Fístulas rectoneovaginales en pacientes tras cirugía de reasignación de sexo. Descripción de nuestra experiencia y revisión de la literatura / Rectoneovaginal fistula after sex reassignment surgery. Description of our experience and literature review

La cirugía de reasignación de sexo masculino a femenino o vaginoplastia incluye aquellos procedimientos quirúrgicos cuyo objetivo es recrear un perineo femenino funcional y estéticamente aceptable con una formación mínima de cicatrices. La técnica de elección en nuestro centro es la vaginoplastia con inversión peneana con o sin injertos escrotales. Presentamos 4 casos diagnosticados con fístulas rectoneovaginales tratados en nuestro centro con evolución favorable. La primera paciente fue diagnosticada en el postoperatorio tardío durante las dilataciones. Se sometió a 2 intentos de reparación vaginal sin éxito. Finalmente, se realizó una colostomía temporal y un colgajo rectal. La segunda paciente fue diagnosticada 2 semanas después de la cirugía inicial debido a una dilatación agresiva, siendo tratada con una colostomía temporal y cierre por segunda intención. La tercera paciente se diagnostica en el quinto día postoperatorio, después de la extracción del taponamiento vaginal. Se indicó restricción dietética y se realizó un colgajo rectal. Una cuarta paciente fue diagnosticada durante el postoperatorio tardío, realizando una exploración quirúrgica y un colgajo de pared rectal. Las fístulas rectoneovaginales después de la cirugía de reasignación de sexo tienen una incidencia de alrededor del 2-17%, siendo el tipo más común de fístulas después de este procedimiento. La causa más común es secundaria a lesiones rectales durante la cirugía inicial. Para el manejo de estas fístulas se plantean opciones que van desde el cierre primario a las colostomías de descarga, pasando por un manejo conservador o la confección de colgajos. Se recomienda el abordaje por un equipo multidisciplinario para el diagnóstico y tratamiento de esta complicación. (AU)

Male-to-female reassignment surgery or vaginoplasty includes those surgical procedures that aim to recreate a functional and cosmetically acceptable female perineum with minimal scarring. The technique of choice at our center is penile inversion vaginoplasty with or without scrotal skin grafts. We present 4 cases diagnosed with rectoneovaginal fistulas treated at our center with favorable evolution. The first patient was diagnosed in the late postoperative period during dilation. She underwent 2 failed vaginal repair attempts. Finally, a temporary colostomy and a rectal flap were performed. The second patient was diagnosed 2 weeks after the initial surgery due to aggressive dilation and was treated with a temporary colostomy and secondary wound closure. The third patient was diagnosed on the fifth post-operative day after removal of the vaginal packing. Dietary restriction was indicated, and a rectal flap was performed. A fourth patient was diagnosed within the late postoperative period; she was submitted to surgical exploration and a rectal wall flap was created. Rectoneovaginal fistulas after sex reassignment surgery has an incidence of about 2-17% and they are the most common type of fistula after this procedure. In most cases, it is secondary to rectal injury during the initial surgery. The management of these fistulas ranges from primary closure, diverting colostomies, conservative management, or the performance of flaps. A multidisciplinary team approach is recommended for the diagnosis and treatment of this complication. (AU)

Rectoneovaginal fistula after sex reassignment surgery. Description of our experience and literature review. / Fístulas rectoneovaginales en pacientes tras cirugía de reasignación de sexo. Descripción de nuestra experiencia y revisión de la literatura.

Male-to-female reassignment surgery or vaginoplasty includes those surgical procedures that aim to recreate a functional and cosmetically acceptable female perineum with minimal scarring. The technique of choice at our center is penile inversion vaginoplasty with or without scrotal skin grafts. We present 4 cases diagnosed with rectoneovaginal fistulas treated at our center with favorable evolution. The first patient was diagnosed in the late postoperative period during dilation. She underwent 2 failed vaginal repair attempts. Finally, a temporary colostomy and a rectal flap were performed. The second patient was diagnosed 2 weeks after the initial surgery due to aggressive dilation and was treated with a temporary colostomy and secondary wound closure. The third patient was diagnosed on the fifth post-operative day after removal of the vaginal packing. Dietary restriction was indicated, and a rectal flap was performed. A fourth patient was diagnosed within the late postoperative period; she was submitted to surgical exploration and a rectal wall flap was created. Rectoneovaginal fistulas after sex reassignment surgery has an incidence of about 2-17% and they are the most common type of fistula after this procedure. In most cases, it is secondary to rectal injury during the initial surgery. The management of these fistulas ranges from primary closure, diverting colostomies, conservative management, or the performance of flaps. A multidisciplinary team approach is recommended for the diagnosis and treatment of this complication.

