Intra-island patterns of cryptic diversity within an oceanic island: Insights from the differentiation of Schizidium Verhoeff, 1901 (Oniscidea, Armadillidiidae) within Cyprus, with descriptions of two new species.

Cyprus is a Mediterranean island of volcanic origin isolated for at least 5.3 Myr from surrounding continental areas. The present study focuses on the diversification of the isopod genus Schizidium within the island, including also specimens from surrounding continental areas. The genus Schizidium sensu lato is probably non monophyletic, comprising 26 species distributed from Greece to Iran. Up to date the only representative of the genus reported from Cyprus was Schizidium fissum. Aiming to investigate the patterns of genetic diversity within the focal island, to evaluate the morphology-based taxonomy of the species in the genus, and at the same time to explore phylogenetic relationships with mainland populations, we applied genome-wide ddRADseq as well as Sanger sequencing targeting three mitochondrial (16S, COI and 12S) and the nuclear NaK loci. Results of phylogenetic analyses support the existence of two distinct epigean Schizidium clades with well-defined geographic boundaries that conform to the known paleogeography of Cyprus, plus one endogean clade with restricted distribution within the island. Genetic data and morphology corroborate the assignment of this latter endogean clade to a new species, Schizidium myrrae n. sp. The two epigean clades are also considered as distinct species, one corresponding to the known S. fissum at the eastern part of the island (Pentadaktylos massif) and the other to the newly described S. christosi n. sp. distributed along the western part of the island (Troodos massif). Even though detailed examination of many specimens could not retrieve any morphological differences among representatives of these two clades, clado-chronological analysis indicates a long isolation between them, estimated at âˆ¼ 9 Mya, as well as the sharing of a common ancestor with S. tiberianum from Israel at âˆ¼ 15 Mya. Hence, we can consider these epigean Schizidium species as one more case of cryptic diversity on Cyprus, exhibiting similar patterns with the recently described case in the genus Armadillo.

Gentlest Ascent Dynamics on Manifolds Defined by Adaptively Sampled Point-Clouds.

Finding saddle points of dynamical systems is an important problem in practical applications, such as the study of rare events of molecular systems. Gentlest ascent dynamics (GAD) (10.1088/0951-7715/24/6/008) is one of a number of algorithms in existence that attempt to find saddle points. It works by deriving a new dynamical system in which saddle points of the original system become stable equilibria. GAD has been recently generalized to the study of dynamical systems on manifolds (differential algebraic equations) described by equality constraints (10.1007/s10915-022-01838-3) and given in an extrinsic formulation. In this paper, we present an extension of GAD to manifolds defined by point-clouds, formulated by using an intrinsic viewpoint. These point-clouds are adaptively sampled during an iterative process that drives the system from the initial conformation (typically in the neighborhood of a stable equilibrium) to a saddle point. Our method requires the reactant (initial conformation), does not require the explicit constraint equations to be specified, and is purely data-driven.

Intrinsic map dynamics exploration for uncharted effective free-energy landscapes.

We describe and implement a computer-assisted approach for accelerating the exploration of uncharted effective free-energy surfaces (FESs). More generally, the aim is the extraction of coarse-grained, macroscopic information from stochastic or atomistic simulations, such as molecular dynamics (MD). The approach functionally links the MD simulator with nonlinear manifold learning techniques. The added value comes from biasing the simulator toward unexplored phase-space regions by exploiting the smoothness of the gradually revealed intrinsic low-dimensional geometry of the FES.

Serum levels of vascular endothelial growth factor in non-alcoholic fatty liver disease.

