Healthy Brother Research Articles

Germline mosaicism in Rubinstein–Taybi syndrome

Rubinstein–Taybi syndrome is an autosomal dominant disorder with multiple congenital anomalies and genetic heterogeneity. Clinical manifestations include mental retardation, postnatal growth deficiency, microcephaly, broad thumbs and halluces, and characteristic facial features. Mutations in the gene encoding the transcriptional coactivator CREB-binding protein (CREBBP; OMIM 600140) on chromosome 16p13, account for about 50% to 70% of patients. Most of CREBBP mutations are de novo and the rate of recurrence in a family is low. Families with several affected children are extremely rare. We report here a Moroccan family with two children with RSTS and apparently unaffected parents. The molecular studies showed a heterozygous mutation c.4361T>A (p.Leu1454His) in exon 26 of the CREBBP gene in the two affected siblings. Neither the parents, nor the healthy brother, carry this mutation in hematologic cells. The mutation was also absent in buccal epithelial cells of both parents. We discuss the hypothesis of germinal mosaicism. This concept is very important because it complicates genetic counseling of this family who has a risk of recurrence of the mutation in subsequent pregnancies.

Identification of New Leukemia Associated Gene Mutations by Whole Genome Sequencing of DNA From Two Siblings with Congenital Neutropenia and Secondary AML

Abstract 12To identify the pattern of genetic aberrations, which may promote leukemia development in patients with severe congenital neutropenia (CN), we have performed a whole genome sequencing (WGS) of DNA samples from myeloid leukemic cells of two affected siblings suffering from CN. Both children harbored ELANE gene mutations. The father of the children demonstrates somatic mosaicism for the ELANE mutation and has no severe neutropenia. For WGS we used Complete Genomics technology (Complete Genomics. Inc, Mountain View, CA.). More than 90 % of genomes were sequenced at high quality with minimum coverage of at least 20-fold. As an example, 3.355.399 single nucleotide variants (SNVs) were identified in DNA isolated from leukemia blasts of one CN patient. The following filters were used to identify mutations in the leukemic cells from the two patients: 1) Non-synonymous SNVs in coding sequences only (9288 SNVs), 2) 54 healthy individuals sequenced by Complete Genomics (557 SNVs), 4) five members of one family from the same ethnic area (healthy parents, one cyclic neutropenia patient and her healthy brother, 471 SNVs), 5) five family members of the affected two children: parents and three healthy siblings (two healthy sisters and one heatlthy brother). Remaining SNVs presented in the two affected children were subsequently analysed using in silico prediction software Polyphen 2, which predicts possible impact of an amino acid substitution on the structure and function of human proteins. Fourteen SNVs with predicted damaging effects on the protein function were used for further analysis. All candidate SNVs were validated by Sanger sequencing. We detected nine inherited candidate SNVs presented in the two affected children but not in healthy siblings. The SNV in the ELANE gene (c.452G>A p.C151Y, dbsnp.129:rs57246956) was inherited from the father. Novel SNVs inherited from the father were as follows: in the TCTE1, FAM135A, M6PR, C20orf144 and PTPN23 genes. Only three SNVs were inherited from the healthy mother (in BLOC1S1, DUS3L and KIAA1543 genes). All SNVs were heterozygous. We also found 5 sporadic SNVs presented in leukemia sample of one CN patient only, but absent in his DNA sample from an earlier time point of CN diagnosis. These are heterozygous SNVs in the CSF3R, ACAP2, GRM1, LASS3, and RUNX1 gene. All five gene mutations might be involved in leukemogenesis. Interestingly, both affected patients had somatic mutation in the RUNX1 gene at the same nucleotide position (c.415C>G, p.R139G in sick brother and c.415C>T, p.R139* in sick sister). In summary, we identified candidate genes that may be relevant for leukemogenesis in CN patients. Our study also establishes WGS as an unbiased method for discovering leukemia-initiating mutations in previously unidentified genes that may respond to targeted therapies Disclosures:No relevant conflicts of interest to declare.

