Role of Antioxidants in Neonatal Hypoxic-Ischemic Brain Injury: New Therapeutic Approaches.

Hypoxic-ischemic brain damage is an alarming health and economic problem in spite of the advances in neonatal care. It can cause mortality or detrimental neurological disorders such as cerebral palsy, motor impairment and cognitive deficits in neonates. When hypoxia-ischemia occurs, a multi-faceted cascade of events starts out, which can eventually cause cell death. Lower levels of oxygen due to reduced blood supply increase the production of reactive oxygen species, which leads to oxidative stress, a higher concentration of free cytosolic calcium and impaired mitochondrial function, triggering the activation of apoptotic pathways, DNA fragmentation and cell death. The high incidence of this type of lesion in newborns can be partly attributed to the fact that the developing brain is particularly vulnerable to oxidative stress. Since antioxidants can safely interact with free radicals and terminate that chain reaction before vital molecules are damaged, exogenous antioxidant therapy may have the potential to diminish cellular damage caused by hypoxia-ischemia. In this review, we focus on the neuroprotective effects of antioxidant treatments against perinatal hypoxic-ischemic brain injury, in the light of the most recent advances.

DLK1 regulates branching morphogenesis and parasympathetic innervation of salivary glands through inhibition of NOTCH signalling.

BACKGROUND INFORMATION: Delta-like proteins 1 and 2 (DLK1, 2) are NOTCH receptor ligands containing epidermal growth factor-like repeats, which regulate NOTCH signalling. We investigated the role of DLK and the NOTCH pathway in the morphogenesis of the submandibular salivary glands (SMGs), using in vitro organotypic cultures. RESULTS: DLK1 and 2 were present in all stages of SMG morphogenesis, where DLK1 inhibited both NOTCH activity and SMG branching. The addition of NOTCH inhibitory agents, either soluble DLK1 (sDLK1) or N-[N-(3, 5-difluorophenacetyl-L-alanyl]-S-phenylglycine t-buthyl ester (DAPT), to the SMG culture medium did not affect the rate of cell proliferation, but induced a strong reduction in SMG branching, increased epithelial apoptosis, and impaired innervation of the epithelial end buds by local parasympathetic ganglion neurons. SMG innervation could be restored by the acetylcholine analog carbachol (CCh), which also rescued cytokeratin 5 (CK5(+))-expressing epithelial progenitor cells. Despite this, CCh failed to restore normal branching morphogenesis in the presence of either sDLK1 or DAPT. However, it improved recovery of branching morphogenesis in SMGs, once DLK1 or DAPT were removed from the medium. CONCLUSIONS: Our data suggest that DLK1 regulates SMGs morphogenesis and parasympathetic nerve fibre outgrowth through inhibition of NOTCH signalling.

Cost-minimisation and cost-effectiveness analysis comparing transoral CO2 laser cordectomy, laryngofissure cordectomy and radiotherapy for the treatment of T1-2, N0, M0 glottic carcinoma.

The financial costs of laryngeal cancer treatment are a notable burden on healthcare budgets. In this study, we assess whether CO2 laser surgery is cheaper than radiotherapy or laryngofissure and cordectomy in the treatment of T1-2, N0, M0 glottic squamous cell carcinoma. 56 patients with a mean age of 65.88 years (SD = 10.04), 53 men and 3 women, with T1-2, N0, M0 glottic squamous cell carcinoma were retrospectively analysed. We conducted a comparative analysis of costs associated with three treatments: carbon dioxide laser cordectomy (n = 21), radiotherapy (n = 20), and laryngofissure cordectomy (n = 15). Complications of the radiotherapy and surgical treatments, need for tracheotomy and its permanence, length of hospital stay, occupation and ability to work and economic costs of treatments were recorded. Cost-minimisation and cost-effectiveness analysis were obtained. The cost of transoral laser cordectomy (2,289.79 euro) is statistically significantly lower than that of radiotherapy (4,804.72 euro) or laryngofissure cordectomy (13,229.75 euro) (p < 0.001). Transoral carbon dioxide laser surgery is the best option in terms of cost-effectiveness for the treatment of T1-2, N0, M0 glottic cancer.

