Mice with renal-specific alterations of stem cell-associated signaling develop symptoms of chronic kidney disease but surprisingly no tumors.

Previously, we found that Wnt and Notch signaling govern stem cells of clear cell kidney cancer (ccRCC) in patients. To mimic stem cell responses in the normal kidney in vitro in a marker-unbiased fashion, we have established tubular organoids (tubuloids) from total single adult mouse kidney epithelial cells in Matrigel and serum-free conditions. Deep proteomic and phosphoproteomic analyses revealed that tubuloids resembled renewal of adult kidney tubular epithelia, since tubuloid cells displayed activity of Wnt and Notch signaling, long-term proliferation and expression of markers of proximal and distal nephron lineages. In our wish to model stem cell-derived human ccRCC, we have generated two types of genetic double kidney mutants in mice: Wnt-ß-catenin-GOF together with Notch-GOF and Wnt-ß-catenin-GOF together with a most common alteration in ccRCC, Vhl-LOF. An inducible Pax8-rtTA-LC1-Cre was used to drive recombination specifically in adult kidney epithelial cells. We confirmed mutagenesis of ß-catenin, Notch and Vhl alleles on DNA, protein and mRNA target gene levels. Surprisingly, we observed symptoms of chronic kidney disease (CKD) in mutant mice, but no increased proliferation and tumorigenesis. Thus, the responses of kidney stem cells in the tubuloid and genetic systems produced different phenotypes, i.e. enhanced renewal versus CKD.

Vitrification and in-straw warming do not affect pregnancy rates of biopsied bovine embryos.

In the cattle industry, in vivo or in vitro embryo production combined with genotyping and cryopreservation technologies allows the selection and conservation of embryos carrying genes for desirable traits. This study aimed to assess the efficiency of a vitrification method suitable for in-straw warming of biopsied in vivo derived (IVD) bovine embryos. Three experiments were carried out using two methodologies: the Cryotop®, the gold standard vitrification and 3-step warming methodology, or the VitTrans, a vitrification/in-straw 1-step warming method that enables direct embryo transfer to the uterus. In experiment 1, intact and biopsied in vitro produced (IVP) day 7 expanded blastocysts were vitrified using the Cryotop® and warmed in 1- or 3-steps. No differences in survival rates were recorded at 24 h after warming for intact or biopsied IVP blastocysts irrespective of the warming procedure. In experiment 2, the effect of the time from trophectoderm (TE) biopsy to vitrification/in-straw warming on post-warming survival rate was assessed. No significant differences in survival were observed when blastocysts were vitrified/in-straw warmed immediately after biopsy or after 3 h in culture when compared to intact blastocysts. In experiment 3, IVD embryos were vitrified 3 h after biopsy using the Cryotop® or the VitTrans method and pregnancy rates were assessed at day 60 after transfer. Fresh, biopsied embryos served as control. Similar pregnancy rates were observed when IVD biopsied embryos were transferred fresh or vitrified/warmed by the Cryotop® or VitTrans method. No significant effect of the embryo quality or developmental stage was detected on the percentage of pregnant recipients when IVD biopsied embryos were transferred fresh or after vitrification. While fresh female IVD embryos produced significantly higher pregnancy rates than male embryos, there were no differences in pregnancy rates when male or female vitrified/warmed embryos were transferred. About 81% from the biopsies analyzed successfully determined the embryo sex, confirming that DNA was there, and it was efficiently amplified. To conclude, our findings indicate that both vitrification methodologies produced similar post-warming outcomes for both intact and biopsied IVP embryos. Besides, vitrification/in-straw warming of biopsied IVD bovine embryos did not affect the viability to originate pregnancy, being a useful option for their direct transfer in field conditions.

Svep1 stabilises developmental vascular anastomosis in reduced flow conditions.

Molecular mechanisms controlling the formation, stabilisation and maintenance of blood vessel connections remain poorly defined. Here, we identify blood flow and the large extracellular protein Svep1 as co-modulators of vessel anastomosis during developmental angiogenesis in zebrafish embryos. Both loss of Svep1 and blood flow reduction contribute to defective anastomosis of intersegmental vessels. The reduced formation and lumenisation of the dorsal longitudinal anastomotic vessel (DLAV) is associated with a compensatory increase in Vegfa/Vegfr pERK signalling, concomittant expansion of apelin-positive tip cells, but reduced expression of klf2a. Experimentally, further increasing Vegfa/Vegfr signalling can rescue the DLAV formation and lumenisation defects, whereas its inhibition dramatically exacerbates the loss of connectivity. Mechanistically, our results suggest that flow and Svep1 co-regulate the stabilisation of vascular connections, in part by modulating the Vegfa/Vegfr signalling pathway.

Macrophages in dermal disease progression of phospholipase D4-deficient Fleckvieh calves.