BACKGROUND: This study aimed to assess the significance of serum levels of vascular endothelial growth factor (VEGF) in non-alcoholic fatty liver disease (NAFLD). METHODS: Sixty-seven consecutive NAFLD patients and 47 healthy controls who visited our liver clinics between May 2008 and December 2010 were included. The NAFLD diagnosis required elevated alanine aminotransferase and/or gamma-glutamyl transpeptidase levels, evidence of hepatic steatosis on ultrasound and/or liver histology, and exclusion of other causes of liver injury. Serum VEGF levels were determined by an enzyme immunoassay. Liver biopsy was obtained in 34 NAFLD patients. Histological lesions were scored by a liver histopathologist. RESULTS: Serum VEGF levels tended to be lower in matched NAFLD patients than in healthy controls (296±146 vs. 365±186 pg/mL, P=0.092); levels in patients with non-alcoholic steatohepatitis (NASH) also tended to be lower than in those with simple fatty liver (FL) (279±149 vs. 359±190 pg/mL, P=0.095); while VEGF levels were significantly lower in NASH patients than in healthy controls (279±149 vs. 365±186 pg/mL, P=0.041). VEGF levels offered poor predictability for the differentiation between NAFLD patients and controls or between NASH and FL patients. However, patients with high VEGF levels (≥300 pg/mL) were significantly more likely to have FL, either in the total NAFLD population (67% vs. 35%, P=0.019) or in the 34 NAFLD patients with liver biopsy (57% vs. 15%, P=0.023), while those with high VEGF levels also had a significantly lower mean fibrosis score (0.7±0.9 vs. 1.6±1.0, P=0.017). CONCLUSION: Our data suggest that serum VEGF levels are equally high in healthy controls and in patients with simple fatty liver, but tend to decrease when NASH develops.

Fibronectin stimulates the osteogenic differentiation of murine embryonic stem cells.

Conditioned medium from human hepatocarcinoma cells (HepG2-CM) has been shown to stimulate the osteogenic/chondrogenic differentiation of murine embryonic stem cells (mESCs). HepG2-CM is considered to contain visceral endoderm (VE)-like signals and attempts have recently been made to characterize it, using proteomic profiling, with fibronectin being identified as one promising candidate. Herein, we investigated whether fibronectin is able to mimic the activities of HepG2-CM during the osteogenic differentiation of mESCs. Specifically, the addition of RGD peptides and heparin in HepG2-CM significantly reduced the growth- and adhesion-promoting effects of HepG2-CM, in addition to suppressing its osteogenic-inductive activity. Furthermore, direct addition of fibronectin to basal medium was able to reproduce, at least partially, the function of HepG2-CM. In particular, fibronectin induced the early onset of osteogenic differentiation in mESCs, as confirmed by gene expression of osteogenic markers, and resulted in the three-fold higher calcium deposition at day 11 of osteogenic culture compared to the control group. These data clearly suggest that fibronectin contributes to the biological activities of HepG2-CM and plays a stimulatory role during the process of osteogenesis in mESCs. Copyright © 2015 John Wiley & Sons, Ltd.

Adherence to follow-up and treatment recommendations in Greek and immigrant patients with chronic hepatitis B in Greece.

BACKGROUND/AIM: Immigrants have multiple barriers to access to health care systems. We evaluated the adherence to follow-up and treatment recommendations of chronic hepatitis B virus (HBV) Greek and immigrant patients. METHODS: In total, 1001 consecutive adult patients with chronic HBV infection who visited our clinics for the first time between 2002 and 2011 were included. All patients born outside Greece were considered immigrants. Diagnosis was considered to be complete if patients could be classified into HBeAg-positive chronic hepatitis B (CHB), inactive carriers, HBeAg-negative CHB, or decompensated cirrhosis. RESULTS: Of the patients, 56% were Greeks and 44% were immigrants. Greeks visited our clinics at a significantly older mean age (50 vs. 35 years, P<0.001) and more frequently with advanced liver disease (11.4 vs. 6.4%, P=0.007). During the first year, Greeks more frequently had several tests and eventually a complete diagnosis (68 vs. 55%, P<0.001). Greeks were more frequently in the phase of HBeAg-negative CHB and less frequently in the phase of inactive carrier or HBeAg-positive CHB, but age was the main determinant for these differences in multivariate analysis. Treatment was initiated more frequently by Greeks than immigrants with treatment indications (86 vs. 65%, P<0.001). Only 30-33% of treated and 4-10% of untreated patients remained under follow-up at year 5, without significant differences between Greeks and immigrants. CONCLUSION: Adherence to follow-up recommendations is rather poor for all chronic HBV patients. Immigrants are lost more frequently during the first year, but only small proportions of treated and particularly untreated Greek or immigrant patients remain under long-term follow-up.