Development of a 64 SNV panel for breed authentication in Iberian pigs and their derived meat products.

Spanish legislation regulates the labelling of Iberian pig meat and dry-cured products, which are labelled as "Ibérico" or "100% Ibérico" when they come from Duroc x Iberian crossbred or Iberian purebred pigs. Although the analytical authentication of breed origin is not mandatory, a genetic diagnostic tool is demanded by producers and consumers. We have designed a 64 Single Nucleotide Variant genotyping panel displaying extreme allelic frequencies between Duroc and Iberian purebred samples. Average proportions of Iberian alleles of 0.99, 0.01, 0.77 and 0.48 were estimated by admixture clustering analysis of known origin samples, for Iberian and Duroc purebred, 75% Iberian and 50% Iberian classes, respectively. A supervised analysis with 1419 samples showed some overlapping between contiguous classes, but the calculated degrees of separability ranged from 0.800 to 0.996, exceeding the threshold value (0.70) for considering suitable for prediction. Therefore, this panel is a useful genetic tool to infer purebred or crossbred Iberian origin of live animals, meat and dry-cured products.

Assessing effective population size, coancestry and inbreeding effects on litter size using the pedigree and SNP data in closed lines of the Iberian pig breed.

The complete pedigree of two closed Iberian pig lines (Gamito and Torbiscal), with 798 and 4077 reproducers, has been used to measure the evolution of coancestry (f) and inbreeding (F) for autosomal and X-linked genes along 16 and 28 respective equivalent discrete generations. At the last generation, the mean values of each line were f = 0.41 and 0.22, F = 0.35 and 0.18, fX  = 0.46 and 0.22 and FX  = 0.47 and 0.19, respectively. Other calculated parameters were the effective number of founders (final values, 6.8 and 35.2) and non-founders (1.5 and 2.4), founder genome equivalents (1.2 and 2.3) and effective population size (16.0 and 57.7). Measures of Torbiscal effective size based on rates of coancestry (66.1), inbreeding (65.0) and linkage disequilibrium (71.0) were estimated from whole-genome SNP genotyping data. Values of new and old inbreeding and their respective rates by generation were computed to detect purging effects of natural selection. The analysis of 6854 Torbiscal litters showed significant negative impacts of new and fast inbreeding on litter size, as expected from the purging hypothesis: -0.20 born piglets per litter by a 10% of new inbreeding, and -0.03 and -0.02 piglets by 1% of total and new inbreeding rates, respectively. The analysis performed on 1274 litters of the Gamito line failed to show purging effects. The only significant results were reductions in -0.91 and -0.17 piglets by a 10% of old and X-linked genes inbreeding, respectively. These results may be useful for some practical issues in conservation programs of farm or captive wild animals.

Effects of dietary fat saturation on fatty acid composition and gene transcription in different tissues of Iberian pigs.

The effect of two diets, respectively enriched with SFA (S) and PUFA (P), on FA tissue composition and gene expression was studied in fattened Iberian pigs. The FA composition of adipose, muscular and liver tissues was affected by dietary treatment. S group showed higher MUFA and MUFA/SFA ratio and lower PUFA and n-6/n-3 ratio than P group in all analyzed tissues. In muscle and liver the extracted lipids were separated into neutral lipids and polar lipid fractions which showed significantly different responses to the dietary treatment, especially in liver where no significant effect of diet was observed in NL fraction. The expression of six candidate genes related to lipogenesis and FA oxidation was analyzed by qPCR. In liver, stearoyl CoA desaturase (SCD), acetyl CoA carboxylase alpha (ACACA) and malic enzyme 1 (ME1) genes showed higher expression in S group. SCD, ACACA, ME1, and fatty acid synthase (FASN) gene expression levels showed a wide variation across the tested tissues, with much higher expression levels observed in adipose tissue than other tissues. Tissue FA profile and gene expression results support the deposition of dietary FA, the lipogenic effect of dietary saturated fat in liver and the employment of saturated dietary fat for endogenous synthesis of MUFA in all the analyzed tissues.

