Pre-and post-surgical non-functional pituitary adenomas and their relationship with high levels of serum glucose.

OBJECTIVES: This clinical, analytical, retro-prospective, auto-controlled, not randomized, and not blinded study, aimed to investigate the association of changes in the serum glucose levels with the pre-and-post changes in the size tumor in mm3 in the Non-Functional pituitary adenomas. METHODS: Pre-and post-surgical MRI, as well as the measurements in the serum glucose levels and immunohistochemical techniques were performed in all the patients in the study, with a mean followed-up until 208.57 days. A comparison was made between the reductions in tumor size of hormonally active pituitary adenomas (HSPAs) vs NFPAs. RESULTS: Seventy-four patients were included in this study, of whom, 46 were NFPAs. The decrease in the NFPAs tumor size after surgery was statistically significant (P ≤ 0.0001). The Mean of the differences of both type of tumors in mm3 were -9552 ± 10287. Pre-surgery, the mean of the HSPAs were 8.923 ± 2.078; and the NFPAs were 14.161 ± 1.912. The differences in the tumor size were statistically significant (p = 0.039). Post-surgical, the mean of the HSPAs were 2.079 ± 971, with a (p = 0.14): and the NFPAs were 4.609 ± 1.205. After surgery of the NFPAs, most of the patients-maintained serum levels ≤ 100 mg/dL, with a statistical significance (P ≤ 0.0003). CONCLUSION: This study demonstrates for the first time the correlation between the presence of pre-and post- surgical changes in the NFPAs, with modifications in the levels of serum glucose, and the comparison, pre- and post-surgical between the tumor size of HSPAs and NFPAs.

Improvement of metabolic syndrome and its components in patients who underwent transsphenoidal resection for pituitary adenoma.

BACKGROUND: Pituitary adenomas (PA) are neoplasms of pituitary adenohypophyseal cell lineage, which are the third most common cause of brain tumors among adults. Due to hormone secretion, PAs are closely related to metabolic syndrome (MetS). However, the relationship between these two entities has been scarcely studied to date. PURPOSE: This paper aims to evaluate changes in the metabolic status of patients with PA before and after surgical treatment and to look for differences in metabolic outcomes among patients according to the adenoma type and the surgery success rate. METHODS: We assessed patients with PA who went through transsphenoidal surgery for its treatment, documenting metabolic parameters before and after surgery, analyzed whole sample changes, and then stratified them according to adenoma type (nonfunctioning, somatotroph, lactotroph, and corticotroph), and surgery success (total resection, near-total resection, partial resection, subtotal resection). RESULTS: A total of 214 patients were enrolled for this study. The prevalence of MetS with AACE criteria went from 51.52% before surgery to 28.99% after surgery (P < 0.001). Hyperglycemia (HG) was the most beneficial component; it went from 56% pre-surgery to 40.51% post-surgery (P = 0.03). The total resection group had the best improvement, with a significant decrease of prevalence in MetS from 83 to 16% (P < 0.001), and every component, except hypoalphalipoproteinemia (HA): obesity went from 96 to 67% (P < 0.001), arterial hypertension (AH) 59 to 24% (P < 0.001), HG 74 to 23% (P < 0.001), and hypertriglyceridemia (HTG) from 81 to 54% (P < 0.001). According to MetS prevalence, there was no difference in the improvement according to PA type. CONCLUSION: Surgical treatment in patients with PA is associated with MetS improvement.

Pre- and post-clinical-radiological and surgical evaluation of patients with pituitary adenoma and metabolic syndrome.

