Assessment of CD200R Activation in Combination with Doxycycline in a Model of Melioidosis.

Antimicrobial resistance continues to be a global issue. Pathogens, such as Burkholderia pseudomallei, have evolved mechanisms to efflux certain antibiotics and manipulate the host response. New treatment strategies are therefore required, such as a layered defense approach. Here, we demonstrate, using biosafety level 2 (BSL-2) and BSL-3 in vivo murine models, that combining the antibiotic doxycycline with an immunomodulatory drug that targets the CD200 axis is superior to antibiotic treatment in combination with an isotype control. CD200-Fc treatment alone significantly reduces bacterial burden in lung tissue in both the BSL-2 and BSL-3 models. When CD200-Fc treatment is combined with doxycycline to treat the acute BSL-3 model of melioidosis, there is a 50% increase in survival compared with relevant controls. This benefit is not due to increasing the area under the concentration-time curve (AUC) of the antibiotic, suggesting the immunomodulatory nature of CD200-Fc treatment is playing an important role by potentially controlling the overactive immune response seen with many lethal bacterial infections. IMPORTANCE Traditional treatments for infectious disease have focused on the use of antimicrobial compounds (e.g. antibiotics) that target the infecting organism. However, timely diagnosis and administration of antibiotics remain crucial to ensure efficacy of these treatments especially for the highly virulent biothreat organisms. The need for early antibiotic treatment, combined with the increasing emergence of antibiotic resistant bacteria, means that new therapeutic strategies are required for organisms that cause rapid, acute infections. Here, we show that a layered defense approach, where an immunomodulatory compound is combined with an antibiotic, is better than an antibiotic combined with a relevant isotype control following infection with the biothreat agent Burkholderia pseudomallei. This approach has the potential to be truly broad spectrum and since the strategy includes manipulation of the host response it's application could be used in the treatment of a wide range of diseases.

Thermomechanical performance of continuous carbon fibre composite materials produced by a modified 3D printer.

First of all, this article aimed to evidence the role of a modified printer developed for continuous carbon fibre reinforced PolyAmide (cCF/PA6-I) together with the use of a fully open slicing step on the printing quality and the longitudinal/transverse tensile and in-plane shear properties. A comprehensive assessment of the microstructure and properties with a similar material (cCF/PA6-I), but produced with a commercial printer (i.e., Markforged® MarkTwo) has been achieved. Our customised printer and the open slicer used have made possible to better control the print conditions (i.e., layer height and distance between filaments), to reduce the porosity from more than 10% to about 2% and improve the mechanical properties. Moreover, the understanding of the behaviour of these 3D printed composites with wide-ranging external temperatures is mandatory for future use in a severe environment and/or development of new thermally active 4D printed composites. The 3D printed cCF/PA6-I composites have been then thermomechanically characterised along different printing directions (0, 90 and ± 45°) from -55 to +100 °C. Unlike the longitudinal properties that hardly change with temperature, the transverse and in-plane shear stiffness and strength of these 3D printed composites were particularly sensitive to temperature variations, with decreases of 25-30% and 30-55%, respectively. This was due to the high sensitivity of the polymer matrix, the fibre/matrix and interfilament interfaces when the composites were loaded along those directions, because damages induced by internal thermal stresses. Fractography has also been carried out to reveal damage mechanisms.

Educational Interventions for Promoting Food Literacy and Patient Engagement in Preventing Complications of Type 2 Diabetes: A Systematic Review.