Mutations in the TNFRSF1A, CEBPE and ELANE genes in a Congenital Cyclic Neutropenia Patient: A New Syndrome?

Abstract 11Cyclic neutropenia (CyN) is a hematologic disorder in which blood cell counts particularly granulocytic neutrophil numbers show cycles at 21 day intervals. The majority of CyN patients (ca 90 %) harbor inherited mutations in the ELANE gene. Intriguingly, same ELANE mutations are present in two different hematologic syndromes: congenital as well as in cyclic neutropenias. It is unclear how mutation in the same gene cause congenital or cyclic neutropenia. We aimed to identify genes which are exclusively mutated in cyclic or in congenital neutropenia additionaly to the ELANE gene mutations. Recently, we found in congenital neutropenia patients additional to ELANE mutations inherited mutations in for example the G6PC3 gene or the HAX1 gene (Germeshausen, M., et al, Haematologica 2010). This suggests cooperating effects of different defective intracellular signaling pathways and excludes that mutated ELANE alone is responsible for the pathogenesis of congenital or cyclic neutropenia.To identify gene mutations causing cyclic neutropenia in association with ELANE mutation, we performed whole genome sequencing using Complete Genomics technology (Complete Genomics. Inc, Mountain View, CA.) of a family with an affected CyN patient. The CyN patient harbors sporadic heterozygous ELANE mutation (c.761C>G p.W241L) and her family consists of a healthy brother and healthy parents. We identified a novel heterozygous point mutation in the tumor necrosis factor receptor superfamily, member 1A (TNFRSF1A) gene (c.664C>T; p.R121Q) in the affected patient and her mother. This mutation was confirmed by Sanger sequencing. The TNFRSF1A gene encodes p55 subunit of the TNFa receptor (TNFR1) and intriguingly this gene is known to be frequently mutated in patients with Tumour necrosis factor receptor-associated periodic fever syndrome (TRAPS), a disease clinically similar to cyclic neutropenia. TRAPS is an autosomal dominant disorder characterized by episodes of fever, inflammation and periodical changes in the neutrophil counts. Functional studies in patients with TRAPS described heterogenous effects of different TNFRSF1A mutations on the surface expression and PMA-induced clevage of TNFR1 on the neutrophilic granulocytes and monocytes. In our CyN patient we measured elevated mRNA levels of TNFR1 on neutrophils, in comparison to her healthy family members and unrelated healthy individuals. However, we detected diminished surface expression of the TNFR1 protein, but elevated PMA-induced receptor shedding in the affected CyN patient, in comparison to healthy individuals, as assessed by estimation of soluble TNFR1 in supernatants of PMA-stimulated neutrophils.We also identified in the same patient and her father a second novel heterozygous point mutation in the CEBPE gene (c.636C>A; p.L155M), which was also confirmed by Sanger sequencing. This C>A substitution changes CTG to ATG creating a new start site for translation of a novel isoform of C/EBPε protein. C/EBPε is a myeloid-specific transcription factor playing an important role in granulopoiesis. Mutations in the CEBPE gene have been described in patients with neutrophil-specific granule deficiency (SGD).In summary, we identified additional to the ELANE mutation two novel mutations in a CyN patient, one in the TNFRSF1A gene inherited from the mother, another in the CEBPE gene inherited from the father. Mutations in both genes are already described in patients with TRAPS (periodic fever syndrome) and granulocyte abnormalities, respectively. These mutations in association with the ELANE gene mutation may contribute to the pathogenesis of cyclic neutropenia in this patient. Whether the combination of these three mutations might be responsible for a subgroup of CyN patients remains to be investigated. Disclosures:No relevant conflicts of interest to declare.