A new gene defect in Fleckvieh calves leads to a syndrome with partial phenotype overlap with bovine hereditary zinc deficiency. A mutation in a gene encoding phospholipase D4 (PLD4), an endosomal exonuclease, causes the disorder. In mice, PLD4 activity indirectly regulates the Toll-like receptor 9 (TLR9) pathway via degradation of microbial DNA. PLD4 absence thus results in visceral macrophage activation comparable to human macrophage activation syndrome. In this study, disease progression and the role of macrophages in affected calves were monitored clinically, clinicopathologically, and histologically over time. Breeding data identified 73 risk matings of heterozygous carriers resulting in 54 potentially PLD4-deficient calves born on farms. PLD4 status was examined via 5'-exonuclease assay, detecting 6 calves carrying the defect. These were purchased and monitored daily until final necropsy. The calves developed progressive skin lesions starting with small scaling areas terminating in severe crusting dermatitis, especially in areas with mechanical exposure. Histological and immunohistochemical analyses indicated that macrophages with cytoplasmic vacuolation increased considerably in skin sections obtained weekly during the disease course. Macrophage increase correlated with increased dermal lesion severity. Macrophage activation was confirmed by prominent phagocytic activity in the superficial dermis using electron microscopy. Dermal mRNA abundance of CCL2 and CCL3 measured by quantitative polymerase chain reaction verified macrophage activation. Further increase in mRNA of downstream molecule MyD88 and cytokine IL12b connected bovine PLD4 deficiency to increased TLR9 pathway activation. In contrast to human macrophage activation syndrome, the main feature of bovine PLD4 deficiency was local disease in organs with contact to microbial DNA (skin, intestine, lungs).

Vasohibin 1 selectively regulates secondary sprouting and lymphangiogenesis in the zebrafish trunk.

Previous studies have shown that Vasohibin 1 (Vash1) is stimulated by VEGFs in endothelial cells and that its overexpression interferes with angiogenesis in vivo Recently, Vash1 was found to mediate tubulin detyrosination, a post-translational modification that is implicated in many cell functions, such as cell division. Here, we used the zebrafish embryo to investigate the cellular and subcellular mechanisms of Vash1 on endothelial microtubules during formation of the trunk vasculature. We show that microtubules within venous-derived secondary sprouts are strongly and selectively detyrosinated in comparison with other endothelial cells, and that this difference is lost upon vash1 knockdown. Vash1 depletion in zebrafish specifically affected secondary sprouting from the posterior cardinal vein, increasing endothelial cell divisions and cell number in the sprouts. We show that altering secondary sprout numbers and structure upon Vash1 depletion leads to defective lymphatic vessel formation and ectopic lymphatic progenitor specification in the zebrafish trunk.

Inhibiting WNT and NOTCH in renal cancer stem cells and the implications for human patients.

Current treatments for clear cell renal cell cancer (ccRCC) are insufficient because two-thirds of patients with metastases progress within two years. Here we report the identification and characterization of a cancer stem cell (CSC) population in ccRCC. CSCs are quantitatively correlated with tumor aggressiveness and metastasis. Transcriptional profiling and single cell sequencing reveal that these CSCs exhibit an activation of WNT and NOTCH signaling. A significant obstacle to the development of rational treatments has been the discrepancy between model systems and the in vivo situation of patients. To address this, we use CSCs to establish non-adherent sphere cultures, 3D tumor organoids, and xenografts. Treatment with WNT and NOTCH inhibitors blocks the proliferation and self-renewal of CSCs in sphere cultures and organoids, and impairs tumor growth in patient-derived xenografts in mice. These findings suggest that our approach is a promising route towards the development of personalized treatments for individual patients.

A missense mutation in TUBD1 is associated with high juvenile mortality in Braunvieh and Fleckvieh cattle.

BACKGROUND: Haplotypes with reduced or missing homozygosity may harbor deleterious alleles that compromise juvenile survival. A scan for homozygous haplotype deficiency revealed a short segment on bovine chromosome 19 (Braunvieh haplotype 2, BH2) that was associated with high juvenile mortality in Braunvieh cattle. However, the molecular genetic underpinnings and the pathophysiology of BH2 remain to be elucidated. RESULTS: The frequency of BH2 was 6.5 % in 8,446 Braunvieh animals from the national bovine genome databases. Both perinatal and juvenile mortality of BH2 homozygous calves were higher than the average in Braunvieh cattle resulting in a depletion of BH2 homozygous adult animals (P = 9.3x10(-12)). The analysis of whole-genome sequence data from 54 Braunvieh animals uncovered a missense mutation in TUBD1 (rs383232842, p.H210R) that was compatible with recessive inheritance of BH2. The availability of sequence data of 236 animals from diverse bovine populations revealed that the missense mutation also segregated at a low frequency (1.7 %) in the Fleckvieh breed. A validation study in 37,314 Fleckvieh animals confirmed high juvenile mortality of homozygous calves (P = 2.2x10(-15)). Our findings show that the putative disease allele is located on an ancestral haplotype that segregates in Braunvieh and Fleckvieh cattle. To unravel the pathophysiology of BH2, six homozygous animals were examined at the animal clinic. Clinical and pathological findings revealed that homozygous calves suffered from chronic airway disease possibly resulting from defective cilia in the respiratory tract. CONCLUSIONS: A missense mutation in TUBD1 is associated with high perinatal and juvenile mortality in Braunvieh and Fleckvieh cattle. The mutation is located on a common haplotype likely originating from an ancient ancestor of Braunvieh and Fleckvieh cattle. Our findings demonstrate for the first time that deleterious alleles may segregate across closed cattle breeds without recent admixture. Homozygous calves suffer from chronic airway disease resulting in poor growth performance and high juvenile mortality. The respiratory manifestations resemble key features of diseases resulting from impaired function of airway cilia.