Changes in serum transforming growth factor-ß1 levels in chronic hepatitis C patients under antiviral therapy.

BACKGROUND: Several cytokines including transforming growth factor (TGF)-ß1 have been suggested to be involved in the pathogenesis of fibrosis in chronic hepatitis C. We examined the changes of TGF-ß1 serum levels and their predictive value in patients with chronic hepatitis C under antiviral therapy. METHODS: We included 84 patients with chronic hepatitis C who were treated with pegylated interferon-α and ribavirin between 2008 and 2009. Treatment was given for 24-48 weeks depending on HCV genotype. Serum TGF-ß1 levels were measured by an ELISA assay at baseline, at the end of therapy (EOT), and at 6 months after the EOT. Liver fibrosis was evaluated by transient elastography. RESULTS: Of the 84 patients, 76.2% achieved sustained virological response (SVR), 8.3% responded at the EOT but relapsed during post-therapy follow up (RR) and 15.5% had no response (NR). In all patients, mean TGF-ß1 levels were 16,980 pg/mL at baseline and decreased significantly at EOT (12,041 pg/mL) and at 6 months of post-treatment follow up (13,254 pg/mL) (P≤0.001). In particular, mean TGF-ß1 levels decreased significantly from baseline to EOT and to six months of post-treatment follow up in patients with SVR and numerically but not significantly in patients with RR or NR. TGF-ß1 levels were not associated with the severity of liver stiffness estimated by transient elastography. CONCLUSION: Our data show that TGF-ß1 serum levels decrease significantly at the EOT and remain decreased 6 months after the EOT mostly in chronic hepatitis C patients who achieve SVR after pegylated interferon-α and ribavirin combination treatment.

Analysis of serum α-fetoprotein-L3% and des-γ carboxyprothrombin markers in cases with misleading hepatocellular carcinoma total α-fetoprotein levels.

Serum fraction of α-fetoprotein L3 (AFP-L3%) and des-γ carboxyprothrombin (DCP) are proposed serum markers for the diagnosis of hepatocellular carcinoma (HCC). We evaluated their performance in two patient populations with total AFP levels non-diagnostic for HCC. From a cohort of 150 consecutive patients with HCC, 60 patients with total AFP <200 ng/ml were identified. Additionally, 50 patients with elevated AFP and no radiological evidence of HCC, for at least one year of follow-up, were included. AFP-L3% and DCP were measured by the Liquid Phase Binding Assay System (LiBASys). In cases where AFP-L3% was undetectable, a more sensitive method based on-chip electrokinetic reaction was applied. AFP-L3% was found to be positive in 22 (36.7%) of patients with HCC and 6 (12%) of non-HCC patients. DCP was found to be positive in 26 patients with HCC (43%) and in none of the non-HCC patients. Thirty-six out of sixty (60%) patients with HCC were positive for either AFP-L3% or DCP. With the on-chip technology, AFP-L3% was found to be positive in 10 patients with HCC and in 5 patients without HCC, who tested negative by LiBASys. The final sensitivity of combined AFP, AFP-L3% and DCP testing, in the entire cohort of patients with HCC, was 84%. The specificity of AFP-L3% and DCP in the studied population was 78.5 and 100%, respectively. The addition of AFP-L3% and DCP increased the sensitivity and specificity of total serum AFP for the diagnosis of HCC. The on chip AFP-L3% assay was more sensitive but less specific compared to LiBASys.

Is HCV core antigen a reliable marker of viral load? An evaluation of HCV core antigen automated immunoassay.