Prenatal diagnostic conundrum involving a novel ATP7A duplication

Prenatal diagnostic conundrum involving a novel <i><scp>ATP7A</scp></i> duplication

Risk evaluation and preimplantation genetic diagnosis in an infertile man with an unbalanced translocation t(10;15) resulting in a healthy baby

Balanced chromosome structural rearrangement, including reciprocal translocation, Robertsonian translocation and inversion, is an important cause of male infertility that results in azoospermia, oligozoospermia or asthenozoospermia [1]. Unbalanced translocation between two autosomes, which usually results in mental retardation or multiple congenital anomalies, is rare in infertile patients. To our knowledge, only the t(15;18) unbalanced translocation has been reported, in two healthy infertile brothers. Although fluorescence in situ hybridization (FISH) analysis showed that chromosomal normal sperms existed in their semen, these two brothers and their partners chose donor insemination rather than preimplantation genetic diagnosis (PGD) [2]. Whether patients with this kind of rare karyotype could deliver a healthy baby is unclear. Here, we report a man with severe oligoasthenoteratozoospermia carrying a derivative chromosome 10 resulting from the rare unbalanced translocation t(10;15). Based on risk evaluation by sperm FISH, we performed PGD for the couple and successfully established a normal pregnancy. Our results show that patients with such an abnormal karyotype can produce their own offspring, and emphasize the importance of risk evaluation using sperm FISH for infertile male patients. Patient data A couple presented to our reproduction center due to primary infertility that had persisted for 5 years. The woman showed no obvious cause of infertility on examinations, which included: (1) clinical findings: physical examination and medical history evaluation,such as menstrual history, obstetrical history, sexual history, family history, personal and lifestyle history; (2) biochemical tests: liver and renal function; blood cholesterol, blood triglycerides and blood glucose; and immunologic function; and (3) hormonal assay: estradiol, testosterone, progesterone, follicle stimulating hormone (FSH), luteinizing hormone, prolactin, 17-hydroxypregnenolone and thyroid stimulating hormone (TSH); (4) other diagnosis test such as hysterosalpingography. Her karyotype was also normal. However, the man was found to have severe oligoasthenoteratozoospermia after three routine sperm analyses and one sperm morphology examination following the WHO guidelines (Table 1). Conventional G-banding (G-bands after trypsin and Giemsa, GTG) analysis showed that he had only 45 chromosomes, with one derivative chromosome comprising chromosomes 10 and 15 (Fig. 1). Table 1 Semen analysis Fig. 1 a Partial karyotype of the patient showing normal chromosome 10,15 and der(10). b ideogram of the normal chromosome 10,15 and rearranged chromosome. Red, green and aqua bands show the position and color of the FISH probes used for preimplantation genetic ... The couple then accepted a genetic counseling and asked for further fertility evaluation. Oligoasthenoteratozoospermia might have been the cause of the primary infertility in this couple. The husband carried a rare unbalanced translocation comprising chromosome 10 and chromosome 15 that had not been reported previously. The genetic risk was unclear for this kind of abnormal karyotype. His chromosomal structure was similar to that of a Robertsonian translocation; therefore, the meiosis pattern might also be similar to a Robertsonian translocation. Sperm chromosome constitution can be obtained by sperm FISH analysis and might predict the probability of producing an embryo with normal karyotype. If chromosomally normal sperms are present in semen at a high prevalence, PGD can be performed to select embryos with balanced chromosomes for transfer and deliver a healthy baby. This study was approved by the Institutional Review Board of Central South University and Reproductive and Genetics Hospital of Citic-Xiangya, and the patients’ written informed consent was obtained.

A novel homozygous mutation in recombination activating gene 2 in 2 relatives with different clinical phenotypes: Omenn syndrome and hyper-IgM syndrome

A novel homozygous mutation in recombination activating gene 2 in 2 relatives with different clinical phenotypes: Omenn syndrome and hyper-IgM syndrome

The impact of childhood sickness on adult socioeconomic outcomes: Evidence from late 19th century America