A frameshift mutation in GON4L is associated with proportionate dwarfism in Fleckvieh cattle.

BACKGROUND: Low birth weight and postnatal growth restriction are the most evident symptoms of dwarfism. Accompanying skeletal aberrations may compromise the general condition and locomotion of affected individuals. Several paternal half-sibs with a low birth weight and a small size were born in 2013 in the Fleckvieh cattle population. RESULTS: Affected calves were strikingly underweight at birth in spite of a normal gestation length and had craniofacial abnormalities such as elongated narrow heads and brachygnathia inferior. In spite of a normal general condition, their growth remained restricted during rearing. We genotyped 27 affected and 10,454 unaffected animals at 44,672 single nucleotide polymorphisms and performed association tests followed by homozygosity mapping, which allowed us to map the locus responsible for growth failure to a 1.85-Mb segment on bovine chromosome 3. Analysis of whole-genome re-sequencing data from one affected and 289 unaffected animals revealed a 1-bp deletion (g.15079217delC, rs723240647) in the coding region of the GON4L gene that segregated with the dwarfism-associated haplotype. We showed that the deletion induces intron retention and premature termination of translation, which can lead to a severely truncated protein that lacks domains that are likely essential to normal protein function. The widespread use of an undetected carrier bull for artificial insemination has resulted in a tenfold increase in the frequency of the deleterious allele in the female population. CONCLUSIONS: A frameshift mutation in GON4L is associated with autosomal recessive proportionate dwarfism in Fleckvieh cattle. The mutation has segregated in the population for more than 50 years without being recognized as a genetic disorder. However, the widespread use of an undetected carrier bull for artificial insemination caused a sudden accumulation of homozygous calves with dwarfism. Our findings provide the basis for genome-based mating strategies to avoid the inadvertent mating of carrier animals and thereby prevent the birth of homozygous calves with impaired growth.

Envelhecimento e morte: percepção de idosas de um grupo de convivência / Aging and death: perception of elderly women from a support group / Envejecimiento y muerte: percepción de ancianas de un grupo de convivencia

O envelhecimento é uma etapa do ciclo vital em que, de acordo com as vivências de cada um, inicia um declínio no funcionamento do indivíduo. O idoso percebe suas possibilidades diminuídas e a probabilidade de morte se amplia. Considerando tais aspectos, objetivou-se conhecer a percepção de 07 (sete) idosas participantes de um grupo de convivência na Região das Hortênsias/Rio Grande do Sul, sobre o envelhecimento e a morte. Na coleta de dados, utilizou-se a técnica de grupo focal com roteiro pré-estabelecido e, para a análise de dados, a metodologia de Bardin (2010), gerando 5 (cinco)categorias e 15 (quinze) subcategorias temáticas. Os resultados evidenciaram a inserção no grupo de convivência como algo positivo, permitindo a ampliação do círculo de amizades e disposição para a realização de atividades. A percepção desta etapa envolveu a sensação de liberdade (tempo para passear, estar perto da família), prazer na descoberta de novas atividades, ambivalência e a presença da negação relacionada ao envelhecimento. Observou-se também que há preocupação com a saúde e apego à religião, que se apresentam como um recurso relevante no enfrentamento dessa etapa. Relacionado à percepção sobre a morte, apesar da negação estar presente, existe também consciência da possibilidade dela, sendo encarada como algo natural, porém misterioso. O desejo de viajar, cuidar da família e trabalhar faz parte dos planos dessas idosas. Espera-se que este estudo promova reflexões, contribuindo na eliminação de tabus que rodeiam os temas envelhecimento e morte, e, portanto, possibilite uma melhora na qualidade de vida nesta etapa do ciclo vital

Aging is a stage of life which, according to an individual's experiences, starts with a decline in his/her functioning. The elderly see their possibilities reducing and the likelihood of their death increasing.Considering such aspects, the aim of this study was to understand the perception of seven elderly women participating in a social group from the Região das Hortênsias, state of Rio Grande do Sul, Brazil, about aging and death. Data was collected through the focus group technique, with a pre-established script, and data was analyzed by means of Bardin's methodology (2010), generating five thematic categories and fifteen thematic subcategories. Results show the inclusion of these women in the social group as something positive, allowing the expansion of their friendship circles and giving them more energy to perform activities. Their perception about this stage included a feeling of freedom (time to go out, to be with their families), pleasure in discovering new activities, ambivalence and presence of denial related to aging. It was also observed that they have a concern about their health and hold on to religiosity, which appears as an important resource for them to face this stage. With regard to their perception of death, although denial is present, they are also aware of the possibility of it happening, which is seen as something natural, but mysterious. A desire to travel, take care of their families and work is part of these elderly women's plans. We expect that this study promotes reflections, contributing to breaking taboos surrounding the matters of aging and death and, therefore, improving their quality of life at this stage

