Search

AbstractLeft Ventricular Assist Devices (LVADs) represent an important therapeutic option in the management of advanced heart failure. We report a case of 74 years-old male with a Jarvik 2000® (Jarvik Heart, New York, USA) LVAD who presented with an infection of the LVAD driveline due to methicillin-resistant Staphylococcus aureus(MRSA) that was treated with dalbavancin as chronic antimicrobial suppression therapy without adverse events and maintaining a good quality of life for more than 37 weeks. Dalbavancin could represent a valid option for the treatment of LVAD infections because of its efficacy, pharmacokinetic/pharmacodynamic proprieties, safety and tolerability.

Metal contamination is a waste-generating and serious issue in the synthesis of chemicals, in particular in the case of products with biological activity. The appropriate selection of operating conditions plays a crucial role in the abatement of metal leaching in solution and associated wastes. Herein we report a waste-minimized continuous flow process for the synthesis of 1,4-disubstituted β-keto 1,2,3-triazoles exploiting the use of a copper tube flow reactor (CTFR). The selection of the proper azeotropic mixture allowed an almost quantitative recovery of the reaction medium greatly influencing the E-factor of the protocol. A thorough understanding of the main parameters affecting the waste generation was given by calculation of the E-factor distribution for different work-up tested under batch and flow conditions. Furthermore, the measurement of different green metrics (AE: Atom Economy, SF: Stoichiometric Factor, RME: Reaction Mass Efficiency, and MRP: Mass Recover Parameter) clearly demonstrated the benefits of the flow scale-up that allowed to perform a low environmental footprint CuAAC reaction.

The aim of this article is to describe the audiological patterns of 71 adult patients presenting severe to profound sensorineural hearing loss, who were rehabilitated by cochlear implants (CIs) and hearing aids. This is a retrospective study in a university setting, where the clinical records of 71 adult patients were reviewed and processed. Speech intelligibility was evaluated at one aided ear (CI) or at both aided ears (double CI or a combination of CI and hearing aid [HA]). Patients with a bilateral CI or with a bimodal hearing setup (CI and HA) performed better than those with a single CI; data from the phonetic matrices test showed that there was a statistically significant difference among patients aided by a single CI versus binaural setup (double CI or CI + HA). In particular, patients aided by a bilateral CI, or by a CI and HA, showed an improvement in the functional results of the speech tests, compared to patients using a single CI. Binaural hearing (either with a bilateral CI or bimodal) allows an improvement in the functional results at the speech tests, compared to the use of a CI only.

•Reconstitution of T cells after transplantation is a determinant of the long-term success of the procedure.•Cytomegalovirus (CMV) infection and disease remain an important cause of morbidity and mortality after allogeneic transplant.•Several studies, from murine model to human, have demonstrated a major role of CMV-specific cytotoxic T cells in the control of viral replication.•Up to now, no data about CD4+and CD8+T cell reconstitution during CMV prophylaxis with letermovir have been reported.

Innate lymphoid cells (ILCs) represent a distinct branch of the lymphoid lineage composed of 3 major subpopulations: ILC1, ILC2, and ILC3. ILCs are mainly described as tissue-resident cells but can be detected at low levels in human blood. However, unlike mouse ILCs, there is still no consistent methodology to purify and culture these cells that enables in-depth analysis of their intrinsic biology. Here, we describe defined culture conditions for ILC2s, which allowed us to dissect the roles of interleukin 2 (IL-2), IL-25, IL-33, and thymic stromal lymphopoietin (TSLP) individually, or in combination, in modulating ILC2 phenotype and function. We show that TSLP is important for ILC2 survival, while ILC2 activation is more dependent on IL-33, especially when in combination with IL-2 or TSLP. We found that activation of ILC2s by IL-33 and TSLP dramatically upregulated their surface expression of c-Kit and downregulated expression of the canonical markers IL-7Ra and CRTH2. IL-2 further amplified ILC2 production of IL-5, IL-13, and granulocyte-macrophage colony-stimulating factor but also induced a more natural killer (NK)–like phenotype in ILC2, with upregulation of granzyme B production by these cells. Furthermore, ILC2 plasticity was observed in serum-free SFEM II media in response to IL-33, IL-25, and TSLP stimulation and independently of IL-12 and IL-1ß. This is the first comprehensive report of an in vitro culture system for human ILC2s, without the use of feeder layers, which additionally evaluates the impact of IL-25, IL-33, and TSLP alone or in combination on ILC2 surface phenotype and activation status.

Innate lymphoid cells (ILCs) represent a distinct branch of the lymphoid lineage composed of 3 major subpopulations: ILC1, ILC2, and ILC3. ILCs are mainly described as tissue-resident cells but can be detected at low levels in human blood. However, unlike mouse ILCs, there is still no consistent methodology to purify and culture these cells that enables in-depth analysis of their intrinsic biology. Here, we describe defined culture conditions for ILC2s, which allowed us to dissect the roles of interleukin 2 (IL-2), IL-25, IL-33, and thymic stromal lymphopoietin (TSLP) individually, or in combination, in modulating ILC2 phenotype and function. We show that TSLP is important for ILC2 survival, while ILC2 activation is more dependent on IL-33, especially when in combination with IL-2 or TSLP. We found that activation of ILC2s by IL-33 and TSLP dramatically upregulated their surface expression of c-Kit and downregulated expression of the canonical markers IL-7Rα and CRTH2. IL-2 further amplified ILC2 production of IL-5, IL-13, and granulocyte-macrophage colony-stimulating factor but also induced a more natural killer (NK)–like phenotype in ILC2, with upregulation of granzyme B production by these cells. Furthermore, ILC2 plasticity was observed in serum-free SFEM II media in response to IL-33, IL-25, and TSLP stimulation and independently of IL-12 and IL-1β. This is the first comprehensive report of an in vitro culture system for human ILC2s, without the use of feeder layers, which additionally evaluates the impact of IL-25, IL-33, and TSLP alone or in combination on ILC2 surface phenotype and activation status.