We use family fixed-effects models to estimate the impact of childhood health on adult literacy, labor force outcomes, and marital status among pairs of white brothers observed as children in the 1880 U.S. Census and then as adults in the 1900–1930 Censuses. Given our focus on the 19th century, we observed a wider array of infectious, chronic, and traumatic health problems than is observed using data that are more recent; our results thus provide some insights into circumstances in modern developing countries where similar health problems are more frequently observed. Compared to their healthy siblings, sick brothers were less likely to be located (and thus more likely to be dead) 20–50 years after their 1880 enumeration. Sick brothers were also less likely to be literate, to have ever been married, and to have reported an occupation. However, among those with occupations, sick and healthy brothers tended to do similar kinds of work. We discuss the implications of our results for research on the impact of childhood health on socioeconomic outcomes in developed and developing countries.

A 14-Year-Old Boy with Vomiting

A 14-year-old boy was transferred from an outside hospital for evaluation of vomiting. He had been well until he traveled to India 3 weeks earlier. Upon return, he developed abdominal pain and recurrent episodes of nonbilious, nonbloody emesis. The day of admission, he ate a slice of pizza and then experienced numerous episodes of vomiting every 15 to 30 minutes. There had been minimal diarrhea at the beginning of the illness, but none over the following 3 weeks. Review of systems was otherwise unremarkable. The patient’s medical history was remarkable for poor weight gain and linear growth without any evaluation. He is in an eighth-grade classroom, but functions at a first-grade level. His parents are aware that his IQ is low, but they can’t describe the extent of his cognitive impairment. All we know of his developmental/intellectual history is that he walked at 2 years of age and spoke his first words at 3 years. The family follows a traditional Jain vegetarian diet (in this case, essentially vegan). They had given him Ensure in the past because of his poor weight gain. The family history is unremarkable. He has one healthy brother of normal stature. There is no known history of consanguinity. Birth history was unremarkable. On exam, the patient was a small, thin, malnourished boy. He was afebrile, pulse 82, respiratory rate 20, and blood pressure 112/70. His height was

A 14‐year‐old girl with lissencephaly and craniofacial dysmorphism

The patient was born at 38 weeks gestation with body weight of 2700 g from non-consanguineous parents. She was the third child with two healthy brothers. At 2 months of age, she had afebrile seizures. She was admitted to hospital because of an epileptic state. Her chromosomal pattern was 46XX by G-band analysis. A deletion in the Miller-Dieker syndrome (MDS) region was confirmed by fluorescence in situ hybridization (FISH) for chromosomal testing. Brain CT/MRI revealed lissencephaly and dysgenesis of corpus callosum: the absence of rostrum and splenium (Fig. 1). Radiologically, no abnormalities were found in the body except the brain. At the age of 4 years, she was hospitalized at a nursing center. Profound mental and motor retardation, seizures and hypertonia were found. She suffered from repeated episodes of bronchitis/ pneumonia, depletion of carnitine, and chronic insufficiency of digestion. Gastrostomy was performed at 6 years of age owing to feeding problems. She died of pneumonia at 14 years of age.

Variable expressivity of osteogenesis imperfecta in a Brazilian family due to p.G1079S mutation in the COL1A1 gene

Osteogenesis imperfecta (OI) is a Mendelian disease with genetic heterogeneity characterized by bone fragility, recurrent fractures, blue sclerae, and short stature, caused mostly by mutations in COL1A1 or COL1A2 genes, which encode the pro-α1(I) and pro-α2(I) chains of type I collagen, respectively. A Brazilian family that showed variable expression of autosomal dominant OI was identified and characterized. Scanning for mutations was carried out using SSCP and DNA sequence analysis. The missense mutation c.3235G>A was identified within exon 45 of the COL1A1 gene in a 16-year-old girl diagnosed as having OI type I; it resulted in substitution of a glycine residue (G) by a serine (S) at codon 1079 (p.G1079S). The proband's mother had the disease signs, but without bone fractures, as did five of nine uncles and aunts of the patient. All of them carried the mutation, which was excluded in four healthy brothers of the patient's mother. This is the first description in a Brazilian family with OI showing variable expression; only one among seven carriers for the c.3235G>A mutation developed bone fractures, the most striking clinical feature of this disease. This finding has a significant implication for prenatal diagnosis in OI disease.

