Clinical biomarkers for cancer recognition and prevention: A novel approach with optical measurements.

Cancer is the most important cause of death worldwide, and early cancer detection is the most fundamental factor for efficacy of treatment, prognosis, and increasing survival rate. Over the years great effort has been devoted to discovering and testing new biomarkers that can improve its diagnosis, especially at an early stage. Here we report the potential usefulness of new, easily applicable, non-invasive and relatively low-cost clinical biomarkers, based on abnormalities of oral mucosa spectral reflectance and fractal geometry of the vascular networks in several different tissues, for identification of hereditary non-polyposis colorectal cancer carriers as well for detection of other tumors, even at an early stage. In the near future the methodology/technology of these procedures should be improved, thus making possible their applicability worldwide as screening tools for early recognition and prevention of cancer.

Survival rate and prevalence of bronchopulmonary dysplasia in extremely low birth weight infants.

BACKGROUND: Bronchopulmonary dysplasia (BPD) continues to represent a significant cause of morbidity among survivors of severe prematurity in the NICU. The increasing prevalence of BPD over the decades has been suggested to be related to the increased survival of extremely low birthweight infants. AIMS: To evaluate differences in prevalence of BPD (BPD28d and BPD36wk) and as a function of survival rate in extremely low birth weight (ELBW) infants over time, and to explore its relationship with known associated risk factors. METHODS: Survival rate and prevalence of oxygen-dependency =28 days (BPD28d) and oxygen-dependency =36 weeks postmenstrual age (BPD36wk) were evaluated in ELBW newborns (mean gestational age: 27.12.2 weeks; mean birth weight: 817142 g) consecutively admitted to the Brindisi NICU over the last 26 years. Two arbitrarily chosen time periods were compared: Period 1: July 1st, 1986 to June 30, 2002 vs. Period 2: July 1st, 2002 to December 31, 2012. Analyzed variables included gestational age, birth weight, intubation time, hours of O2 administration, NCPAP, and use of surfactant. Differences between the time periods were assessed by chi-square statistics, Fisher's tests or Mann-Whitney test, as appropriate. A two-tailed p value <0.05 was considered to indicate statistical significance. RESULTS: Survival rate of ELBW infants over the examined time periods dramatically improved from 42.3% to 72.6% (p < 0.0001), whereas changes in the prevalence of BPD28d and BPD36wk were not statistically significant (30.5% vs. 39.3%, p = 0.2137 and 5.5% vs. 13.1%, p = 0.1452, respectively). Likewise, BPD severity was not significantly different between the two time periods (p = 0.1635). Gestational age and birth weight of surviving neonates did not significantly change between the two time periods (p = 0.8050 and p = 0.6986, respectively), whereas significantly increased intubation time (median values: 144 hours vs. 33 hours, p <0.0001) and use of exogenous surfactant (89.3% vs. 48.6%, p < 0.0001) was evidenced for the second time period, as well as NCPAP (median values: 600 hours vs. 377 hours, p = 0.0005). A statistically non-significant trend for a prolonged O2 administration in period 2 (p = 0.0850) was also observed. CONCLUSION: Our findings indicate that a significantly increased survival is not necessarily associated with a significant difference in the prevalence of BPD among ELBW infants.

Oral mucosal color changes as a clinical biomarker for cancer detection.

Screening is a key tool for early cancer detection/prevention and potentially saves lives. Oral mucosal vascular aberrations and color changes have been reported in hereditary nonpolyposis colorectal cancer patients, possibly reflecting a subclinical extracellular matrix abnormality implicated in the general process of cancer development. Reasoning that physicochemical changes of a tissue should affect its optical properties, we investigated the diagnostic ability of oral mucosal color to identify patients with several types of cancer. A total of 67 patients with several histologically proven malignancies at different stages were enrolled along with a group of 60 healthy controls of comparable age and sex ratio. Oral mucosal color was measured in selected areas, and then univariate, cluster, and principal component analyses were carried out. Lower red and green and higher blue values were significantly associated with evidence of cancer (all P<0.0001), and efficiently discriminated patients from controls. The blue color coordinate showed significantly higher sensitivity and specificity (96.66±2.77 and 97.16±3.46%, respectively) compared with the red and green coordinates. Likewise, the second principal component coordinate of the red-green clusters discriminated patients from controls with 98.2% sensitivity and 95% specificity (cut-off criterion≤0.4547; P=0.0001). The scatterplots of the chrominances revealed the formation of two well separated clusters, separating cancer patients from controls with a 99.4% probability of correct classification. These findings highlight the ability of oral color to encode clinically relevant biophysical information. In the near future, this low-cost and noninvasive method may become a useful tool for early cancer detection.

