Modeling the Iatrogenic Pancreatic Cancer Risk After Islet Autotransplantation in Mouse.

Iatrogenic pancreatic cancer metastasis after islet infusion is a potential risk of islet autotransplantation performed after pancreatectomy. To model this risk, islets and/or pancreatic exocrine clusters obtained from a genetically engineered mouse model for pancreatic ductal adenocarcinoma (the LSL-KrasG12D/+ ;LSL-Trp53R172H/+ ;Pdx-1-Cre, termed KPC mouse) were transplanted via the portal vein in syngeneic wild type (WT) severely diabetic recipients in the following treatment groups: group A (n = 11) received KPC exocrine clusters in volume equal to 250 islet equivalents (IEQs); group B (n = 12) received 250 WT IEQs mixed with KPC exocrine clusters (1:1 volume ratio); group C (n = 5) received 250 KPC IEQs, and group D (n = 7) received 250 WT IEQs. The incidence of hepatic metastasis was assessed by magnetic resonance imaging and histology over the 13 months of follow-up. Overall survival was not different in the four groups. No mice developed liver metastases during the follow-up. Two mice developed spontaneous tumors: a liver hepatocellular tumor in group A and a malignant lymphoma in group D. Islets and/or exocrine clusters obtained by KPC mouse, a model that develops pancreatic cancer with 100% penetrance, do not retain the same risk of tumor development when transplanted via the portal vein in a syngeneic diabetic recipient.

Stem cells to restore insulin production and cure diabetes.

BACKGROUND: The advancement of knowledge in the field of regenerative medicine is increasing the therapeutic expectations of patients and clinicians on cell therapy approaches. Within these, stem cell therapies are often evoked as a possible therapeutic option for diabetes, already ongoing or possible in the near future. AIM: The purpose of this document is to make a point of the situation on existing knowledge and therapies with stem cells to treat patients with diabetes by focusing on some of the aspects that most frequently raise curiosity and discussion in clinical practice and in the interaction with the patient. In fact, at present there are no clinically approved treatments based on the use of stem cells for the treatment of diabetes, but several therapeutic approaches have already been evaluated or are being evaluated in clinical trials. DATA SYNTHESIS: It is possible to identify three large potential application fields: 1) the reconstruction of the ß cell mass; 2) the immunomodulation in type 1 diabetes (T1D); 3) the treatment of complications. In this study we will limit the discussion to approaches that have the potential for clinical translation, deliberately omitting aspects of basic biology and preclinical data. Also, we intentionally omit the treatment of the complications that will be the subject of a future document. Finally, an overview of the Italian situation regarding the storage of cord blood cells for the therapy of diabetes will be given.

Physical Exercise Improves Total Antioxidant Capacity and Gene Expression in Rat Hippocampal Tissue.

Exercise may exert beneficial effects on cognitive functions and play an important role in the prevention of neurodegenerative diseases. Such effects seem to be mediated by changes in anti-oxidative status, but limited information is available on the nature of molecular pathways supporting the antioxidant effects of exercise in the brain. In this study 3-5-month-old male Wistar albino rats were subjected to three times/week moderate intensity exercise on a rodent treadmill for a period of 6 weeks. The tissue antioxidant activity towards various reactive oxygen species (ROS) was determined in the hippocampus. In addition, to identify the molecular pathways that may be involved in ROS metabolism, the expression of nerve growth factor (NGF) and sirtuins (SIRT1 and SIRT3) were measured. Our results showed a higher antioxidant activity in the hippocampus of physically trained rats compared to sedentary controls. Furthermore, exercise induced an up-regulation of NGF, possibly related to an improved redox balance in the hippocampus. These results suggest that physical exercise might prevent age-induced oxidative damage in the hippocampus.

Harm aversion explains utilitarian choices in moral decision-making in males but not in females.