An evaluation of an innovative screening program based on risk criteria for early diagnosis of head and neck cancers.

Introduction: Head and neck cancer represents 3% of all cancers and is the cause of 5% of the deaths caused by cancer. The purpose of this study is to evaluate the implementation of a screening program to diagnose the early phase of the head and neck oncological processes. Methods: We have studied 324 asymptomatic patients who had at least one major risk factor (habitual consumption of tobacco or alcohol) or two minor risk factors: family history of head and neck cancer of the upper aerodigestive tract, occupational exposure, poor oral hygiene and history of Human Papillomavirus or chronic inflammatory processes of the aerodigestive tract. Family and personal head and neck oncological medical history, ENT exploration, performance of CT scans or biopsies and program procedures were analyzed. Results: The most usual referral criteria for being sent to a specialist was being a smoker (98.1%). 10.5% reported family histories of head and neck cancer, 9.9% reported occupational exposure, 7.1% were referred due to poor oral hygiene and 5.9% were referred for gastroesophageal reflux disease. Although being asymptomatic was a requirement for inclusion, we verified that, after the anamnesis, 9.6% of the patients had some symptom to which they did not give importance to 119 patients (36.7%) presented a lesion that potentially could become malignant, located in the larynx and hypopharynx (25%) and in the oral cavity and oropharynx (10.8%). Eighteen patients (5.56%) presented more than one lesion. The detection rate of neoplasia was 1.2% and the detection rate of pre-neoplastic lesions was 4.6%. There did exist a statistically significant ratio between the detection of pre-neoplastic lesions and occupational exposure to carcinogenic agents (p = 0.006), poor oral hygiene (p = 0.01) and gastroesophageal reflux disease (p = 0.007). Samples were taken for a pathological anatomy study in 30 patients (9.25%). In order to follow up the patients, 22.8% were controlled at hospital medical consultations, 11.1% were examined at outpatient consultation and 66% were given appointments for follow-up visits. Conclusions: The use of this screening program could be a tool for the early diagnosis of malignant head and neck tumors and to foster healthy habits for cancer prevention.

Mutational profiling can identify laryngeal dysplasia at risk of progression to invasive carcinoma.

Early diagnosis of laryngeal squamous cell carcinoma (LSCC) at the stage of dysplasia could greatly improve the outcome of affected patients. For the first time we compared the mutational landscape of non-progressing dysplasia (NPD; n = 42) with progressing dysplasia (PD; n = 24), along with patient-matched LSCC biopsies; a total of 90 samples. Using targeted next-generation sequencing identified non-synonymous mutations in six genes (PIK3CA, FGFR3, TP53, JAK3, MET, FBXW7), and mutations were validated by Sanger sequencing and/or qPCR. Analysis was extended in silico to 530 head and neck (HNSCC) cases using TCGA data. Mutations in PIK3CA and FGFR3 were detected in PD and LSCC cases, as well as other HNSCC cases, but absent in NPD cases. In contrast, mutations in JAK3, MET and FBXW7 were found in NPD cases but not PD, LSCC or other HNSCC cases. TP53 was the most frequently mutated gene in both PD and NPD cases. With the exception of R248W, mutations were mutually exclusive. Moreover, five of seven PD mutations were located in motif H2 of p53, whereas none of the NPD mutations were. In summary, we propose that the mutational profile of laryngeal dysplasia has utility for the early detection of patients at risk of progression.

A comparative morphologic and morphometric study about geniculate ganglion development in man and in rat.