Copy number variants in a highly inbred Iberian porcine strain.

We carried out a comprehensive genomic analysis of porcine copy number variants (CNVs) based on whole-genome SNP genotyping data and provided new measures of genomic diversity (number, length and distribution of CNV events) for a highly inbred strain (the Guadyerbas strain). This strain represents one of the most ancient surviving populations of the Iberian breed, and it is currently in serious danger of extinction. CNV detection was conducted on the complete Guadyerbas population, adjusted for genomic waves, and used strict quality criteria, pedigree information and the latest porcine genome annotation. The analysis led to the detection of 65 CNV regions (CNVRs). These regions cover 0.33% of the autosomal genome of this particular strain. Twenty-nine of these CNVRs were identified here for the first time. The relatively low number of detected CNVRs is in line with the low variability and high inbreeding estimated previously for this Iberian strain using pedigree, microsatellite or SNP data. A comparison across different porcine studies has revealed that more than half of these regions overlap with previously identified CNVRs or multicopy regions. Also, a preliminary analysis of CNV detection using whole-genome sequence data for four Guadyerbas pigs showed overlapping for 16 of the CNVRs, supporting their reliability. Some of the identified CNVRs contain relevant functional genes (e.g., the SCD and USP15 genes), which are worth being further investigated because of their importance in determining the quality of Iberian pig products. The CNVR data generated could be useful for improving the porcine genome annotation.

Measuring inbreeding and inbreeding depression on pig growth from pedigree or SNP-derived metrics.

Multilocus homozygosity, measured as the proportion of the autosomal genome in homozygous genotypes or in runs of homozygosity, was compared with the respective pedigree inbreeding coefficients in 64 Iberian pigs genotyped using the Porcine SNP60 Beadchip. Pigs were sampled from a set of experimental animals with a large inbreeding variation born in a closed strain with a completely recorded multi-generation genealogy. Individual inbreeding coefficients calculated from pedigree were strongly correlated with the different SNP-derived metrics of homozygosity (r = 0.814-0.919). However, unequal correlations between molecular and pedigree inbreeding were observed at chromosomal level being mainly dependent on the number of SNPs and on the correlation between heterozygosities measured across different loci. A panel of 192 SNPs of intermediate frequencies was selected for genotyping 322 piglets to test inbreeding depression on postweaning growth performance (daily gain and weight at 90 days). The negative effects on these traits of homozygosities calculated from the genotypes of 168 quality-checked SNPs were similar to those of inbreeding coefficients. The results support that few hundreds of SNPs may be useful for measuring inbreeding and inbreeding depression, when the population structure or the mating system causes a large variance of inbreeding.

Quantitative trait loci for fatness at growing and reproductive stages in Iberian × Meishan F(2) sows.

A considerable number of fatness QTL have been identified in growing pigs, but there is a lack of knowledge about the genetic architecture of this trait in gilts and sows. We have performed a genome scan, in 255 Iberian × Meishan F(2) sows, for backfat thickness (BF) at 150 (BF(150) ) and 210 (BF(210)) days of age, 30 days after conception (BF(30)) and 7-10 days before farrowing (BF(bf)). We have found one BF150 QTL in SSC6 (120 cM) that was highly significant (P < 0.001) at the chromosome-wide level and suggestive at the genome-wide level (P < 0.1). Ten additional chromosome-wide significant QTL were found for sow BF(150) (SSC1, SSC13), BF(210) (SSC6, SSC8, SSC15), BF(30) (SSC5, SSC6) and BF(bf) (SSC1, SSC6, SSC13). The location of several of the BF QTL varied depending on the growing and reproductive status of the sow, suggesting that part of these genetic effects may have a temporal pattern of phenotypic expression.

Effects of porcine MC4R and LEPR polymorphisms, gender and Duroc sire line on economic traits in Duroc × Iberian crossbred pigs.

