Identification of telocytes in the oviduct of the mare.

Telocytes (TCs), a recently discovered special type of stromal cells, have been identified in many organs of many species, including the female and male reproductive system, with proposed multiple potential bio-functions such as homeostasis, immunomodulation, tissue remodeling and regeneration, embryogenesis, angiogenesis and even tumorigenesis. The aim of this study was to investigate the existence, and characteristics of telocytes in normal equine oviduct. To identify them, we used routine light microscopy, non-conventional light microscopy (NCLM), transmission electron microscopy (TEM), and immunohistochemistry. We found that telocytes of the equine oviduct can be recognized in fixed specimens by light microscopy (methylene blue staining), with more details on Epon semi-thin sections (toluidine blue staining) by NCLM, and that they showed positive immunostaining for CD34. The telocytes, with their typical long and moniliform prolongations, formed networks in the stromal space of the submucosa, muscular and serosa layers, particularly in the lamina propia where they were observed in greater quantity. By TEM we have also confirmed the presence of cells ultrastructurally identifiable as telocytes (cells with telopodes alternating podomers and podoms) in the aforementioned locations. Direct intercellular contacts between epithelial cells and neighboring telocytes were evidenced. EIn conclusion, we demonstrated that telocytes are present in the equine oviduct as previously reported in other species. The potential implication of telocytes in multiple potential functions of physiological and pathological processes deserves further investigation.

Improved clinical utility of preimplantation genetic testing through the integration of ploidy and common pathogenic microdeletions analyses.