BACKGROUND: Hepatitis C viral (HCV) load detection and quantification is routinely accomplished by HCV RNA measurement, an expensive but essential test, both for the diagnosis and treatment of chronic hepatitis C (CHC). HCV core antigen (Ag) testing has been suggested as an attractive alternative to molecular diagnostics. The aim of the study was to evaluate an automated chemiluminescent immunoassay (CLIA) for HCV core Ag measurement in comparison to quantitative HCV RNA determination. METHODS: HCV Ag was measured in 105 anti-HCV positive patients, from which 89 were HCV RNA positive with CHC and 16 HCV RNA negative after spontaneous HCV clearance. Viral load was quantified with branched DNA (bDNA, Versant, Siemens). Sera were stored at -70°C and then tested with the Architect HCV Ag test (Abbott Laboratories), a two-step CLIA assay, with high throughput and minimal handling of the specimens. Statistical analysis was performed on logarithmically transformed values. RESULTS: HCV-Ag was detectable and quantifiable in 83/89 and in grey zone in 4/89 HCV RNA positive sera. HCV-Ag was undetectable in all 16 HCV RNA negative samples. The sample with the lowest viral load that tested positive for HCV-Ag contained 1200 IU/mL HCV RNA. There was a positive correlation between HCV RNA and HCV-Ag (r=0.89). The HCV RNA/ HCV Ag ratio varied from 1.5 to 3.25. CONCLUSION: The HCV core Ag is an easy test with comparable sensitivity (>90%) and satisfactory correlation with the HCV RNA bDNA assay. Its role in diagnostics and other clinical applications has to be determined based on cost effectiveness.

Efficacy of tissue autofluorescence imaging (VELScope) in the visualization of oral mucosal lesions.

BACKGROUND: Technology that highlights potentially malignant oral lesions in a highly sensitive and specific manner will aid clinicians in early diagnosis of these conditions. This study assessed the efficacy of direct tissue autofluorescence imaging Visually Enhanced Lesion Scope (VELScope) in the detection of oral mucosal lesions. METHODS: One hundred twelve patients referred with a potentially malignant oral mucosal lesion were examined under routine incandescent light, and then with VELScope, noting loss of autofluorescence and presence of blanching. Incisional biopsies were performed to provide definitive histopathological diagnoses. RESULTS: VELScope enhanced the visibility of 41 lesions and helped uncover 5 clinically undetected lesions. VELScope examination alone showed a sensitivity of 30% and a specificity of 63%. Its accuracy at identifying dysplasia was 55%. CONCLUSION: VELScope examination cannot provide a definitive diagnosis regarding the presence of epithelial dysplasia. Loss of autofluorescence is not useful in diagnosing epithelial dysplasia in its own right without relevant clinical interpretation.

Changes of serum levels of keratin-18 fragments in hepatitis B e antigen-negative chronic hepatitis B patients under oral antiviral therapy.

BACKGROUND: The effect of chronic hepatitis B (CHB) treatment on hepatocyte apoptosis has not been evaluated. We investigated the changes in serum levels of the apoptotic caspase-generated fragments of keratin-18 (K-18) in hepatitis B e antigen (HBeAg)-negative CHB patients under nucleoside/nucleotide analogue(s). METHODS: We included 46 patients with HBeAg-negative CHB who had received lamivudine or adefovir monotherapy for ≥12 months. Of the 42 lamivudine treated patients, 23 developed virological breakthroughs and received add-on adefovir therapy for ≥12 months. Levels of K-18 fragments were measured blindly in stored serum samples using the M30-Apoptosense(®) ELISA Kit (PEVIVA, Alexis, Grünwald, Germany). RESULTS: During initial therapy, the median serum K-18 fragment levels were significantly reduced from baseline (280 U/l [149-2,461]) to 6 months (164 U/l [110-454]; P<0.001) and from 6 to 12 months (145 U/l [113-280]; P=0.031). K-18 fragment levels increased at breakthroughs (187 U/l [137-1,063]) compared to last previous visits (145 U/l [118-389]; P=0.015) but remained lower than baseline (P=0.036). Changes of K-18 fragment levels correlated positively with changes of alanine aminotransferase (ALT) and HBV DNA from baseline to 6 or 12 months of initial therapy. In patients with breakthroughs, K-18 fragment levels had only a trend towards a decrease from adefovir addition (198 U/l [124-1,219]) to 6 months (170 U/l [122-1,576]; P=0.109) and mainly to 12 months (156 U/l [122-878]; P=0.055). CONCLUSIONS: Serum levels of K-18 fragments decrease significantly during oral antiviral therapy in HBeAg-negative CHB in parallel with the improvements in ALT and HBV DNA levels. They increased again during breakthroughs and improved after an effective rescue therapy, perhaps at a slower rate compared to the initial therapy.