BACKGROUND: Pituitary adenomas are benign tumors located in the anterior hypophysis. Its appearance is associated with the development of parameters related to metabolic syndrome; therefore, surgical treatment could reduce associated morbimortality. METHODS: Pre- and post-surgical MRI, using the Hardy-Wilson and Knosp classification, and clinical data according to the American Association of Clinical Endocrinology (AACE) criteria for metabolic syndrome: all the patients were followed-up until 208.57 days were reviewed on 217 consecutive patients with pituitary surgery. RESULTS: Seventy-four patients were included in this study. There was a significant reduction in tumor size in mm3 [average pre- and post-surgery respectively: 12,362 mm3 (±12,397); 3,910 mm3 (±7,160)], (p < 0.0001). This was confirmed by the Hardy-Wilson and Knosp classification, where most patients went from grade IV C (33.7%) to grade 0 (12.1%), IA (20.2%), IIB (21.36%), and IIC (16.2%); as well as from grade 4 (24.3%) to grade 0 (45.9%), respectively. After surgery, there were statistically significant reductions in total serum levels of glucose [average pre- and post-surgery, respectively: 116 mg/dL (±26.9); 90 mg/dL (±10.2)], (p < 0.001), triglycerides [average pre- and post-surgery, 240 mg/dL (±102); 171 mg/dL (±60.5)], (p = 0.001); and HDL-c [average pre- and post-surgery, respectively: 39 mg/dL (±11.8); 44.6 mg/dL (±8.4)], (p = 0.029). The other parameters remained unchanged. CONCLUSION: This is the first study to demonstrate the relationship between the presence of pituitary adenoma and significative changes in serum glucose, triglycerides and c-HDL related to metabolic syndrome.

The percentage of CD39+ monocytes is higher in pregnant COVID-19+ patients than in nonpregnant COVID-19+ patients.

Current medical guidelines consider pregnant women with COVID-19 to be a high-risk group. Since physiological gestation downregulates the immunological response to maintain "maternal-fetal tolerance", SARS-CoV-2 infection may constitute a potentially threatening condition to both the mother and the fetus. To establish the immune profile in pregnant COVID-19+ patients, a cross-sectional study was conducted. Pregnant women with COVID-19 (P-COVID-19+; n = 15) were analyzed and compared with nonpregnant women with COVID-19 (NP-COVID-19+; n = 15) or those with physiological pregnancy (P-COVID-19-; n = 13). Serological cytokine and chemokine concentrations, leucocyte immunophenotypes, and mononuclear leucocyte responses to polyclonal stimuli were analyzed in all groups. Higher concentrations of serological TNF-α, IL-6, MIP1b and IL-4 were observed within the P-COVID-19+ group, while cytokines and chemokines secreted by peripheral leucocytes in response to LPS, IL-6 or PMA-ionomicin were similar among the groups. Immunophenotype analysis showed a lower percentage of HLA-DR+ monocytes in P-COVID-19+ than in P-COVID-19- and a higher percentage of CD39+ monocytes in P-COVID-19+ than in NP-COVID-19+. After whole blood polyclonal stimulation, similar percentages of T cells and TNF+ monocytes between groups were observed. Our results suggest that P-COVID-19+ elicits a strong inflammatory response similar to NP-COVID19+ but also displays an anti-inflammatory response that controls the ATP/adenosine balance and prevents hyperinflammatory damage in COVID-19.

Chlorination for low-cost household water disinfection - A critical review and status in three Latin American countries.

Chlorination has historically provided microbiologically safe drinking water in public water supplies. Likewise, chlorine has also been introduced as a low-cost disinfection method in rural and marginalized communities, both at community and household level, as well as during emergencies. Although this practice is common and well established for use as a household water treatment technology in the Global South, several challenges in effective and efficient implementation still need to be addressed. Here, we explored these issues by a literature review and narrowed them to the status of three Latin American countries (Mexico, Colombia, and Brazil). Overall, it was found that although guidance on household-based chlorination includes information on health risks and hygiene, this may not create enough incentive for the user to adapt the method satisfactorily. Physicochemical quality of the water influences chlorination efficiency and it is found that variations in quality are rarely considered when recommending chlorine doses during implementation. These are far more often based on a few measurements of turbidity, thereby not considering dissolved organic matter, or seasonal and day-to-day variations. Other factors such as user preferences, chlorine product quality and availability also represent potential barriers to the sustainable use of chlorination. For chlorination to become a sustainable household water treatment, more focus should therefore be given to local conditions prior to the intervention, as well as support and maintenance of behavioural changes during and after the intervention.