STUDY QUESTION: Can chromosomal abnormalities beyond copy-number aneuploidies (i.e. ploidy level and microdeletions (MDs)) be detected using a preimplantation genetic testing (PGT) platform? SUMMARY ANSWER: The proposed integrated approach accurately assesses ploidy level and the most common pathogenic microdeletions causative of genomic disorders, expanding the clinical utility of PGT. WHAT IS KNOWN ALREADY: Standard methodologies employed in preimplantation genetic testing for aneuploidy (PGT-A) identify chromosomal aneuploidies but cannot determine ploidy level nor the presence of recurrent pathogenic MDs responsible for genomic disorders. Transferring embryos carrying these abnormalities can result in miscarriage, molar pregnancy, and intellectual disabilities and developmental delay in offspring. The development of a testing strategy that integrates their assessment can resolve current limitations and add valuable information regarding the genetic constitution of embryos, which is not evaluated in PGT providing new level of clinical utility and valuable knowledge for further understanding of the genomic causes of implantation failure and early pregnancy loss. To the best of our knowledge, MDs have never been studied in preimplantation human embryos up to date. STUDY DESIGN, SIZE, DURATION: This is a retrospective cohort analysis including blastocyst biopsies collected between February 2018 and November 2021 at multiple collaborating IVF clinics from prospective parents of European ancestry below the age of 45, using autologous gametes and undergoing ICSI for all oocytes. Ploidy level determination was validated using 164 embryonic samples of known ploidy status (147 diploids, 9 triploids, and 8 haploids). Detection of nine common MD syndromes (-4p=Wolf-Hirschhorn, -8q=Langer-Giedion, -1p=1p36 deletion, -22q=DiGeorge, -5p=Cri-du-Chat, -15q=Prader-Willi/Angelman, -11q=Jacobsen, -17p=Smith-Magenis) was developed and tested using 28 positive controls and 97 negative controls. Later, the methodology was blindly applied in the analysis of: (i) 100 two pronuclei (2PN)-derived blastocysts that were previously defined as uniformly euploid by standard PGT-A; (ii) 99 euploid embryos whose transfer resulted in pregnancy loss. PARTICIPANTS/MATERIALS, SETTING, METHODS: The methodology is based on targeted next-generation sequencing of selected polymorphisms across the genome and enriched within critical regions of included MD syndromes. Sequencing data (i.e. allelic frequencies) were analyzed by a probabilistic model which estimated the likelihood of ploidy level and MD presence, accounting for both sequencing noise and population genetics patterns (i.e. linkage disequilibrium, LD, correlations) observed in 2504 whole-genome sequencing data from the 1000 Genome Project database. Analysis of phased parental haplotypes obtained by single-nucleotide polymorphism (SNP)-array genotyping was performed to confirm the presence of MD. MAIN RESULTS AND THE ROLE OF CHANCE: In the analytical validation phase, this strategy showed extremely high accuracy both in ploidy classification (100%, CI: 98.1-100%) and in the identification of six out of eight MDs (99.2%, CI: 98.5-99.8%). To improve MD detection based on loss of heterozygosity (LOH), common haploblocks were analyzed based on haplotype frequency and LOH occurrence in a reference population, thus developing two further mathematical models. As a result, chr1p36 and chr4p16.3 regions were excluded from MD identification due to their poor reliability, whilst a clinical workflow which incorporated parental DNA information was developed to enhance the identification of MDs. During the clinical application phase, one case of triploidy was detected among 2PN-derived blastocysts (i) and one pathogenic MD (-22q11.21) was retrospectively identified among the biopsy specimens of transferred embryos that resulted in miscarriage (ii). For the latter case, family-based analysis revealed the same MD in different sibling embryos (n = 2/5) from non-carrier parents, suggesting the presence of germline mosaicism in the female partner. When embryos are selected for transfer based on their genetic constitution, this strategy can identify embryos with ploidy abnormalities and/or MDs beyond aneuploidies, with an estimated incidence of 1.5% (n = 3/202, 95% CI: 0.5-4.5%) among euploid embryos. LIMITATIONS, REASONS FOR CAUTION: Epidemiological studies will be required to accurately assess the incidence of ploidy alterations and MDs in preimplantation embryos and particularly in euploid miscarriages. Despite the high accuracy of the assay developed, the use of parental DNA to support diagnostic calling can further increase the precision of the assay. WIDER IMPLICATIONS OF THE FINDINGS: This novel assay significantly expands the clinical utility of PGT-A by integrating the most common pathogenic MDs (both de novo and inherited ones) responsible for genomic disorders, which are usually evaluated at a later stage through invasive prenatal testing. From a basic research standpoint, this approach will help to elucidate fundamental biological and clinical questions related to the genetics of implantation failure and pregnancy loss of otherwise euploid embryos. STUDY FUNDING/COMPETING INTEREST(S): No external funding was used for this study. S.C., M.F., F.C., P.Z., I.P., L.G., C.P., M.P., D.B., J.J.-A., D.B.-J., J.M.-V., and C.R. are employees of Igenomix and C.S. is the head of the scientific board of Igenomix. A.C. and L.P. are employees of JUNO GENETICS. Igenomix and JUNO GENETICS are companies providing reproductive genetic services. TRIAL REGISTRATION NUMBER: N/A.

Wrist kinetics after scaphoidectomy.

BACKGROUND: The scaphoid cannot be excised without generating substantial carpal dysfunction. The extent and nature of such a destabilizing procedure, however, has never been properly studied in the laboratory. METHODS: We used a six-degrees-of-freedom motion tracking device to quantify the changes in carpal alignment produced by isometric simultaneous loading of five wrist motor tendons in 12 fresh normal cadaver arms, before and after excising the entire scaphoid. FINDINGS: In the intact wrist, tendon loading consistently extended and supinated the capitate while flexing the triquetrum. After scaphoidectomy, the opposite rotations were always found: the capitate collapsed into flexion and pronation, whereas the triquetrum migrated proximally, while extending and radial deviating. All these changes were statistically significant. INTERPRETATION: Unless it is supplemented by some sort of midcarpal stabilization, scaphoidectomy alone is much too aggressive as a procedure to be considered a treatment option for wrist osteoarthritis. LEVEL OF EVIDENCE: Laboratory study. Not applicable.

Effect of mifepristone on the transcriptomic signature of endometrial receptivity.