Bax, cytochrome c, and caspase-8 staining in parotid cancer patients: markers of susceptibility in radiotherapy?

OBJECTIVE: Negative bcl-2 and HLA-DR protein expression have been associated with responsiveness to adjuvant radiotherapy in surgically treated parotid cancer patients. The aim of this study was to investigate the prognostic significance of bax, cytochrome c, and caspase-8 protein expression in a group of surgically treated patients to determine whether they also suggest markers of responsiveness to adjuvant radiotherapy. STUDY DESIGN: Historical cohort study. SETTING: Otolaryngology department in a university hospital. SUBJECTS AND METHODS: The immunohistochemical expression of bax, cytochrome c, and caspase-8 were studied in paraffin-embedded tissue specimens originating from 27 surgically treated parotid cancer patients and nine patients with Warthin parotid tumors (control group) and correlated with the patients' clinicopathological characteristics and clinical outcome. RESULTS: Caspase-8 negative staining was more frequently observed in higher TNM stages and in tumors measuring more than 4 cm (P = 0.009 and P = 0.018, respectively). Caspase-8 (-)/cytochrome c (-) patients carried low-grade lesions without nodal involvement (P = 0.01 and P = 0.05, respectively). Caspase-8 (-) patients who received postoperative radiotherapy presented a significantly increased disease-free survival compared to those who did not (P = 0.04). Patients bearing bax (-) tumors who received postoperative radiotherapy presented an improved four-year disease-free survival compared to bax (-) patients who did not receive any type of adjuvant radiotherapy (P = 0.017). CONCLUSION: Bax, cytochrome c, and caspase-8 protein expression failed to independently predict survival in parotid cancer patients. However, patients with bax (-) or caspase-8 (-) tumors should be considered as candidates for adjuvant radiotherapy in order to achieve better local disease control.

Serum apoptotic caspase activity in chronic hepatitis C and nonalcoholic Fatty liver disease.

BACKGROUND: Apoptotic caspases are substantially activated in liver and serum caspase activity has been suggested as a marker of early liver injury. AIM: To investigate whether serum levels of caspase-generated fragments of cytokeratin-18 are associated with the severity of histologic lesions in chronic hepatitis C virus (HCV) infection and nonalcoholic fatty liver disease (NAFLD). METHODS: We included 134 patients with chronic HCV infection and 58 patients with NAFLD, who consecutively underwent liver biopsy, and 40 healthy controls. Caspase-generated cytokeratin-18 fragment levels were blindly measured in stored serum samples. RESULTS: Median cytokeratin-18 fragment levels were lower in HCV-positive patients with minimal/mild than patients with moderate/severe histologic lesions (174 U/L vs. 223 U/L, P<0.001) offering moderate accuracy for differentiation between the 2 groups (c-statistic: 0.74). Cytokeratin-18 fragments levels were lower in healthy subjects (148 U/L) than patients with simple fatty liver (174 U/L, P=0.013) than patients with nonalcoholic steatohepatitis (355 U/L, P<0.001) offering excellent diagnostic accuracy for differentiation between the 2 latter groups (c-statistic: 0.87). CONCLUSIONS: Serum apoptotic caspase activity is associated with the severity of liver histologic lesions in both chronic HCV infection and NAFLD, but it has excellent diagnostic accuracy in NAFLD and moderate accuracy in chronic HCV patients.

