative and histopatologic factors) have been investigated to evaluate their role in predicting long term survival. In this single-institution study, immediate and long-term outcome after pancreatic resection in patients with pancreatic adenocarcinoma was retrospectively evaluated, focusing attention on the possible impact of different clinico-pathologic factors on long-term survival.Methods Sixty-six patients with a confirmed histologic diagnosis of adenocarcinoma of the pancreas, treated by pancreatic resection at the Department of Surgery of the Catholic University of Rome in the years 1988-1997, were retrospectively analyzed. Morbidity and survival data were reviewed and potential prognostic factors were compared statistically by univariate analysis.Results There was no postoperative mortality. Twenty-five patients (38%) developed major operative complications. Pancreatic fistula was the most common complication, and occurred in 7 patients (11%). The actuarial overall and disease-specific survival for all 66 patients were respectively 58% and 59% at 1 year, 27% and 31% at 3 years, and 13% and 20% at 5 years, with a median survival time of 13.4 months. Nodal status was the only single factor significantly affecting survival by univariate analysis. The 3-and 5-year survival rates were respectively 35% and 19% for node-negative patients and 7% and 0% for node-positive patients (P = .04). A positive correlation with improved survival, even if not of statistical significance, was shown for other pathologic or intraoperative factors. Among the former, 5-year survival rates were better for patients with negative resection margins as compared to patients with positive margins (12% vs 7%, P = ns). Among the latter, a better actuarial 5-year survival rate was shown for patients with shorter operative time (<4 hours, 21% survival vs >4 hours 5%, P = ns) and for patients that received fewer transfusions (0-2 blood units, 14% survival vs 3 or more blood units, 0%; P = ns). Age, gender, tumor diameter and tumor grading showed no influence on survival in this series.Conclusions Our series confirmed that nodal status is the strongest independent predictor of survival. Limited intraoperative transfusion, reduced operative time and clear margins could also yeald a prognostic significance, and require further confirmation in larger series.

CD28, expressed by T cells, plays a central role in providing costimulatory signals to T cells. Thecd28gene is organized into 4 exons. An alternatively spliced CD28 mRNA lacking most of the exon 2 has been previously evidenced. We report here that non stimulated human T cells express three additional alternatively spliced variants of CD28 mRNA (CD28a-c) in. The CD28a variant, expressed at similar levels to that of the full length CD28 mRNA encoding for the membrane form, lacks exon 3. This deletion introduces (i) a frame shift resulting in the addition of two extra amino acids and a premature stop codon and, (ii) induces the loss of the transmembrane region, suggesting that it could encodes for a soluble monomeric molecule which conserves the binding sites of CD28. The CD28b and CD28c variants, expressed at a low level compared with CD28a, are generated by deletion of most of the 3′ end of exon 2 plus exon 3 and exon 2 plus exon 3, respectively. Activated T cells express only the membrane CD28 mRNA. These results suggest that resting human T cells may constitutively express both membrane and soluble CD28 which can differentially regulate the outcome of the T cell response.

An interview with clarinetist Gervase de Peyer is presented. He shares his perspectives regarding the young generation of clarinet players in Italy where he gives master classes after performing in Trapani and in Rome. He offers his views on the importance of master classes to students and the substantial advice he can give to help students. He states his reason on why he decided to become a solo clarinet player after he had been playing for some orchestras in London, England.

• The medical records of 135 consecutive patients (74 women and 61 men) who underwent surgery for hydatid liver disease were reviewed. The patients ranged in age from 4 to 81 years. Twenty-seven patients had undergone previous surgery for hydatid liver disease. Cysts were solitary in 100 patients and multiple in 35 patients. Seventeen patients had concomitant extrahepatic disease. Conservative procedures were used in 71 patients (capitonnage in 50 patients and partial pericystectomy in 21 patients). Radical procedures were used in 64 patients (total pericystectomy in 35 patients, subtotal pericystectomy in 16 patients, and wedge or major liver resection in 13 patients). Operative mortality was 2.2% and morbidity rate was 23.7%. Recurrent disease was found in 13 patients at a mean interval of 3 years from the first operation. Better short- and long-term results were obtained with the use of radical procedures.(Arch Surg. 1991;126:518-523)

The PTP35/IA-2 protein shows high homology to protein tyrosine phosphatases (PTPases) but harbours a few changes in invariant PTPase residues. Accordingly, PTP35/IA-2 has been reported to lack catalytic activityin vitro,and itsin vivobiological function remains to be determined. We investigated if reversion of selected amino acids to the PTPase consensus could reconstitute enzymatic activity. Substitution of aspartic acid 911 in the putative active site with alanine resulted in the appearance of low but reproducible activity onpNPP dephosphorylation. Moreover, contemporary replacement of alanine 877 with aspartic acid greatly increased the catalytic efficiency of the D911A mutant. The A877D/D911A double mutant protein was also found to specifically dephosphorylate myelin basic protein phosphorylated on tyrosine. These results suggest that the general scaffold of the PTP35 protein is compatible with a common catalytic mechanism shared by PTPases and argue against an intrinsic enzymatic function of the wild type form.