The present review aims to map the current literature on educational interventions to promote food literacy in type 2 diabetes, with a particular focus on the concept of patient engagement. The systematic review was implemented on five databases with no restrictions on the publication year. The studies selected for the review were focused on patients with type 2 diabetes, ranging from 2003 to 2021 and published in 13 countries (44% USA). Thirty-three articles were analyzed. Twenty-seven articles targeted singular patients; fifteen articles conceptualized patient engagement as self-management. In seven articles, the provider is a multidisciplinary team. Twenty articles did not report a theoretical framework in the intervention development, and eleven did not use an intervention material. Twenty-six articles did not use a technology proxy. Outcome categories were narratively mapped into four areas: clinical, psychological, behavioral, and literacy. To date, most of the interventions are heterogeneous in the adopted methodology, measures, and outcomes considered. More attention should be given to the psychosocial characterization of patient engagement as well as the technological support. High-quality, randomized controlled trials and longitudinal studies are lacking and need to be conducted to verify the efficacy of these insights.

CD200R deletion promotes a neutrophil niche for Francisella tularensis and increases infectious burden and mortality.

Pulmonary immune control is crucial for protection against pathogens. Here we identify a pathway that promotes host responses during pulmonary bacterial infection; the expression of CD200 receptor (CD200R), which is known to dampen pulmonary immune responses, promotes effective clearance of the lethal intracellular bacterium Francisella tularensis. We show that depletion of CD200R in mice increases in vitro and in vivo infectious burden. In vivo, CD200R deficiency leads to enhanced bacterial burden in neutrophils, suggesting CD200R normally limits the neutrophil niche for infection. Indeed, depletion of this neutrophil niche in CD200R-/- mice restores F. tularensis infection to levels seen in wild-type mice. Mechanistically, CD200R-deficient neutrophils display significantly reduced reactive oxygen species production (ROS), suggesting that CD200R-mediated ROS production in neutrophils is necessary for limiting F. tularensis colonisation and proliferation. Overall, our data show that CD200R promotes the antimicrobial properties of neutrophils and may represent a novel antibacterial therapeutic target.

Mitogen-activated protein kinases (MAPKs) are modulated during in vitro and in vivo infection with the intracellular bacterium Burkholderia pseudomallei.

Burkholderia pseudomallei is a Gram-negative intracellular bacterium that causes the disease melioidosis. The disease can be fatal if left untreated or when antibiotic therapy is delayed and total clearance of the pathogen from the host is often not accomplished with current therapies. Thus, new therapeutic approaches for the treatment of infections caused by B. pseudomallei are required. To better understand host responses to B. pseudomallei infection, the activation of key proteins involved in the TLR inflammatory cascade was measured by western blotting. Activation of the mitogen-activated protein kinases (MAPKs) p38 and ERK were both significantly altered during both in vitro and in vivo infection. In considering an approach for therapy of B. pseudomallei infection the inhibition of ERK was achieved in vitro using the inhibitor PD0325901, along with decreased TNF-α production. However, the reduction in phosphorylated ERK and TNF-α release did not correspond with decreased bacterial replication or enhance clearance from infected macrophages. Despite this apparent lack of effect on the intracellular growth of B. pseudomallei in vitro, it is not clear what effect inhibition of ERK activation might have on outcome of disease in vivo. It may be that decreasing the levels of TNF-α in vivo could aid in reducing the overactive immune response that is known to ensue following B. pseudomallei infection, thereby increasing host survival.

Mitogen-activated protein kinases (MAPKs) are modulated during Francisella tularensis infection, but inhibition of extracellular-signal-regulated kinases (ERKs) is of limited therapeutic benefit.

