Oligodendrocyte gene expression is reduced by and influences effects of chronic social stress in mice.

Oligodendrocyte gene expression is downregulated in stress-related neuropsychiatric disorders, including depression. In mice, chronic social stress (CSS) leads to depression-relevant changes in brain and emotional behavior, and the present study shows the involvement of oligodendrocytes in this model. In C57BL/6 (BL/6) mice, RNA-sequencing (RNA-Seq) was conducted with prefrontal cortex, amygdala and hippocampus from CSS and controls; a gene enrichment database for neurons, astrocytes and oligodendrocytes was used to identify cell origin of deregulated genes, and cell deconvolution was applied. To assess the potential causal contribution of reduced oligodendrocyte gene expression to CSS effects, mice heterozygous for the oligodendrocyte gene cyclic nucleotide phosphodiesterase (Cnp1) on a BL/6 background were studied; a 2 genotype (wildtype, Cnp1+/- ) × 2 environment (control, CSS) design was used to investigate effects on emotional behavior and amygdala microglia. In BL/6 mice, in prefrontal cortex and amygdala tissue comprising gray and white matter, CSS downregulated expression of multiple oligodendroycte genes encoding myelin and myelin-axon-integrity proteins, and cell deconvolution identified a lower proportion of oligodendrocytes in amygdala. Quantification of oligodendrocyte proteins in amygdala gray matter did not yield evidence for reduced translation, suggesting that CSS impacts primarily on white matter oligodendrocytes or the myelin transcriptome. In Cnp1 mice, social interaction was reduced by CSS in Cnp1+/- mice specifically; using ionized calcium-binding adaptor molecule 1 (IBA1) expression, microglia activity was increased additively by Cnp1+/- and CSS in amygdala gray and white matter. This study provides back-translational evidence that oligodendrocyte changes are relevant to the pathophysiology and potentially the treatment of stress-related neuropsychiatric disorders.

Results of Early Versus Intermediate or Delayed Voice Therapy in Patients With Unilateral Vocal Fold Paralysis: Our Experience in 171 Patients.

OBJECTIVES: Vocal fold paralysis can have an important impact on a patient's quality of life. The goal of this study was to compare, in terms of vocal improvement and motility recovery, the post-vocal treatment results of our patients with unilateral vocal fold paralysis (UVFP) when treatment was started early (within 4 weeks from injury) versus intermediate (from 4 to 8 weeks) or delayed (at least 8 weeks after injury) treatment. STUDY DESIGN: An 11-year retrospective study of patients with UVFP who underwent multidimensional diagnostic-therapeutic assessment. METHODS: In total, 171 patients with UVFP were included in our study, divided into three groups who underwent early (first group), intermediate (second group), or delayed (third group) voice treatment. All patients underwent voice therapy based on forcible exercises supplemented by manipulations and maneuvers. RESULTS: Of the 171 patients with UVFP, 106 (62%) recovered vocal fold motility. Of these 106 patients, 51/78 (65%) were in the first group, 30/49 (61%) in the second group, and 25/44 (56%) in the third group. A significant (P < 0.0001) reduction in fundamental frequency (Fo) was present in the first group with a manifest improvement in the mean values of Jitter (Jitt%; P = 0.001), Shimmer (Shim%; P < 0.0001), and noise-to-harmonic ratio (NHR; P < 0.0001). A significant (P < 0.0001) reduction in Fo was found in the second group with a manifest improvement in Jitt% (P < 0.001), Shim% (P < 0.0001), and NHR (P < 0.0001). For the third group, no values were statistically significant apart from the improvement in NHR (P < 0.001). CONCLUSIONS: This study confirms the importance of early rehabilitation underlining the non-functional vocal recovery in patients who started treatment later than 8 weeks after injury.

Human cytomegalovirus product UL44 downregulates the transactivation of HIV-1 long terminal repeat.