Autoimmune Hepatitis Type 2 in a Child With IPEX Syndrome

Autoimmune Hepatitis Type 2 in a Child With IPEX Syndrome

Compound Heterozygous Mutations in Vitamin D Receptor Gene in Two Sisters With Hereditary Vitamin D Resistant Rickets Type II

Background: Hereditary vitamin D resistant rickets (HVDRR) type II , is a rare autosomal recessive disorder with known heterogeneity in clinical features, response to treatment and presence or absence of vitamin D receptor (VDR) gene mutation. Here, we described VDR gene mutations in 2 sisters with HVDRR with correlation to clinical features, and response to treatment. Methods: Vitamin D receptor gene sequence analysis was performed for exons 2 and 3, in addition to polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) for exons 7 and 8. Patients, their parents, their elder sister and brother were studied. Treatment with high dose vitamin D, response to therapy, and 30 months follow up were recorded for both patients. Results: A novel heterozygous point mutation (c.A155G) at the initiation codon of exon 3 was found in the 2 patients, their father and their healthy brother. This point mutation changed the amino acid (aa) at position 51 of the VDR from serine to glycine (p.Ser51Gly) at the start of the second zinc finger of the DNA-binding domain of the VDR. Exon 7 showed a heterozygous mutation in both patients and their mother. The mutation was located at the 970 base pair (bp) of VDR. Patients responded well to high doses oral calcium and Alfacalcidol without relapse of their rickets for the whole follow up period. Conclusion: Compound heterozygous mutation was identified in the VDR gene in 2 sisters with HVDRR. This mutation resulted in HVDRR without alopecia that was responsive to high dose vitamin D therapy. doi:10.4021/jem41w

A new case of rare proximal 3q13 interstitial deletion

Abstract We present the case of a 20-year-old man referred to the clinical geneticist because of mental retardation and dysmorphic features because of concerns about hereditability when his older, healthy brother was expecting a child. Deletion of proximal 3q arm was found with standard G-banding, and array comparative genomic hybridisation (array-CGH) was used to further locate the breakpoints. A unique interstitial deletion del 3q13.11q13.33 was confirmed. The first clinical symptoms in the 20-year-old were described at the age of 4 months when the pediatrician reported muscle hypertonia of the lower limbs, which later evolved into hypotonia. Later clinical observations revealed that the patient’s psychomotor development was delayed: he exhibited craniofacial abnormalities, cryptorchidism, thoracic kyphosis, and tapering fingers. Interstitial deletions of the proximal long arm of chromosome 3 have rarely been reported:; there are only 12 previously reported cases. The breakpoints and sizes of described deletions vary greatly, which makes definite genotype-phenotype conclusions impossible at this time. Developmental delay is one of the common features described in the majority of reported cases. The BTB-zinc finger gene ZBTB20 might be a potential candidate gene: it was shown in the mouse hippocampus to be expressed during the important period of neurogenesis of pyramidal neurons. Also, four of patients reported to date had agenesis of the corpus callosum and one, holoprosencephaly. We suggest that the GAP43 gene is involved in the development of structural neurological abnormalities in patients with 3q deletion.

NMD and microRNA expression profiling of the HPCX1 locus reveal MAGEC1 as a candidate prostate cancer predisposition gene

