BAC array CGH in patients with Velocardiofacial syndrome-like features reveals genomic aberrations on chromosome region 1q21.1

<p>Abstract</p> <p>Background</p> <p>Microdeletion of the chromosome 22q11.2 region is the most common genetic aberration among patients with velocardiofacial syndrome (VCFS) but a subset of subjects do not show alterations of this chromosome region.</p> <p>Methods</p> <p>We analyzed 18 patients with VCFS-like features by comparative genomic hybridisation (aCGH) array and performed a face-to-face slide hybridization with two different arrays: a whole genome and a chromosome 22-specific BAC array. Putative rearrangements were confirmed by FISH and MLPA assays.</p> <p>Results</p> <p>One patient carried a combination of rearrangements on 1q21.1, consisting in a microduplication of 212 kb and a close microdeletion of 1.15 Mb, previously reported in patients with variable phenotypes, including mental retardation, congenital heart defects (CHD) and schizophrenia. While 326 control samples were negative for both 1q21.1 rearrangements, one of 73 patients carried the same 212-kb microduplication, reciprocal to TAR microdeletion syndrome. Also, we detected four copy number variants (CNVs) inherited from one parent (a 744-kb duplication on 10q11.22; a 160 kb duplication and deletion on 22q11.21 in two cases; and a gain of 140 kb on 22q13.2), not present in control subjects, raising the potential role of these CNVs in the VCFS-like phenotype.</p> <p>Conclusions</p> <p>Our results confirmed aCGH as a successful strategy in order to characterize additional submicroscopic aberrations in patients with VCF-like features that fail to show alterations in 22q11.2 region. We report a 212-kb microduplication on 1q21.1, detected in two patients, which may contribute to CHD.</p

Pichia pastoris regulates its gene-specific response to different carbon sources at the transcriptional, rather than the translational, level

Background: The methylotrophic, Crabtree-negative yeast Pichia pastoris is widely used as a heterologous protein production host. Strong inducible promoters derived from methanol utilization genes or constitutive glycolytic promoters are typically used to drive gene expression. Notably, genes involved in methanol utilization are not only repressed by the presence of glucose, but also by glycerol. This unusual regulatory behavior prompted us to study the regulation of carbon substrate utilization in different bioprocess conditions on a genome wide scale. Results: We performed microarray analysis on the total mRNA population as well as mRNA that had been fractionated according to ribosome occupancy. Translationally quiescent mRNAs were defined as being associated with single ribosomes (monosomes) and highly-translated mRNAs with multiple ribosomes (polysomes). We found that despite their lower growth rates, global translation was most active in methanol-grown P. pastoris cells, followed by excess glycerol- or glucose-grown cells. Transcript-specific translational responses were found to be minimal, while extensive transcriptional regulation was observed for cells grown on different carbon sources. Due to their respiratory metabolism, cells grown in excess glucose or glycerol had very similar expression profiles. Genes subject to glucose repression were mainly involved in the metabolism of alternative carbon sources including the control of glycerol uptake and metabolism. Peroxisomal and methanol utilization genes were confirmed to be subject to carbon substrate repression in excess glucose or glycerol, but were found to be strongly de-repressed in limiting glucose-conditions (as are often applied in fed batch cultivations) in addition to induction by methanol. Conclusions: P. pastoris cells grown in excess glycerol or glucose have similar transcript profiles in contrast to S. cerevisiae cells, in which the transcriptional response to these carbon sources is very different. The main response to different growth conditions in P. pastoris is transcriptional; translational regulation was not transcript-specific. The high proportion of mRNAs associated with polysomes in methanol-grown cells is a major finding of this study; it reveals that high productivity during methanol induction is directly linked to the growth condition and not only to promoter strength

Experimental infection of swine and cat central nervous systems by the pig paramyxovirus of the blue eye disease.