Di-(2-ethylhexyl)-phthalate migration from irradiated poly(vinyl chloride) blood bags for graft-vs-host disease prevention.

Irradiation with 20-25 kGy is a process commonly used for sterilizing poly(vinyl chloride) (PVC) medical devices. Moreover, whole blood and blood components undergo additional irradiation with 25-50 Gy to inhibit the proliferative capacity of lymphocytes and reduce the risk of transfusion-associated graft-vs-host disease (GVHD). Di-(2-ethylhexyl)-phthalate (DEHP) plasticized PVC is extensively used for the production of flexible medical devices including blood bags, but since DEHP is not covalently bound to PVC, it tends to migrate and leach out of the medical device, with harmful consequences for the patients. In this study, the effects of different doses of gamma irradiation on DEHP migration from PVC blood bags was investigated using differential scanning calorimetry (DSC) analysis. Our findings indicate that irradiation with 25-100 Gy reduces the ability of DEHP to migrate from the blood bags, and in the case of a primary container a correlation between the doses of gamma ray irradiation was also observed. In particular, a decrease in DEHP leachability was obtained by increasing the dose of gamma ray irradiation.

Ultrasound mass screening for congenital anomalies of the kidney and urinary tract.

BACKGROUND: There is a high incidence of congenital anomalies of the kidneys and urinary tract (CAKUT). Early diagnosis of these defects may allow the best medical and/or surgical treatment to be implemented as rapidly as possible, preventing or at least slowing down an evolution toward chronic kidney disease. METHODS: Ultrasound mass screening for kidney and urinary tract abnormalities in infants at 2 months of age was carried out in Salento, Italy. The centers involved in the study examined a total of 17,783 infants between January 1992 and December 2010. RESULTS: A total of 171 CAKUT were identified in the course of the mass screening. The frequency of CAKUT was 0.96%. Vesicoureteral reflux (n = 39) was the most frequent renal abnormality found, followed by ureteropelvic junction obstruction (n = 33), ectopic kidney (n = 26), and renal dysplasia (n = 19). In addition, nephrogenic rests (n = 2), as well as several extra-renal pathologies, including abdominal neuroblastoma (n = 3), were diagnosed incidentally. CONCLUSION: Ultrasound has been effective for early detection of renal and urinary tract anomalies. In addition, this screening has proved to be very useful for the early identification and management of both renal and extra-renal precancerous as well as cancerous lesions. However, most patients requiring surgery in this study (0.24%) would probably have been symptomatic and come to medical attention without routine screening. On the basis of our results screening is not justified.

Systematic underestimation of oxygen delivery in ventilated preterm infants.

BACKGROUND: Emerging evidence indicates that hyperoxia is a risk factor for bronchopulmonary dysplasia, a common multifactorial long-term complication of prematurity. To date, the equivalence between set and delivered oxygen (O(2)) in ventilated preterm infants has not been rigorously studied. OBJECTIVES: To test the hypothesis of systematic underestimation of O(2) delivery in extremely low birth weight (ELBW) infants during long-term ventilation. METHODS: Actually achieved O(2) concentrations were measured and compared to the set inspired oxygen fraction (FiO(2)). A total of 108 O(2) measurements were carried out during the ventilation of 54 ELBW infants: O(2)-Delta error (i.e., the difference between O(2) concentrations achieved by the ventilator and set FiO(2)) was the main study outcome measure. RESULTS: Systematic O(2)-Delta errors were found, with mean values of +9.52% (FiO(2) 0.21-0.40), +2.10 (FiO(2) 0.41-0.60), +2.86% (FiO(2) 0.61-0.80), and +0.016% (FiO(2) 0.81-1.0; p < 0.0001). Theoretical simulations from the observed data indicate that, if not corrected, systematic O2-Delta errors would lead to a non-intentional total O(2) load of 1,202.9 (FiO(2) 0.21-0.40), 252.46 (FiO(2) 0.41-0.60), 342.85 (FiO(2) 0.61-0.80), and 2 (FiO(2) 0.81-1.0) extra liters/kg body weight/100 ventilation hours. CONCLUSIONS: Systematic underestimation of the O(2) delivered by infant ventilators can potentially lead to surprisingly large increases in total O(2) load during long-term ventilation of ELBW infants, especially in the lower FiO(2) range (i.e., 0.21-0.40). Underestimation of true O(2) delivery can potentially lead to unrecognized high O(2) loads, and more pronounced and prolonged hyperoxia.

