A Full Set of In Vitro Assays in Chitosan/Tween 80 Microspheres Loaded with Magnetite Nanoparticles.

Microspheres have been proposed for different medical applications, such as the delivery of therapeutic proteins. The first step, before evaluating the functionality of a protein delivery system, is to evaluate their biological safety. In this work, we developed chitosan/Tween 80 microspheres loaded with magnetite nanoparticles and evaluated cell damage. The formation and physical-chemical properties of the microspheres were determined by FT-IR, Raman, thermogravimetric analysis (TGA), energy-dispersive X-ray spectroscopy (EDS), dynamic light scattering (DLS), and SEM. Cell damage was evaluated by a full set of in vitro assays using a non-cancerous cell line, human erythrocytes, and human lymphocytes. At the same time, to know if these microspheres can load proteins over their surface, bovine serum albumin (BSA) immobilization was measured. Results showed 7 nm magnetite nanoparticles loaded into chitosan/Tween 80 microspheres with average sizes of 1.431 µm. At concentrations from 1 to 100 µg/mL, there was no evidence of changes in mitochondrial metabolism, cell morphology, membrane rupture, cell cycle, nor sister chromatid exchange formation. For each microgram of microspheres 1.8 µg of BSA was immobilized. The result provides the fundamental understanding of the in vitro biological behavior, and safety, of developed microspheres. Additionally, this set of assays can be helpful for researchers to evaluate different nano and microparticles.

Dosage of Sex Chromosomal Genes in Blood Deposited on Filter Paper for Neonatal Screening of Sex Chromosome Aneuploidy.

AIMS: In this study, we examined the doses of the stature homeobox (SHOX), vesicle-associated membrane protein 7 (VAMP7), and SRY genes to establish a protocol for using peripheral blood samples deposited on filter paper for the screening of sex chromosome aneuploidy in neonates. We also measured correlations with karyotypes to assess this method as a neonatal screening strategy. MATERIALS AND METHODS: This was an observational, descriptive, comparative blind study. Thirty-two healthy young adults (17 women, 15 men; age, ≥18 years), four patients with known sex chromosome aneuploidy (positive control group), and 1000 healthy newborns were included. Gene dosages were determined using quantitative real-time polymerase chain reaction (RT-PCR). Values with standard deviations (SDs) of three or more were considered abnormal. RESULTS: Men and women differed in the gene dosage of the SRY gene. Cases with Turner syndrome showed values below 3 SDs for SHOX and VAMP7 genes, and cases with Klinefelter syndrome showed values above 3 SDs for SHOX and VAMP7 genes. Two suspected cases of sex chromosome aneuploidy were diagnosed using our neonatal screening strategy; these cases were confirmed as Turner syndrome and 47,XYY syndrome by karyotyping. CONCLUSIONS: Our data establish a basis for the determination of chromosomal sex and neonatal screening of sex chromosome aneuploidy using RT-PCR.

[Chromosomal abnormalities in patients from Obstetrics and Gynaecology hospital]. / Anormalidades cromosómicas en pacientes de un hospital de ginecoobstetricia.

BACKGROUND: Chromosomal abnormalities are present in 2-4 % of all newborns, and they cause 20 % of deaths in the first year of life. The estimated prevalence of chromosomal abnormalities is one for each 500-1000 newborns. These abnormalities can be numerical or structural, and they can affect autosomal or sexual chromosomes. They affect from 1 to 3 % of general population, and from 6 to 7 % of individuals with congenital anomalies. METHODS: Descriptive study, which included all the registries of cytogenetic analysis (of adults and newborns) made in a genetic laboratory in a period of 14 years. The prevalence of polymorphisms and chromosomal abnormalities in the patients from the Hospital de Ginecoobstetricia 23, Instituto Mexicano del Seguro Social (Monterrey, Nuevo León) was assessed. RESULTS: Of 4006 cytogenetic studies, 253 (6.3 %) did not show in vitro growth, 2667 (66.5 %) were normal, and 1175 (29.3 %) were abnormal. Of these, 614 (52.2 %) had polymorphisms, and 561 (47.7 %) structural or numerical chromosomal abnormalities. In regards to these chromosomopathies (561), trisomy 21 was observed in 429 (36.5 %); Turner's syndrome, in 84 (7.1 %); trisomy 18, in 57 (4.8 %); and trisomy 13, in 32 (2.7 %). With G-band technique, we found 93 % of in vitro cell growth. CONCLUSIONS: Of these studies, 55 % was performed due to non-numerical abnormalities; 14.4 %, due to structural abnormalities; and the rest, due to polymorphisms.

INTRODUCCIÓN: las anormalidades cromosómicas se presentan en 2 a 4 % de los recién nacidos y causan 20 % de las muertes en el primer año de vida. Su prevalencia es de uno por cada 500 a 1000 recién nacidos vivos. Pueden ser numéricas o estructurales y afectar a los cromosomas autosómicos o sexuales. Se presentan en 1 a 3 % de la población general y en 6 a 7 % de los individuos con anomalías congénitas. MÉTODOS: estudio descriptivo en el que se incluyeron todos los resultados citogenéticos de cariotipos tomados de sangre periférica de adultos y neonatos. Se evaluó la prevalencia de polimorfismos y alteraciones cromosómicas en derechohabientes del Hospital de Ginecoobstetricia 23 del Instituto Mexicano del Seguro Social, en Monterrey, Nuevo León. RESULTADOS: de 4006 estudios citogenéticos, en 253 no se obtuvo crecimiento de linfocitos (6.3 %), 2667 fueron normales (66.5 %) y 1175, anormales (29.3 %); de estos últimos, en 614 (52.2 %) se identificaron polimorfismos cromosómicos y en 561 (47.7 %), aberraciones cromosómicas estructurales o numéricas. De las cromosomopatías (561, 47.7 %), la trisomía 21 se observó en 429 (36.5 %), el síndrome de Turner en 84 (7.1 %), la trisomía 18 en 57 (4.8 %) y la trisomía 13 en 32 (2.7 %). Con la técnica de bandeo G se obtuvo un crecimiento celular in vitro de 93 %. CONCLUSIONES: 55 % de los estudios se realizó por anormalidades diferentes a las numéricas, 14.4 % por alteraciones estructurales y el resto se debió a polimorfismos.