In recent years, a great deal of research has relied on hypothetical sacrificial dilemmas to investigate decision-making processes involved in pro-social utilitarian choices. Recent evidence, however, has suggested that moral sacrificial choices may actually reflect reduced harm aversion and antisocial dispositions rather than an utilitarian inclination. Here, we used moral dilemmas to confront healthy volunteers with controversial action choices. We measured impulsiveness and venturesomeness personality traits, which have been shown to influence harm aversion, to test their role in utilitarian action and evaluation of moral acceptability. The results of the present study show that, in males, venturesomeness drives engagement in actions and increases moral acceptability. In contrast, in females no effects of venturesomeness were observed on moral action and evaluation. Rather, in females empathetic concern and personal distress, elicited by the vicarious experience of the other's emotional states, exerted an inhibitory effect on action. Taken together, these findings indicate that the "harm aversion hypothesis" may contribute to explain utilitarian choices in males but not in females. In both genders, no association was observed between impulsiveness and moral action.

Brain connectivity is altered by extreme physical exercise during non-REM sleep and wakefulness: indications from EEG and fMRI studies.

Brain connectivity is associated to behavioral states (e.g. wake, sleep) and modified by physical activity although, to date, it is not clear which components (e.g. hypothalamus-pituitary-adrenal axis hormones, cytokines) associated to the exercise are involved. In this pilot study, we used extreme exercise (UltraTriathlon) as a model to investigate physical-activity-related changes of brain connectivity. We studied post-race brain synchronization during wakefulness and sleep as well as possible correlations between exercise-related cytokines/hormones and synchronization features. For wakefulness, global synchronization was evaluated by estimating from fMRI data (12 athletes) the brain global connectivity (GC). GC increased in several brain regions, mainly related to sensory-motor activity, emotional modulation and response to stress that may foster rapid exchange of information across regions, and reflect post-race internally-focused mental activity or disengagement from previous motor programs. No significant correlations between cytokines/hormones and GC were found. For sleep (8 athletes), synchronization was evaluated by estimating the local-(cortical) and global-related (thalamo- cortical) EEG features associated to the phenomenon of Sleep Slow Oscillations (SSO) of NREM sleep. Results showed that: power of fast rhythms in the baseline preceding the SSO increased in midline and parietal regions; amplitude and duration of SSOs increased, mainly in posterior areas; sigma modulation in the SSO up state decreased. In the post race, IL-10 positively correlated with fast rhythms baseline, SSO rate and positive slope; IL-1ra and cortisol inversely correlated with SSO duration; TNF-α and C-reactive protein positively correlated with fast rhythm modulation in the SSO up state. Sleep results suggest that: arousal during sleep, estimated by baseline fast rhythms, is increased; SSO may be sustained by cortical excitability, linked to anti-inflammatory markers (IL-10); thalamo-cortical entrainment, (sigma modulation), is impaired in athletes with higher inflammatory markers.

Quantification of CatSper1 expression in human spermatozoa and relation to functional parameters.