Persistent pulmonary hypertension of the newborn: therapeutical approach.

Persistent pulmonary hypertension of the newborn (PPHN), is defined as a failure of the pulmonary vasculature to relax at birth and consequently of the normal adaptation to extra uterine life of the fetal heart/lung system, resulting in hypoxemia. This condition, occurs in about 1-2 newborns per 1000 live births and despite significant improvements in treatment it is associated with substantial infant mortality and morbidity. Over the years wider application of inhaled nitric oxide (iNO) therapy and improved ventilation strategies including surfactant, high-frequency oscillatory ventilation has led to a decrease in the need for invasive life-sustaining therapies such as extracorporeal membrane oxygenation (ECMO). Mortality rate varies from 10 to 20 % of affected newborns in developed countries, but it is much higher when PPHN is refractory to the above reported therapies or when they are not available. As a consequence, development of new therapeutic strategies for severe PPHN is crucial. In particular, recent studies seem to show that sildenafil, a phosphodiesterase inhibitor type 5 that selectively reduces pulmonary vascular resistance may be a useful therapeutic adjunct to critically ill neonates with PPHN.

Recurrent otitis media with effusion in preterm infants with histologic chorioamnionitis--a 3 years follow-up study.

BACKGROUND: Recurrent otitis media with effusion (OME) is a leading cause of acquired hearing loss in childhood. Histological chorioamnionitis (HCA) is an important cause of preterm delivery and neonatal morbidity and mortality. Here, we tested the hypothesis of an association between recurrent OME during the first 3 years of life and HCA in very low birth weight (VLBW) infants. METHODS: A total of 110 randomly selected VLBW preterm newborns with HCA and 135 gestational age and gender-matched, HCA-negative VLBW infants were evaluated prospectively during the first 3 years of life for the presence of OME, as diagnosed on the basis of otoscopy, type B or C tympanogram, ipsilateral absence of transient evoked otoacoustic emissions responses, and ipsilaterally increased threshold at diagnostic auditory brain responses evaluation. Potential risk factors for OME were also examined in the two groups. RESULTS: The HCA-positive infants showed a approximately six times higher frequency of recurrent OME (P<0.0001), increased frequency (>5/yr) of clinical otitis media episodes (P=0.000020), approximately five times higher frequency of adenoid hypertrophy (P<0.00001), a significant seasonal pattern of birth with autumn predominance (P<0.00001), and the first OME occurred earlier (P<0.0001), as compared to the HCA-negative counterparts. Recurrent OME was significantly associated with HCA (O.R.=17.76, 95% CI: 8.98-35.13, P<0.00001), adenoid hypertrophy (O.R.=9.96, 95% CI: 5.17-19.18, P<0.00001), frequency of acute otitis episodes >5/yr (O.R.=8.91, 95% CI: 1.96-40.41, P=0.0005), and birth in autumn (O.R.=5.58, 95% CI: 2.79-11.12, P<0.00001). CONCLUSIONS: These findings indicate that HCA is a previously unrecognized risk factor for the development of recurrent bilateral OME in VLBW preterm infants during the first 3 years of life.

Nephrogenic rests: their frequency and their fate.

Nephrogenic rests (NRs) are considered to be precursor lesions of Wilms tumor, the most common malignant neoplasm of the urinary tract in children. We have previously reported on 2 cases of NRs, incidentally diagnosed at 2 to 3 months of age during an ultrasound mass screening for urinary tract malformations between 1992 and 2006. As the screened population consisted of 17,065 infants, the observed prevalence of NRs in our area in the examined time period was of 1.17/10,000. This is the first reliable estimate of the frequency of clinically appreciable NRs in infants. Microscopic NRs have been found at autopsy in about 1% of infants. Our data are, therefore, helpful in the assessment of the proportion of NRs that disappear spontaneously in the childhood age group. To the best of our knowledge, no false-negative cases were found. Our observations indicate that our policy of "wait and see" is appropriate when NRs are identified incidentally during ultrasonographic screening done for whatever purpose.

Abnormal oral mucosal light reflectance in achondroplasia.

