De novo 5q35.5 duplication with clinical presentation of Sotos syndrome

We report on a girl with developmental delay and a de novo 264 kb interstitial duplication in the region of Sotos syndrome at 5q35.3 in the immediate vicinity of critical NSD1 gene, but manifesting the phenotype, of overgrowth both prenatal stage and postnatal, macrocephaly, developmental delay, and resembling that of Sotos syndrome, rather than the recently reported syndrome of reciprocal duplication. The duplication is located right downstream from the NSD1 gene, a region which appears critical for the expression of the gene as regulatory elements might be disrupted or the expression of a not amplified critical gene might be otherwise affected by the duplicated region. Thus,in the process of evaluating identified CNVs attention should be drawn to the possible influence of chromosomal rearrangement on distant genes, which could add additional diversity to genomic disorders. Our case demonstrates that evaluation of the size of chromosomal alteration and gene content are not sufficient for assessment of CNV's pathogenicity and the context of adjacent genes should be considered.     

A single gene deletion on 4q28.3: PCDH18 – A new candidate gene for intellectual disability?

We report a boy with severe developmental delay, seizures, microcephaly, hypoplastic corpus callosum, internal hydrocephalus and dysmorphic features (narrow forehead, round face, deep-set eyes, blue sclerae, large and prominent ears, nose with anteverted nares, thin upper lip, small and wide-spaced teeth, hyperextensibility of the elbows, wrists, and fingers, fingertip pads, broad hallux, sacral dimple), carrying a 1.53 Mb interstitial deletion at 4q28.3. The deletion was detected by Agilent 105K oligo-array genome hybridization and involves the genomic region between 137,417,338 and 138,947,282 base pairs on chromosome 4 (NCBI build 36). The alteration was inherited from a healthy mother and contains a single gene, PCDH18 which encodes a cadherin-related neuronal receptor thought to play a role in the establishment and function of cell-cell connections in the brain. Thus, haploinsufficiency of this gene may contribute to altered brain development and associated malformations. We found that this deletion is a private inherited copy number variation that is associated with specific clinical findings in our patient and propose the PCDH18 gene as a possible candidate gene for intellectual disability.     

Differences on the prevalence of cervical HPV between Lithuania and Belarus

In some countries the cervical cancer incidence and mortality rates are much higher compared to the European average. The differences of HPV and its type prevalence between countries and regions influence cervical cancer incidence and mortality. Regarding the differences in cervical cancer incidence and mortality in Lithuania and Belarus, the aim of this study was to describe HPV infection level and HPVs type distribution among two study groups of patients with moderate or severe cervical intraepithelial neoplasia (CIN2-3) and cervical cancer. Our data shows that 74.2% [95% CI: 63.64÷84.76] of Lithuanian patients with cervical cancer and 85.6% [95% CI: 85.53÷92.85] of the study group with CIN2-3 were HPV positive, while in the study groups of Belarusian patients HPV infection was detected in 92.6% [95% CI, 74.25÷98.71] and 65.4% [95% CI, 44.36÷82.06] cases respectively. HPV 16 was the most prevalent type in Lithuanian as well as in Belarusian patients of the study groups. HPV 18 in Lithuanian patients of the study group with cervical cancer was identified in 10.2% [95% CI: 1.73÷18.67] and in the study group with CIN2-3 — in 2.6% [95% CI: 0.95÷6.15] of cases. HPV 18 was not detected in Belarusian patients of both groups.     

Biologically active ADAMTS13 is expressed in renal tubular epithelial cells

ADAMTS13 mRNA, which encodes the von Willebrand factor-cleaving protease, has been detected in a variety of tissues, including the kidney. The aim of our study was to characterize tubular expression and bioactivity of ADAMTS13. ADAMTS13 mRNA was detected in cultured primary human renal tubular epithelial cells (HRTEC) and in A498 cells, a human renal carcinoma cell line, by real-time PCR. Protein was detected using immunofluorescence and immunoblotting. Immunoblots demonstrated that the protein was secreted. The protease was proteolytically active in both cell lysates and cleaved the FRETS–VWF73 substrate. ADAMTS13 was demonstrated in situ in the renal cortex by immunohistochemistry. Protease was detected in both the proximal and distal renal tubules in normal renal tissue (n = 3) as well as in patients with tubular disorders (n = 3). Immunoblotting revealed that ADAMTS13 was present in the urine of patients with tubulopathy (n = 5) but not in normal urine. ADAMTS13 in urine had a molecular size similar to that in plasma, which indicates that the protease originates in the tubuli because such large proteins do not normally pass the glomerular filter. In conclusion, human renal tubular epithelial cells synthesize biologically active ADAMTS13 which may, after release from tubuli, regulate hemostasis in the local microenvironment.     

Impact of quantitative parameters,such as number of retrieved oocytes,number of transferred embryos and availability of surplus embryos for cryopreservation,on clinical pregnancy rates in ART

Abstract

Objective: To estimate the impact of quantitative parameters such as number of retrieved oocytes, number of transferred embryos and availability of surplus embryos for cryopreservation on clinical pregnancies in assisted reproductive technology (ART).

Design: We used the database of fertility clinic on in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) treatment cycles from year 2000 to 2010. Follicular fluids containing oocyte–cumulus complexes were recovered by a single lumen follicle aspiration needle, and IVF or ICSI procedure was used depending on infertility indications. Fertilization determination was performed on the second day. Zygotes were transferred into fresh medium and cultivated until the embryo transfer procedure. According to morphological criteria, the best quality embryos were transferred into uterus on the second or third day. Surplus embryos were frozen by slow freezing or by vitrification. We evaluated only clinical pregnancies that were diagnosed by ultrasonography.

Patients: We allocated all 1161 women cohort into three age groups: 671 women were <35; 397 were 35–40 years old and 93 women were >40. Indications for IVF treatment were as follows: mechanical factor (436 cases), male infertility (361 cases), idiopathic infertility (129 cases), endometriosis (78 cases), immunological infertility (14 cases), anovulation (28 cases), and other indications (28 cases). There were no data about the cause of infertility of 87 patients. The stimulation protocol was composed from gonadotropin hormone stimulation with antagonist or agonist supplementation. The triggering of luteinizing hormone (LH) surge was performed by recombinant human chorionic gonadotropin (hCG). The clinical pregnancy rates were: 42.2% (283/671) for women <35, 31.2% (124/397) for women 35–40-year-old and 16.1% (15/93) for women >40.

Interventions: All materials and methods that we used were based on results from our daily practice in IVF clinic and have no experimental design.

Main outcome measures: We evaluated clinical pregnancies in terms of the number of obtained oocytes, the amount of transferred embryos and the availability of surplus embryos for freezing. The number of retrieved oocytes was divided as follows: 1–5 (group I); 6–10 (group II); ≥11 (group III). The number of transferred embryos was from 1 to 3. Cases at which cryopreservation of embryos was not performed were evaluated as well.

Results: For women <35 and 35–40 years old, the highest percentage of pregnancies was achieved when 11 and more oocytes were obtained. Differences in women under 35 between groups were statistically significant. Statistically significant differences in clinical pregnancies depend on the number of transferred embryos in the age groups of <35 and 35–40. Women <35 and 35–40 years old, with the availability of surplus embryos for freezing, had better chances for pregnancy compared with women, who had no excess embryos for freezing.

Conclusions: The number of obtained oocytes, the number of transferred embryos and the availability of surplus embryos for freezing may serve as predictors of pregnancy rates in ART.