El envejecimiento es una etapa del ciclo vital en la cual, de acuerdo a las vivencias de cada uno, se inicia un declive en el funcionamiento del individuo. El anciano percibe sus posibilidades disminuidas y un aumento en la probabilidad de muerte. Considerando estos aspectos, el objetivo es conocer la percepción de siete (7) ancianos integrantes de un grupo de convivencia en la Región de Las Hortensias / Rio Grande do Sul, sobre el envejecimiento y la muerte. En la recolección de datos, se utilizó la técnica de grupo focal como guía preestablecida y análisis de datos, la metodología Bardin (2010); lo cual generó cinco (5) categorías y quince (15) subcategorías temáticas. Los resultados evidenciaron la inclusión en el grupo de convivencia como algo positivo, permitiendo la ampliación del círculo de amistades y mejor disposición para la realización de actividades. La percepción en esta etapa involucra el sentido de libertad (tiempo para pasear, estar cerca de la familia), placer en el descubrimiento de nuevas actividades, ambivalencia y presencia de negación relacionada alenvejecimiento. También se observó la preocupación por la salud y el apego a la religión, que se presenta como recurso importante en el enfrentamiento de esta etapa. En relación a la percepción de la muerte, a pesar de estar presente la negación, también existe conciencia de posibilidad de ella, siendo encuadrada como algo natural, sin embargo, misterioso. El deseo de viajar, de cuidar a la familia y de trabajar, son parte de los planes de los ancianos encuestados. Se espera que este estudio promueva reflexiones, contribuyendo a la eliminación de tabúes que rodean los temas de envejecimiento y muerte y, por tanto, mejoren la calidad de vida en esta etapa del ciclo vital.

Envelhecimento e morte: percepção de idosas de um grupo de convivência / Aging and death: perception of elderly women from a support group / Envejecimiento y muerte: percepción de ancianas de un grupo de convivencia

O envelhecimento é uma etapa do ciclo vital em que, de acordo com as vivênciasde cada um, inicia um declínio no funcionamento do indivíduo. O idoso percebe suas possibilidades diminuídas e a probabilidade de morte se amplia. Considerando tais aspectos, objetivou-se conhecer a percepção de 07 (sete) idosas participantes de um grupo de convivência na Região das Hortênsias/Rio Grande do Sul, sobre o envelhecimento e a morte. Na coleta de dados, utilizou-se a técnica de grupo focal com roteiro pré-estabelecido e, para a análise de dados, a metodologia de Bardin (2010), gerando 5 (cinco)categorias e 15 (quinze) subcategorias temáticas. Os resultados evidenciaram a inserção no grupo de convivência como algo positivo, permitindo a ampliação do círculo de amizades e disposição para a realização de atividades. A percepção desta etapa envolveu a sensação de liberdade (tempo para passear, estar perto da família), prazer na descoberta de novas atividades, ambivalência e a presença da negação relacionada ao envelhecimento. Observou-se também que há preocupação com a saúde e apego à religião, que se apresentam como um recurso relevante no enfrentamento dessa etapa. Relacionado à percepção sobre a morte, apesar da negação estar presente, existe também consciência da possibilidade dela, sendo encarada como algo natural, porém misterioso. O desejo de viajar, cuidar da família e trabalhar faz parte dos planos dessas idosas. Espera-se que este estudo promova reflexões, contribuindo na eliminação de tabus que rodeiam os temas envelhecimento e morte, e, portanto, possibilite uma melhora na qualidade de vida nesta etapa do ciclo vital. (AU)

Aging is a stage of life which, according to an individual’s experiences, starts with a decline in his/her functioning. The elderly see their possibilities reducing and the likelihood of their death increasing.Considering such aspects, the aim of this study was to understand the perception of seven elderly women participating in a social group from the Região das Hortênsias, state of Rio Grande do Sul, Brazil, about aging and death. Data was collected through the focus group technique, with a pre-established script, and data was analyzed by means of Bardin’s methodology (2010), generating five thematic categories and fifteen thematic subcategories. Results show the inclusion of these women in the social group as something positive, allowing the expansion of their friendship circles and giving them more energy to perform activities. Their perception about this stage included a feeling of freedom (time to go out, to be with their families), pleasure in discovering new activities, ambivalence and presence of denial related to aging. It was also observed that they have a concern about their health and hold on to religiosity, which appears as an important resource for them to face this stage. With regard to their perception of death, although denial is present, they are also aware of the possibility of it happening, which is seen as something natural, but mysterious. A desire to travel, take care of their families and work is part of these elderly women’s plans. We expect that this study promotes reflections, contributing to breaking taboos surrounding the matters of aging and death and, therefore, improving their quality of life at this stage. (AU)

El envejecimiento es una etapa del ciclo vital en la cual, de acuerdo a las vivencias de cada uno, se inicia un declive en el funcionamiento del individuo. El anciano percibe sus posibilidades disminuidas y un aumento en la probabilidad de muerte. Considerando estos aspectos, el objetivo es conocer la percepción de siete (7) ancianos integrantes de un grupo de convivencia en la Región de Las Hortensias / Rio Grande do Sul, sobre el envejecimiento y la muerte. En la recolección de datos, se utilizó la técnica de grupo focal como guía preestablecida y análisis de datos, la metodología Bardin (2010); lo cual generó cinco (5) categorías y quince (15) subcategorías temáticas. Los resultados evidenciaron la inclusión en el grupo de convivencia como algo positivo, permitiendo la ampliación del círculo de amistades y mejor disposición para la realización de actividades. La percepción en esta etapa involucra el sentido de libertad (tiempo para pasear, estar cerca de la familia), placer en el descubrimiento de nuevas actividades, ambivalencia y presencia de negación relacionada alenvejecimiento. También se observó la preocupación por la salud y el apego a la religión, que se presenta como recurso importante en el enfrentamiento de esta etapa. En relación a la percepción de la muerte, a pesar de estar presente la negación, también existe conciencia de posibilidad de ella, siendo encuadrada como algo natural, sin embargo, misterioso. El deseo de viajar, de cuidar a la familia y de trabajar, son parte de los planes de los ancianos encuestados. Se espera que este estudio promueva reflexiones, contribuyendo a la eliminación de tabúes que rodean los temas de envejecimiento y muerte y, por tanto, mejoren la calidad de vida en esta etapa del ciclo vital. (AU)