STUDY QUESTION: How does a single dose of mifepristone on Day 2 after the LH peak (LH + 2) affect the endometrial receptivity transcriptome as assessed by the receptive signature established by the endometrial receptivity analysis (ERA)? SUMMARY ANSWER: A single dose of mifepristone on day LH + 2 renders the endometrium non-receptive by altering the transcriptome associated with endometrial receptivity. WHAT IS KNOWN ALREADY: Mifepristone is a progesterone receptor modulator that has been shown to alter endometrial receptivity. The ERA is a computational predictor that utilizes gene expression data of 248 genes from next generation sequencing to identify endometrial receptivity status. STUDY DESIGN, SIZE, DURATION: Endometrial biopsies were collected on day LH + 7 from controls (n = 11) and from women treated with mifepristone (n = 7). For further comparative analysis, samples were also obtained from women in the proliferative phase (n = 7). PARTICIPANTS/MATERIALS, SETTING, METHODS: Mifepristone treatment consisted of 200 mg administered on day LH + 2. Endometrial biopsies were treated for RNA isolation and cDNA conversion and sequencing. Endometrial receptivity status was assessed by the ERA computational predictor. Differential gene expression between groups was also assessed. Ingenuity Pathway Analysis was used for network analysis. Validation of gene expression results was done by qPCR. MAIN RESULTS AND THE ROLE OF CHANCE: Control samples were all staged around 'receptive' as would be clinically expected for LH + 7. Treatment samples were all staged as non-receptive (all but one was classified as 'proliferative' and the last as 'pre-receptive'). Differential gene expression analysis yielded 60 differentially expressed genes between the control and treatment groups. Bioinformatic pathway analysis for differential expression showed inactivation of the progesterone and glucocorticoid receptors, consistent with mifepristone action. LIMITATIONS, REASONS FOR CAUTION: The primary limitations are the relative small number of subjects and the use of a limited gene panel. WIDER IMPLICATIONS OF THE FINDINGS: This study sheds further light on the endometrial receptivity altering effects of mifepristone and on progesterone action. It further shows the capacity of the ERA to identify pharmacologically induced non-receptive endometrium, which expands its possible use clinically and in research. STUDY FUNDING/COMPETING INTEREST(S): C.v.G. and N.R.B. have no conflicts of interest. P.G.L. reports honorarium from University of HK/Shenzhen, other from NIF, India, outside the submitted work. K.G.D. reports consultancy for Bayer AG, Exelgyn, HRA-Pharma, Gedeon Richter, MSD, Mithra, Exeltis and Natural cycles, payment for lectures from Bayer AG, NSD, Ferring, HRA-Pharma, Exelgyn and Exeltis and clinical trials for Bayer AG, MSD, Exeltis, Mithra, HRA-Pharma and Sun Pharma. C.S. has a patent gene expression profile (ERA) issued to Igenomix and is scientific director of Igenomix S.L. M.R., R.N. and J.M.V. are employees of Igenomix S.L. TRIAL REGISTRATION NUMBER: N/A.

Estudio prospectivo aleatorizado sobre la influencia del bloqueo distal del clavo Gamma 3 en el tratamiento de las fracturas pertrocantéreas de fémur / Randomized prospective study on the influence distal block and Gamma 3 nail on the treatment of intertrochanteric fractures of femur

Objetivo. El propósito de este estudio es valorar la necesidad de bloquear distalmente los clavos Gamma 3 (Stryker. Mahwah, New Jersey. USA) en fracturas pertrocantéreas de fémur 31-A1 y 31-A2 de la AO. Material y métodos. Desde junio de 2011 hasta enero de 2013 se recoge una muestra formada por 177 pacientes con fractura pertrocantérea de fémur tratados en nuestro centro mediante osteosíntesis con clavo Gamma 3 estándar. Es un estudio prospectivo y aleatorizado según el año de nacimiento de cada paciente, par con bloqueo o impar sin bloqueo distal del clavo, formando dos grupos de 90 y 87 fracturas respectivamente. Resultados. En los pacientes intervenidos mediante clavo con bloqueo distal se observó una mayor incidencia de complicaciones médicas, una menor incidencia de complicaciones biomecánicas y un aumento en el colapso del foco de fractura en comparación con el grupo control, siendo estas diferencias estadísticamente significativas (p < 0,05). También se observa en el grupo con bloqueo distal un mayor requerimiento transfusional y una mayor tasa de éxitus presentando diferencias estadísticamente significativas (p < 0,05), sin embargo esta significación desaparece al ajustar los resultados por otras características relacionadas con los pacientes. Conclusiones. Basándonos en los resultados hallados en este trabajo, el uso del tornillo de bloqueo distal en los clavos Gamma 3 debe restringirse a fracturas pertrocantéreas inestables tras reducción donde se requiera una estabilidad adicional al clavo intramedular, pudiendo así disminuir el riesgo de complicaciones derivadas de su uso (AU)