Polymorphisms MC4R c.1426 A>G (p.Asp298Asn) and LEPR c.2002 C>T (p.Leu663Phe) have been associated in pigs with feed intake, growth and fatness. Our aims were to analyze how these polymorphisms influence production and quality traits in Duroc × Iberian crossbred pigs, and moreover evaluate how the effects of gender and Duroc line were affecting these traits. Significant effects of gender, Duroc sire line and LEPR c.2002 C>T mutation, were detected on the main traits. LEPR c.2002 T allele increased body weight (2.5 ± 0.7 kg), backfat thickness (1.5 ± 0.4mm), loin intramuscular fat (0.7 ± 0.2 %), and lightness (1.0 ± 0.3); however it was decreasing premium cuts yield (-0.9 ± 0.2%). Allele c.2002 T increased SFA (0.60 ± 0.12 %), and decreased MUFA (-0.35 ± 0.12%) and PUFA (-0.25 ± 0.06%). No significant effects of MC4R c.1426 A>G SNP were identified. A genetic test based on the LEPR c.2002 C>T SNP may be useful to select Duroc sires to reduce undesirable heterogeneity of carcasses and premium cuts of crossbred animals.

Catéter en vena cava superior para hemodiálisis entre los últimos recursos en hemitórax superior / Catheter in the superior vena cava for hemodialysis as a last resort in superior hemithorax

Se presentan 4 pacientes con enfermedad renal crónica en terapia hemodialítica en quienes se habían agotado los accesos venosos clásicos (yugular interno, subclavio)y no clásicos (axilar e innominado) en el hemitórax superior para hemodiálisis, debido principalmente a trombosis de los mismos por cateterismos anteriores, y que no eran candidatos a diálisis peritoneal. En ellos, con la técnica recomendada por Archundia et al., se implantaron4 catéteres permanentes directamente en la vena cava superior, con posterior tunelización subcutánea. Los catéteres funcionaron adecuadamente y están permeables actualmente después de un período de utilización promedio de 19 meses (AU)

We report four patients with chronic kidney disease undergoing haemodialysis therapy, which had exhausted conventional venous access (internal jugular, subclavian) and non-conventional access (axillary, innominate) in the upper hemithorax for haemodialysis. This was primarily due tothrombosis of these veins caused by previous catheterisation. These patients did not qualify for peritoneal dialysis. Using the technique recommended by Archundia et al., 4 indwelling catheters were implanted directly in the superior vena cava in each of the patients with subsequent subcutaneous tunneling. The catheters operated correctly and are currently permeable after being used for an average of 19 months (AU)

[Catheter in the superior vena cava for hemodialysis as a last resort in superior hemithorax]. / Catéter en vena cava superior para hemodiálisis entre los últimos recursos en hemitórax superior.

We report four patients with chronic kidney disease undergoing haemodialysis therapy, which had exhausted conventional venous access (internal jugular, subclavian) and non-conventional access (axillary, innominate) in the upper hemithorax for haemodialysis. This was primarily due to thrombosis of these veins caused by previous catheterisation. These patients did not qualify for peritoneal dialysis. Using the technique recommended by Archundia et al., 4 indwelling catheters were implanted directly in the superior vena cava in each of the patients with subsequent subcutaneous tunneling. The catheters operated correctly and are currently permeable after being used for an average of 19 months.

[Implanting haemodialysis catheters in the brachiocephalic vein: a little-used approach]. / Implantación de catéteres para hemodiálisis en vena innominada, una ruta poco utilizada.

We report three patients with chronic renal failure in hemodialysis therapy, which had exhausted conventional venous access in the upper hemitórax (internal jugular, subclavian, axillary) for hemodialysis, primarily due to thrombosis of these veins by previous catheterization. We chose the technique of Rao et al. and cols for puncture of innominate vein, resulting in the implantation of catheters and their subsequent subcutaneous tunneling. Permanent catheters3 work properly and are permeable to the date after a period averaging 8 months.