CONCLUSIONS: Human-rat geniculate ganglion (GG) have multiple origins: (1) An initial proximity (20 µm) to the endocranial foramen of the IAM, suggests neural crest induction; and (2) The influence of epibranchial placodes: the tensor tympani muscle (TTM) and the otic apical coil. OBJECTIVES: This study was undertaken to determine the comparative development of human-rat GG. MATERIALS AND METHODS: A light microscopic study of the GG in human material obtained from spontaneous abortions at 9, 13, 14, 17, 18, and 30 weeks, and one neonate was done. This study examined Webster rat embryos and a post-natal series. Specimens were fixed in Bouin fluid, embedded in paraffin, cut, and stained with H&E. The histomorphometric data were obtained with image analysis software. RESULTS: The human fetus of 9 weeks presents two neuronal groups in the VII nerve: one near (20 µm) the IAM endocranial foramen, foraminal, and the other, tympanic. Neonate GG is located between the TTM and the cochlear apex (inwards). In the 16 day old rat embryo GG is placed within a canal containing the stapedial artery. In the adult rat the GG and the stapedial artery are placed within the IAM.

Recent advances in cochlear hair cell regeneration-A promising opportunity for the treatment of age-related hearing loss.

The objective of this paper is to review current information regarding the treatment of age-related hearing loss by using cochlear hair cell regeneration. Recent advances in the regeneration of the inner ear, including the usefulness of stem cells, are also presented. Based on the current literature, cochlear cell regeneration may well be possible in the short term and cochlear gene therapy may also be useful for the treatment of hearing loss associated with ageing. The present review provide further insight into the pathogenesis of Inner Ear senescence and aged-related hearing loss and facilitate the development of therapeutic strategies to repair hair cells damaged by ageing. More research will be needed in order to translate them into an effective treatment for deafness linked to cochlear senescence in humans.

Biomolecular bases of the senescence process and cancer. A new approach to oncological treatment linked to ageing.

Human ageing is associated with a gradual decline in the physiological functions of the body at multiple levels and it is a key risk factor for many diseases, including cancer. Ageing process is intimately related to widespread cellular senescence, characterised by an irreversible loss of proliferative capacity and altered functioning associated with telomere attrition, accumulation of DNA damage and compromised mitochondrial and metabolic function. Tumour and senescent cells may be generated in response to the same stimuli, where either cellular senescence or transformation would constitute two opposite outcomes of the same degenerative process. This paper aims to review the state of knowledge on the biomolecular relationship between cellular senescence, ageing and cancer. Importantly, many of the cell signalling pathways that are found to be altered during both cellular senescence and tumourigenesis are regulated through shared epigenetic mechanisms and, therefore, they are potentially reversible. MicroRNAs are emerging as pivotal players linking ageing and cancer. These small RNA molecules have generated great interest from the point of view of future clinical therapy for cancer because successful experimental results have been obtained in animal models. Micro-RNA therapies for cancer are already being tested in clinical phase trials. These findings have potential importance in cancer treatment in aged people although further research-based knowledge is needed to convert them into an effective molecular therapies for cancer linked to ageing.

Effect of neonatal asphyxia on the impairment of the auditory pathway by recording auditory brainstem responses in newborn piglets: a new experimentation model to study the perinatal hypoxic-ischemic damage on the auditory system.

INTRODUCTION: Hypoxia-ischemia (HI) is a major perinatal problem that results in severe damage to the brain impairing the normal development of the auditory system. The purpose of the present study is to study the effect of perinatal asphyxia on the auditory pathway by recording auditory brain responses in a novel animal experimentation model in newborn piglets. METHOD: Hypoxia-ischemia was induced to 1.3 day-old piglets by clamping 30 minutes both carotid arteries by vascular occluders and lowering the fraction of inspired oxygen. We compared the Auditory Brain Responses (ABRs) of newborn piglets exposed to acute hypoxia/ischemia (n = 6) and a control group with no such exposure (n = 10). ABRs were recorded for both ears before the start of the experiment (baseline), after 30 minutes of HI injury, and every 30 minutes during 6 h after the HI injury. RESULTS: Auditory brain responses were altered during the hypoxic-ischemic insult but recovered 30-60 minutes later. Hypoxia/ischemia seemed to induce auditory functional damage by increasing I-V latencies and decreasing wave I, III and V amplitudes, although differences were not significant. CONCLUSION: The described experimental model of hypoxia-ischemia in newborn piglets may be useful for studying the effect of perinatal asphyxia on the impairment of the auditory pathway.