A critical overview of household slow sand filters for water treatment.

Household, or point-of-use (POU), water treatments are effective alternatives to provide safe drinking water in locations isolated from a water treatment and distribution network. The household slow sand filter (HSSF) is amongst the most effective and promising POU alternatives available today. Since the development of the patented biosand filter in the early 1990s, the HSSF has undergone a number of modifications and adaptations to improve its performance, making it easier to operate and increase users' acceptability. Consequently, several HSSF models are currently available, including those with alternative designs and constant operation, in addition to the patented ones. In this scenario, the present paper aims to provide a comprehensive overview from the earliest to the most recent publications on the HSSF design, operational parameters, removal mechanisms, efficiency, and field experiences. Based on a critical discussion, this paper will contribute to expanding the knowledge of HSSF in the peer-reviewed literature.

Reliability and validity in Mexico of the Spanish version of the Children's Fear Survey Schedule - Dental Subsca.

AIM: Low attendance and abandonment of dental treatment caused by dental fear varies from 6% to 20%. Various specific scales have been used to evaluate and measure dental fear, like Children´s Fear Survey Schedule - Dental Subscale, which is considered the "gold standard", due to its efficacy and validity. However, not all the studies that have been conducted have performed the Exploratory Factorial and Confirmation Analysis to infer the validity of the scale's content, which is why this study has as its main objective to analyse the confidentiality and validation of this scale. MATERIALS AND METHODS: In a sample of 163 school age children, from 6 to 12 years old, of both genders, from a public school, a Children's Fear Survey Schedule - Dental Subscale was applied to measure dental fear. The internal consistency of the scale was estimated with the Cronbach's alpha, we also performed the Exploratory Factorial Analysis with the method of the main component with Varimax rotation and the Confirmatory Factorial Analysis to confirm the structure of the factors. RESULTS: The Cronbach's alpha was obtained with a .837 (p?.05). The adequate coefficients were determined with the Kaiser-Meyer-Olkin (KMO = .843) and the Bartlett's test of specificity (?2 = 565,325, p= .000), in which there was a good adequate data, expressing an appropriate and intercorrelation between items or strongly related. The factorial structure showed four factors extracted, 4 factors explain the 53.57% of the accumulated variation; 14 items were above the .40. The statistics of the second and third model obtained adequate values in the statistics of goodness of fit index. CONCLUSION: The Mexican version of the CFSS-DS in the present study gives us data about the adaptation and validation of the dental fear scale of CFSSS-DS by the Exploratory Factorial Analysis through a model of structural equations. This means that this instrument is a trusting and valid tool to measure dental fear in the children's population of Mexico.

Curcumin induces p53-independent inactivation of Nrf2 during oxidative stress-induced apoptosis.

The transcription factor Nrf2 (nuclear factor erythroid 2-related factor 2) is a master regulator of a battery of antioxidant and detoxificant genes with cytoprotective function. Since Nrf2 inactivation is necessary for the complete execution of apoptosis in the presence of extensive cellular damage caused by oxidative stress, constant activation of Nrf2 may protect tumoral cells from apoptosis. The tumor suppressor gene p53 has been suggested to participate in apoptosis-related repression of Nrf2. Thus, we studied the inactivation of Nrf2 during oxidant-induced apoptosis in a p53 dysfunctional cellular model. Using curcumin dose-response assay and time-response assay in an immortalized lymphoblastoid cell line (control line 45), we observed a time-dependent increase in apoptotic markers such as deoxyribonucleic acid (DNA) fragmentation, phosphatidylserine exposure, and caspase-3, caspase-9 and poly (ADP-ribose) polymerases (PARP) cleavage. Interestingly, at early times of exposure to a proapoptotic dose of curcumin (15 µM), we observed nuclear accumulation of Nrf2 and the expression of Nrf2 target genes, whereas at late exposure times we found a reduction of total and nuclear protein levels of Nrf2 as well as downregulation of Nrf2 target genes in the absence of p53 activation. These data suggest that apoptosis-related inactivation of Nrf2 could occur in a p53 dysfunctional background, opening the possible occurrence of p53-independent mechanism to explain Nrf2 inactivation during apoptosis.