This study analyses whether the pig paramyxovirus of blue eye disease (PPBED) infects the central nervous system (CNS) utilizing anterograde and retrograde peripheral nerve transport systems. The virus was injected into muscle and skin, and inoculated per nasum. The presence of PPBED was detected by an immunohistochemical method using polyclonal mouse antibodies against the whole inactivated virus, and was revealed with polyclonal rabbit antibodies against mouse immunoglobulin G (IgG) labelled with peroxidase. The PPBED injected into the pig medial gastrocnemius (MG) muscle was detected in a terminal branch innervating the MG muscle, in neural fibres of the sciatic nerve, in fibres of the ventral and dorsal spinal roots and in ventral horn neurones of the spinal cord. When PPBED was injected into the skin area innervated by the sural nerve, it was detected in neural fibres of the sural and sciatic nerves and in spinal cord dorsal horn neurones. The per nasum inoculum rapidly invaded the CNS through the olfactory nerve. The study concluded that, in order to invade the CNS, PPBED was transported retrogradely by peripheral cutaneous and muscular nerves, and anterogradely by the olfactory nerve. No PPBED was detected in either cat peripheral nerves or in cat CNS

Síndrome de Silver-Russell: relato de caso Silver-Russell Syndrome: case report

OBJETIVO: descrever o fenótipo da síndrome de Silver-Russell (SSR) e apresentar um caso diagnosticado com esta afecção genética, abordando aspectos genéticos, psicológicos e fonoaudiológicos. MÉTODOS: trata-se de relato de caso de uma criança do gênero feminino, sete anos e onze meses, portadora da síndrome de Silver-Russel. Foram realizadas avaliação genética médica e molecular, avaliação psicológica, avaliação fonoaudiológica e aplicação de testes complementares. RESULTADOS: a análise molecular da região 7p11 excluiu a dissomia uniparental para este caso. No exame físico foram constatados os principais sinais clínicos da SSR que incluiu retardo no crescimento de origem pré-natal, fácies típica, assimetrias ósseas e clinodactilia do 5º dedo. A avaliação cognitiva e fonoaudiológica mostraram deficiência mental, distúrbio de linguagem oral e comprometimento das funções orais. CONCLUSÃO: o estudo deste caso possibilitou a divulgação do fenótipo da SSR com suas manifestações físicas, cognitivas e fonoaudiológicas. Embora o teste molecular não tenha confirmado um dos possíveis mecanismos etiológicos da síndrome, a avaliação genética médica constatou a presença dos principais sinais clínicos que foram correlacionados à literatura. A avaliação psicológica e fonoaudiológica apontaram para comprometimento cognitivo e de comunicação, funções orais , sugerindo que importantes alterações fonoaudiológicas podem fazer parte do fenótipo desta síndrome, ainda pouco difundida para fonoaudiólogos.<br>PURPOSE: to describe Silver-Russell syndrome (SRS) phenotype and report a case with this condition, concerning genetics, psychological and speech/language and oral function evaluation. METHODS: we will submit the case of a girl, seven years and eleven months with Silver-Russell syndrome. Medical genetic evaluation, molecular test, psychological and speech/language evaluation was done. Complementary tests were applied. RESULTS: molecular test excluded a maternal uniparental disomy (UPD) of chromosome 7 in this case. In physical examination identified the most important signs of SSR phenotype that included prenatal growth retardation, typical face, skeletal asymmetry, and fifth finger clinodactily. Cognitive and speech language evaluation showed mental retardation, speech/language and oral functions disorders. CONCLUSIONS: the study of this case allowed for spreading the phenotype of SRS with its physical manifestations, cognitive, language and oral functions disorders. Although the molecular test has not confirmed one of the possible etiological mechanisms of the syndrome, the medical genetic evaluation evidenced the presence of the main signals that had been correlated to literature. The psychological and speech/language evaluation pointed out to cognitive, communication impairment and oral functions disorders suggesting that these findings can be part of phenotype for this syndrome, still little known among speech-pathologists