STUDY QUESTION: Is CatSper1 expression in human spermatozoa related to semen parameter values and sperm functions? SUMMARY ANSWER: CatSper1 expression is positively related to progressive and hyperactivated (HA) motility, [Ca(2+)]i responsiveness to progesterone but not the acrosome reaction (AR). WHAT IS KNOWN ALREADY: The role of cationic channel of sperm (CatSper) in sperm functions is clear in animal models but less defined in human sperm cells. Current knowledge is mostly based on low specificity CatSper inhibitors showing agonistic and toxic effects on human spermatozoa and is thus of little help in clarifying the role of the CatSper channel in human sperm functions. STUDY DESIGN, SIZE, DURATION: CatSper1 protein expression was evaluated in 115 men undergoing semen analysis for couple infertility. CatSper1 expression was related to routine semen parameters, motility kinematic parameters and basal and progesterone-stimulated [Ca(2+)]i and the AR. PARTICIPANTS/MATERIALS, SETTING, METHODS: CatSper1 expression was evaluated (n = 85 normozoospermic, n = 30 asthenozoospermic patients) by immunofluorescence coupled to flow cytometry leading to quantitative measurement of the percentage of ejaculated sperm cells expressing the protein. Semen analysis was evaluated according to World Health Organization guidelines. Kinematic parameters were evaluated by a computer-aided sperm analysis system. [Ca(2+)]i was measured by a spectrofluorimetric method in fura-2-loaded spermatozoa. The AR was evaluated in live sperm cells by fluorescent-labeled lectin. MAIN RESULTS AND THE ROLE OF CHANCE: CatSper1 protein expression in spermatozoa was reduced in asthenozoospermic men (mean ± SD: 53.0 ± 15.5%, n = 30 versus 67.9 ± 17.1% in normozoospermic, n = 85, P < 0.01) and was significantly correlated with progressive (r = 0.36, P < 0.001), total (r = 0.35, P < 0.001) and HA (r = 0.41, P < 0.005) motility. In addition to a higher percentage of spermatozoa not expressing CatSper1, asthenozoospermic men showed a large number of spermatozoa with immunofluorescent signal localized outside the principal piece compared with those in normozoospermia. A significant positive correlation was found between CatSper1 protein expression and the increase of [Ca(2+)]i in response to progesterone (r = 0.36, P < 0.05, n = 40) but not with basal [Ca(2+)]i. No correlation was found with the AR, either basal or in response to progesterone. LIMITATIONS, REASONS FOR CAUTION: The study is partly descriptive. Furthermore, we cannot rule out the possibility that some round cells remain after a single round of 40% density gradient centrifugation or that this step may have removed some defective or slow swimming sperm, and therefore this preparation may not be representative of the entire sperm sample. Although our data suggest that CatSper1 may be a useful marker for infertility, and a possible contraceptive target, any clinical application is limited without further research. WIDER IMPLICATIONS OF THE FINDINGS: Our results demonstrate an association of CatSper1 expression with human sperm progressive and HA motility and provide preliminary evidence that lack of expression or mislocalization of CatSper1 in spermatozoa may be involved in the pathogenesis of asthenozoospermia. However, mechanistic studies are needed to confirm that the correlations between CatSper1 expression and sperm functions are causative. STUDY FUNDING/COMPETING INTERESTS: Supported by grants from Ministry of University and Scientific Research (PRIN project to E.B. and FIRB project to S.M.) and by Regione Toscana (to G.F.). L.T. was recipient of a grant from Accademia dei Lincei (Rome, Italy). The authors have no conflicts of interest to declare.

Chlamydia abortus in Cows Oviducts, Occasional Event or Causal Connection?

Fifty-seven genital tracts of regularly slaughtered culled Piedmontese cows, aged 7.4 ± 4.3 years (mean ± SD), range: 2.6-15.6 years, were grossly and microscopically examined. DNA extracted from oviducts was subjected to PCR to evaluate the presence of Chlamydia spp. The 15 PCR-positive oviducts were subjected to Sanger sequencing and showed the presence of Chamydia abortus, with an identity range between 99 and 100%. Nine of the PCR-positive samples belonged to the 24 animals with a normal macroscopic appearance of the whole genital tract (percentage of positive oviducts in normal genital tracts 9/24 = 37.5%), while six belonged to the 33 genital tracts with lesions in one or more organs (percentage of positive oviducts in pathological genital tracts 6/33 = 18.1%); of these, a single animal had salpingitis. The detection of C. abortus in bovine oviducts is of particular interest because it has never been previously investigated or reported.

An individually fitted physical barrier device as a tool to restrict the birds' spatial access: can their use alter behavioral responses?