Effects of 5-HT5A receptor blockade on amnesia or forgetting.

Previously the effects (0.01-3.0 mg/kg) of post-training SB-699551 (a 5-HT5A receptor antagonist) were reported in the associative learning task of autoshaping, showing that SB-699551 (0.1 mg/kg) decreased lever-press conditioned responses (CR) during short-term (STM; 1.5-h) or (3.0 mg/kg) long-term memory (LTM; 24-h); relative to the vehicle animals. Moreover, as pro-cognitive efficacy of SB-699551 was reported in the ketamine-model of schizophrenia. Hence, firstly aiming improving performance (conditioned response, CR), in this work autoshaping lever-press vs. nose-poke response was compared; secondly, new set of animals were randomly assigned to SB-699551 plus forgetting or amnesia protocols. Results show that the nose-poke operandum reduced inter-individual variance, increased CR and produced a progressive CR until 48-h. After one week of no training/testing sessions (i.e., interruption of 216 h), the forgetting was observed; i.e., the CR% of control-saline group significantly decreased. In contrast, SB-699551 at 0.3 and 3.0 mg/kg prevents forgetting. Additionally, as previously reported the non-competitive NMDA receptor antagonist dizocilpine (0.2 mg/kg) or the non-selective cholinergic antagonist scopolamine (0.3 mg/kg) decreased CR in STM. SB-699551 (0.3 mg/kg) alone also produced amnesia-like effect. Co-administration of SB-699551-dizocilpine or SB-699551-scopolamine reversed the SB-699551 induced-amnesic effects in LTM (24-h). Nose-poke seems to be a reliable operandum. The anti-amnesic and anti-forgetting mechanisms of amnesic SB-699551-dose remain unclear. The present findings are consistent with the notion that low doses of 5-HT5A receptor antagonists might be useful for reversing memory deficits associated to forgetting and amnesia. Of course, further experiments are necessary.

Microcosm environment models for studying the stability of adenovirus and murine norovirus in water and sediment.

Microcosms are useful tools for understanding the survival and fate of enteric viruses in aquatic environments. This study set out to determine the stability of infectious enteric viruses in an aquatic environment using a laboratory scale microcosm. Sediment and overlaying water were collected from a lagoon and inoculated with known concentrations of recombinant adenovirus (AdV-GFP) and murine norovirus (MNV-1). Infectious particles of these viruses were measured using fluorescence microscopy (AdV-GFP) or the plaque assay method (MNV-1), over 85 days in two different conditions: under natural sunlight and in fully darkened environments. The time required to reach one log reduction in viral titres (T90) of viable viruses in a natural condition microcosm for AdV-GFP and MNV-1 was shorter than in a dark condition microcosm. There was also a negative correlation between the temperature and infectivity of these viruses in both water and sediment samples. Considering that microcosms aim to mimic natural environment conditions and that AdV-GFP and MNV-1 are excellent surrogates for measuring the infectivity of the respective viruses strains, the results presented here have the potential to be applied in future health hazard studies and also would be useful for future climate scenarios.

Effect of immunocastration with Bopriva on carcass characteristics and meat quality of feedlot Holstein bulls.