No markers for the unaffected parents of children with achondroplasia (ACH) are known to date. Here, the value of oral mucosal reflectance in identifying unaffected parents of ACH patients with sporadic mutations was assessed. The study involved ACH children (n = 30), unaffected ACH parents (n = 60), and a sex- and age-matched controls (control children, n = 30; control parents, n = 60). Light reflectance of the lower gingival and vestibular oral mucosa was measured using an imaging reflectance system in the optical spectrum (400-700 nm wavelengths). Significantly higher reflectance values in the 410 to 630 nm wavelength range were observed in ACH patients (P < or = .0063), whereas significantly lower reflectance values over the whole optical spectrum were observed in ACH parents (P < or = .0001). A reflectance cutoff value < or =4.71% at the 430 nm wavelength identified ACH parents with 100% sensitivity and 96.4% specificity. These findings indicate the presence of previously unrecognized oral mucosal reflectance abnormalities in unaffected ACH parents of ACH children.

Recent advances on neural tube defects with special reference to Valproic Acid.

Epilepsy is a common medical problem and many studies have demonstrated that infants of women with epilepsy (WWE) have a two to threefold higher risk of congenital malformations compared with the background population. The majority of WWE have normal, healthy children. However, WWE have an increased risk of congenital malformations. Congenital malformations are twice as common in infants exposed to antiepileptic drugs in utero. A variety of congenital malformations have been reported, with a particular preponderance of orofacial clefts. Valproate is often associated with the development of neural tube defects. In this review, we analyse the problem of neural tube defects and report in detail the main pathogenetic theories about the onset of this type of congenital malformation. There is strong evidence for a protective effect of adequate folate consumption.

Low serum concentrations of di-(2-ethylhexyl)phthalate in women with uterine fibromatosis.

Di-(2-ethylhexyl)phthalate (DEHP) is the most commonly used plasticizer in flexible polyvinylchloride formulations, and is a widespread ubiquitous environmental contaminant. A potential role of exposure to DEHP and its primary metabolite, monoethylhexylphthalate (MEHP), on women's reproductive function is suggested in the current study. The aim of the study was to test serum concentrations of DEHP and/or MEHP in women with uterine fibromatosis. Two groups of women were enrolled in the study: (i) women with uterine fibromatosis undergoing surgical menopause (n = 15) and (ii) healthy women (n = 20). Serum DEHP and MEHP concentrations were measured by high-performance liquid chromatography. Serum MEHP distribution was found to be non-Gaussian (p = 0.001) while serum DEHP distribution was compatible with a normal curve (p = 0.141). Patients with uterine fibromatosis showed significantly lower serum MEHP concentrations (median [interquartile range]: 0 [0-0] microg/ml, range: 0-0.57 microg/ml) than controls (0.42 [0-0.51] microg/ml, range: 0-1.20 microg/ml, z = -2.93, p = 0.0034). Likewise, serum DEHP concentrations in women with fibromatosis were found to be significantly lower than in controls (patients: 0.27 +/- 0.096 microg/ml (mean +/- standard deviation (SD)), range: 0.14-0.59 microg/ml vs. controls: 0.30 +/- 0.14 microg/ml (mean +/- SD), range: 0-0.63 microg/ml; t = 3.212, df = 33, difference: -0.325 (95% confidence interval: -0.5309, -0.1191), p = 0.0029). In conclusion, the present findings indicate for the first time that serum DEHP and MEHP concentration are lower in women with uterine fibromatosis, suggesting a possible correlation between phthalate esters and fibromatosis pathology.

Abnormal oral mucosal light reflectance in bronchopulmonary dysplasia.

BACKGROUND: Bronchopulmonary dysplasia (BPD) is an important cause of mortality and morbidity in preterm infants. A disordered vascular development and a decreased production of angiogenic factors have been recently reported in the condition. Extracellular matrix (ECM) is known to play an important role on angiogenesis and blood vessel geometry and changes in ECM components have been previously reported in experimental models and patients with BPD. Here, we aimed to assess the potential value of light reflectance on the oral mucosa in detecting infants who will develop BPD. METHODS: A total of 75 preterm newborns (gestational age: 27.7 +/- 2.8 weeks, birth weight: 870 +/- 145 g) were recruited to the study, of whom 25 developed BPD (gestational age: 26.9 +/- 3.0 weeks, birth weight: 855 +/- 150 g). Reflectance was measured on the postnatal days 1 and 28, using high-resolution photographs of the lower gingival and vestibular oral mucosa, using imaging spectrophotometry in the 400-700 nm wavelength electromagnetic spectral range. The median of artefact- and vessel-free areas was n = 78 (interquartile range: 59-88). Median range values were comparable for both groups: BPD-positive infants, median 77 (interquartile: 60-90) vs. control infants, median 74 (interquartile: 62-92). The predictive accuracy of oral spectrophotometry was calculated using receiver operating characteristic curve analysis. RESULTS: BPD patients showed significantly lower light reflectance values in the red (610-700 nm, P < 0.0001), with higher values in the violet (400 nm, P = 0.0056; 430 nm, P=0.014), and blue-green (480-500 nm, P < or = 0.024) sections of the spectrum already on the first day of life. A low reflectance value in the 640-700 nm wavelengths interval was found to identify BPD patients with 100% sensitivity and 100% specificity (640 nm: cutoff < or = 44.91%; 650 nm: < or = 45.64%; 660 nm: < or = 46.56%; 670 nm: < or = 47.14%; 680 nm: < or = 47.56%; 690 nm: < or = 48.95%; 700 nm: < or = 50.81%). CONCLUSIONS: These findings indicate the presence of previously unrecognised, early abnormalities in the average optical properties of the oral mucosa from infants developing BPD.