Homozygous haplotype deficiency reveals deleterious mutations compromising reproductive and rearing success in cattle.

BACKGROUND: Cattle breeding populations are susceptible to the propagation of recessive diseases. Individual sires generate tens of thousands of progeny via artificial insemination. The frequency of deleterious alleles carried by such sires may increase considerably within few generations. Deleterious alleles manifest themselves often by missing homozygosity resulting from embryonic/fetal, perinatal or juvenile lethality of homozygotes. RESULTS: A scan for homozygous haplotype deficiency in 25,544 Fleckvieh cattle uncovered four haplotypes affecting reproductive and rearing success. Exploiting whole-genome resequencing data from 263 animals facilitated to pinpoint putatively causal mutations in two of these haplotypes. A mutation causing an evolutionarily unlikely substitution in SUGT1 was perfectly associated with a haplotype compromising insemination success. The mutation was not found in homozygous state in 10,363 animals (P=1.79×10(-5)) and is thus likely to cause lethality of homozygous embryos. A frameshift mutation in SLC2A2 encoding glucose transporter 2 (GLUT2) compromises calf survival. The mutation leads to premature termination of translation and activates cryptic splice sites resulting in multiple exon variants also with premature translation termination. The affected calves exhibit stunted growth, resembling the phenotypic appearance of Fanconi-Bickel syndrome in humans (OMIM 227810), which is also caused by mutations in SLC2A2. CONCLUSIONS: Exploiting comprehensive genotype and sequence data enabled us to reveal two deleterious alleles in SLC2A2 and SUGT1 that compromise pre- and postnatal survival in homozygous state. Our results provide the basis for genome-assisted approaches to avoiding inadvertent carrier matings and to improving reproductive and rearing success in Fleckvieh cattle.

Desistência e Conclusão em Psicoterapia Psicanalítica, um estudo qualitativo de pacientes de Porto Alegre, Brasil / Dropout and completion in Psychoanalytic therapy: A qualitative study of patients from Porto Alegre, Brazil

Este estudo compara o início e o fim do tratamento de pacientes classificados por seus psicoterapeutas como pacientes desistentes (D) e pacientes que concluíram a psicoterapia psicanalítica (PP). O objetivo do estudo é entender os fatores associados com o término unilateral e a conclusão da psicoterapia em uma clínica de psicoterapia psicanalítica comunitária ligada a um curso de pós-graduação em PP. Trata-se de um estudo qualitativo que examina o conteúdo de entrevistas iniciais e pós-tratamento de 10 casos de PP, cinco desistentes e cinco que concluíram o tratamento. O corpus analisado contava 236 páginas e gerou 672 unidades de registro, agrupadas em oito categorias, cinco das entrevistas iniciais e três das entrevistas pós-tratamento. Em comparação com o grupo C, no início da terapia, pacientes do grupo D mostravam objetivos menos amplos, menor disposição para a mudança e menos capacidade de insight, maior percepção negativa de tratamentos anteriores, mais transferência negativa e mais resistência. As entrevistas pós-tratamento indicaram que, durante a terapia, os pacientes que concluíram o tratamento tinham menor resistência que os desistentes. Além disso, após a terapia, pacientes que concluíram o tratamento se mostraram mais satisfeitos com os resultados e exibiram benefícios mais efetivos, como a habilidade de continuar a trabalhar questões psicológicas por si próprios. Tomados juntos, os resultados oferecem hipóteses para os fenômenos complexos da desistência e conclusão em PP. Essas hipóteses devem ser consideradas levando-se em conta as limitações metodológicas do estudo. Outros estudos são necessários para uma melhor compreensão das razões da interrupção e conclusão da terapia psicanalítica.(AU)

This paper compares beginning and end of treatment of patients who were classified by their therapists either as dropouts (D) or completers (C) in psychoanalytic therapy (PT). The study aims to understand factors associated with unilateral termination and completion of therapy in a community PT clinic related to a post-graduation PT course. This is a qualitative study that examined the content of initial and post-treatment interviews of 10 PT cases, five dropouts and five completers. The analyzed corpus comprised 236 pages and generated 672 record units, grouped into 8 categories, 5 from initial and 3 from after-treatment interviews. In comparison to C, at the beginning of therapy, D patients showed less broad objectives, less willingness to change and less insight, more negative perception of previous treatments, more negative transference and more resistance. Post treatment interviews indicated that during therapy completers were less resistant than dropouts. Also, after therapy C were more satisfied with results and exhibited more effective benefits, like the ability to continue working thought psychological issues by their own. Taking together, results offer hypotheses for the complex phenomena of the dropout and completion in PT. These hypotheses may be considered taking into account the methodological limitations of the study. Others studies are needed to better understand reasons for interruption or completion of psychoanalytic therapy.(AU)

A nonsense mutation in PLD4 is associated with a zinc deficiency-like syndrome in Fleckvieh cattle.