BackgroundSeveral predisposition loci for hereditary prostate cancer (HPC) have been suggested, including HPCX1 at Xq27-q28, but due to the complex structure of the region, the susceptibility gene has not yet been identified.MethodsIn this study, nonsense-mediated mRNA decay (NMD) inhibition was used for the discovery of truncating mutations. Six prostate cancer (PC) patients and their healthy brothers were selected from a group of HPCX1-linked families. Expression analyses were done using Agilent 44 K oligoarrays, and selected genes were screened for mutations by direct sequencing. In addition, microRNA expression levels in the lymphoblastic cells were analyzed to trace variants that might alter miRNA expression and explain partly an inherited genetic predisposion to PC.ResultsSeventeen genes were selected for resequencing based on the NMD array, but no truncating mutations were found. The most interesting variant was MAGEC1 p.Met1?. An association was seen between the variant and unselected PC (OR = 2.35, 95% CI = 1.10-5.02) and HPC (OR = 3.38, 95% CI = 1.10-10.40). miRNA analysis revealed altogether 29 miRNAs with altered expression between the PC cases and controls. miRNA target analysis revealed that 12 of them also had possible target sites in the MAGEC1 gene. These miRNAs were selected for validation process including four miRNAs located in the X chromosome. The expressions of 14 miRNAs were validated in families that contributed to the significant signal differences in Agilent arrays.ConclusionsFurther functional studies are needed to fully understand the possible contribution of these miRNAs and MAGEC1 start codon variant to PC.

Familial intellectual disability and autistic behavior caused by a small FMR2 gene deletion

Alterations of the Fragile Mental Retardation 2 gene (FMR2, synonym AFF2) can result in non-specific, mild to borderline X-linked intellectual disability (XLID), and behavioral problems. The well-known molecular pathomechanism of this condition, also referred to as FRAXE, is a (CCG)(n) trinucleotide repeat expansion which leads to silencing of the FMR2 gene. However, deletions within the FMR2 gene may also be causative of the disorder. Here, we report on two brothers diagnosed with FRAXE in whom a small deletion in the FMR2 gene was detected by whole genome array comparative genomic hybridization (CGH). The deletion was also present in their clinically healthy mother and maternal uncle who was similarly affected, but not in a healthy older brother of the two patients. Our observation demonstrates that FMR2 gene deletions may contribute to the FRAXE phenotype. Therefore, we suggest that screening for FMR2 gene deletions using array CGH should be considered in patients with non-specific XLID and absent trinucleotide expansion.

Enzyme replacement therapy for mucopolysaccharidosis II from 3 months of age: a 3‐year follow‐up

We present a 3-year follow-up of a boy with mucopolysaccharidosis type II (MPS II) who had idursulfase therapy initiated at the age of 3 months and compare his clinical course to his healthy twin brother. Detailed anthropometric features, ultrasound studies of liver and spleen volumes, echocardiography and audiological examinations, psychological tests, joint range of motion (ROM) and skeletal radiographs were monitored. After 3 years of treatment, the patient has not developed any clinical manifestations of MPS II. He did not develop coarse facial features, joint disease, or organomegaly, and his cardiac function remained normal. There were no pronounced signs of dysostosis multiplex on radiographs. The only difference when compared with his healthy twin brother was lower IQ (Termann-Merrill 98 vs. 118) and mild deformity of one vertebrae. Our study suggests that early initiation of enzyme replacement therapy may significantly slow or prevent the development of irreversible disease manifestations and therefore modify the natural history of MPS II.

The GH/IGF-I Axis and Pituitary Function and Size in Adults with Prader-Willi Syndrome

Background: In adults with Prader-Willi syndrome (PWS), limited information is available about pituitary function, more specifically the prevalence of growth hormone deficiency (GHD). The aim of this study was to gain more insight into endocrine function in PWS adults, with emphasis on GH secretion. Methods: 15 randomly selected adult PWS individuals were included and 14 healthy brothers and sisters served as a control group. Main outcome measures were IGF-I, IGFBP-3 and peak GH level after a combined GHRH-arginine test. Other pituitary hormone deficits are diagnosed based on serum levels of the concerning hormones. The size of the pituitary gland was measured on MRI images. Results: In PWS adults, IGF-I levels were low and IGFBP-3 levels normal when compared to healthy controls. GHD was diagnosed in 8–38% of the PWS patients, depending on the criteria used. Hypogonadism was present in 87% of the patients. Hypothyroidism and adrenal insufficiency could also be demonstrated. Anterior pituitary size was lower in PWS individuals when compared to healthy controls. Conclusion: In this study, pituitary hormone deficiencies are demonstrated in a considerable number of adults with PWS, hypogonadism and GHD being most prominent. Furthermore, the anterior pituitary is smaller in comparison with healthy controls.