Social interactions have been extensively studied in poultry in a variety of environmental situations. Many studies allow full social contacts between birds, but there are others in which the interactions are tested through barriers (wire mesh or glass). Thus a situation where, according to their needs, some birds can get access to physical contact with conspecifics while others cannot, would be useful to expand the testing options for social interaction studies. We developed an individual physical barrier device (IPB) that is fitted on the birds to delimit their ambulation areas by preventing them from passing across metal mesh boundaries that IPB free counterparts can easily overcome. The prototypes showing greater efficacy consisted of a small metal bar placed in the bird's back perpendicular to the sagittal plane that slightly exceeds body width, held with a harness fitted by 2 elastic fabric bands around the wings' base. To be useful, the IPB should allow natural movements and not affect the expression of behaviors (non-invasive). This study assessed whether the IPB may alter adult Japanese quail behavioral responses using 4 classical test situations: Open-Field, Runway, Time Budget in Home Box, and Mating Interactions. Open-field ambulatory behaviors were affected 1 h, but not 7 d, after IPB was fitted, suggesting that 7 d (or less) are required to habituate to the device. After that time period, IPB fitted birds showed no differences in any of the behaviors registered in the other 3 tests situations when compared to non IPB fitted birds. Findings suggest that after habituation, the IPB does not affect main behaviors in adult quail. Its application could be expanded if an IPB device is also found suitable for other poultry species.

Effects of diet, time since defecation, and drying process of the droppings on corticosterone metabolite measurements in Japanese quail.

The use of noninvasive methods for measuring fecal glucocorticoid metabolites is a useful tool for endocrine assessment particularly in studies where animals cannot be captured, when they should be sampled without disturbing their activities, and/or when welfare needs to be maximized. However, still no complete standardization exists for the methodology, and some confounding variables may play an important role affecting measurements and interpretation of results. The present study focused on whether two different diets (laying feed or seed mixture), the time since defecation (0, 4, 24, or 48 h) and the drying method of those samples (oven-dried or naturally nonoven-dried) may affect concentrations of corticosterone metabolites (CM) measured in male Japanese quail. Half of the birds were provided with plain water (control) and the other half received a corticosterone solution. Birds fed with a seed mixture exhibited higher values of CM (nanogram/gram) in droppings than quail that received a laying feed diet suggesting that diet should be carefully considered as a potential source of variation. As expected both groups exhibited higher CM concentration after corticosterone treatment. While CM concentrations increased significantly in nonoven-dried samples over time (0 < 4 < 24 = 48 h), oven-dried samples exhibited similar high CM values. At 24 and 48 h postdefecation, nonoven-dried samples had similar CM concentrations as all oven-dried samples. Drying of samples may be considered a reliable method to reduce variations due to water loss over time, facilitating comparisons up to 48 h postdefecation. This finding would allow to enhance the range of application of this noninvasive and welfare friendly method to situations where samples cannot be collected or frozen shortly after defecation.

New risks from ancient food dyes: cochineal red allergy.

This study reports an unusual case of IgE-mediated hypersensitivity to Cochineal red or Carmine red, a coloring agent of natural origin. Although the risk of anaphylactic reactions is well known, since the nineties the use of this additive seems to be nowadays on the rise. The problem of labeling of additives used in handmade food products is highlighted.

Induction therapy in adult intestinal transplantation: reduced incidence of rejection with "2-dose" alemtuzumab protocol.

The incidence of early rejection after intestinal transplantation correlates with heightened risk of graft loss and mortality. Many different induction or pre-conditioning protocols have been reported in the last 10 yr to improve outcomes; however, sepsis remains prevalent and diminishes long-term results. We recently began a "2-dose" alemtuzumab trial protocol - 15 mg at day 0 and 15 mg repeated on day 7 - with the hope of reducing our infection rate. We compared three different protocols used at our institution (daclizumab, conventional "4-dose" alemtuzumab, and "2-dose" alemtuzumab). There was a significantly lower rate of early rejection with the "2-dose" alemtuzumab protocol in our study group of mainly (88%) intestinal grafts without accompanying liver engraftment with its protective immunologic effect. Sepsis remained low. Longer follow-up will be required to evaluate the effects of this new protocol on longer-term outcomes.

Social interaction of juvenile Japanese quail classified by their permanence in proximity to a high or low density of conspecifics.