OBJECTIVE: Human cytomegalovirus (HCMV) is often isolated from HIV-1-infected patients and the two viruses can infect the same cell type giving rise to direct bidirectional interactions. Whereas the long terminal repeat (LTR) transactivation ability of HCMV immediate early gene (IE1/IE2) is well documented, no information is available on the possible role of other HCMV proteins. In this study, the activity of ppUL44, an early DNA-binding protein, on HIV LTR transactivation was investigated. METHODS: HIV LTR transactivation by ppUL44 in presence or absence of HIV-1 Tat and HCMV IE1/IE2 was determined in J-Jhan and U973 cells through transient transfection experiments with a series of different expression vectors. Some experiments were also performed on U373-MG astrocytoma cells permanently transfected with UL44 or with another HCMV gene used as a control (UL55). RESULTS: The basal transactivation activity of the HIV LTR was not influenced by the presence of ppUL44. On the contrary, the transactivation observed in the presence of Tat, IE1/IE2 or both factors in synergy was strongly downregulated by ppUL44 in a dose-dependent manner. Deletion constructs of ppUL44 demonstrated that the region of the molecule responsible for the inhibition of the LTR is located within the last 114 amino acids at the carboxyl-terminal region. CONCLUSIONS: The results obtained indicate that within the last 114 amino acids of ppUL44 there is a domain that has a negative effect on the ability of HIV-1 LTR to be activated by both its autologous transactivator Tat and the heterologous transactivator HCMV IE1/IE2 functioning individually or synergistically.

Age-dependent remodeling of rat thymus. Morphological and cytofluorimetric analysis from birth up to one year of age.

Structural and phenotypic modifications of rat thymocytes from birth up to one year of age, i.e. during maturation and at the beginning of the involutive process of the thymus are described. Since the biological significance and the mechanisms of thymic involution are still a matter of debate, this study aims at clarifying the complexity of the compensatory events occurring during this relatively neglected period of time. Thymuses from Sprague-Dawley rats were analyzed morphologically and morphometrically by light and electron microscopy. At the same time, thymocyte subsets, isolated from the same animals, were characterized by flow cytometry according to physical parameters and phenotypic markers. Results indicate that major changes occur during the first month from birth and from six months onward. In particular, already during the first weeks after birth, thymocytes undergo a slight reduction of mitoses associated with an increased number of apoptoses. Moreover, during the same period of time, flow cytometry revealed an expansion of small thymocytes and changes in thymocyte subsets such as increase of CD4+CD8+ and CD5+alpha(beta)TCR- and a decrease of CD4-CD8-, CD4-CD8+ cells. The thymus of adult rats was characterized by time-dependent decrease of both mitoses and apoptoses, progressive physical disconnection among cells, increase of necrotic areas and fibrosis. Around one year of age tissue changes were associated with a dramatic reduction of the population of large thymocytes and the rise of numerous small thymocytes that were unexpectedly negative for all tested markers. By contrast, medium-size thymocytes exhibited a marked decrease of CD4+CD8+ and CD5+alpha(beta)TCR- subsets. In conclusion, our data indicate that thymus undergoes, with time, a complex remodeling and suggest that thymic involution is not only a simple shrinkage of the organ but rather the result of a series of compensatory mechanisms among different cell populations in a setting of progressive involution.

Intracellular production of a major cytomegalovirus antigenic protein in the methylotrophic yeast Pichia pastoris.

We have previously shown that single or multiple epitopes of the major human cytomegalovirus (HCMV) antigens, produced as fusion proteins in prokaryotes can be valuable diagnostic material in the serology of HCMV infection. In this work we moved to a eukaryotic system, to produce one of the most immunogenic HCMV antigens, ppUL44 (also called pp52 due to its apparent molecular size on acrylamide gels), as a non-fusion protein, in an attempt to eliminate some non-specific reactivity of human sera with bacterial carrier proteins. We expressed the DNA encoding ppUL44 in a highly efficient expression system based on the methylotrophic yeast, Pichia pastoris. Good levels of intracellular, soluble pp52 were produced. We observed an indistinguishable pattern of the yeast pp52 from the viral native protein in immunoblotting and a good reactivity with human sera.

Avascular tympanojugular paraganglioma.

The avascular paraganglioma described in this article appears to be the second such tumor reported in the international literature and the first to be reported in the tympanojugular region. Despite a highly suggestive history and clinical appearance, the tumor showed no signs of vascularization on radiologic studies. The pathologic postoperative study confirmed the diagnosis of paraganglioma with extensive stromal fibrosclerosis and without the typical well-vascularized thin fibrous septa. In the authors' opinion, this observation is notable because of the difficulties encountered in the correct diagnostic interpretation of an avascular mass in the tympanojugular region. In such cases, the possibility of a paraganglioma should always be considered.