-TCRαβ/CD19-cell depleted haploHSCT is characterized by low TRM and acute/chronic GVHD, with OS and DFS similar to other transplants-Use of TBI in the conditioning regimen, transplant in CR1/2 and low/negative levels of MRD are associated with improved outcome

F095Allogeneic, donor-derived CD19-CAR T cells can be readily generated for the treatment of pediatric patients with relapsed/refractory BCP-ALLF095Allogeneic CD19-CAR T cells are effective and, compared to autologous CAR, they neither increase toxicity, nor induce high GvHD incidence

Transabdominal ultrasound examination carried out at 11.3 weeks' gestation suggested the diagnosis of holoprosencephaly (HPE). Transvaginal three-dimensional (3D) scan performed using the niche-mode technique enabled diagnosis of HPE, hypotelorism, and cleft lip (CL). The fetus was diagnosed with trisomy 18 by means of transvaginal celocentesis at the time of pregnancy termination. Although prenatal diagnosis of orofacial cleft can be enhanced by 3D ultrasound, only a few cases have been detected early in pregnancy. Here, we report a first-trimester case in which 3D ultrasound in niche mode improved the antenatal diagnosis of CL. Early fetal karyotyping can be accomplished by celocentesis in these cases.

To describe the value of hysteroscopic fetoscopy (virtuopsy) at the time of uterine suction in a case of early diagnosis of congenital anomalies in parents refusing conventional full autopsy examination.

Abstract: Background: Surgical resection is the mainstay treatment for resectable esophageal cancer. Minimally invasive esophagectomy is performed with increasing frequency and proves to be a safe and effective surgical alternative to the open technique. Minimally invasive esophagectomy using thoracoscopic esophageal mobilization with the patient in prone position seems to offer some advantages with regard to surgeon ergonomics and clinical outcome. Methods: Between July 2005 and September 2010, 46 patients (35 men and 11 women) underwent minimally invasive esophagectomy in the prone position at the authors’ institution. Three patients had previously undergone a thoracic intervention (one patient had previously undergone left pneumonectomy because of lung cancer). The preoperative indication was squamous cell carcinoma for 35 patients and adenocarcinoma for 11 patients. In one case, the histology of the biopsy samples showed a squamous cell carcinoma with neuroendocrine differentiation. Neoadjuvant treatment was administered to 15 patients. Results: All 46 patients underwent esophagectomy using minimally invasive thoracic mobilization of the esophagus with the patient in prone position. The abdominal stage of intervention was performed by laparoscopy for 37 patients and by laparotomy for 9 patients. No thoracotomic conversion was performed. In all cases, a cervical end-to-side anastomosis was performed using a circular stapler. The mean operative time was 263 min. The median intensive care unit stay was 2 days, and the median postoperative hospital stay was 15 days. The mean number of procured lymph nodes was 13. The perioperative morbidity rate was 37%, and the perioperative mortality rate was 4.4%. Conclusions: Minimally invasive esophagectomy is safe and technically feasible. It entails a lower mortality rate and a shorter hospital stay than those reported in most open series. Thoracoscopy with the patient in prone position offers results comparable with those obtained using other minimally invasive techniques regarding the number of procured lymph nodes. This technique shows considerable advantages such as improved surgeon ergonomics, increased operative field exposure, and satisfactory respiratory results.

Rosacea is a chronic inflammatory skin disease of unknown etiology. Although described centuries ago, the pathophysiology of this disease is still poorly understood. Epidemiological studies indicate a genetic component, but a rosacea gene has not been identified yet. Four subtypes and several variants of rosacea have been described. It is still unclear whether these subtypes represent a “developmental march” of different stages or are merely part of a syndrome that develops independently but overlaps clinically. Clinical and histopathological characteristics of rosacea make it a fascinating “human disease model” for learning about the connection between the cutaneous vascular, nervous, and immune systems. Innate immune mechanisms and dysregulation of the neurovascular system are involved in rosacea initiation and perpetuation, although the complex network of primary induction and secondary reaction of neuroimmune communication is still unclear. Later, rosacea may result in fibrotic facial changes, suggesting a strong connection between chronic inflammatory processes and skin fibrosis development. This review highlights recent molecular (gene array) and cellular findings and aims to integrate the different body defense mechanisms into a modern concept of rosacea pathophysiology.

Construction of the 24,8 km Modena Viaducts, which form part of the Milan–Naples High Speed Railway Project in Italy, required full-span precasting of 755 simply supported spans and in-place casting of 9 three-span continuous bridges in only 30 months under global warranty on time, costs, and quality. In this paper, details are given on how such a record-breaking goal has been met.Custom-designed special construction equipment was used for most of the project. Defining the QA/QC qualifications for design, fabrication, and site operations of equipment took almost 1 year. Analysis of methods, risks, and their mitigations was performed for every major activity. Contingency plans were identified and also prequalified.Performance requirements, technical specifications, and design criteria were identified for special equipment. Equipment design was subjected to full independent checking. Fabrication of equipment—traceability of materials welding, dimensional control, systems and plants, controls, site assembly, and load testing—and site operations were also ruled by specific QA/QC procedures.