One challenge faced by ethologists in poultry welfare is helping birds to form functionally successful groups over time through the expression of appropriate behaviors. Searching for phenotypic variability, we developed in Japanese quail a density-related permanence (DRP) test that enables us to classify young birds (while in groups) according to their individual permanence in boxes containing a high or low density of confined conspecifics (HD or LD, respectively). This study addressed the question of whether contrasting DRP quail behavior may reflect underlying differences in social responses. Birds were classified at 11 d of age in an apparatus consisting of 2 boxes interconnected by a central region delimited by sliding doors. Each box contained at its distal end either 12 or 3 conspecifics confined behind a glass (high or low density, respectively). The doors were closed 9 times every 1 h, and positioning of 36 experimental birds was registered. If birds were found in the box containing high density, low density, or in the central region, they received a 1, -1, or a 0 score, respectively. Birds with final summed scores of ≥3 or ≤-3 were categorized as HD or LD, respectively. Same category groups (HD or LD) were evaluated in their home box (undisturbed) and in a resident/intruder test when 38 d old. A higher proportion of LD than HD groups (5/6 vs. 1/6, respectively) showed at least one aggressive pecking event during a 1-h trial. The LD groups also showed a higher number of aggressive pecking events than HP groups. When an unfamiliar intruder (either HD or LD) was incorporated during 5 min in the HD or LD box, LD resident quail showed shorter latencies and a higher number of aggressive pecking events toward the intruder bird than their HD counterparts. The early individual permanence in the DRP test could be considered a consequence of a different adaptability strategy for group living. This novel test could be relevant for selection programs aiming to obtain birds better suited for rearing in high-density conditions.

The insemination status and social context influences quail laying and social behavior: A novel experimental set up.

Japanese quails in wild life live in small groups with females being even solitary during the laying period. Although it is a poultry species widely used for egg production, information regarding laying behavior motivations or influencing variables is scarce. Our study focuses on evaluating along 7 d the quail laying behavior in a novel environmental set up. This set up allows the female to choose between remaining separated from a conspecific in one side of the apparatus or to voluntarily enter their space (box-mate side) and interact with it. We evaluated whether the female insemination status prior to enter the environmental set up, and the presence of a female or a male partner in the box-mate side can influence their laying and social behavior. Thus, 4 experimental groups were established. Females spent a higher (P < 0.05) percentage of time in the box-mate side than in their separated sector in all groups. In 3 of the 4 experimental groups (non-inseminated females interacting with a female or a male box-mate, and inseminated females interacting with a male box-mate) females also laid a greater percentage (≥65%, P < 0.05, in all cases) of eggs in the box-mate sector than in their separated sector. However, the group of inseminated females that interacted with a female box-mate shifted their egg distribution and laid equally between both sides of the apparatus. Aggressive social interactions were reduced (P < 0.05) throughout the testing days but this was depending upon the female insemination status and the sex of their box-mate. Findings suggest that females can change their laying side choice when they are inseminated but depending on the sex identity of their box-mate partners. Thus, providing quail female breeders with the option of laying their eggs in separated enclosures from conspecifics could be key to favor their well-being.

Cellular consequences of small supernumerary marker chromosome derived from chromosome 12: mosaicism in daughter and father.

Constitutional genomic imbalances are known to cause malformations, disabilities, neurodevelopmental delay, and dysmorphia and can lead to dysfunctions in the cell cycle. In extremely rare genetic conditions such as small supernumerary marker chromosomes (sSMC), it is important to understand the cellular consequences of this extra marker, as well the factors that contribute to their maintenance or elimination through successive cell cycles and phenotypic impact. The study of chromosomal mosaicism provides a natural model to characterize the effect of aneuploidy on genome stability and compare cells with the same genetic background and environment exposure, but differing in the presence of sSMC. Here, we report the functional characterization of different cell lines from two familial patients with mosaic sSMC derived from chromosome 12. We performed studies of proliferation dynamics, stability, and variability of these cells using fluorescent in situ hybridization (FISH), sister chromatid exchanges (SCE), and conventional staining. We also quantified the telomere-related genomic instability of sSMC cells using 3D telomeric profile analysis by quantitative-FISH. sSMC cells exhibited differences in the cell cycle dynamics compared to normal cells. First, the sSMC cells exhibited lower proliferation index and higher frequency of SCE than normal cells, associated with a higher level of chromosomal instability. Second, sSMC cells exhibited more telomeric-related genomic instability. Lastly, the differences of sSMC cells distribution among tissues could explain different phenotypic repercussions observed in patients. These results will help in our understanding of the sSMC stability, maintenance during cell cycle, and the cell cycle variables involved in the different phenotypic manifestations.

