Role of Recent and Remote Context Exposures on Incubation of Fear Memories

Studying learning and memory through the methods of Pavlovian fear conditioning has been a topic of behavioral neuroscience research for decades. Anxiety disorders such as post-traumatic stress disorder can be modeled in rodents through fear conditioning. This study took on a different approach to study the learning and recall of a fear memory by using a within subjects group. This group was tested at both a recent and remote interval. The recent timepoint was three days after conditioning and the remote timepoint was thirty-one days after conditioning. The timepoints are used to observe the effect the passage of time has on a fear memory, and multiple tests are used to model exposures someone with PTSD might experience. The time spent between acquisition and testing and the number of tests did have an effect on the percent freezing of each group. Remotely tested animals had higher freezing levels than the recently tested group. The group tested twice had some animals increasing in freezing and others decreasing in freezing at the later test point. To investigate the neural correlates of these differences c-Fos was stained for in the Basolateral amygdala complex to signify neuronal activation, and GAD-65 for neuronal inhibition

Remarkable stability of an instability-prone lentiviral vector plasmid in Escherichia coli Stbl3

Large-scale production of plasmid DNA to prepare therapeutic gene vectors or DNA-based vaccines requires a suitable bacterial host, which can stably maintain the plasmid DNA during industrial cultivation. Plasmid loss during bacterial cell divisions and structural changes in the plasmid DNA can dramatically reduce the yield of the desired recombinant plasmid DNA. While generating an HIV-based gene vector containing a bicistronic expression cassette 5′-Olig2cDNA-IRES-dsRed2-3′, we encountered plasmid DNA instability, which occurred in homologous recombination deficient recA1 Escherichia coli strain Stbl2 specifically during large-scale bacterial cultivation. Unexpectedly, the new recombinant plasmid was structurally changed or completely lost in 0.5 L liquid cultures but not in the preceding 5 mL cultures. Neither the employment of an array of alternative recA1 E. coli plasmid hosts, nor the lowering of the culture incubation temperature prevented the instability. However, after the introduction of this instability-prone plasmid into the recA13E. coli strain Stbl3, the transformed bacteria grew without being overrun by plasmid-free cells, reduction in the plasmid DNA yield or structural changes in plasmid DNA. Thus, E. coli strain Stbl3 conferred structural and maintenance stability to the otherwise instability-prone lentivirus-based recombinant plasmid, suggesting that this strain can be used for the faithful maintenance of similar stability-compromised plasmids in large-scale bacterial cultivations. In contrast to Stbl2, which is derived wholly from the wild type isolate E. coli K12, E. coli Stbl3 is a hybrid strain of mixed E. coli K12 and E. coli B parentage. Therefore, we speculate that genetic determinants for the benevolent properties of E. coli Stbl3 for safe plasmid propagation originate from its E. coli B ancestor

Gankyrin: An intriguing name for a novel regulator of p53 and RB

SummaryThe RB and p53 tumor suppressors lie at the heart of cancer biology, and inactivation of both pathways is seemingly essential for tumor development. Previous studies identified gankyrin as a component of the 26S proteasome that is consistently overexpressed in liver cancer and promotes cell transformation by binding RB. In the current issue of Cancer Cell, Fujita and colleagues (Higashitsuji et al., 2005) show that gankyrin also binds MDM2 and facilitates its destruction of p53. These important findings implicate gankyrin as a dual-purpose negative regulator of RB and p53, thereby identifying gankyrin as a rational cancer therapeutic target

Taste and olfaction in middle ear surgery

OBJECTIVE: The aim of this study was to assess pre- and postoperative taste ability in patients undergoing middle ear surgery for otosclerosis or chronic otitis media. Olfactory function was also evaluated to rule out taste deficits due to concomitant nasal pathology. METHODS: All patients underwent ear, nose, and throat examination, otomicroscopy, nasal endoscopy, anterior rhinomanometry, taste testing, and olfactory testing. Patients were evaluated at 1 to 5 days preoperatively (T0), and at 1 (T1), 6 (T6), and 12 (T12) months postoperatively. RESULTS: Both groups of patients experienced worsening of the mean taste threshold postoperatively. This phenomenon was more serious in poststapedotomy patients. Follow-up showed progressive improvement in both groups. All values of olfactory testing were within the normal range for otosclerosis patients. Patients with chronic otitis media showed variable postoperative findings. CONCLUSION: Chorda tympani function can be negatively affected by middle ear surgery. Deficits may be more marked in stapedotomy patients than in those undergoing tympanoplasty. Postoperative recovery of taste is satisfactory, although with different timelines for the 2 types of patholog