Quantitative real-time polymerase chain reaction (PCR) has become an important tool for Porcine circovirus-2(PCV-2) research and diagnosis. However, significant differences in detection limit and quantification data, among laboratories and quantitative real-time PCR methods, have been demonstrated. New efforts are required for providing more accurate and comparable results. The current study is an evaluation of the effects of DNA extraction procedures on PCV-2 quantification in lymph node samples. Differences, greater than 1 log10copies/g, were shown among PCV-2 loads detected after different extraction procedures. The work highlighted the critical role of the DNA extraction method in PCV-2 quantification by quantitative real-time PCR. This important aspect should be evaluated when comparing data from different laboratories or different studies. The PCV-2 quantification data should not be considered comparable before demonstrating the equivalence of the DNA extraction methods performed.

AbstractConstruction of the 24,8 km Modena Viaducts, which form part of the Milan–Naples High Speed Railway Project in Italy, required full-span precasting of 755 simply supported spans and in-place casting of 9 three-span continuous bridges in on ly 30 months under global warranty on time, costs, and quality. In this paper, details are given on how such a record-breaking goal has been met.Custom-designed special construction equipment was used for most of the project. Defining the QA/QC qualifications for design, fabrication, and site operations of equipment took almost 1 year. Analysis of methods, risks, and their mitigations was performed for every major activity. Contingency plans were identified and also prequalified.Performance requirements, technical specifications, and design criteria were identified for special equipment. Equipment design was subjected to full independent checking. Fabrication of equipment–traceability of materials welding, dimensional control, systems and plants, controls, site assembly, and load testing–and site operations were also ruled by specific QA/QC procedures.

Construction of the 24,8 km Modena Viaducts, which form part of the Milan–Naples High Speed Railway Project in Italy, required full-span precasting of 755 simply supported spans and in-place casting of 9 three-span continuous bridges in only 30 months under global warranty on time, costs, and quality. In this paper, details are given on how such a record-breaking goal has been met.Custom-designed special construction equipment was used for most of the project. Defining the QA/QC qualifications for design, fabrication, and site operations of equipment took almost 1 year. Analysis of methods, risks, and their mitigations was performed for every major activity. Contingency plans were identified and also prequalified.Performance requirements, technical specifications, and design criteria were identified for special equipment. Equipment design was subjected to full independent checking. Fabrication of equipment—traceability of materials welding, dimensional control, systems and plants, controls, site assembly, and load testing—and site operations were also ruled by specific QA/QC procedures.

Pituitary adenylate cyclase-activating peptide (PACAP) is an important neuropeptide and immunomodulator in various tissues. Although this peptide and its receptors (ie, VPAC1R, VPAC2R, and PAC1R) are expressed in human skin, their biological roles are unknown. Therefore, we tested whether PACAP regulates vascular responses in human skin in vivo. When injected intravenously, PACAP induced a significant, concentration-dependent vascular response (ie, flush, erythema, edema) and mediated a significant and concentration-dependent increase in intrarectal body temperature that peaked at 2.7°C. Topical application of PACAP induced marked concentration-dependent edema. Immunohistochemistry revealed a close association of PACAP-immunoreactive nerve fibers with mast cells and dermal blood vessels. VPAC1R was expressed by dermal endothelial cells, CD4+and CD8+T cells, mast cells, and keratinocytes, whereas VPAC2R was expressed only in keratinocytes. VPAC1R protein and mRNA were also detected in human dermal microvascular endothelial cells. The PACAP-induced change in cAMP production in these cells demonstrated VPAC1R to be functional. PACAP treatment of organ-cultured human skin strongly increased the number of CD31+vessel cross-sections. Taken together, these results suggest that PACAP directly induces vascular responses that may be associated with neurogenic inflammation, indicating for the first time that PACAP may be a crucial vascular regulator in human skin in vivo. Antagonists to PACAP function may be beneficial for the treatment of inflammatory skin diseases with a neurogenic component.

A prenatal ultrasound diagnosis of epignathus in a dichorionic-diamniotic twin pregnancy is reported. A complex mass protruding from the fetal face was seen at week 19. Amniocentesis resulted in a 46,XX fetus with elevated alpha-fetoprotein (a-FP). An increase in tumor size and severe polyhydramnios ensued. Selective feticide performed at 22 weeks led to untreatable uterine contractions with iatrogenic abortion and early neonatal mortality of the healthy cotwin. Without development of polyhydramnios and tumor growth, weekly scan and transvaginal cervical assessment would have been carried out and cesarean section planned at around 32 weeks. Necroscopy and histology aided the ultrasound-based prenatal diagnosis.

Cystic fibrosis (CF), a disease caused by mutations in the CF transmembrane conductance regulator (CFTR) gene, is characterized by chronic bacterial infections and inflammation in the lung. Having previously shown that deletion of CFTR is associated with lower expression of the endogenous anti-inflammatory protein Annexin A1 (AnxA1), we investigated further this possible functional connection using a validated CFTR inhibitor. Treatment of mice with the CFTR inhibitor-172 (CFTR172) augmented the acute peritonitis promoted by zymosan, an effect associated with lower AnxA1 levels in peritoneal cells. Similar results were obtained with another, chemically distinct, CFTR inhibitor. The pro-inflammatory effect of CFTR172was lost in AnxA1−/−, as well as CFTR−/−mice. Importantly, administration of hrAnxA1 and its peptido-mimetic to CFTR−/−animals or to animals treated with CFTR172corrected the exaggerated leukocyte migration seen in these animals. In vitroassays with human Polymorphonuclear leukocyte (PMN) demonstrated that CFTR172reduced cell-associated AnxA1 by promoting release of the protein in microparticles. We propose that the reduced impact of the counterregulatory properties of AnxA1 in CF cells contributes to the inflammatory phenotype characteristic of this disease. Thus, these findings provide an important insight into the mechanism underlying the inflammatory disease associated with CFTR inhibition while, at the same time, providing a novel pharmacological target for controlling the inflammatory phenotype of CF.