To evaluate the effect of immunocastration on carcass and meat characteristics, Holstein bulls aged between 7 and 8months with a live weight of 232±1.19kg were given two separate treatments, placebo (intact bulls) versus Bopriva, and then slaughtered after approximately 239days of fattening. While the testosterone levels in intact bulls remained at 0.42ng/ml throughout the study, by day 181, differences (P<0.05) were observed in immunized bulls, with values of 0.21ng/ml. The carcasses of animals treated with Bopriva recorded both a higher hot carcass weight (HCW) and a cold carcass weight (CCW), as well as higher dorsal fat density, marbling and KPH (P<0.05); however, no differences (P>0.05) were observed in the Longissimus lumborum area. No significant differences (P>0.05) were recorded between the treatments for pH, L*, a*, b* C* and H*. The carcasses of the animals treated with Bopriva were heavier, with higher dorsal fat density and marbling score.

Critical limb ischemia and its treatments: a review.

Critical limb ischemia (CLI) encompasses the most extreme end of the peripheral artery disease (PAD) spectrum leading to significant morbidity and mortality. CLI is defined as greater than 2 weeks of extremity rest pain, ulcers or extremity gangrene, secondary to objectively proven peripheral artery disease. Corresponding to Fontaine Stages III/IV and Rutherford category IV through VI, CLI is a complex disease comprising of both macrovascular and microvascular systems with inconsistent historical data on optimal treatment. CLI is distinct from intermittent claudication with different goals of treatment, however in both groups risk factor modification is of the utmost importance involving tobacco cessation, and treatment of underlying conditions like diabetes mellitus, hyperlipidemia and hypertension. In CLI, medical therapy involves wound care and also consists of antiplatelet therapy, anti-inflammatory therapy including statin use or ACE inhibitors. Surgical therapies include distal bypass surgery, thromboendartectomy and amputation. Endovascular techniques include percutaneous transluminal angioplasty, bare metal stents, atherectomy, drug-coated balloon and drug-eluting stents. CLI is considered the end-stage of PAD, requiring a thoughtful and multidisciplinary approach, risk-benefit analysis and treatment of comorbid conditions. Conservative and surgical treatments, along with endovascular techniques, have allowed excellent opportunities for treating complicated patients for wound healing and limb salvage.

Paternity identification in sugarcane polycrosses by using microsatellite markers.

Although polycrosses have been used to test the potential of cross-combination of a large number of sugarcane parents, the male parent of the half-sib progenies produced is unknown. The present study aimed to integrate the molecular marker technology to the sugarcane polycross approach by the application of microsatellite markers to identify the male parent of 41 elite clones derived from polycross families. Ten microsatellite [single sequence repeats (SSRs)] primer pairs were used to identify the most likely male parent considering markers present in the selected clone but absent in the female parent. The number of alleles generated by the 10 microsatellite primer pairs ranged from 102 (cross-pollination lantern 4) to 120 (cross-pollination lantern 2) with an average of 113.25 alleles per SSR. The average genetic similarity among the involved parents in the polycrosses was 45.9%. The results of the analysis of the SSR markers absent in the female parent and present only in the selected clone as well as the genetic similarity values allowed the identification of the most likely male parent in 73% of the total clones evaluated and also to detect probable contaminations. The obtained results highlight the importance of using molecular marker technology in the identification and confirmation of the male parent of high-performance clones derived from polycrosses in the sugarcane breeding programs.

Stability of human enteric viruses in seawater samples from mollusc depuration tanks coupled with ultraviolet irradiation.