Predictors of germline status for hereditary melanoma: 5 years of multi-gene panel testing within the Italian Melanoma Intergroup.

BACKGROUND: The incidence of cutaneous melanoma is increasing in Italy, in parallel with the implementation of gene panels. Therefore, a revision of national genetic assessment criteria for hereditary melanoma may be needed. The aim of this study was to identify predictors of susceptibility variants in the largest prospective cohort of Italian high-risk melanoma cases studied to date. MATERIALS AND METHODS: From 25 Italian centers, we recruited 1044 family members and germline sequenced 940 cutaneous melanoma index cases through a shared gene panel, which included the following genes: CDKN2A, CDK4, BAP1, POT1, ACD, TERF2IP, MITF and ATM. We assessed detection rate according to familial status, region of origin, number of melanomas and presence and type of non-melanoma tumors. RESULTS: The overall detection rate was 9.47% (5.53% analyzing CDKN2A alone), ranging from 5.14% in sporadic multiple melanoma cases (spoMPM) with two cutaneous melanomas to 13.9% in familial cases with at least three affected members. Three or more cutaneous melanomas in spoMPM cases, pancreatic cancer and region of origin predicted germline status [odds ratio (OR) = 3.23, 3.15, 2.43, P < 0.05]. Conversely, age > 60 years was a negative independent predictor (OR = 0.13, P = 0.008), and was the age category with the lowest detection rate, especially for CDKN2A. Detection rate was 19% when cutaneous melanoma and pancreatic cancer clustered together. CONCLUSIONS: Gene panel doubled the detection rate given by CDKN2A alone. National genetic testing criteria may need a revision, especially regarding age cut-off (60) in the absence of strong family history, pancreatic cancer and/or a high number of cutaneous melanomas.

Effects of mometasone furoate on the quality of life: a randomized placebo-controlled trial in persistent allergic rhinitis and intermittent asthma using the Rhinasthma questionnaire.

BACKGROUND: Allergic rhinitis, especially when persistent (PER) and associated with asthma heavily impairs patients' quality of life (QoL). OBJECTIVE: This study assessed the effect of mometasone furoate nasal spray (MFNS) on the QoL of patients with PER and asthma, using the Rhinasthma questionnaire (EUDRACT n. 2007-004683-45). METHODS: Patients with moderate/severe PER and intermittent asthma were randomized to MFNS (alcohol-free) 200 µg/day or placebo for 28 days. Rhinasthma was completed at baseline and at weeks 2 and 4. The total five symptom score (T5SS) for rhinitis, the asthma symptom score and the sum of the two [global symptoms score (GSS)] were recorded daily. The primary outcome was the change in the Rhinasthma global summary (GS) at the end of treatment. Secondary end-points were (a) the change from baseline to end of treatment of each Rhinasthma factor: upper airways (UAs), lower airways (LAs) and respiratory allergy impact; (b) the change from baseline to end of treatment of the T5SS and of the GSS and (c) the use of rescue medication. RESULTS: Fifty-two adults were randomized. Compared with placebo, MFNS produced a significant change in the Rhinasthma GS (-10.4 vs. 0.4; P<0.01). MFNS also achieved a significant improvement of the UA (-16.6 vs. 0.1; P<0.001), LA (-10.8 vs. 1.1; P<0.001) and GSS (-6.7 vs. -3.1; P=0.019). The change of the T5SS was greater in the MFNS group but did not reach statistical significance. CONCLUSION: In patients with PER rhinitis and intermittent asthma, MFNS improves the QoL and the burden of respiratory symptoms. Treating rhinitis may affect the asthma-related QoL.