The problem considered in this paper is the optimisation of the gear-set of a race car in a virtual (software) environment. This task has been split into three sub-tasks: first, the maximum achievable speed (as a function of the local curvature radius) in every position of the circuit has been computed; then, the lap-time has been computed with an innovative forward?backward integration procedure of a simple and numerically efficient dynamic simulator; finally, using the lap-time computation algorithm, the best gear-set is obtained by a multi-dimensional optimisation procedure. All the results have been developed and tested on real data, and the gear-set optimiser has been implemented in a complete and stand-alone software package.

Carbon monoxide (CO) generated by the enzyme heme oxygenase during the breakdown of heme is known to mediate a number of biological effects. Here, we investigated whether CO liberated from a water-soluble CO-releasing molecule (CO-RM) is capable of modulating leukocyte-endothelial interactions. Tricarbonylchoro(glycinato)ruthenium (II) (CORM-3), a fast CO releaser, proved to be anti-inflammatory in two distinct models of acute inflammation in vivo. In both cases, a significant reduction in neutrophil extravasation was observed. Subsequent in vitro static experiments showed that CORM-3 produced a direct effect on neutrophil (polymorphonuclear neutrophil; PMN) adhesion molecule expression; dose-dependently inhibiting platelet-activating factor stimulated CD11b up-regulation and L-selectin shedding, whereas no effect was observed on up-regulation of human umbilical vein endothelial cell (HUVEC) adhesion molecules intercellular adhesion molecule-1 or E-selectin nor on interleukin-8 chemokine production. In addition, when PMN interaction with HUVECs was studied, an inhibitory effect of CORM-3 on cell capture and rolling was observed. The effect of CORM-3 on PMN CD11b expression was mimicked by the incubation of PMN with the selective large potassium channel opener 1,3-dihydro-1-(2-hydroxy-5-(trifluoromethyl)-phenyl)-5-(trifluoromethyl)-2H-benzimidazol-2-one (NS-1619), which suggests that CORM-3 actions in this instance are mediated, at least in part, via opening of this channel. In conclusion, we have reported that CORM-3 possesses acute anti-inflammatory effects in vivo and that these are probably the result of targeting PMN activation and rolling upon the endothelium.

Launching gantries are frequently used in the erection of precast segmental and I-beam bridges, while movable shuttering systems (MSS) are used for in situ span-by-span and balanced-cantilever construction. The use of launching units is therefore frequent in most types of medium-span, prestressed concrete bridges.Launching gantries and MSS are complex and delicate structures. They resist huge loads during bridge erection and overtake long spans during self-launching. They are light–i.e. designed for high stress levels in different load and support conditions, which make them prone to instability. And they are reused many times, on different projects and by different crews.As a matter of fact, a number of launching units have collapsed in recent years, with many fatalities and extreme delays on the project schedule. As a consequence, the launching units are almost always leased or purchased on the basis of stringent technical specifications, and their design is subjected to independent checking.Design and operational failures detected by the author within numerous independent design check assignments are discussed in the paper and recommendations and suggestions are provided to prevent failures and assure adequate robustness of launching units. Owing to the legal proceedings surrounding such events, only general causes of collapse are discussed in this paper and no photographs of gantry collapses are included, and the photographs do not refer to collapsed units.

Launching gantries are frequently used in the erection of precast segmental and I-beam bridges, while movable shuttering systems (MSS) are used for in situ span-by-span and balanced-cantilever construction. The use of launching units is therefore frequent in most types of medium-span, prestressed concrete bridges.Launching gantries and MSS are complex and delicate structures. They resist huge loads during bridge erection and overtake long spans during self-launching. They are light—i.e. designed for high stress levels in different load and support conditions, which make them prone to instability. And they are reused many times, on different projects and by different crews.As a matter of fact, a number of launching units have collapsed in recent years, with many fatalities and extreme delays on the project schedule. As a consequence, the launching units are almost always leased or purchased on the basis of stringent technical specifications, and their design is subjected to independent checking.Design and operational failures detected by the author within numerous independent design check assignments are discussed in the paper and recommendations and suggestions are provided to prevent failures and assure adequate robustness of launching units. Owing to the legal proceedings surrounding such events, only general causes of collapse are discussed in this paper and no photographs of gantry collapses are included, and the photographs do not refer to collapsed units.

We tested here the hypothesis that calcitonin and glucocorticoids, known to modulate bone metabolism, could have opposite actions on bone cells regulating expression of cytokine receptor activator of nuclear factor-κB ligand (RANKL) and osteoprotegerin (OPG). In the U2OS osteosarcoma cell line, calcitonin (10−11to 10−9mol/L) reduced RANKL and augmented OPG both at the mRNA and protein levels. Cell incubation with prednisolone (10−8to 10−6mol/L), the glucocorticoid chosen for this study, produced opposite results. These molecular studies prompted more functional analyses whereby osteoclast bone resorptive activity was determined. Calcitonin (10−10mol/L) abrogated the stimulating effect of 10 ng/ml RANKL or 10−9mol/L prednisolone; similar results were obtained with OPG. Assessment of calcitonin and prednisolone effects in an in vivomodel of rheumatoid arthritis revealed partially surprising results. In fact, calcitonin not only preserved bone morphology (as assessed on day 18) in rats subjected to arthritis and treated with prednisolone (0.8 to 4 mg/kg daily from day 13) but also synergized with the steroid to elicit its antiarthritic effects. These results suggest that calcitonin could be used as a novel cotreatment to augment efficacy and reduce side effects associated with the prolonged use of steroids.