AIMS: To evaluate the stability in seawater of human adenovirus (HAdV2), murine norovirus (MNV-1) and hepatitis A virus (HAV) in a shellfish depuration system with and without ultraviolet (UV) treatment. METHODS AND RESULTS: Seawater was seeded with viruses and disinfected using a 36 W lamp. Samples were collected at 24, 48, 72, 96 and 120 h; viruses were concentrated and the viral decay was evaluated using molecular and cell culture methods. Based on the molecular results, at 120 h of disinfection, there was a reduction of more than 3 log(10) for HAdV2 and HAV; MNV-1, a 4.5 log(10) reduction was observed at 72 h. Infectious MNV-1 was not detected after 72 h of treatment; while HAdV2 remained infectious. Seawater not treated demonstrated a progressive viral reduction for the three viruses tested. CONCLUSIONS: The UV reduced the number of viral particles, and the results indicate there is natural and gradual decrease of viral load and viability in seawater. SIGNIFICANCE AND IMPACT OF THE STUDY: UV irradiation is the method of choice for shellfish depuration in many countries; this work showed useful information about the viral stability in seawater and application of UV to water disinfection to be used in shellfish depuration tanks.

Surveillance of human and swine adenovirus, human norovirus and swine circovirus in water samples in Santa Catarina, Brazil.

Animal and human wastewater can potentially contaminate water sources and the treatment of drinking water may not effectively remove all contaminants, especially viruses. The purpose of the present study was to evaluate the viral contamination of water used for human and animal consumption in the city of Concórdia, located in southern Brazil. Porcine circovirus type 2 (PCV2), porcine adenovirus (PAdV), human adenovirus (HAdV) and human norovirus (NoV) were searched for using quantitative polymerase chain reaction (qPCR). HAdV-positive samples were tested for viral infectivity by plaque assay. The qPCR results showed that PAdV, PCV2 and HAdV genetic material were present in all sampling sites. NoV was absent in all samples. The presence of genetic material from PAdV and PCV2 was detected in 30% and 45% of the 36 analyzed samples, respectively, with an average of 10(2) gc mL(-1) for PAdV and 10(4) gc mL(-1) for PCV2. HAdV was present in 100% of the samples, with an average of 10(4) gc mL(-1). However, in plaque assay, only 36% of the samples were positive. As viable particles of HAdV were found in drinking water, these results confirm that swine manure and human sewage impact surface water and groundwater, endangering water quality and indicating a potential risk to public health.

Culturing and molecular methods to assess the infectivity of porcine circovirus from treated effluent of swine manure.

Samples were collected at the effluent of two swine manure treatment systems and were analyzed by qPCR to determine the presence and amounts of porcine circovirus (PCV2) genetic material. ST cells were inoculated with the positive samples to evaluate virus viability and for viral genotyping. Twenty-five water samples were collected monthly from treated effluent (March 2009 to December 2010). The PCV2 genome was identified by qPCR in 60% of the samples, and all of the positive samples were able to infect ST cells in vitro. Positive samples were genotyped and 60% of them were positive for both PCV2a and PCV2b, 20% were positive for genotype 2a, and 20% were positive for genotype 2b. Our results suggest that these viruses were able to resist the regular wastewater treatment, and this finding demonstrates the necessity of adding a virus inactivation step to the treatment system to guarantee the safety of water reuse.

Detection of circoviruses and porcine adenoviruses in water samples collected from swine manure treatment systems.

Samples collected from two swine manure treatment systems including: swine manure treatment system and demonstrative unit (SMTS and DU), were analyzed by qPCR to quantify the amount of porcine adenovirus (PAdV) and porcine circovirus (PCV2) present. Positive samples were tested for virus integrity using DNase assay. Fifty-six water samples were collected monthly from March 2009 to May 2010. PAdV genome was found 66% of the samples in the SMTS and in 78% of the samples in the DU system. PCV2 was detected in 96% of samples collected from the SMTS system and in 86% of samples from DU. DNase assay revealed that there were undamaged virus particles of both PAdV and PCV2 in all sampling sites in the SMTS. However, undamaged particles of both viruses were detected in samples from the DU system in the affluent and middle sites, though undamaged PCV2 was absent in the effluent samples.

Microbiological and physicochemical analysis of the coastal waters of southern Brazil.