Francisella tularensis is a Gram-negative intracellular bacterium that causes the disease tularemia. The disease can be fatal if left untreated and there is currently no licenced vaccine available; the identification of new therapeutic targets is therefore required. Toll-like receptors represent an interesting target for therapeutic modulation due to their essential role in generating immune responses. In this study, we analysed the in vitro expression of the key mitogen-activated protein kinases (MAPKs) p38, JNK and ERK in murine alveolar macrophages during infection with F. tularensis. The phosphorylation profile of ERK highlighted its potential as a target for therapeutic modulation and subsequently the effect of ERK manipulation was measured in a lethal intranasal F. tularensis in vivo model of infection. The selective ERK1/2 inhibitor PD0325901 was administered orally to mice either pre- or post-challenge with F. tularensis strain LVS. Both treatment regimens selectively reduced ERK expression, but only the pre-exposure treatment produced decreased bacterial burden in the spleen and liver, which correlated with a significant reduction in the pro-inflammatory cytokines IFN-γ, MCP-1, IL-6, and TNF-α. However, no overall improvements in survival were observed for treated animals in this study. ERK may represent a useful therapeutic target where selective dampening of the immune response (to control the damaging pathology seen during infection) is combined with antibiotic treatment required to eradicate bacterial infection. This combination treatment strategy has been shown to be effective in other models of tularemia.

"FoxP3 Hunting" during infection with Francisella tularensis.

Francisella tularensis is a Gram-negative intracellular bacterium that can cause acute disease in mouse models of infection when administered via the inhalational route. The immune response to a pulmonary infection is typified by an initial lack of pro-inflammatory cytokines, followed by hypercytokinemia prior to host death. It remains unclear what causes this delay in the host immune response. In this study we determine the presence of FoxP3 regulatory T cells in the lung, liver and spleen following intranasal infection with F. tularensis SCHU S4. In the lung, the site of initial infection, there is an increase in FoxP3+ cells during the first few days of infection and a notable absence of these cells at the point of cytokine storm and death (day 4 post-infection). This coincides with a decrease in the anti-inflammatory cytokine TGF-ß and increases of chemokines MIP-1α, MIP-1ß and RANTES. In our model, we also observed an overall decrease in the number of regulatory T cells in the spleen, which was not as evident in the liver. Overall, this data suggests that early on in an acute F. tularensis SCHUS4 infection regulatory T cells contribute to a dampening of the pro-inflammatory response, allowing for bacterial replication and spread.

In vitro antigen presenting cell-derived IL-10 and IL-6 correlate with Trichuris muris isolate-specific survival.

Trichuris muris, the mouse whipworm, is used as a laboratory model of the human parasite T. trichiura. Three laboratory isolates of T. muris exist - the E, J and S isolates. Previous data have shown that the S isolate survives to chronicity in C57BL/6 mice unlike the E and J isolates, which are expelled. The ability of the S isolate to persist is thought to be due to it secreting unique excretory/secretory antigens, which interact with APCs such that protective T cell responses do not develop. To determine whether APCs respond differently to E/S antigens from the three isolates we cultured isolate-specific E/S with bone marrow-derived macrophages (BMMPhi) and dendritic cells (BMDCs) in vitro. Markers of co-stimulation and levels of MHC-II were analysed by FACS and cytokine levels in supernatants quantified. E/S antigens from the S isolate consistently stimulated significantly higher levels of IL-10 and IL-6 from both macrophages (F4/80(+)CD11b(+)CD11c(-)) and dendritic cells (CD11c(+)CD11b(+)F4/80(-)) compared to J and E isolate E/S. If these in vitro differences in APC-derived cytokines, particularly IL-10, are biologically significant in vivo, they may contribute to the S isolate survival, by creating a regulatory cytokine environment in which protective immune responses are less effective.

Invasive pneumococcal infections in infants up to three years of age: results of a longitudinal surveillance in North-East Italy.

Pneumococcus is considered one of the main causes of the infections acquired in the community setting and also seems to be the most frequent cause of community-acquired pneumonia in children under 5 years of age. To establish suitable preventive measures as vaccination policy, it would be important to document the incidence of IPD. The main feature of this study was that it demanded a cooperative effort between family pediatricians and those working in the hospitals to estimate the real burden of IPD in children aged 0-36. From 1 January 2003 to 31 December 2003, a prospective active surveillance of clinical cases due to S. pneumoniae was conducted by 87 specifically-trained sentinel pediatricians [all family pediatricians] randomly selected from among those working in North-East Italy. Suspected pneumococcal infections were confirmed by blood cultures at the laboratories of the hospitals involved in the study. 32 cases were suspected, 12 of those proved positive on blood culture and 6 of these 12 confirmed cases were hospitalized. 2 were cases of meningitis, 1 of pneumonia and 9 of bacteremia. The cumulative annual incidence was 58.9 cases/100,000 infants aged 0-36 months (95% CI 30.38-102.71), meaning that North-East Italy can be classified as a mesoendemic area. This study demonstrated that the incidence of IPD in infants aged (0-36 months) is often under-estimated, documenting the importance of prospective active surveillance for assisting rational choices for public health issues.