Annexin-1 is an anti-inflammatory protein that plays an important homeostatic role in innate immunity; however, its potential actions in the modulation of adaptive immunity have never been explored. Although inactive by itself, addition of annexin-1 to stimulated T cells augmented anti-CD3/CD28-mediated CD25 and CD69 expression and cell proliferation. This effect was paralleled by increased nuclear factor-κB (NF-κB), nuclear factor of activated T cells (NFATs), and activator protein-1 (AP-1) activation and preceded by a rapid T-cell receptor (TCR)–induced externalization of the annexin-1 receptor. Interestingly, differentiation of naive T cells in the presence of annexin-1 increased skewing in Th1 cells; in the collagen-induced arthritis model, treatment of mice with annexin-1 during the immunization phase exacerbated signs and symptoms at disease onset. Consistent with these findings, blood CD4+ cells from patients with rheumatoid arthritis showed a marked up-regulation of annexin-1 expression. Together these results demonstrate that annexin-1 is a molecular “tuner” of TCR signaling and suggest this protein might represent a new target for the development of drugs directed to pathologies where an unbalanced Th1/Th2 response or an aberrant activation of T cells is the major etiologic factor.

Annexin-1 is an anti-inflammatory protein that plays an important homeostatic role in innate immunity; however, its potential actions in the modulation of adaptive immunity have never been explored. Although inactive by itself, addition of annexin-1 to stimulated T cells augmented anti-CD3/CD28-mediated CD25 and CD69 expression and cell proliferation. This effect was paralleled by increased nuclear factor-κB (NF-κB), nuclear factor of activated T cells (NFATs), and activator protein-1 (AP-1) activation and preceded by a rapid T-cell receptor (TCR)–induced externalization of the annexin-1 receptor. Interestingly, differentiation of naive T cells in the presence of annexin-1 increased skewing in Th1 cells; in the collagen-induced arthritis model, treatment of mice with annexin-1 during the immunization phase exacerbated signs and symptoms at disease onset. Consistent with these findings, blood CD4+cells from patients with rheumatoid arthritis showed a marked up-regulation of annexin-1 expression. Together these results demonstrate that annexin-1 is a molecular “tuner” of TCR signaling and suggest this protein might represent a new target for the development of drugs directed to pathologies where an unbalanced Th1/Th2 response or an aberrant activation of T cells is the major etiologic factor.

Efficacious protection for future generations from HIV-1 infection through the development of prophylactic vaccines is the best hope for the millions of individuals living with the threat of HIV-1 infection. Advances in the development of non-curative chemotherapy for those already infected have changed the course of the epidemic for those with access to the drugs. However in the ten years since the advent of highly active anti-retroviral therapy, the expectancy of curative chemotherapy has been quashed, and the constant need for a next generation of drugs is evident. As our understanding of HIV-1 pathogenesis increases, it is becoming apparent that novel approaches and strategies will be required to halt the global progression of HIV-1. Immune-based therapies are being considered in the context of effective antiretroviral therapy. Such immunebased therapy must allow the induction or regeneration of HIV-1-specific T-cell responses with the potential to control viremia and purge viral reservoirs. Studies of therapy substitution, treatment interruption, therapeutic vaccines and/or cytokines and/or hormones have been carried out and are briefly summarised in this review.

Reinforced concrete structures are often conceived for a certain time span of serviceability. Due to the superposition of different kinds of loads and particularly due to the presence of aggressive substances the resistance of construction materials is insufficient in numerous cases. Hence, many structures have to be repaired before the end of their designed life span. In case of reinforced concrete structures these repair measures are not only very expensive but they also consume high amounts of energy and materials which causes strong environmental impacts. The main challenge in developing reliable concrete technologies is the capability to enhance the life span of new and already repaired structures to a reasonable maximum. When aiming this objective not only durability related material properties have to be accomplished but their environmental impact has to be minimized simultaneously. This paper evaluates different concrete technologies and materials from diverse perspectives: Durability (simulating expected life span using numerical analyses), ecology (product life cycle and environmental impact assessments) and economy (estimating life cycle costs by investment appraisals). This kind of combined analysis facilitates the efficient design of structural elements and repair measures and provides the possibility to significantly increase the life span of new and repaired concrete structures.

The role of the endogenous anti‐inflammatory mediator annexin 1 (AnxA1) in controlling polymorphonuclear leukocyte (PMN) trafficking and activation was addressed using the recently generated AnxA1 null mouse. In the zymosan peritonitis model, AnxA1 null mice displayed a higher degree (50–70%) of PMN recruitment compared with wild‐type littermate mice, and this was associated with reduced numbers of F4/80+cells. Intravital microscopy analysis of the cremaster microcirculation inflamed by zymosan (6 h time‐point) indicated a greater extent of leukocyte emigration, but not rolling or adhesion, in AnxA1 null mice. Real‐time analysis of the cremaster microcirculation did not show spontaneous activation in the absence of AnxA1; however, superfusion with a direct‐acting PMN activator (1 nM platelet‐activating factor) revealed a subtle yet significant increase in leukocyte emigration, but not rolling or adhesion, in this genotype. Changes in the microcirculation were not secondary to alterations in hemodynamic parameters. The phenotype of the AnxA1 null PMN was investigated in two in vitro assays of cell activation (CD11b membrane expression and chemotaxis): the data obtained indicated a higher degree of cellular responses irrespective of the stimulus used. In conclusion, we have used a combination of inflammatory protocols and in vitro assays to address the specific counter‐regulatory role of endogenous AnxA1, demonstrating its inhibitory control on PMN activation and the consequent impact on the inflamed microcirculation.