Nasal histamine responses in nonallergic rhinitis with eosinophilic syndrome

Background: Nonallergic rhinitis with eosinophilic syndrome (NARES) is persistent, without atopy, but with ≥25% nasal eosinophilia. Hypereosinophilia seems to contribute to nasal mucosa dysfunction. Objectives: This analytical case-control study aimed at assessing the presence and severity of nonspecific nasal hyperactivity and at finding out whether eosinophilia may be correlated with the respiratory and mucociliary clearance functions. Materials: The symptom score was assessed in 38 patients and 15 controls whose nasal smear was also tested for eosinophils and mucociliary transport (MCT). Nonspecific nasal provocation tests (NSNPT) with histamine were also carried out, and total nasal resistance (TNR) was determined. Results: The symptom score of NARES after NSNPT were not significantly different from the control group, and there was poor or no correlation among the single symptoms and the differences studied for every nasal reactivity class. This correlation improved when using the composite symptom score. The most severe eosinophilia was observed in high reactivity groups, and it was correlated with an increase in TNR. MCT worsened as eosinophilia and nasal reactivity increased. Unlike controls, a significant correlation was observed between the increase in MCT and TNR. Conclusions: In NARES, nonspecific nasal hyperreactivity is the result of epithelial damage produced by eosinophilic inflammation, which causes MCT slow down, an increase in TNR, and nasal reactivity classes, with possible impact on classification, prognosis, and treatment control

Pathological and cytological changes of the nasal mucosa in acute rhinosinusitis: the role of hyaluronic acid as supportive therapy

OBJECTIVE: The aim of this study was to evaluate the reparative role of hyaluronic acid in acute rhinosinusitis (ARS). PATIENTS AND METHODS: 48 patients affected by ARS were submitted to nasal endoscopy, nasal cytology, mucociliary transport evaluation (MCTt) and visual analogue scale questionnaire (VAS) at T0, after 14-18 days (T1) and after 30-35 days (T2). The patients were randomized into two groups, A and B, and received Levofloxacin and Prednisone. Moreover, using a nebulizer ampoule for nasal douche, Group A received high molecular weight Sodium Hyaluronate (3%) plus saline solution (NaCl 0.9%) twice a day for 30 days; Group B received saline solution twice a day for 30 days. RESULTS: At T0 only the VAS score showed differences regarding nasal discharge and post-nasal drip. At T1, in Group A MCTt and the number of bacteria were significantly lower than in Group B. The VAS score showed improvement in Group A. At T2 in Group A, MCTt and number of neutrophils were significantly lower than in Group B. The VAS score showed statistically significant differences between the two groups regarding nasal discharge. CONCLUSIONS: In ARS patients sodium hyaluronate plus saline solution significantly improved symptoms, MCT time and reduced neutrophil count on nasal cytology