Efficacy and tolerability of brodimoprim in bacterial otitis media in children. Controlled study versus cefaclor.

78 pediatric patients affected by acute otitis media were selected and randomized into two balanced groups of treatment: brodimoprim, at the dosage of 200 mg once-a-day on the first day and of 100 mg once-a-day on the following days, and cefaclor at a dosage of 40 mg/Kg/day in three doses. Brodimoprim resulted more efficacious in the reduction of symptoms, especially hypoacusis and tinnitus (p < 0.05 between treatments); tympanometry showed a higher number of normalizations in the brodimoprim group, without significant differences between treatments. Both drugs resulted active against most of isolated bacterial strains. Side effects were reported in 4 patients treated with brodimoprim and in 6 patients in the control group.

Descending necrotizing mediastinitis. Diagnosis and surgical treatment.

AIM: Descending necrotizing mediastinitis (DNM) is an unusual and severe disease with a high mortality rate. Surgical management remains controversial. Our investigations reviews the most effective surgical treatment in the management of this rare pathology. METHODS: Seven patients with DNM and treated over a 20-year period are reported. All patients were evaluated according to the classification suggested by Endo et al. of the degree of mediastinal diffusion, based on CT scan findings. Five patients underwent combined cervical drainage and thoracotomy, 2 patients were treated with cervical drainage alone. RESULTS: The outcome was favorable in 5 patients, 4 treated with a combined cervical and thoracic approach and 1 with a cervical approach alone. Two patients that underwent a combinated cervical and thoracic approach alone, died of septic shock. Overall mortality rate was 28.5%. CONCLUSION: Early diagnosis and early, aggressive surgical treatment are required to improve the poor prognosis of DNM. Although a unique surgical management is still not completely accepted, we state, in agreement with other authors, a wide approach consisting of a cervical drainage and mediastinotomy in case of upper mediastinitis and a combined cervical and thoracic approach in case of lower mediastinitis. In the course of thoracotomy a wide excision of necrotic and particularly fat mediastinal tissue is needed, to avoid a recurrent infection. A continuous cervico-mediastinal irrigation system is suggested during the postoperative period.

Nasal cytology in allergic processes and other syndromes caused by hyperreactivity.

The spontaneous nasal secretions from about 128 patients with the symptomatic triad of rhinorrhea, sneezing, and nasal obstruction were studied. The cytological examination consisted of a morphological evaluation and a semiquantitative evaluation. Noncellular and nonliving presences in the smears have been considered, including mucus, Charcot-Layden crystals, and pollen grains, as well as fungi and bacilli. We found a net increase in eosinophils, goblet cells, and cellular debris in secretions of atopic patients. Pollen grains and vegetative fragments were present in the nasal smears of pollinosis. The presence of fungi was observed in five cases of allergic rhinopathy. The exfoliative cytology represents a valuable means for the differential diagnosis of rhinitis. In particular, the presence of noncellular elements has value regarding rhinocytology.

Integrin expression in middle ear cholesteatoma.

Cholesteatoma is lined by a squamous keratinizing epithelium exhibiting most of the features of normal epidermis. In this study, we investigated by immunohistochemistry the expression of integrin adhesion molecules in primary acquired and recurrent cholesteatomas, and compared it with common epidermal cysts and normal human skin. The results showed that cholesteatoma epithelium exhibited a markedly augmented expression of alpha v integrin subunit and a corresponding increased deposition of vitronectin (alpha v ligand) in the surrounding stroma as compared to epidermal cyst and normal human skin. In contrast, the expression pattern of alpha 2 beta 1, alpha 3 beta 1, and alpha 6 beta 4 integrins as well as the distribution of laminin, collagen IV and fibronectin were similar in cholesteatomas, epidermal cysts and normal human skin. Similar staining pattern was observed in primary acquired and recurrent cholesteatoma.

Construction of a polyepitope fusion antigen of human cytomegalovirus ppUL32 and detection of specific antibodies by ELISA.