Cardiovascular autonomic neuropathy in HIV-positive African patients.

AIM: HIV infection causes cardiac autonomic neuropathy (CAN); little is known about the relevance of CAN in sub-Sahara African patients, in spite of the highest prevalence of AIDS in that population. The authors assessed prevalence rates of CAN in HIV-positive treatment-naïve African patients and investigated the correlation between degree of immunodeficiency and CAN. METHODS: Thirty HIV-positive patients and 11 HIV-negative controls underwent a battery of cardiovascular autonomic function tests; the Ewing-Clarke score was calculated along with the stage of severity of CAN. The patients' immunological status was evaluated by CD4 T-lymphocytes counts. RESULTS: During paced respiration of normal depth, the patients showed shorter baseline RR intervals (739.2+/-136.0 vs 846.2+/-88.7 ms; P<0.05), with an inverse correlation with CD4 counts, and lower heart rate variability (85.3+/-73.0 vs 123.0+/-46.2 ms; P<0.02). Although patients with lower CD4 counts tended to present blunted response to hand-grip and cold-face tests, no linear correlation was found between results of cardiovascular reflex tests and CD4 counts. Eight patients (27%) obtained borderline Ewing-Clarke scores; 9 patients resulted affected by early (6 pts, 20%) or intermediate (3 pts, 10%) stage of CAN. CONCLUSION: Signs of HIV-related CAN are present in 30% of the African HIV+ patients observed, with no direct correlation to their immunological status. Based on the relevance of the problem and the presence of signs of CAN even in newly diagnosed and treatment-naïve patients, the authors suggest that all HIV-patients should be screened for the presence of the complication, in view of the possible serious events associated with it.

Prospective study of the burden of acute gastroenteritis and rotavirus gastroenteritis in children less than 5 years of age, in Padova, Italy.

BACKGROUND: Data on the burden of rotavirus gastroenteritis in Europe are needed to help understand the potential impact of introducing new rotavirus vaccines. MATERIALS AND METHODS: As part of prospective observational study (Rotavirus gastroenteritis Epidemiology and Viral types in Europe Accounting for Losses in Public Health and Society Study, REVEAL) conducted in 2004--2005 in seven European countries, we studied, the characteristics of acute gastroenteritis and rotavirus gastroenteritis in children less than 5 years in primary care, emergency room and hospital settings (Padova, Italy). RESULTS: A total of 757 children with acute gastroenteritis were included and enzyme-linked immunoabsorbent assay (ELISA) results were available for 725 cases. The overall estimated annual incidence for rotavirus gastroenteritis was 4.7%. Overall, rotavirus gastroenteritis was estimated to account for 43.6% of acute gastroenteritis cases. Among children with acute gastroenteritis (AGE) aged 6-23 months, 61.2% were rotavirus positive. Rotavirus gastroenteritis (RVGE) was responsible for 68.8% of hospitalizations, 61% of emergency consultations, and 33% of primary care consultations. The most prevalent serotype was G9 (84.4%) followed by G1 (11.8%). The relative risk for rotavirus gastroenteritis of being referred to hospital after an initial consultation in primary care was 3.37 (95% CI: 1.77-6.43) and 3.38 (95% CI: 2.28-5.01) for emergency room referral. Children with rotavirus gastroenteritis generally had more severe disease than children with rotavirus-negative gastroenteritis. CONCLUSION: Rotavirus accounts for a significant proportion of acute gastroenteritis cases in children less than 5 years in Italy, many of whom require frequent primary care consultations, or care in emergency room or hospital settings.