Toll‐like receptor 2 (TLR2) and ‐4 mediate signals from a great variety of bacterial gut products, giving the host a panel of microbe‐recognizing receptors. Under homeostatic conditions, TLRs act as protective receptors of the intestinal epithelium. When homeostasis is disrupted in diseases such as inflammatory bowel disease, TLR2 and ‐4 are deregulated. Our study demonstrates, by using a trinitrobenzene sulfonic acid‐induced colitis model of Crohn’s disease, the constitutive expression and the up‐regulation of TLR2 and ‐4 at messenger and protein levels in colon extracts, as well as in macrophages, dendritic cells, and lymphocytes from mesenteric lymphoid nodes. Vasoactive intestinal peptide (VIP) treatment induced a decrease of TLR2 and ‐4 expressions approaching ethanol control levels. Our results suggest that VIP modulation of TLR2 and ‐4 could be explained by two possible mechanisms. The first one would be the secondary reduction of TLR2 and ‐4 caused by the VIP‐mediated decrease of inflammatory mediators such as interleukin‐1β and interferon‐ γ, which synergize with bacterial products, contributing to the amplification of TLR presence in the intestine. The other possible mechanism would involve a VIP‐mediated decrease of nuclear factor‐κB, which would cause a direct down‐regulation of TLR expression. In summary, the resultant physiological effect is the decrease of TLR2 and ‐4 expressions to homeostatic levels. Our study describes for the first time the role of a peptide present in the gut microenvironment as an effective modulator of the initial steps of acute inflammation, acting at local and systemic levels and leading to the restoration of the homeostasis lost after an established inflammatory/autoimmune disease.

Infections caused by Gram‐negative bacteria constitute one of the major causes of septic shock, which results from the inability of the immune system to limit bacterial spread during the ongoing infection. In the last decade, it has been demonstrated that vasoactive intestinal peptide (VIP) and pituitary adenylate cyclase‐activing polypeptide (PACAP) are two endogenous immunopeptides, which together with three G protein‐coupled receptors (VPAC1, VPAC2, and PAC1) exert a significant, therapeutic effect attenuating the deleterious consequences of septic shock by balancing pro‐ and anti‐inflammatory factors. We have recently shown PAC1 receptor involvement in vivo as an anti‐inflammatory receptor, at least in part, by attenuating lipopolysaccharide‐induced production of proinflammatory interleukin‐6. The present study deepens in the protective role of PAC1 receptor in septic shock, elucidating its involvement in the modulation of neutrophil recruitment and in the expression of different molecular sensors such as intercellular adhesion molecule‐1, vascular cell adhesion molecule‐1, fibrinogen, serum amyloid A, and nitric oxide as important, systemic players of the development of septic shock. Our results, using a mice deficient in PAC1 and a PAC1 antagonist, show that VIP and PACAP as well as the PAC1 receptor are involved in neutrophil recruitment in different target organs, in adhesion molecules expression, and in coagulation‐related molecule fibrinogen synthesis. Thus, this study provides some important insights with respect to the involvement of PAC1 into the complexities of sepsis and represents an advantage for the design of more specific drugs complementing standard intensive care therapy in severe sepsis, confirming VIP and PACAP as candidates for multitarget therapy of septic shock.

Objective: To report in singleton pregnancies the post-procedure safety and maternal complications of early amniocenteses performed between 13 + 0 and 14 + 6 weeks of gestation and mid-trimester amniocenteses performed between 15 + 0 and 18 + 6 weeks of gestation. Methods: The study was carried out at the Prenatal Diagnosis Center, Siena University, Italy, during a 10-year period, following the Regional Protocol for Prenatal Diagnosis. Our study population included 3769 amniocenteses, 475 early and 3294 mid-trimester. Complications considered included miscarriage (immediately after the procedure and until 24 weeks of gestation), blood-stained amniotic fluid, failed cell culture, amniotic fluid leakage, preterm premature rupture of the membranes (PROM), preterm delivery and presence of neonatal talipes equinovarous. Results: Cytogenetic anomalies were found in 111 cases (2.9%), 18 occurring early and the other 93 in mid-trimester. Miscarriage occurred in two cases in the early amniocentesis group (0.4%) and in ten cases among the mid-trimester group (0.3%). The overall loss of pregnancies due to amniocentesis in this study was 0.3%. Amniotic fluid was stained in 1.2% in the early group and 0.9% in the mid-trimester group. Amniotic fluid leakage was noted in 1.4% and 1.2%, preterm PROM was noted in 3.3% and 3%, and preterm delivery occurred in 8% and 7.6%, respectively. There were no cases of failed amniotic culture and no cases of talipes equinovarous documented. Conclusions: The risks of early amniocentesis performed between 13 + 0 and 14 + 6 weeks appear to be comparable to those of mid-trimester amniocentesis and thus early amniocentesis could be offered to the parents, as an alternative to chorionic villus sampling, in order to obtain cytogenetic results earlier in pregnancy without a significantly increased risk for both mother and fetus. Further operators' experience with the method, based on long and accurate follow-up, and further studies are necessary to assess the safety of the method.