Oral mucosal microvascular abnormalities: an early marker of bronchopulmonary dysplasia.

An abnormal pulmonary vasculature has been reported as an important component of bronchopulmonary dysplasia (BPD). We tested the hypothesis of an early abnormal vascular network pattern in infants with BPD. Fifteen infants with BPD (nine boys and six girls; gestational age 27.5 +/- 2.0 wk; birth weight 850 +/- 125 g) and 15 sex- and gestational age-matched infants (nine boys and six girls; gestational age 27.6 +/- 2.6 wk; birth weight 865 +/- 135 g) were examined on postnatal days 1 and 28. BPD infants showed a significantly higher prevalence of histologic chorioamnionitis (p = 0.009), as well as higher intubation duration (p = 0.0004), oxygen supplementation (p < 0.0001), and initial illness severity (p = 0.0002) than the BPD-negative population. The lower gingival and vestibular oral mucosa was chosen as the study area. The blood vessel area was determined, and the oral vascular networks were characterized by analyzing their complexity (D, at two scales: D 1-46, D 1-15), tortuosity (Dmin), and randomness (L-Z) of the vascular loops. Infants with BPD showed a significantly lower blood vessel area as well as a higher vascular network complexity (D 1-46, D 1-15, and L-Z) than control subjects (p < 0.0001). Our findings provide a new early clinical sign in BPD and stress the importance of an early disorder in the oral mucosal vascularization process in the disease pathogenesis.

Abnormal oral vascular network geometric complexity in Ehlers-Danlos syndrome.

Ehlers-Danlos syndrome (EDS) leads to impaired extracellular matrix (ECM) structure. Since ECM plays a major role in blood vessel geometry, we tested the hypothesis of an abnormal geometrical complexity of the visible microvascular network in EDS. Twelve patients with EDS-II or EDS-III and a control population (n=12) were examined. The geometry characteristics for the networks on the lower vestibular oral mucosa were analyzed using high-resolution photographic images. Fractal dimension, D, at 2 scales (D 1-46 and D 1-15), blood vessel tortuosity (minimum-path fractal dimension , Dmin ), and relative Lempel-Ziv complexity (L-Z) values were determined. EDS vascular networks exhibited significantly higher D 1-46 (P <.00001) and D 1-15 (P <.00001), as well as L-Z complexity (P <.00001), together with lower Dmin values (P=.0001) than controls. These findings indicate the presence of a previously unrecognized microvascular network abnormality on the oral mucosa in EDS patients, and provide an additional phenotypical marker for the condition.

Abnormal oral mucosal light reflectance: a new clinical sign of Ehlers-Danlos syndrome.

Ehlers-Danlos syndrome (EDS) is a major inherited connective tissue disorder leading to an impaired extracellular matrix structure. Although several odontostomatologic signs have been reported, their diagnostic accuracy remains to be ascertained. We tested the hypothesis that EDS is associated with an abnormal reflectance of the oral mucosa. Twelve patients with EDS-II or EDS-III and 12 age- and gender-matched controls were examined. Reflectance of the lower gingival and vestibular oral mucosa in the optical spectrum was measured using an imaging spectrophotometer. EDS patients showed significantly higher reflectance values in the 400-590 nm wavelengths (P10.51% at the 400 nm wavelength identified the EDS patients with 100% sensitivity and 100% specificity. These findings indicate that an abnormal oral mucosal reflectance is a previously unrecognized clinical marker of EDS.