Search for Neuro-Endocrine Markers (Chromogranin A,Synaptophysin and VGF) in Breast Cancers. An integrated Approach Using Immunohistochemistry and Gene Expression Profiling

Discordant data are reported in the literature on the definition, incidence and clinical features of neuroendocrine (NE) carcinomas of the breast. This tumour entity is currently assessed by immunohistochemistry (IHC) detecting “general” NE markers such as chromogranin A (CHGA) and synaptophysin (SYP), but other markers have been considered as well. In the present study, in addition to CHGA and SYP, we investigated the expression of VGF, a neurotrophin-inducible gene, which is emerging as a new specific NE marker. In order to evaluate the differential expression of these neuro-endocrine markers in breast cancers, we conducted parallel immunohistochemical and gene expression analyses, using PCR, gene array and real-time quantitative PCR procedures. Data obtained in 28 cases were further validated with a meta-analysis of published datasets of 103 breast cancer cases. The value of IHC positivity (irrespective of the percentage of positive cells) was confirmed by over-expression of the related gene. However, the genetic approach emerged as more sensitive, showing over-expression of NE markers in a subset of IHC-negative carcinomas. In conclusion, the present study confirms, by a novel approach, the occurrence of NE differentiation in breast cancers. Over-expression of one or more NE marker (CHGA and/or SYP and/or VGF) characterizes a significant fraction (approximately 10 %) of infiltrative breast cancers.     

Trypanosoma cruzi nucleoside triphosphate diphosphohydrolase 1 (TcNTPDase-1) biochemical characterization,immunolocalization and possible role in host cell adhesion

Previous work has suggested that Trypanosoma cruzi diphosphohydrolase 1 (TcNTPDase-1) may be involved in the infection of mammalian cells and serve as a potential target for rational drug design. In this work, we produced recombinant TcNTPDase-1 and evaluated its nucleotidase activity, cellular localization and role in parasite adhesion to mammalian host cells. TcNTPDase-1 was able to utilize a broad range of triphosphate and diphosphate nucleosides. The enzyme's K_m for ATP (0.096 mM) suggested a capability to influence the host's ATP-dependent purinergic signaling. The use of specific polyclonal antibodies allowed us to confirm the presence of TcNTPDase-1 at the surface of parasites by confocal and electron microscopy. In addition, electron microscopy revealed that TcNTPDase-1 was also found in the flagellum, flagellum insertion region, kinetoplast, nucleus and intracellular vesicles. The presence of this enzyme in the flagellum insertion region and vesicles suggests that it may have a role in nutrient acquisition, and the widespread distribution of TcNTPDase-1 within the parasite suggests that it may be involved in other biological process. Adhesion assays using anti-TcNTPDase-1 polyclonal antibodies as a blocker or purified recombinant TcNTPDase-1 as a competitor revealed that the enzyme has a role in parasite–host cell adhesion. These data open new frontiers to future studies on this specific parasite–host interaction and other unknown functions of TcNTPDase-1 related to its ubiquitous localization.     

One year postoperative hard and soft tissue volumetric changes after a BSSO mandibular advancement

In this study, cone beam computed tomography (CBCT) and three dimensional (3D) stereophotogrammetry are used to compare the 3D skeletal and soft tissue changes caused by a bilateral sagittal split osteotomy (BSSO) 1 year after a mandibular advancement. Eighteen consecutive patients with a hypoplastic mandible were treated with a BSSO according to the Hunsuck modification. Preoperatively and 1 year postoperatively, a CBCT scan was acquired and a 3D photograph. The pre- and postoperative CBCT scans were matched using voxel based registration. After registration, the mandible could be segmented in the pre- and postoperative scans. The preoperative scan was subtracted from the postoperative scan, resulting in the hard tissue difference. To investigate the soft tissue changes, the pre- and postoperative 3D photographs were registered using surface based registration. After registration the preoperative surface could be subtracted from the postoperative surface, resulting in the overall volumetric difference. As expected, a correlation between mandibular advancent and volumetric changes of the hard tissues was found. The correlation between advancement and soft tissues was weak. The labial mental fold stretched after surgery. This study proved that using 3D imaging techniques it is possible to document volumetric surgical changes accurately and objectively.     