Purpose On the basis of intraocular pressure measurements and fluorophotometry we assessed the effects of 2% ibopamine eye drops on aqueous humor production in normal and glaucomatous eyes.Methods Thirty subjects (15 healthy volunteers and 15 open-angle glaucoma patients with ocular hypertension) were included in a placebo-controlled study with random assignment of treatment from masked containers. All subjects underwent ophthalmologic examinations and intraocular pressure (IOP) measurements. Fluorophotometry was done in both eyes at baseline (without treatment) and during treatment. Each subject was treated with 1 drop of 2% ibopamine in one eye and 1 drop of placebo in the fellow eye 30 minutes before fluorophotometric scans and every hour after the first instillation (for a total of 4 times). Safety was evaluated by recording adverse events and ocular symptoms and signs. Aqueous humor flow data were analyzed using the paired t-test, comparing ibopamine and placebo-treated eyes.Results No changes in IOP were detected in normal eyes, whereas glaucomatous eyes showed a mean increase of 4 mmHg (95% CI 3.46–4.51) from baseline. The difference in IOP between healthy eyes and those with glaucoma was significant (p<0.0001). In normal eyes and patients with glaucoma ibopamine led to a significant increase in aqueous humor flow compared with placebo-treated eyes (p<0.01). The safety profile of ibopamine was very good.Conclusions The results seem to confirm that ibopamine increases aqueous humor production in normal and glaucomatous eyes, raising IOP only in eyes with glaucoma.

AbstractBackground:Recently the role of bacteria in acute exacerbations of chronic bronchitis (AECB) as well as antibiotic treatment with selected drugs, especially fluoroquinolones, have been better defined. Objective:To assess the efficacy and safety in patients with AECB of prulifloxacin in comparison with ciprofloxacin. Methods:AECB was defined according to the guidelines for the evaluation of new anti-infective drugs for the treatment of respiratory tract infections (1992). 235 patients took part in the trial; 117 (88 males and 29 females, mean age 64.8 years) received 600 mg prulifloxacin once daily and 118 (91 males and 27 females, mean age 64.5 years) 500 mg ciprofloxacin twice a day, for a duration of 10 days. The study design was randomized, multicenter, double-blind, double-dummy. Efficacy evaluations were performed by comparing pretreatment and posttreatment assessments. The clinical response was determined by 4-point rating scores on cough, dyspnea, and expectoration (volume and appearance). The microbiological response was assessed on sputum specimen. Results:Clinical success was observed in 84.7 and 85% of patients in the prulifloxacin and ciprofloxacin groups, respectively. The 95% confidence interval proved the equivalence of treatments. Both drugs successfully eradicated the most commonly isolated strains, including Haemophilus influenzae, Streptococcus pneumoniae, Klebsiella pneumoniae andPseudomonas aeruginosa. Both treatments were well tolerated. Adverse drug reactions were always of mild or moderate intensity. Conclusion:The study showed that a 10-day course of prulifloxacin is as effective and safe as ciprofloxacin in the treatment of patients with AECB.Copyright © 2002 S. Karger AG, Basel

The non-obese diabetic (NOD) mouse model of autoimmune sialadenitis offers the possibility of studying the l-arginine/nitric oxide signaling pathway in salivary glands in basal and neurotransmitter-stimulated conditions and, thus, of analyzing the neural control of the secretory process in the target organ. The purpose of this study was to explore putative alterations in the activity and expression of nitric oxide synthase (NOS) in submandibular glands of NOD mice in relation to parotid glands and unrelated tissues. Here we report that NOD mice with incipient signs of secretory dysfunction presented a marked decrease in basal and vasoactive intestinal peptide (VIP)-stimulated NOS activity and a differential expression of NOS I in submandibular glands compared to control BALB/c mice. Similar alterations in NOS I were found in parotid glands but not in brain or spleen of NOD mice. No differences between NOD and controls appeared in NOS II and NOS III expression in any of the tissues studied.

Epidemiological studies support the involvement of short-chain fatty acids (SCFA) in colon physiology and the protective role of butyrate on colon carcinogenesis. Among the possible mechanisms by which butyrate may exert its anti-carcinogenicity an antioxidant activity has been recently suggested. We investigated the effects of butyrate and mixtures of SCFA (butyrate, propionate and acetate) on DNA damage induced by H2O2 in isolated human colonocytes and in two human colon tumour cell lines (HT29 and HT29 19A). Human colonocytes were isolated from endoscopically obtained samples and the DNA damage was assessed by the comet assay. H2O2 induced DNA damage in normal colonocytes in a dose-dependent manner which was statistically significant at concentrations over 10 μM. At 15 μM H2O2 DNA damage in HT29 and HT29 19A cells was significantly lower than that observed in normal colonocytes (P < 0.01). Pre-incubation of the cells with physiological concentrations of butyrate (6.25 and 12.5 mM) reduced H2O2 (15 μM) induced damage by 33 and 51% in human colonocytes, 45 and 75% in HT29 and 30 and 80% in HT29 19A, respectively. Treatment of cells with a mixture of 25 mM acetate + 10.4 mM propionate + 6.25 mM butyrate did not induce DNA damage, while a mixture of 50 mM acetate + 20.8 mM propionate + 12.5 mM butyrate was weakly genotoxic only towards normal colonocytes. However, both mixtures were able to reduce the H2O2-induced DNA damage by about 50% in all cell types. The reported protective effect of butyrate might be important in pathogenetic mechanisms mediated by reactive oxygen species, and aids understanding of the apparent protection toward colorectal cancer exerted by dietary fibres, which enhance the butyrate bioavailability in the colonic mucosa.