[Health promotion and education in the secondary schools]. / Educazione sanitaria nelle scuole medie: conoscenze e informazioni in tema di AIDS.

To prevent the risks of HIV infection, some meetings have been carried with the students of secondary school; actually the school is the best and the most appropriate place to conduct these health promotion and education meetings. Two questionnaires have been given to the students, a pre-test before the interview, to evaluate their knowledge about drugs, and a re-test after the interview to evaluate whether the knowledge objectives suggested had been reached. After the meeting the students appear to be more informed; differences were statistically significant between the percentages of the correct answers of the pre-test and the re-test.

[Osteoporosis: problems and considerations of economical nature]. / Osteoporosi: problematiche e considerazioni di carattere economico.

In 1991 WHO classified osteoporosis as a major social disease, on the basis of its high prevalence, expected to rise in the future, its physical and psychological consequences and its economical costs for both the society and the individual. In the past, costs, especially the undirected and intangible ones, have been largely underestimated, due to their complex quantification. A more accurate and a deeper evaluation of the pathology and its economical burden on National Health Services will lead to a better planning of prevention strategies.

Environmental risk factors for type 1 diabetes in Rome and province.

BACKGROUND: In subjects genetically susceptible to type 1 diabetes, exposure to environmental factors during the gestational period, the neonatal period, and the first years of life is thought to play an important role in triggering the immune process leading to beta cell destruction. AIMS: To investigate risk factors for inhabitants of continental Italy. METHODS: A case-control study of 150 type 1 diabetes cases and 750 control subjects (age range 6-18 years) was carried out in Rome and its province, measuring the exposure to environmental risk factors. RESULTS: Three environmental factors were found to occur significantly more in the diabetic group than in the controls. During the mothers' pregnancies, the one risk factor which proved to be higher in diabetics than in controls was maternal infectious disease. During the neonatal period, no risk factors associated with the disease were detected. During early life, eczema and a short duration of breast feeding (less than three months), occurred significantly more in diabetic cases than controls. CONCLUSION: Eczema and breast feeding for less than three months are risk factors for type 1 diabetes in a southern European population. The type, duration, and mode of treatment for infectious diseases during pregnancy need additional investigation as risk factors for type 1 diabetes.

[Health promotion and education in secondary schools: knowledge on drugs abuse]. / Educazione sanitaria nelle scuole medie: conoscenze e informazioni sulle sostanze da abuso.

To prevent behaviours at risk for the use of substances that induce dependence, some meetings have been planned with children primary school, age in which it's still possible to prevent the contact with drugs and children are more receptive. Two questionnaires have been given to the students, a pre-test before the interview, to value their knowledge about drugs, and a re-test after the interview, to value if the knowledge objectives suggested had been reached. After meeting the students seemed to be more informed because there were statistically significative differences between the percentages of the correct answers of the pre-test and the re-test.

HIV: mother to child transmission, current knowledge and on-going studies.

It is estimated that approximately 6000 women of childbearing age, mostly living in the developing world, acquire HIV infection every day. Taking into account that approximately 98% of HIV infected children have acquired HIV from the mother, during pregnancy, at delivery or through breastfeeding, therefore, prevention of mother-to-child transmission (MTCT) is a major health priority. Several studies have showed how MTCT of HIV may be prevented using antiretrovirals. Results from a study conducted in Thailand have also recently showed how a short oral zidovudine course during pregnancy and labor may reduce the risk of HIV transmission by approximately 50%. These findings represent a major challenge for the International Health Agencies and Organizations that will have the major obligation to provide HIV tests, counseling and antiviral drugs in settings with high HIV prevalence.