Clinic manifestations in granulomatosis with polyangiitis

Granulomatosis with polyangiitis (GPA), formerly Wegener's granulomatosis (WG), is an uncommon immunologically mediated systemic small-vessel vasculitis that is pathologically characterised by an inflammatory reaction pattern (necrosis, granulomatous inflammation and vasculitis) that occurs in the upper and lower respiratory tracts and kidneys. Although the aetiology of GPA remains largely unknown, it is believed to be autoimmune in origin and triggered by environmental events on a background of genetic susceptibility.In Europe, the prevalence of GPA is five cases per 100,000 population, with greater incidence in Northern Europe. GPA can occur in all racial groups but predominantly affects Caucasians. Both sexes are affected equally. GPA affects a wide age range (age range, 8-99 years).Granulomatosis with polyangiitis is characterised by necrotising granulomatous lesions of the respiratory tract, vasculitis and glomerulonephritis. Classically, the acronym ELK is used to describe the clinical involvement of the ear, nose and throat (ENT); lungs; and kidneys. Because the upper respiratory tract is involved in 70-100% of cases of GPA, classic otorhinolaryngologic symptoms may be the first clinical manifestation of disease. The nasal cavity and the paranasal sinuses are the most common sites of involvement in the head and neck area (85-100%), whereas otological disease is found in approximately 35% (range, 19-61%) of cases.Diagnosis of GPA is achieved through clinical assessment, serological tests for anti-neutrophil cytoplasmic antibodies (ANCA) and histological analysis. The 10-year survival rate is estimated to be 40% when the kidneys are involved and 60-70% when there is no kidney involvement.The standard therapy for GPA is a combination of glucocorticoids and cyclophosphamide. In young patients, cyclophosphamide should be switched to azathioprine in the maintenance phase.A multidisciplinary approach, involving otorhinolaryngologists, oral and maxillofacial surgeons, oral physicians, rheumatologists, renal and respiratory physicians, and ophthalmologists, is necessary for the diagnosis and therapeutic treatment of GPA. ENT physicians have a determining role in recognising the early onset of the disease and starting an appropriate therapy

Preparation and characterization of engineered antigen presenting cells (APCs) for the generation of cytotoxic response for transplantation-related immunotherapy

Expansion and activation of cytotoxic T lymphocytes (CTL) is one of the major goals of immunotherapy but the hurdles in generating a large number of antigen-specific CTL are manifold. One of the limiting factors is the scarce availability of antigen presenting cell (APC) for T cell priming; dendritic cells (DCs) are the best APCs but their generation from adult progenitors is difficult and expensive so there is currently an active research into suitable alternatives. In this project, three different APCs models, cellular and acellular, were prepared and characterized and their potential use for clinical immunotherapy was explored. One setup was acellular, liposome-based, prepared from material currently used in in vivo applications and was tested in vitro both in human and murine settings. The other two APC systems were cellular-based; in both cases the antigen-presenting potential of a specific cord blood (CB)-derived cell population was evaluated in vitro. In the first case, a DCs-autologous responder culture was set up using whole frozen CB samples as starting material and tested in a peptide-specific model. In the second case, the antigenpresenting potential of γδ T cell in CB samples was evaluated and compared to the same population in adult, as γδ T cells have been recently shown to express antigen presentation-related markers and promote T cell proliferation in adult systems. All the three models were assessed for their practicality of preparation and use, antigen presenting capability and overall feasibility of employment in clinical settings

Nasal hypersensitivity in purulent middle ear effusion

The existence of a physiopathologic connection between nose and middle ear is widely accepted so that chronic purulent middle ear effusion (CPMEE) could be expected to be usually associated with nasal chronic disease or impaired function. Nevertheless such association is less frequently observed in clinical practice than one could expect, possibly because of inadequate nasal function evaluation. Thirty-five patients affected by CPMEE were included in this study in order to assess the incidence of nasal disorders. E.N.T. clinical history was obtained and E.N.T. physical examination, nasal endoscopy by fiberoptics, anterior rhinorheomanometry, non-specific nasal provocation test with histamine, mucoliary transport test, and allergic skin tests were performed. In the clinical history assessment 26 patients were affected by chronic rhinopathies, 16 by chronic pharyngitis, and 20 by frequent headache. At rhinoscopy we registered nasal septum deviation in 24 cases and mean and inferior turbinates hypertrophy in 31 cases. CPMEE and nasal septum deviation or turbinates hypertrophy were more frequently omolateral (p < .001 and p < .05, respectively). Total nasal resistance was 0.99 ± 0.49; it was abnormally high in 11 subjects bilaterally and in 4 subjects monolaterally and increased significantly in 32 patients following nasal provocation test. Mucociliary transport time was longer in CPMEE subjects than in 10 healthy subjects (18 ± 5 vs 13 ± 4 min; p < .05). Finally 10 patients presented positive skin tests. On the whole, 96% of non allergic patients included in this study showed signs of non-specific nasal hypersensitivity which could theoretically cause purulent middle ear effusion to chronicize. Indeed recurrent histamine release in response to specific and/or aspecific stimuli could cause the obstruction of the Eustachian tube and consequently inadequate middle ear ventilation