We have previously shown that single linear epitopes of the major human cytomegalovirus (HCMV) antigens, expressed as fusion proteins or synthesized as oligopeptides can be valuable diagnostic material in the serology of HCMV infection (5, 6, 13). In this work we fused sequences expressing two different epitopes (aa 1005-1048 and aa 595-614) contained in the basic phosphoprotein of 150 KD coded by UL32 (1, 2), (ppUL32), which has repeatedly been shown to be the strongest immunogen present in the viral particle. The fusion protein was tested by ELISA with HCMV-positive human sera in comparison with other fusion proteins of ppUL32. We found that the double epitope fusion protein was recognised by IgM present in a larger number of sera and with more intense reactions than all the other ppUL32 fusion proteins. The double epitope reacted positively with 81.3% and, when denatured, with 94.7% of IgM-positive sera respectively. IgG reactivity was also high, reaching a percentage of 90.7.

Connective tissue in skin biopsies from patients suffering systemic sclerosis.

Little information is available on elastin during systemic sclerosis since biochemical and morphological data have primarily focused on collagen and glycosaminoglycan alterations of connective tissues in this pathological process. We performed ultrastructural, morphometric, biochemical and in situ hybridisation analyses on skin biopsies from patients affected by scleroderma and from site and age-matched control subjects. Affected skin revealed alterations in the distribution and organisation of collagen bundles and fibrils together with zonal increase of the microfibrillar component. Elastic fibres were significantly more numerous than in control skin, were more frequently vacuolated and characterised by electron-dense inclusions; moreover, morphometric analysis provided evidence for a significant increase of the percentage of both collagen bundles and elastin fibres in the measured tissue, compared to normal skin. Biochemical analysis seemed to confirm the increased elastin content per unit of dried weight tissue in sclerodermic skin. Differences observed among patients were only partially associated with disease duration and/or to severity of clinical manifestations. The abnormal amount of elastic fibres observed in skin biopsies from patients, and data from in situ hybridisation suggest the presence of a deregulation of the whole extracellular matrix that might be related to the role of cytokines such as TGF-beta, which has already been suggested to be involved in systemic sclerosis and in enhanced collagen and elastin production.

[Pre- and post-operative information for the laryngectomized patient: verification of the validity of a brochure]. / L'informazione pre e post-operatoria del laringectomizzato: verifica della validità di un opuscolo.

For those patients who are candidates for total laryngectomy, therapy is not terminated upon surgery and subsequent speech therapy. Treatment must, instead, take into consideration numerous, important psychological problems which arise upon learning the diagnosis and the need for radical surgery. Therefore, all those who come into contact with the patient during the different phases of disease (i.e. the surgeon, ward nurses, speech therapists) must all be made sensitive to such problems. At times the participation of a psychologist is required. Detailed information on both the temporary and permanent modifications due to surgery and on the possibility of obviating the changed physical status are essential to this "global" therapeutic strategy. The authors have, therefore, prepared an explanatory handbook which briefly analyzes the different problems linked to the total laryngectomy in clear, understandable language. Furthermore, they supply information regarding the new behaviour required after surgery. In order to check how valid this type of instrument is as a graphic support to the irreplaceable information given by the health care workers, a questionnaire was sent to 123 members of the Modena Section of the Laryngectomee Association. This questionnaire dealt with the handbook, the information and advice they gained from it both before and after surgery. Questions also involved what sort of speech therapy they had undergone, whether they had an electronic larynx and, if so, was it used. The judgement given on the brochure by most of those interviewed and on the timeliness of its uses from the preoperative stages on was favorable. This favorable judgment stems from the often insufficient information and psychological support provided patients who have undergone, or who will undergo, a total laryngectomy. Such conclusion is clearly drawn from the responses given to the questionnaire. Moreover, the statements given by some of the laryngectomees (even some who had undergone surgery quite some time ago) indicate that they did not know some of the information and practical advice given. Another fact that emerged from the responses is that the esophageal voice is the means of communication of choice and that the electronic larynx was only used in cases where speech therapy had failed.

[The experience of early rehabilitation]. / Esperienze di riabilitazione precoce.