The aim of this study was to assess the impact of sewage discharge on coastal waters by evaluating the influence of physicochemical parameters on the presence of enteric microorganisms in seawater samples collected from 11 beaches in Florianopolis, Santa Catarina, Brazil, over a one-year period (August 2009 to July 2010). Samples were assessed for the presence of human adenoviruses (HAdV), polyomavirus (JCPyV), hepatitis A virus (HAV), and noroviruses (HuNoV GI and GII). Escherichia coli and physicochemical parameters (salinity, temperature, pH and dissolved oxygen) were also evaluated. From the 132 samples analyzed, 55% were positive for HAdV, 51.5% for HAV, 7.5% for HuNoV GI, 4.5% for HuNoV GII, and 3% for JCPyV. E. coli levels ranged from 8 to 1325 CFU/100mL at all sites. The overall results highlight the problem of sewage discharge into coastal waters and confirm that there is no correlation between viral presence and bacterial contamination.

Hippocampal abnormalities in an MR imaging series of patients with tuberous sclerosis.

BACKGROUND AND PURPOSE: Hippocampal abnormalities are known as highly epileptogenic precursor lesions in the general population, primarily manifesting as MTS. The purpose of this study was to evaluate the occurrence of hippocampal abnormalities on MR imaging in patients with TS to explore the possible underlying mechanisms of the abnormalities and to identify the relationship between an abnormal HF and epilepsy. MATERIALS AND METHODS: We studied MR images and clinical data from 31 patients with TS. The MR imaging protocol was identical for all patients and included tilted coronal images of their temporal lobes. The diagnosis of TSC was made according to established criteria. The HFs of the patients were evaluated from coronal images according to size, morphology, and signal intensity. The data were submitted to statistical analysis, and P values < or = .05 were considered significant. RESULTS: We found HF abnormalities in 5 patients. Four had typical MTS, and 1 had HIMAL. We found a positive correlation between childhood febrile seizures and MTS in patients with TS. We also confirmed that patients with altered hippocampi had a tendency to exhibit more cortical tubers. CONCLUSIONS: Analysis of this series of patients demonstrated the presence of HF abnormalities, mainly MTS in patients with TS. We also found that the occurrence of febrile seizures during the first year of life appears to be one of the determining factors for MTS development in these patients.

Distinctive clinicopathological associations of amplification of the cortactin gene at 11q13 in head and neck squamous cell carcinomas.

Amplification of the 11q13 region is a prevalent genetic alteration in head and neck squamous cell carcinoma (HNSCC). We investigated the clinical significance of cortactin (CTTN) and cyclin D1 (CCND1) amplification in both malignant transformation and tumour progression. CTTN and CCND1 amplification was analysed by differential and real-time PCR in a prospective series of laryngeal/pharyngeal carcinomas and archival premalignant tissues. CTTN mRNA and protein expression were respectively determined by real-time RT-PCR and immunohistochemistry, and correlated with gene status. Molecular alterations were associated with clinicopathological parameters and disease outcome. CTTN and CCND1 amplifications were respectively found in 75 (37%) and 90 (45%) tumours. Both correlated with advanced disease; however, only CTTN amplification was associated with recurrence and reduced disease-specific survival (p = 0.0022). Strikingly, CTTN amplification differentially influenced survival depending on tumour site (p = 0.0001 larynx versus p = 0.68 pharynx) and was an independent predictor of reduced survival in the larynx (p = 0.04). CCND1 amplification was detected in early tumourigenesis and increased with the severity of dysplasia. Importantly, CTTN amplification was only found in high-grade dysplasias that progressed to invasive carcinoma. CTTN gene status strongly correlated with mRNA and protein expression. Furthermore, CTTN overexpression correlated significantly with reduced disease-specific survival (p = 0.018). Taken together, these data indicate that CTTN may serve as a valuable biomarker to identify patients with laryngeal tumours at high risk of recurrence and poor outcome.