BCL-2, p53 and HLA-DR antigen expression in surgically treated parotid cancer patients.

Our objective was to investigate the prognostic significance of bcl-2 protein, p53 protein and HLA-DR antigen expression in a group of surgically treated parotid cancer patients. We studied bcl-2, p53 and HLA-DR immunohistochemical expression in paraffin-embedded surgically removed tissue specimens derived from 26 patients with parotid cancer and 9 patients with Warthin parotid tumors operated between 2000 and 2006 at the Hippokration Hospital of Athens. The staining results were correlated with the patients' clinicopathological characteristics and clinical outcome. Bcl-2 expression was associated with a significantly decreased survival in patients with advanced tumor stage (P = 0.04), high grade lesions (P = 0.02), or cervical node involvement (P = 0.03). Radiotherapy was associated with a significantly improved recurrence-free survival among patients with negative tumor staining for either bcl-2, or both HLA-DR and bcl-2 [HLA-DR(-)/Bcl-2(-)] (P = 0.04 for both comparisons). Classical clinicopathologic factors failed to show prognostic value both in the univariate and the multivariate analyses performed. Our results suggest that bcl-2 can be used to identify locally advanced or histologically aggressive tumors with a lower survival probability following the application of standard treatment modalities. Furthermore, bcl-2(+) patients should be considered for more aggressive adjuvant treatment protocols, since conventional radiotherapy often fails to decrease relapse rates in this setting of patients.

Serum adipokine levels in chronic liver diseases: association of resistin levels with fibrosis severity.

OBJECTIVE: Leptin and adiponectin have been implicated in the pathogenesis and progression of non-alcoholic steatohepatitis (NASH) and chronic hepatitis C (CHC), but little is known about the role of resistin in chronic liver diseases. The objective of this study was to investigate serum levels of the above three adipokines in relation to the etiology of liver disease and to determine their associations with histological severity. MATERIAL AND METHODS: We prospectively evaluated 146 patients (HBeAg-negative chronic hepatitis B (CHB): 52, CHC: 70, NASH: 24) who consecutively underwent liver biopsy. Detailed epidemiological, anthropometric and laboratory data were recorded. Histological lesions were evaluated blindly according to the Ishak and the Brunt classifications for CHB/CHC and NASH, respectively. RESULTS: Serum adipokine levels were similar between CHB and CHC patients, while CHB/CHC patients had significantly lower leptin levels compared with NASH patients (8.3+/-7.3 versus 17.6+/-16.6 ng/ml, p=0.012) and higher adiponectin (10.2+/-5.1 versus 7.5+/-4 microg/ml, p=0.018) and resistin levels (7.1+/-2.5 versus 5.7+/-2.8 ng/ml, p=0.016). In CHB/CHC, there was no significant association between steatosis or necroinflammation and levels of adipokines, while the presence of moderate/severe fibrosis (stages 4-6) was associated with higher leptin and adiponectin levels in male but not in female patients and with lower resistin levels irrespective of gender or other factors (adjusted odds ratio=0.788, p=0.035). CONCLUSIONS: Serum adipokine levels depend on the etiology of liver disease differing between chronic viral hepatitis and NASH, but not between CHB and CHC. In CHB/CHC, resistin levels are independently associated with fibrosis severity, whereas in the association of leptin and adiponectin levels with fibrosis, it seems to be a gender effect.

Performance characteristics of microparticle enzyme and chemiluminescence immunoassays for measurement of anti-HBc immunoglobulin M in sera of patients with HBeAg-negative chronic hepatitis B virus infection.

The IMx, AxSym, and Architect immunoglobulin M anti-HBc assay systems for detecting hepatitis B virus e antigen-negative chronic hepatitis B virus infection were compared. Despite good intra- and interassay coefficients of variation, significantly different values and low correlation (overestimation by AxSym and underestimation by Architect) were observed. Association and cutoff values for distinguishing patients with viral replication should be established for all methods.