Functional laryngectomies permit a more or less ideal preservation of laryngeal functions whose recovery, especially in les conservative operations, occurs very slowly and depends on several conditions: post operative course, sensitivity and motility of the hypopharynx, patient's ability to restore swallowing mechanisms. The Authors relate their experience concerning use of a rehabilitative program partially based on the experiences of some French logopedic schools and partially original. They illustrate the steps and goals of this program which starts on the fifth post-operative day with respiration exercises immediately followed by eight days of exercises to re-establish arytenoid mobilization and swallowing movements. If deglutition is not completely recovered and important inhalation problems persist, the logopedic approach is integrated with surgical rehabilitation consisting of one or more injection of gax-collagen. It is possible to use the same surgical technique later, after hospital discharge, if a slight dysphagia is still present in spite of continuous logopedic rehabilitation. Voice restoration exercises are introduced in the last days of the hospital stay when the patient is tube-free and continues at the office or outpatient clinic for two or three times every week. Concerning removal priority (tracheotomy tube followed by nasogastric tube or vice versa), we propose a diversified strategy for each patient, depending on the anatomicofunctional postoperative situation. Up to now 25 patients have taken part in this rehabilitation program (14 cricohyoidopexy, 6 Cricohyoidoepiglottopexy, 5 supraglottic laryngectomies). The results with regard to the amount of time that nasogastric feeding as well as tracheal tube are kept and the length of the hospital stay, were compared to those ones of a similar number of consecutive cases operated at our institution (ENT Department of Modena University) before February 1990 but not rehabilitated. In the early rehabilitated group, we observe a quicker functional recovery with a shorter hospital stay (about a week).

[Reconstructive subtotal laryngectomy in the treatment of laryngeal cancer]. / Le laringectomie sub-totali ricostruttive nel trattamento del cancro della laringe.

Surgical techniques of sub-total reconstructive laryngectomies can often prevent the serious impairment of total laryngectomy without having to relinquish oncological radicality. The aim of the present work has been to report on the experience in this field accrued in the ENT Department of the University of Modena from 1987 to 1992. During this period 54 subtotal laryngectomies were performed. Of these, 13 were crico-hyoido-epiglotto-pexies (C.H.E.P.) and the remaining 41 were crico-hyoido-pexies (C.H.P.). The criteria suggested in the literature was adopted for tumor evaluation, surgical indications and contraindications. All the patients had a follow-up of at least 2 years and 31 of them have had at least 5 years of follow-up. There were 9 deaths: 3 due to intervening illnesses, 2 from second primary tumors and 4 from tumor and/or node recurrences. The overall survival was 83.3% at 2 years and 77.6% at 5 years. Determinate survival (ruling out those who had died because of intervening illnesses) were 88.2% and 80%, respectively. There were 11 neoplastic repetitions of which 2 were of the primary tumor, 2 of the primary tumor plus cervical metastases, and 7 of cervical metastases alone. Recovery surgery was performed in 9 patients, 5 of whom are still alive and disease free. Functional recovery (respiration, deglutition) took place slightly earlier in C.H.E.P. than in C.H.P. but in both cases this could be shortened, particularly by introducing a rehabilitative protocol during the immediate post-operative period. In no case did it prove necessary to perform a total laryngectomy to avoid "ab ingestis" problems and only one patient has a permanent tracheostomy.

[Logopedic rehabilitation of laryngeal granulomas]. / La rieducazione logopedica dei granulomi laringei.