Epidemiological studies support the involvement of short-chain fatty acids (SCFA) in colon physiology and the protective role of butyrate on colon carcinogenesis. Among the possible mechanisms by which butyrate may exert its anti-carcinogenicity an antioxidant activity has been recently suggested. We investigated the effects of butyrate and mixtures of SCFA (butyrate, propionate and acetate) on DNA damage induced by H(2)O(2) in isolated human colonocytes and in two human colon tumour cell lines (HT29 and HT29 19A). Human colonocytes were isolated from endoscopically obtained samples and the DNA damage was assessed by the comet assay. H(2)O(2) induced DNA damage in normal colonocytes in a dose-dependent manner which was statistically significant at concentrations over 10 microM. At 15 microM H(2)O(2) DNA damage in HT29 and HT29 19A cells was significantly lower than that observed in normal colonocytes (P < 0.01). Pre-incubation of the cells with physiological concentrations of butyrate (6.25 and 12.5 mM) reduced H(2)O(2) (15 microM) induced damage by 33 and 51% in human colonocytes, 45 and 75% in HT29 and 30 and 80% in HT29 19A, respectively. Treatment of cells with a mixture of 25 mM acetate + 10.4 mM propionate + 6.25 mM butyrate did not induce DNA damage, while a mixture of 50 mM acetate + 20.8 mM propionate + 12.5 mM butyrate was weakly genotoxic only towards normal colonocytes. However, both mixtures were able to reduce the H(2)O(2)-induced DNA damage by about 50% in all cell types. The reported protective effect of butyrate might be important in pathogenetic mechanisms mediated by reactive oxygen species, and aids understanding of the apparent protection toward colorectal cancer exerted by dietary fibres, which enhance the butyrate bioavailability in the colonic mucosa.

An epidemiological study comparing speech audiometry with self-assessed hearing disability and an analysis of other factors influencing the quality of life was conducted. In the Veneto region (Italy), a representative sample of 2700 independently living individuals of 65 years of age and older was selected for the study. All participants were administered a comprehensive questionnaire and a brief examination at their home, including a general physical examination, speech audiometry, Sanders' Speech Disability test, part I and 111, Mini Mental State Examination, CES-D scale for depression, visual acuity, self-reported diseases and physical function. Auditory function was worst in the older individuals: auditory performance was within acceptable limits up to the 75-79 age group, while it rapidly deteriorates in the older groups. This trend is consistent with self-reported auditory disability (Sanders' test). A detailed analysis of the type of errors made in the speech audiometry was conducted for each subject. Speech audiometry is a good indicator of real hearing difficulties faced by the elderly, and it might be preferred to pure-tone audiometry, since hearing deficits with age are not always limited to an increased detection threshold, but include other aspects of hearing such as distortion of sounds, comprehension of speech and noise discrimination.

The present study has evaluated the use of distortion product otoacoustic emission (DPOAE) responses in the detection of cisplatin-induced ototoxicity in a Sprague Dawley rat animal model. The cisplatin was administered as a 16 mg/kg, dose introduced by a slow 30-min intraperitoneal infusion. Data from three DP-gram protocols, DPOAE input-output responses at 8 kHz, and auditory brainstem responses (ABRs) at 8, 12 and 16 kHz were collected before and 72 h after treatment. The post-treatment ABRs at 16 kHz showed the greatest mean threshold shift of 33.6 dB. The post-treatment DP-gram data showed significant reduction of the signal to noise ratios in the majority of the frequencies tested, across all tested protocols. The data suggest that the most sensitive DPOAE procedure for the early detection of the cisplatin-induced ototoxic damage is the DPOAE I/O protocol. Morphological analyses indicated that the inner hair cells remained intact, while several types of alterations were observed in the arrangement of the stereocilia in the outer hair cells.

The DNA-damaging ability of benzene and its metabolites on peripheral blood mononuclear cells (PBMC) has been investigated by using the alkaline comet assay. The PBMC were incubated with different compounds in two different media for 2 and 24 hr at concentrations that did not affect cell viability and the DNA damage was quantified by a computerized image analysis system. Benzene and phenol (5 mM) did not show any genotoxic activity after 2 hr of incubation in the two media tested, phosphate-buffered saline (PBS) and RPMI containing 5% of heat-inactivated fetal calf serum (RPMI + 5% FCS), whereas phenol was genotoxic and cytotoxic at 10 mM after 24 hr of incubation in RPMI + 5% FCS. All other benzene metabolites were genotoxic at micromolar concentrations when incubated in PBS with the following decreasing order of potency: benzenetriol, catechol, hydroquinone, and benzoquinone. When the PBMC were incubated in RPMI + 5% FCS, the effect of catechol (200–600 μM) and benzenetriol (10 μM) was reduced, whereas the genotoxicity of benzenetriol at high concentrations (50–100 μM) and hydroquinone (150–2500 μM) was not affected. In contrast, the effect of benzoquinone at 5 and 10 μM was greatly enhanced when the cells were incubated in RPMI + 5% FCS. This effect resulted mainly from the presence of serum in the medium and it was almost completely inhibited by boiling the serum (100°C, 5 min) and was partially reduced by extensive dialysis. Benzoquinone was the most damaging compound when tested under more physiological conditions, thereby supporting the general observation that it is the most myelotoxic benzene metabolite. Environ. Mol. Mutagen. 37:1–6, 2001 © 2001 Wiley-Liss, Inc.