BACKGROUND: Bovine hereditary zinc deficiency (BHZD) is an autosomal recessive disorder of cattle, first described in Holstein-Friesian animals. Affected calves suffer from severe skin lesions and show a poor general health status. Recently, eight calves with the phenotypic appearance of BHZD have been reported in the Fleckvieh cattle population. RESULTS: In spite of the similar disease phenotypes, SLC39A4, the gene responsible for BHZD in Holstein-Friesian was excluded as underlying gene for the disorder in the affected Fleckvieh calves. In order to identify the disease-associated region, genotypes of eight affected calves obtained with the Illumina BovineHD BeadChip comprising 777,962 SNPs were contrasted with the genotypes of 1,339 unaffected animals. A strong association signal was observed on chromosome 21 (P = 5.87 × 10(-89)). Autozygosity mapping in the eight affected animals revealed a common segment of extended homozygosity encompassing 1,023 kb (BTA 21: 70,550,045 - 71,573,501). This region contains 17 genes/transcripts, among them two genes encoding gastro-intestinal zinc transporters (CRIP1, CRIP2). However, no mutation that was compatible with recessive inheritance could be detected in these candidate genes. One of the affected calves was re-sequenced together with 42 unaffected Fleckvieh animals. Analysis of the sequencing data revealed a nonsense mutation (p.W215X) in a phospholipase encoding gene (PLD4) as candidate causal polymorphism. To confirm the causality, genotypes of the p.W215X-mutation were obtained from 3,650 animals representing three different breeds. None of the unaffected animals was homozygous for the defect allele, while all eight affected calves were homozygous. The deleterious effect of the mutation is manifested in a significantly lower survival rate of descendants from risk matings when compared with the survival rate of descendants from non-risk matings. The deleterious allele has an estimated frequency of 1.1% in the Fleckvieh population. CONCLUSION: Our results provide strong evidence that a newly identified recessive disorder in the Fleckvieh population is caused by a nonsense mutation in PLD4, most likely resulting in an impaired function of the encoded protein. Although the phenotype of affected calves strongly resembles BHZD, a zinc deficiency resulting from malabsorption is unlikely to be responsible for the diseased Fleckvieh calves.

Momentos distintos no abandono da psicoterapia psicanalítica / Different moments in the dropout of psychoanalytic psychotherapy

Objetivo Identificar características de início e término de tratamento de pacientes que abandonaram a psicoterapia psicanalítica (PP) em momentos distintos: entre 2 e 11 meses (abandono médio = AM) ou com mais de um ano (abandono tardio = AT) de psicoterapia. Métodos Entrevistas iniciais e de pós-tratamento de 14 adultos (sete de AM e sete de AT), classificados por seus psicoterapeutas como pacientes que abandonaram a psicoterapia, foram analisadas utilizando-se o método de análise de conteúdo de Bardin. Resultados As entrevistas iniciais geraram cinco categorias (motivo, objetivos, disposição para mudança, tratamento anterior, transferência) e as de pós-tratamento geraram três categorias (processo de mudança, avaliação de resultados, término). Conclusão Em comparação com os pacientes de AT, os pacientes de AM apresentaram maior resistência, possuíam expectativas de mais apoio, apresentaram menor transferência positiva e mais queixas depressivas e relataram experiências negativas com tratamentos anteriores. A maior parte dos pacientes de AM iniciou o tratamento por indicação de terceiros, enquanto a maioria dos de AT buscou a psicoterapia por conta própria. Na entrevista pós-tratamento, os pacientes de AM revelaram mais resistência durante o processo de psicoterapia, demonstraram menor capacidade de insight e fizeram avaliações mais negativas do tratamento, tanto nos aspectos gerais como nos específicos. A diferenciação dos grupos de AM e AT é tênue, sendo necessárias mais investigações sobre a temática. .

Objective To identify characteristics of beginning and end of treatment of patients who dropped out of psychoanalytic psychotherapy (PP) at different times: between 2 and 11 months (middle dropout = MD) or with more than one year (late dropout = LD) from the beginning of psychotherapy. Methods Initial and post-treatment interviews of 14 adults (seven MD and seven LD) considered as dropouts by their therapists were analyzed using Bardin’s content analysis method. Results The analysis of initial interviews generated five categories (reason, goals, willingness to change, previous treatment, transference) and three post-treatment categories (process of change, evaluation of results, termination). Conclusion In comparison with LD patients, MD patients have more resistances, expected more support, showed less positive transferences, more depressive complaints and reported more negative experiences with previous treatments. The majority of MD patients began treatment by indication of others while most LD patients sought psychotherapy by themselves. In post-treatment interview, MD patients revealed more resistance during psychotherapy, showed less capacity for insight and made more negative evaluations of treatment, both in general and in specific ways. The differentiation MD and LD patients is tenuous and there is a need of more research on the subject. .