Posterior laryngeal granuloma is an infrequent pathology of multidisciplinary interest. Actually, its real prevalence is difficult to quantify because in some cases it is asymptomatic and in other instances it may either be reabsorbed or eliminated spontaneously. It is located at the vocal apophysis of the arytenoid or, less frequently, above it or on the laryngeal side of the arytenoid. The many etiologic factors (laryngeal intubation, gastro-esophageal refluxes, blunt trauma of the larynx, vocal dysfunction), sometimes concomitant and with the possible addition of enhancing circumstances (upper aerodigestive tract inflammation, naso-gastric tube, smoking and alcohol abuse), converge to a single pathogenetic mechanism: an ulceration of the mucosa and the pericondrium, sometimes complicated by an infection, which does not heal but instead produces a typical granulation tissue with capillaries oriented radially from the center of the lesion. Post intubation granulomas, extremely rare in children, are more frequent in females. It appears that there is no correlation with duration of intubation in that granulomas, can also occur after short general anesthesia. Idiopathic or contact granulomas are more frequent in the males. They are the result of vocal laryngeal hyperfunction, habitual throat clearing or cough-like throat clearing. Gastro-esophageal reflux of gastric juice, coughing or throat clearing may injure the mucosa. A blunt trauma of the larynx may cause a granuloma if the cartilage of the vocal process is exposed. Symptoms, when present, are dysphonia, tiredness during or after voicing, bolus, laryngeal unilateral pain, sensation of something in the throat which is mobile during breathing and swallowing, traces of blood in the expectoration. Therapeutic options are surgical, medical or logopedic. Surgery, although followed by frequent recurrences, is mandatory when the granuloma causes dispnea or if a pathologic essay is needed. Medical treatment aims at solving gastroesophageal reflux and/or inflammations of the district. Logopedic rehabilitation is the most successful therapy. Since January 1992 the Authors have been adopting the rehabilitation protocol planned by the French phoniatrician Brigitte Arnoux-Sindt for post-intubation granulomas, which, moreover, is utilyzed for all type of granulomas, including those arising during the early postoperative period after cordectomy. This protocol is analytically presented and discussed. In the cases of contact granulomas, and when there is concomitant vocal dysfunction, logopedic treatment is prolonged after granuloma dissapearance with some sessions aiming at restoring correct vocal behaviour. In all the ten patients rehabilitated up to now, granulomas disappeared after a mean of 16.3 sessions held twice a week. After several months of follow-up we had no recurrences. This clinical experience, while limited in number, seems to confirm the good results already reported in French Literature.

Partial laryngectomies: when the problem is the pexy.

To analyse the complications related to pexy, the main clinical manifestations that may raise suspicions of a pexy line rupture/detachment, the most suitable diagnostic technique and the optimum treatment to resolve this complication. This is a retrospective chart review in tertiary university referral centre. Medical charts of patients with oncological laryngeal pathologies admitted to the Otolaryngology Department of the University Hospital of Modena between May 2003 and March 2012 were analysed. Ten patients with rupture of the pexy were identified and included in the present study. The clinical manifestations were dysphagia, alteration of sensitivity of hypopharyngeallaryngeal structures, fever, infection and diastasis of surgical wounds, bleeding, dysphonia and aspiration pneumonia. Rupture of the pexy was diagnosed through endoscopic evaluations, radiological techniques or directly in the operating room during revision surgery of the earlier operation. Surgical treatments, coupled with effective swallowing rehabilitation, allowed progressive functional recovery. Patients were hospitalised until recovery of laryngeal functions was complete. In conclusion, pexy line rupture is one of the complications in the post-operative period of partial laryngectomies. Certain clinical manifestations may indicate this complication, helping the surgeon to establish an early diagnosis and administer prompt treatment.

Therapy of unilateral vocal fold paralysis with polydimethylsiloxane injection laryngoplasty: our experience.

The objective of this study was to document functional results and to compare objective and subjective voice measures after endoscopic laryngoplasty with injection of polydimethylsiloxane (PDMS) for the treatment of unilateral vocal fold paralysis, and to verify PDMS biocompatibility in vocal fold. The design used was a longitudinal prospective study. Fifteen patients with unilateral vocal fold paralysis underwent endoscopic injection of PDMS in general anesthesia. Accurate voice evaluation protocol (acoustic and aerodynamics analyses, GIRBAS [Grade, Instability, Roughness, Breathiness, Asthenia, and Strain] scale, videostrobolaryngoscopy, and Voice Handicap Index test) before, after surgery, and at follow-up time was performed. The median follow-up was 21.7 months (range, 6-35). Data obtained were statistically significant. All acoustic, aerodynamics, perceptive, and subjective evaluations showed a significant improvement. No complications due to PDMS were reported. Functional results were found comparable to framework surgery. Endoscopic injection laryngoplasty with PDMS is a safe and long-term option for treatment of unilateral vocal fold paralysis.