LH supplementation of ovarian stimulation protocols influences follicular fluid steroid composition contributing to the improvement of ovarian response in poor responder women.

In this prospective study, we evaluated the steroid levels in 111 follicular fluids (FF) collected from 13 women stimulated with FSH monotherapy and 205 FF collected from 28 women stimulated with FSH + LH because of a previous history of hypo-responsiveness to FSH. Steroid levels were measured by HPLC/MS-MS and related to ovarian stimulation protocol, oocyte maturity, fertilization and quality of blastocysts, after individually tracking the fate of all retrieved oocytes. 17-Hydroxy-Progesterone, Androstenedione, Estradiol and Estrone were significantly higher in the FSH + LH protocol. Progesterone, 17-Hydroxy-Progesterone and Estradiol were more expressed in FF yielding a mature oocyte (p < 0.01) in the FSH + LH protocol. FF Progesterone concentration was correlated with the rate of normal fertilization in the FSH protocol. None of the FF steroids measured were associated with blastocyst quality and achievement of pregnancy. Our results indicate that LH supplementation in hypo-responsive women modifies ovarian steroid production, mimicking physiological production better and likely contributing to an improved ovarian response. Employing a correct methodological procedure to evaluate the relationship between FF steroid hormones and assisted reproduction outcomes, our study reveals that some steroids in single follicles may be helpful in predicting oocyte maturity and fertilization.

Interferon alpha inhibits a Src-mediated pathway necessary for Shigella-induced cytoskeletal rearrangements in epithelial cells.

Shigella flexneri, the causative agent of bacillary dysentery, has the ability to enter nonphagocytic cells. The interferon (IFN) family of cytokines was found to inhibit Shigella invasion of cultured epithelial cells. We show here that IFN-alpha inhibits a Src-dependent signaling cascade triggered by Shigella that leads to the reorganization of the host cell cytoskeleton. Immunofluorescence studies showed that IFN-alpha inhibits Shigella-induced actin polymerization required for bacterial entry into cells. Phosphorylation of cortactin, a Src-substrate specifically tyrosyl-phosphorylated during Shigella entry, was inhibited by IFN-alpha. Overexpression of a dominant interfering form of pp60c-src led to inhibition of Shigella-induced cytoskeletal rearrangements and decreased cortactin phosphorylation indicating a role for Src in Shigella entry. Also, Shigella uptake in cells that expressed constitutively active Src was unaffected by IFN-alpha treatment. We conclude that Src kinase activity is necessary for Shigella invasion of epithelial cells and that IFN-alpha inhibits this Src-dependent signaling pathway.

Association and activation of Jak-Tyk kinases by CNTF-LIF-OSM-IL-6 beta receptor components.

A recently defined family of cytokines, consisting of ciliary neurotrophic factor (CNTF), leukemia inhibitory factor (LIF), oncostatin M (OSM), and interleukin-6 (IL-6), utilize the Jak-Tyk family of cytoplasmic tyrosine kinases. The beta receptor components for this cytokine family, gp130 and LIF receptor beta, constitutively associate with Jak-Tyk kinases. Activation of these kinases occurs as a result of ligand-induced dimerization of the receptor beta components. Unlike other cytokine receptors studied to date, the receptors for the CNTF cytokine family utilize all known members of the Jak-Tyk family, but induce distinct patterns of Jak-Tyk phosphorylation in different cell lines.

Phenobarbital-induced DiHS and ceftriaxone hypersensitivity reaction: a case of multiple drug allergy.

Patients with DiHS show an increased risk of sensitization to multiple drugs. We report a case of a young woman who developed cutaneous rash, lymphoadenopathy, malaise and fever after the introduction of phenobarbitale. Because of these symptoms, she was treated with ceftriaxone and she experienced a severe flare-up of the cutaneous and general reaction. Allergological work-up, by cutaneous and lymphocyte transformation test, confirmed a double sensitization to phenobarbital and ceftriaxone. In conclusion, the high risk of DiHS during anticonvulsive therapy should suggest caution in using additional drugs, because of an increased risk of multiple reactions.