Usefulness of the BAST-24 smell and taste test in the study of diabetic patients: a new approach to the determination of renal function.

CONCLUSION: A reduction in the percentage of correct responses in the olfactory test indirectly indicated increased albuminuria and worse glomerular filtration rate (GFR) in diabetic patients. The olfactory function test is an indirect indicator of early microvascular complications in diabetic patients. OBJECTIVES: Diabetes mellitus is a highly prevalent disease that causes numerous complications. The aim of this study was to determine whether olfactory and taste sensations are related to renal failure in diabetic patients. METHODS: We studied 61 patients diagnosed with diabetes mellitus, mean age = 65.9 years (SD = 16.8), 54.1% male. We evaluated olfactory and taste sensations by determining the capacity of detection, identification and percentage of correct responses of the 29 components of the Barcelona Smell-taste Test-24 (BAST-24). We determined the relationship between these results and glycosylated haemoglobin (HbA1c), creatinine, albumin/creatinine, albuminuria and GFR (normal = GFR ≥ 60 ml/min/1.73 m(2); impaired renal function = GFR < 60 ml/min/1.73 m(2)). RESULTS: There was no significant relationship between HbA1c and olfactory and taste sensations. There was a significant relationship between the percentage of correct responses and albuminuria (p = 0.03) and between identification of odours through the olfactory nerve and GFR (p = 0.029), and the percentage of correct responses and GFR (p = 0.03). There was no significant relationship between taste and renal failure.

Inner ear hair cell regeneration: A look from the past to the future.

Most recent studies on regeneration of inner ear hair cells focus on use of stem cells, gene therapy and neurotrophic factors. Cochlear gene therapy has been successfully used in the treatment of neurosensory hearing loss. This suggests that cochlear hair cell regeneration is possible. The objective of this paper is to review research and clinical application of inner near hair cell regeneration.

Altered peptidase activities in thyroid neoplasia and hyperplasia.

BACKGROUND: Papillary thyroid carcinoma (PTC), follicular thyroid adenoma (FTA), and thyroid nodular hyperplasia (TNH) are the most frequent diseases of the thyroid gland. Previous studies described the involvement of dipeptidyl-peptidase IV (DPPIV/CD26) in the development of thyroid neoplasia and proposed it as an additional tool in the diagnosis/prognosis of these diseases. However, very little is known about the involvement of other peptidases in neoplastic and hyperplastic processes of this gland. METHODS: The catalytic activity of 10 peptidases in a series of 30 PTC, 10 FTA, and 14 TNH was measured fluorimetrically in tumour and nontumour adjacent tissues. RESULTS: The activity of DPPIV/CD26 was markedly higher in PTC than in FTA, TNH, and nontumour tissues. Aspartyl aminopeptidase (AspAP), alanyl aminopeptidase (AlaAP), prolyl endopeptidase, pyroglutamyl peptidase I, and aminopeptidase B activities were significantly increased in thyroid neoplasms when compared to nontumour tissues. AspAP and AlaAP activities were also significantly higher in PTC than in FTA and TNH. CONCLUSIONS: These data suggest the involvement of DPPIV/CD26 and some cytosolic peptidases in the neoplastic development of PTC and FTA. Further studies will help to define the possible clinical usefulness of AlaAP and AspAP in the diagnosis/prognosis of thyroid neoplasms.

Altered dipeptidyl peptidase IV and prolyl endopeptidase activities in chronic tonsillitis, tonsillar hyperplasia and adenoid hyperplasia.