Influence of Er,Cr:YSGG laser irradiation on enamel caries prevention

The objective of this study was to evaluate the effects of chromium:yttrium-scandium-gallium-garnet (Er,Cr:YSGG) laser irradiation on the acid resistance of dental enamel. Forty human enamel samples were divided into four groups. They were manually irradiated with an Er,Cr:YSGG laser device (λ?=?2.78 µm, 20 Hz, 20 s), in a scanning mode, with and without water cooling, according to the following parameters: Group 1: 0.25 W, 62.5 J/cm², no water cooling; group 2: 0.25 W, 62.5 J/cm², 5.0 ml/min; group 3: 0.5 W, 125 J/cm², no water cooling; group 4: 0.5 W, 125 J/cm², 5.0 ml/min. No airflow was used. Afterwards, the samples were submitted to an acid challenge and assessed by cross-sectional Knoop microhardness at different depths (20, 40, 60, 80, and 100 µm) from the outer enamel surface. Average values were obtained for both irradiated and control areas in each sample and they were compared to obtain a percentage of microhardness increase. Data were analyzed by analysis of variance and Fisher’s exact test (α?=?5%). The percentage of microhardness increase observed in group 1 (+23.58%) was similar to group 3 (+19.12%), but higher than groups 2 (+3.61%) and 4 (10.9%) (p?<?0.05). The comparison of the depths showed that the Er,Cr:YSGG laser acted in the superficial layers of the dental enamel. The findings of the present study suggest that the energy densities of 62.5 and 125 J/cm² were capable of increasing the acid resistance of human enamel. The presence of water during irradiation makes it difficult to obtain an enamel surface more resistant to acids.     

A Two-Way Road: Rates of HIV Infection and Behavioral Risk Factors Among Deported Mexican Labor Migrants

A large number of Mexican migrants are deported to Mexico and released in the North Mexican border region every year. Despite their volume and high vulnerability, little is known about the level of HIV infection and related risk behaviors among this hard-to-reach population. We conducted a cross-sectional, probability survey with deported Mexican migrants in Tijuana, Mexico (N = 693) and estimated levels of HIV infection and behavioral risk factors among this migrant flow. The sample and population estimated rates of HIV for deported males were 1.23 and 0.80 %, respectively. No positive cases were found among the female sample. We found high lifetime rates of reported sexually transmitted infections (22.3 %) and last 12-months rates of unprotected sex (63.0 %), sex with multiple sexual partners (18.1 %), casual partners (25.7 %), and sex workers (8.6 %), compared to U.S. and Mexico adults. HIV prevention, testing, and treatment programs for this large, vulnerable, and transnational population need to be implemented in both the U.S. and Mexico.     

Assessment of nitrite oxide and maternal–fetal Doppler parameters during pregnancy

Objective: The objective was to evaluate and compare the whole blood nitrite concentration in the three trimesters of pregnancy. Additionally, we investigate whether there is any relation between nitrite concentrations and Doppler ultrasound analysis of some maternal and fetal vessels.

Methods: Thirty-three healthy pregnant women were examined at the first (11–14 weeks), second (20–24 weeks) and third trimester (34–36 weeks) of pregnancy. In the three exams, we determined the maternal whole blood nitrite concentration and uterine arteries Doppler analysis to determine pulsatility index (PI), and resistance index (RI). In the second and third trimester we also performed fetal umbilical and middle cerebral arteries PI and RI. We compared the concentrations of nitrite in three trimesters and correlated with Doppler parameters.

Results: No difference was observed in the whole blood nitrite concentrations across trimesters: 151.70?±?77.90?nmol/ml, 142.10?±?73.50?nmol/ml and 147.10?±?87.30?nmol/ml; first, second and third trimesters, respectively. We found no difference in correlation between whole blood nitrite concentration and Doppler parameters from the evaluated vessels.

Conclusions: In healthy pregnant women, the nitrite concentrations did not change across gestational trimesters and there was also no strong correlation with Doppler impedance indices from maternal uterine arteries and fetal umbilical and middle cerebral arteries.