Beginning and end of treatment of patients who dropped out of psychoanalytic psychotherapy.

INTRODUCTION: Even though it is expected that patients and therapist will agree on when to terminate psychotherapy, quite often patients discontinue treatment on their own. This study aims to better understand dropout in psychoanalytic psychotherapy (PP). METHOD: Baseline and post-treatment interviews were conducted with six adult women classified by their therapists as dropout patients at a PP outpatient clinic. Results were analyzed using Bardin's content analysis. RESULTS: Content analysis generated five categories (reasons for treatment, goals, readiness to change, previous treatment, transference) for the baseline interviews and three categories (change process, results, interruption) for post-treatment interviews. Unclear treatment goals, low readiness to change, early signs of negative transference and resistance, and absence of recognition of their own participation in problems emerged in baseline interviews. In most cases, the patient's evaluation of the therapeutic relationship was not determinant of dropout. Therapeutic gains, dissatisfaction, and resistance during the psychotherapeutic process seemed to be associated with noncompliance. CONCLUSIONS: Decisions to initiate, continue, or quit PP depend on multiple factors, such as initial goals, readiness to change, insight capacity, awareness of personal problems, resistance, and transference. However, these factors should be considered in view of the methodological limitations of this study. Further investigation of PP dropout is still necessary.

Beginning and end of treatment of patients who dropped out of psychoanalytic psychotherapy / Início e fim de tratamento de pacientes que abandonaram a psicoterapia psicanalítica

Introduction: Even though it is expected that patients and therapist will agree on when to terminate psychotherapy, quite often patients discontinue treatment on their own. This study aims to better understand dropout in psychoanalytic psychotherapy (PP). Method: Baseline and post-treatment interviews were conducted with six adult women classified by their therapists as dropout patients at a PP outpatient clinic. Results were analyzed using Bardin's content analysis. Results: Content analysis generated five categories (reasons for treatment, goals, readiness to change, previous treatment, transference) for the baseline interviews and three categories (change process, results, interruption) for post-treatment interviews. Unclear treatment goals, low readiness to change, early signs of negative transference and resistance, and absence of recognition of their own participation in problems emerged in baseline interviews. In most cases, the patient's evaluation of the therapeutic relationship was not determinant of dropout. Therapeutic gains, dissatisfaction, and resistance during the psychotherapeutic process seemed to be associated with noncompliance. Conclusions: Decisions to initiate, continue, or quit PP depend on multiple factors, such as initial goals, readiness to change, insight capacity, awareness of personal problems, resistance, and transference. However, these factors should be considered in view of the methodological limitations of this study. Further investigation of PP dropout is still necessary.

Introdução: Embora se espere que pacientes e terapeuta concordem sobre o momento da alta ou fim do tratamento, a interrupção por conta própria do paciente é bem comum. O objetivo deste estudo é compreender melhor o abandono de tratamento em psicoterapia psicanalítica (PP). Método: Entrevistas foram conduzidas no início e após o tratamento com seis mulheres adultas classificadas por seus terapeutas como tendo abandonado o tratamento em uma clínica de PP. Os resultados foram analisados utilizando-se o método de análise de conteúdo de Bardin. Resultados: A análise de conteúdo deu origem a cinco categorias (razões para o tratamento, objetivos, disposição para mudar, tratamento prévio, transferência) nas entrevistas iniciais e três categorias (processo de mudança, resultados, interrupção) nas entrevistas pós-tratamento. Objetivos terapêuticos obscuros, pouca disposição para mudar, sinais de transferência negativa e resistência, e o não reconhecimento da própria participação nos problemas surgiram nas entrevistas iniciais. Na maioria dos casos, a avaliação da paciente sobre a relação terapêutica não foi determinante para o abandono. Ganhos terapêuticos, insatisfação e resistência durante o processo psicoterapêutico pareceram estar associados com a não aderência. Conclusões: As decisões de iniciar, continuar ou interromper a PP dependem de múltiplos fatores, como objetivos iniciais, disposição para mudar, capacidade de insight, consciência de problemas pessoais, resistência e transferência. No entanto, esses fatores devem ser considerados em vista das limitações metodológicas deste estudo. Mais investigações com foco no abandono da PP são necessários.

Identification and dissection of four major QTL affecting milk fat content in the German Holstein-Friesian population.

Milk composition traits exhibit a complex genetic architecture with a small number of major quantitative trait loci (QTL) explaining a large fraction of the genetic variation and numerous QTL with minor effects. In order to identify QTL for milk fat percentage (FP) in the German Holstein-Friesian (HF) population, a genome-wide association study (GWAS) was performed. The study population consisted of 2327 progeny-tested bulls. Genotypes were available for 44,280 SNPs. Phenotypes in the form of estimated breeding values (EBVs) for FP were used as highly heritable traits. A variance components-based approach was used to account for population stratification. The GWAS identified four major QTL regions explaining 46.18% of the FP EBV variance. Besides two previously known FP QTL on BTA14 (P = 8.91×10-(198)) and BTA20 (P = 7.03×10(-12)) within DGAT1 and GHR, respectively, we uncovered two additional QTL regions on BTA5 (P = 2.00×10(-13)) and BTA27 (P = 9.83×10(-5)) encompassing EPS8 and GPAT4, respectively. EPS8 and GPAT4 are involved in lipid metabolism in mammals. Re-sequencing of EPS8 and GPAT4 revealed 50 polymorphisms. Genotypes for five of them were inferred for the entire study population. Two polymorphisms affecting potential transcription factor binding sites of EPS8 (P = 1.40×10(-12)) and GPAT4 (P = 5.18×10(-5)), respectively, were highly significantly associated with the FP EBV. Our results provide evidence that alteration of regulatory sites is an important aspect of genetic variation of complex traits in cattle.