Primary small cell undifferentiated (neuroendocrine) carcinoma of the maxillary sinus.

Primary small cell undifferentiated (neuroendocrine) carcinomas of the paranasal sinuses are extremely uncommon neoplasms. This tumor was first reported in this site in 1965, and since then there have been only 61 documented cases in the literature. The median age at presentation is 53 years, with no gender predilection. There is no reported association of occurrence of this tumor with either tobacco use or form of occupation, and case outcome is usually poor. We report a case in a 25-year-old man, initially treated as an odontogenic infection and thus delaying institution of appropriate management. Further investigation identified a locally advanced neuroendocrine carcinoma of the left maxilla. Despite radiotherapy and chemotherapy, the patient exhibited rapid tumor dissemination and died.

Otoacoustic emissions in universal hearing screening: which day after birth should we examine the newborns?

Transiently evoked otoacoustic emissions (TEOAEs) have been widely used in universal newborn hearing screening programs. Although there is consensus with regard to the avoidance of early screening, especially during the first hours after birth, the optimum testing day is not yet unanimously accepted. The aim of the present study was to compare the 'pass-refer' results between 4 groups of newborns tested during the 4 postbirth days and determine the most appropriate day for assessing newborn hearing. Our results suggest that, although TEOAEs can be recorded in very high rates from the first 24 h of life, 'refer' scores are lower on the third and fourth days after birth. It may be thus concluded that the optimum time of assessing newborn hearing in universal hearing screening programs seems to be the third or fourth postbirth day, provided that other social or financial reasons do not suggest an earlier discharge from the hospital.

Cell kinetics and apoptosis in laryngeal carcinoma patients.

Cellular proliferation and apoptosis are both implicated in the process of carcinogenesis. The objective of this study was to access the prognostic significance of the expression of proliferating cell nuclear antigen (PCNA) and the apoptosis-related genes (bax, bcl-2, and p53) in laryngeal carcinoma patients. Thirty consecutive patients with stage I to IV squamous cell laryngeal carcinoma were treated in our department from 1992 to 1994. We immunohistochemically studied the expression of PCNA and bax, bcl-2, and p53 genes in their tumor specimens. Five healthy men were used as the control group. The staining results were correlated with clinicopathologic data. The PCNA protein expression was correlated with a significantly worse survival in those patients who were bax-negative (0% versus 42.86%, p = .0445). Similarly, the presence of PCNA led to an unfavorable clinical outcome in those patients who were bax-negative, bcl-2-negative, and p53-negative (0% versus 50%, p = .0278). Expression of bcl-2 protein was found to be an independent prognostic factor related to an unfavorable clinical outcome (p = .0262). The expression of bcl-2 protein appears to predict survival in laryngeal carcinoma patients. Furthermore, the combined study of proliferation markers and apoptosis-related genes helped us to identify a high-risk group of patients who may benefit from a more aggressive treatment protocol.

The effect of different 'pass-fail' criteria on the results of a newborn hearing screening program.

'Pass' criteria in newborn hearing screening programs are important, since they affect the operating characteristics of the programs. In the present study, we intended to compare the results of two screening procedures, using different 'pass' criteria, in two samples from the same pool of screened newborns. The subjects were divided into two study groups, screened consecutively during 6 months. Testing and all procedures were exactly the same in both groups, differing only in the 'pass' criteria. In the first group a signal-to-noise ratio of at least 3 dB in the frequency bands of 1-2, 2-3 and 3-4 kHz was considered necessary for a 'pass', whereas a signal-to-noise ratio > or =6 dB was used in the second group, at the same frequency bands. During the period of the study, no other minor or major modification of the protocol was applied. The comparison of the screening predischarge results between the two groups showed no statistically significant differences in the 'pass-refer' results. Thus, it appears that the 3-dB signal-to-noise ratio is as valid as the 6-dB criterion, and it may be confidently used, especially in settings where rescreening is not available.