Placement of vascular access catheters for haemodialysis in the innominate vein: a little-used approach

We report three patients with chronic renal failure in hemodialysis therapy, which had exhausted conventional venous access in the upper hemitórax(internal jugular, subclavian, axillary) for hemodialysis, primarily due to thrombosis of these veins by previous catheterization. We chose the technique of Raoandcols for puncture of in nominate vein, resulting in the implantation of catheter s and their subsequent subcutaneous tunneling. Permanent catheters3 work properly and are permeable to the date after a period averaging 8 months (AU)

Se estudian tres pacientes con enfermedad renal crónica en terapia hemodialítica, en los cuales se habían agotado los accesos venosos clásicos en el hemitórax superior (yugular interno, subclavio, axilar) para hemodiálisis, debido principalmente a trombosis de los mismos por cateterismos anteriores. En ellos se optó, mediante la técnica de Raoetal., por puncionar la vena innominada, lográndose la implantación posterior de catéteres y su tunelización subcutánea. Los catéteres permanentes3funcionaron adecuadamente y están permeables a la fecha después de un periodo promedio de ocho meses (AU)

Pig melatonin receptor 1a (MTNR1A) genotype is associated with seasonal variation of sow litter size.

The duration of nocturnal melatonin secretion reaches its minimum in summer, a physiological event that is likely related with the diminished sow fertility and delayed puberty typically observed in this season. Melatonin exerts its function by binding two different receptors named as MTNR1A and MTNR1B. Interestingly, the MTNR1A gene is located on a chromosome SSC17 region where QTL for prolificacy traits have been detected in previous studies. In this work, we have found a synonymous T162C polymorphism at exon 2 of the pig MTNR1A gene. An association analysis between this polymorphism and sow prolificacy in an Iberianx Meishan intercross was performed. The utilization of four statistical models of increasing complexity demonstrated that the MTNR1A gene has both additive and dominant effects on total number of born piglets (TNB) and number of piglets born alive (NBA). Additive effects were significant in summer (TNB, P<0.01; NBA, P<0.001), whereas dominant effects reached significance both in fall (TNB, P<0.01; NBA, P<0.05) and in winter (TNB, P<0.001; NBA, P<0.05). The seasonal variation observed for MTNR1A additive and dominant effects might be produced by the influence of photoperiod on the pattern and duration of melatonin secretion. These results illustrate that the complex interaction between genotype and environment can be an important source of phenotypic variation of reproductive traits.

Empirical Bayes factor analyses of quantitative trait loci for gestation length in Iberian × Meishan F2 sows.

The aim of this study was to investigate chromosomal regions affecting gestation length in sows. An experimental F2 cross between Iberian and Meishan pig breeds was used for this purpose and we genotyped 119 markers covering the 18 porcine autosomal chromosomes. Within this context, we have developed a new empirical Bayes factor (BF) approach to compare between nested models, with and without the quantitative trait loci (QTL) effect, and after including the location of the QTL as an unknown parameter in the model. This empirical BF can be easily calculated from the output of a Markov chain Monte Carlo sampling by averaging conditional densities at the null QTL effects. Linkage analyses were performed in each chromosome using an animal model to account for infinitesimal genetic effects. Initially, three QTL were detected at chromosomes 6, 8 and 11 although, after correcting for multiple testing, only the additive QTL located in cM 110 of chromosome 8 remained. For this QTL, the allelic effect of substitution of the Iberian allele increased gestation length in 0.521 days, with a highest posterior density region at 95% ranged between 0.121 and 0.972 days. Although future studies are necessary to confirm if detected QTL is relevant and segregating in commercial pig populations, a hot-spot on the genetic regulation of gestation length in pigs seems to be located in chromosome 8.

QTL detection on porcine chromosome 12 for fatty-acid composition and association analyses of the fatty acid synthase, gastric inhibitory polypeptide and acetyl-coenzyme A carboxylase alpha genes.

Refinement of previous QTL on porcine chromosome 12 for fatty-acid composition and a candidate gene association analysis were conducted using an Iberian x Landrace cross. The concentrations of ten fatty acids were assayed in backfat tissue from which four metabolic ratios were calculated for 403 F2 animals. Linkage analysis identified two significant QTL. The first QTL was associated with the average chain length ratio and the percentages of myristic, palmitic and gadoleic acids. The second QTL was associated with percentages of palmitoleic, stearic and vaccenic acids. Based upon its position on SSC12, fatty acid synthase was tested as a candidate gene for the first QTL and no significant effects were found. Similarly, gastric inhibitory polypeptide (GIP) and acetyl-coenzyme A carboxylase alpha (ACACA) were tested as candidate genes for the second QTL using three SNPs in GIP and 15 synonymous SNPs in ACACA cDNA sequences. Two missense SNPs in GIP showed significant effects with palmitoleic and stearic fatty-acid concentration. Highly significant associations were found for two SNPs in ACACA with stearic, palmitoleic and vaccenic fatty-acid concentrations. These associations could be due to linkage disequilibrium with the causal mutations.