Identification of QTL for UV-protective eye area pigmentation in cattle by progeny phenotyping and genome-wide association analysis.

Pigmentation patterns allow for the differentiation of cattle breeds. A dominantly inherited white head is characteristic for animals of the Fleckvieh (FV) breed. However, a minority of the FV animals exhibits peculiar pigmentation surrounding the eyes (ambilateral circumocular pigmentation, ACOP). In areas where animals are exposed to increased solar ultraviolet radiation, ACOP is associated with a reduced susceptibility to bovine ocular squamous cell carcinoma (BOSCC, eye cancer). Eye cancer is the most prevalent malignant tumour affecting cattle. Selection for animals with ACOP rapidly reduces the incidence of BOSCC. To identify quantitative trait loci (QTL) underlying ACOP, we performed a genome-wide association study using 658,385 single nucleotide polymorphisms (SNPs). The study population consisted of 3579 bulls of the FV breed with a total of 320,186 progeny with phenotypes for ACOP. The proportion of progeny with ACOP was used as a quantitative trait with high heritability (h(2) = 0.79). A variance component based approach to account for population stratification uncovered twelve QTL regions on seven chromosomes. The identified QTL point to MCM6, PAX3, ERBB3, KITLG, LEF1, DKK2, KIT, CRIM1, ATRN, GSDMC, MITF and NBEAL2 as underlying genes for eye area pigmentation in cattle. The twelve QTL regions explain 44.96% of the phenotypic variance of the proportion of daughters with ACOP. The chromosomes harbouring significantly associated SNPs account for 54.13% of the phenotypic variance, while another 19.51% of the phenotypic variance is attributable to chromosomes without identified QTL. Thus, the missing heritability amounts to 7% only. Our results support a polygenic inheritance pattern of ACOP in cattle and provide the basis for efficient genomic selection of animals that are less susceptible to serious eye diseases.

Genome-wide association study uncovers four QTL predisposing to supernumerary teats in cattle.

Supernumerary teats (hyperthelia, SNTs) are a common abnormality of the bovine udder with a medium to high heritability and a postulated oligogenic or polygenic inheritance pattern. SNTs not only negatively affect machine milking ability but also act as a reservoir for bacteria. A genome-wide association study was carried out to identify genes involved in the development of SNTs in the dual-purpose Fleckvieh breed. A total of 2467 progeny-tested bulls were genotyped at 43 698 single nucleotide polymorphisms, and daughter yield deviations (DYDs) for 'udder clearness' (UC) were used as high-heritability phenotypes. Massive structuring of the study population was accounted for by principal components analysis-based and mixed model-based approaches. Four loci on BTA5, BTA6, BTA11 and BTA17 were significantly associated with the UC DYD. Three associated regions contain genes of the highly conserved Wnt signalling pathway. The four QTL together account for 10.7% of the variance of the UC DYD, whereas the major fraction of the DYD variance is attributable to chromosomes with no identified QTL. Our results support both an oligogenic and a polygenic inheritance pattern of SNTs in cattle. The identified candidate genes permit insights into the genetic architecture of teat malformations in cattle and provide clues to unravel the molecular mechanisms of mammary gland alterations in cattle and other species.

Genome-wide association study identifies two major loci affecting calving ease and growth-related traits in cattle.

Identifying quantitative trait loci (QTL) underlying complex, low-heritability traits is notoriously difficult. Prototypical for such traits, calving ease is an important breeding objective of cattle (Bos taurus)-improving programs. To identify QTL underlying calving ease, we performed a genome-wide association study using estimated breeding values (EBVs) as highly heritable phenotypes for paternal calving ease (pCE) and related traits. The massively structured study population consisted of 1800 bulls of the German Fleckvieh (FV) breed. Two pCE-associated regions on bovine chromosomes (BTA) 14 and 21 (P = 5.72 × 10(-15) and P = 2.27 × 10(-8), respectively) were identified using principal components analysis to correct for population stratification. The two most significantly associated SNPs explain 10% of the EBV variation. Since marker alleles with negative effect on pCE have positive effects on growth-related traits, the QTL may exert their effects on the birthing process through fetal growth traits. The QTL region on BTA14 corresponds to a human chromosome (HSA) region that is associated with growth characteristics. The HSA region corresponding to the BTA21 pCE QTL is maternally imprinted and involved in the Prader-Willi and Angelman syndromes. Resequencing of positional candidate genes on BTA14 revealed a highly significantly (P = 1.96 × 10(-14)) associated polymorphism ablating a polyadenylation signal of the gene encoding ribosomal protein S20 (RPS20). Our study demonstrates the leverage potential of EBVs in unraveling the genetic architecture of lowly heritable traits.