Compound Heterozygous Mutations in Vitamin D Receptor Gene in Two Sisters With Hereditary Vitamin D Resistant Rickets Type II

Background: Hereditary vitamin D resistant rickets (HVDRR) type II , is a rare autosomal recessive disorder with known heterogeneity in clinical features, response to treatment and presence or absence of vitamin D receptor (VDR) gene mutation. Here, we described VDR gene mutations in 2 sisters with HVDRR with correlation to clinical features, and response to treatment. Methods: Vitamin D receptor gene sequence analysis was performed for exons 2 and 3, in addition to polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) for exons 7 and 8. Patients, their parents, their elder sister and brother were studied. Treatment with high dose vitamin D, response to therapy, and 30 months follow up were recorded for both patients. Results: A novel heterozygous point mutation (c.A155G) at the initiation codon of exon 3 was found in the 2 patients, their father and their healthy brother. This point mutation changed the amino acid (aa) at position 51 of the VDR from serine to glycine (p.Ser51Gly) at the start of the second zinc finger of the DNA-binding domain of the VDR. Exon 7 showed a heterozygous mutation in both patients and their mother. The mutation was located at the 970 base pair (bp) of VDR. Patients responded well to high doses oral calcium and Alfacalcidol without relapse of their rickets for the whole follow up period. Conclusion: Compound heterozygous mutation was identified in the VDR gene in 2 sisters with HVDRR. This mutation resulted in HVDRR without alopecia that was responsive to high dose vitamin D therapy. J Endocrinol Metab. 2011;1(4):174-181 doi: https://doi.org/10.4021/jem41w

Experiência do irmão sadio em relação à doença e hospitalização do irmão com câncer

OBJETIVO: Compreender o significado da experiência de ter um irmão com câncer na perspectiva do irmão sadio. MÉTODOS: Pesquisa descritiva de abordagem qualitativa. Foram realizadas entrevistas semiestruturadas com cinco irmãos de crianças com câncer assistidas em instituição filantrópica localizada no Município de São Paulo. RESULTADOS: O tema central tornando-se amadurecido com o sofrimento permitiu revelar que o irmão sadio torna-se amadurecido em razão do sofrimento que lhe é imposto no decorrer dos acontecimentos. Passa a valorizar a convivência familiar e os pequenos atos do cotidiano que lhe eram despercebidos. CONCLUSÃO: A equipe de saúde precisa reconhecer a experiência do irmão e incluí-lo no cuidado prestado em todas as fases da doença.

The WST survival assay: an easy and reliable method to screen radiation-sensitive individuals

An easy, fast and reliable method was developed to screen hundreds of Epstein-Barr virus-transformed cell lines (lymphoblastoid cell lines, LCLs) for radiation sensitivity that were generated from lymphocytes isolated from young lung cancer patients. The WST-1 test explores the metabolic activity of the mitochondria as an indicator for the vital status of cells. Cell proliferation as well as indirect cell death can be quantified by this method on a large scale in microtiter plates. Cell survival was measured at 24- and 48-h post-irradiation with 10 Gy ((137)Cs source) by the WST-1 assay and Trypan blue staining. To set up the experimental screening conditions and to establish a positive and a negative control, an ATM-mutated cell line from a radiation-sensitive ATM patient and an ATM proficient cell line from a healthy brother were compared. An optimal differentiation between the two cell lines was demonstrated for 10 Gy and 24- and 48-h cell growth after irradiation. Upon screening 120 LCLs of young lung cancer patients under these conditions, 5 of them were found to be radiation sensitive to a high degree of statistical significance. The results have been confirmed by a second laboratory by means of Trypan blue testing. The WST-1 test represents an efficient and reliable method by means of screening for radiation-sensitive cell lines.