OBJECTIVE: To analyse peptidase activities in the removed tonsils and adenoids from patients with chronic tonsillitis, tonsillar hyperplasia and adenoid hyperplasia. METHODS: We have analyzed 48 tissue samples from patients undergoing tonsillectomy and adenoidectomy for chronic tonsillitis, tonsillar hyperplasia or adenoid hyperplasia. Tonsillectomy and adenoidectomy samples were collected and frozen for later enzyme analysis. The catalytic activity of a pool of peptidases (dipeptidyl peptidase IV, prolyl endopeptidase, aminopeptidase A, aminopeptidase N, aspartyl aminopeptidase, aminopeptidase B, neutral endopeptidase, pyroglutamyl peptidase I, puromycin-sensitive aminopeptidase and cystinyl aminopeptidase) was measured fluorometrically. RESULTS: The activity of prolyl endopeptidase was higher in tonsillar hyperplasia and adenoid hyperplasia than in chronic tonsillitis. On the contrary, dipeptidyl peptidase IV activity was higher in chronic tonsillitis than in hypertrophic tissues. When data were stratified by age and gender, dipeptidyl peptidase IV was also found to be more active in adult and male chronic tonsillitis tissues. Inversely, dipeptidyl peptidase IV activity was higher in tissues of females with tonsillar hyperplasia. CONCLUSIONS: These data indicate the involvement of dipeptidyl peptidase IV and prolyl endopeptidase in the mechanisms underlying chronic tonsillitis, tonsillar hyperplasia and adenoid hyperplasia.

Activity of soluble aminopeptidase A and dipeptidyl peptidase IV and membrane-bound aminopeptidase B and pyroglutamyl peptidase I in adenoid hyperplasia, tonsillar hyperplasia and chronic tonsillitis.

OBJECTIVE: To analyze soluble and membrane-bound peptidase activities in the tonsils and adenoids removed from patients with adenoid hyperplasia, tonsillar hyperplasia and chronic tonsillitis. METHODS: A total of 48 tissue samples from patients undergoing adenoidectomy and tonsillectomy for adenoid hyperplasia, tonsillar hyperplasia or chronic tonsillitis were analyzed. The catalytic activity of a pool of peptidases in the soluble (dipeptidyl peptidase IV, aminopeptidase A, aminopeptidase N and cystinyl aminopeptidase) and membrane-bound (prolyl endopeptidase, aspartyl aminopeptidase, aminopeptidase B and pyroglutamyl peptidase I) fractions was measured fluorometrically. RESULTS: The activity of membrane-bound aminopeptidase B was higher in cases of chronic tonsillitis and adenoid hyperplasia than in tonsillar hyperplasia, p=0.004. Soluble dipeptidyl peptidase IV and membrane-bound pyroglutamyl peptidase I were found to be more active in tissues from male chronic tonsillitis tissues, p<0.05, while membrane-bound aminopeptidase B activity was higher in tissues of females with tonsillar hyperplasia, p<0.001. In the case of chronic tonsillitis, soluble aminopeptidase A was found to have a higher level of activity in tissues from children than those from adults, p=0.005. CONCLUSIONS: Our results suggest a potential role of soluble aminopeptidase A, soluble dipeptidyl peptidase IV, membrane-bound aminopeptidase B and membrane-bound pyroglutamyl peptidase I in the pathobiology of adenoid hyperplasia, tonsillar hyperplasia and chronic tonsillitis that is differently regulated as a function of gender. These finfings may modify in the future the clinical approach to these diseases.

Increased prolyl endopeptidase activity in human neoplasia.

Prolyl endopeptidase (EC 3.4.21.26) (PEP) is a serine peptidase that converts several biologically active peptides. This enzyme has been linked to several neurological, digestive, cardiovascular and infectous disorders. However, little is known about its involvement in neoplastic processes. This study analyzes fluorimetrically cytosolic and membrane-bound PEP activity in a large series (n=122) of normal and neoplastic tissues from the kidney, colon, oral cavity, larynx, thyroid gland and testis. Cytosolic PEP activity significantly increased in clear cell renal cell carcinoma, urothelial carcinoma of the renal pelvis and head and neck squamous cell carcinoma. Both cytosolic and membrane-bound PEP activity were also increased in colorectal adenomatous polyps. These data suggest the involvement of PEP in some mechanisms that underlie neoplastic processes.