QTL mapping for teat number in an Iberian-by-Meishan pig intercross.

The aim of this study was to investigate chromosomal regions affecting the number of teats in pigs and possible epistatic interactions between the identified quantitative trait loci (QTL). An experimental F2 cross between Iberian and Chinese Meishan lines was used for this purpose. A genomic scan was conducted with 117 markers covering the 18 porcine autosomes. Linkage analyses were performed by interval mapping using an animal model to estimate QTL and additive polygenic effects. Complementary analyses with models fitting two QTL were also carried out. The results showed three genomewide significant QTL mapping on chromosomes 5, 10 and 12, whose joint action control up to 30% of the phenotypic variance of the trait. Meishan alleles had a positive additive effect on teat number, and a positive-additive x additive-epistatic interaction was detected between QTL on chromosomes 10 and 12.

Fine mapping of porcine chromosome 6 QTL and LEPR effects on body composition in multiple generations of an Iberian by Landrace intercross.

The leptin receptor gene (LEPR) is a candidate for traits related to growth and body composition, and is located on SSC6 in a region where fatness and meat composition quantitative trait loci (QTL) have previously been detected in several F2 experimental designs. The aims of this work were: (i) to fine map these QTL on a larger sample of animals and generations (F3 and backcross) of an Iberian x Landrace intercross and (ii) to examine the effects of LEPR alleles on body composition traits. Eleven single nucleotide polymorphisms (SNPs) were detected by sequencing LEPR coding regions in Iberian and Landrace pig samples. Three missense polymorphisms were genotyped by pyrosequencing in 33 F0, 70 F1, 418 F2, 86 F3 and 128 individuals coming from the backcross of four F2 males with 24 Landrace females. Thirteen microsatellites and one SNP were also genotyped. Traits analysed were: backfat thickness at different locations (BF(T)), intramuscular fat percentage (IMF(P)), eye muscle area (EM(A)), loin depth (LO(D)), weight of shoulder (SH(W)), weight of ribs (RIB(W)) and weight of belly bacon (BB(W)). Different statistical models were applied in order to evaluate the number and effects of QTL on chromosome 6 and the possible causality of the LEPR gene variants with respect to the QTL. The results support the presence of two QTL on SSC6. One, at position 60-100 cM, affects BF(T) and RIB(W). The other and more significant maps in a narrow region (130-132 cM) and affects BF(T), IMF(P), EM(A), LO(D), SH(W), RIB(W) and BB(W). Results also support the association between LEPR alleles and BF(T) traits. The possible functional implications of the analysed polymorphisms are considered.

Analysis of genetic diversity and conservation priorities in Iberian pigs based on microsatellite markers.

The Iberian breed is the most important pig population of the Mediterranean type. The genetic structure of two strains (Torbiscal and Guadyerbas) and three varieties (Retinto, Entrepelado and Lampiño) of this breed was studied using 173 pigs genotyped for 36 microsatellites. In addition, 40 pigs of the related Duroc breed were also analysed. In the 1960s, the Iberian breed's numbers were severely reduced by disease, due to economic change and to crossbreeding. Varieties are in danger of disappearance or blending. A new conservation strategy is required. An analysis was performed that allows us to ascertain the loss or gain of genetic diversity if one or several subpopulations are removed. The results are compared with those using the Weitzman method. The two methodologies produce conservation priorities that are completely different, the reason being that the Weitzman method does not take into account the within-population genetic diversity. We apply optimal contribution theory and a new procedure for cluster analysis, and discuss their value in the general framework of the problems of setting of priorities and tactics for the conservation of genetic resources.

Estimation of genetic variability of the founder population in a conservation scheme using microsatellites.

In a conservation programme with genealogical records it is possible to estimate the amount of variability of the founder population from a measure of the similarity among the individuals in the current population based on microsatellite markers. Here we compare three available methods and we shown that the one based on the molecular coancestry coefficient should be preferred.