Mandibular Metastasis of a Signet Ring Cell Carcinoma of the Breast in a Patient who Underwent Bilateral Mastectomy more than 25 Years Earlier.

SUMMARY: BACKGROUND: Metastatic tumors account for less than 1% of all malignant tumors occurring in the oral cavity. CASE REPORT: The clinical case of a 94-year-old patient with a mandibular tumor is reported here. The patient had undergone bilateral mastectomy more than 25 years before. An immunohistochemical study found hormone receptors in signet ring cells, suggesting a diagnosis of breast cancer metastasis. CONCLUSION: Immunohistochemical diagnosis and antineoplastic hormone therapy is the cornerstone in the management of this clinical case.

Increased APN/CD13 and acid aminopeptidase activities in head and neck squamous cell carcinoma.

BACKGROUND: Involvement of peptidases in carcinogenic processes of several tumor types has been investigated in recent years. Head and neck squamous cell carcinoma (HNSCC) is one of the most common cancers worldwide and accounts for more than 90% of all head and neck cancers. Increased understanding of its pathophysiology has led to implication of several proteinases, specially matrix metalloproteinases, in its genesis, growth, and dissemination. However, very little is known about involvement of peptidases in this neoplasm. METHODS: Seventeen HNSCC tissue samples were selected for the study. Tumor and normal tissue samples were frozen for enzymatic study. The catalytic activity for a pool of peptidases (PSA, APN/CD13, APB, APA, Asp-AP, CAP, DPPIV/CD26, NEP/CD10, and PGI) was measured fluorometrically. RESULTS: The activity of 2 cell surface aminopeptidases (APN/CD13 and APA) and a cytosolic aminopeptidase (Asp-AP) was significantly increased in HNSCC tissues. CONCLUSIONS: These data show the involvement of cell surface and cytosolic peptidases in the mechanisms underlying HNSCC.

Efficacy of selective percutaneous embolization for the treatment of intractable posterior epistaxis and juvenile nasopharyngeal angiofibroma (JNA).

CONCLUSION: Percutaneous embolization reduces the reappearance of epistaxis and the mean length of hospital stay for patients with intractable epistaxis or juvenile nasopharyngeal angiofibroma (JNA). OBJECTIVES: To assess whether percutaneous embolization is effective for the treatment of intractable epistaxis and JNA. PATIENTS AND METHODS: Twenty-eight patients with intractable posterior epistaxis treated by embolization (25 males; m = 59.78 years; SD = 14.3) and 28 unembolized patients (24 males; m = 59.21 years; SD = 15.13) were studied retrospectively. Eight patients with JNA (all males; mean = 16.5 years; SD = 2.35; four embolized before surgery and four unembolized) were also analyzed. RESULTS: Embolization was bilateral in 71.4% of subjects, blood transfusion was required in 28.57%, incidence of complications was 53.57%, and seven of the embolized patients with intractable epistaxis suffered from recurrence of the epistaxis. There were no statistically significant differences between the embolized and unembolized groups. The mean hospital stay was longer in unembolized patients (4.46 days; SD = 2.42) than for the embolized patients (3.78 days; SD = 3.9), p=0.394. The most serious complications occurred in patients embolized with idiopathic epistaxis and the mean post-embolization hospital stay was longer in this subgroup (4.14 days; SD = 4.39) than in patients with Rendu-Osler-Weber syndrome (2.40 days; SD = 1.140), p=0.395. Unembolized patients with JNA presented greater hemorrhage (m = 2025 ml; SD = 325) and a longer mean hospital stay (m = 18 days; SD = 3) than the group of embolized patients (m = 360 ml; SD = 185 and m = 9 days; SD = 1, respectively).