Objective. The purpose of this study is to assess the need to lock the Gamma 3 nail (Stryker, Mahwah New Jersey USA) distally for intertrochanteric fractures of femur 31-A1 and 31-A2 of the AO. Material and methods. Details were recorded on a sample of 177 patients with intertrochanteric femoral fractures treated in our hospital by a standard Gamma nail between June 2011 and January 2013. A prospective study was conducted by randomizing patients by year of birth, even numbers with, or odd number without, distal locking, forming two groups of 90 and 87 fractures, respectively. Results. The patients treated with a distal locking nail had an increased incidence of medical complications, a lower incidence of biomechanical complications, and an increase in the fracture collapse compared with the control group, with statistical significance (p < 0 .05). It is also observed in the group with distal locking increased transfusion requirement and a higher death rate, with statistically significant differences (p < 0 .05), but this significance disappears when adjusting for other patient-related characteristics. Conclusions. Based on the results found in this work, the use of distal locking screw in the Gamma 3 nails should be restricted to unstable trochanteric fractures after reduction where additional stability to the intramedullary nail is required, and may decrease the risk of complications from use (AU)

[Randomized prospective study on the influence distal block and Gamma 3 nail on the treatment of intertrochanteric fractures of femur]. / Estudio prospectivo aleatorizado sobre la influencia del bloqueo distal del clavo Gamma 3 en el tratamiento de las fracturas pertrocantéreas de fémur.

OBJECTIVE: The purpose of this study is to assess the need to lock the Gamma 3 nail (Stryker, Mahwah New Jersey USA) distally for intertrochanteric fractures of femur 31-A1 and 31-A2 of the AO. MATERIAL AND METHODS: Details were recorded on a sample of 177 patients with intertrochanteric femoral fractures treated in our hospital by a standard Gamma nail between June 2011 and January 2013. A prospective study was conducted by randomizing patients by year of birth, even numbers with, or odd number without, distal locking, forming two groups of 90 and 87 fractures, respectively. RESULTS: The patients treated with a distal locking nail had an increased incidence of medical complications, a lower incidence of biomechanical complications, and an increase in the fracture collapse compared with the control group, with statistical significance (p < 0.05). It is also observed in the group with distal locking increased transfusion requirement and a higher death rate, with statistically significant differences (p < 0.05), but this significance disappears when adjusting for other patient-related characteristics. CONCLUSIONS: Based on the results found in this work, the use of distal locking screw in the Gamma 3 nails should be restricted to unstable trochanteric fractures after reduction where additional stability to the intramedullary nail is required, and may decrease the risk of complications from use.

[Amebic liver abscess. Three years experience]. / Abscesos amebianos de hígado. Tres años de experiencia.

BACKGROUND: amebic liver abscess is frequently seen in endemic regions, and has a poor prognosis when diagnosis and treatment are inappropriate. AIM: to evaluate and compare our own results; to propose a new classification and therapeutic algorithm. DESIGN: an observational and retrospective study. METHOD: medical records were reviewed for sex, age, signs and symptoms, images, laboratory tests, size, location, treatment, hospital stay, and morbidity-mortality. RESULTS: sixteen patients with amebic liver abscess had been treated -9 were males, mean age was 30.56 years, all abscesses were solitary, 14 were in the right hepatic lobe, average size was 63.25, and 10 were of the collected type according to N Gbesso s classification. Seven patients had a good response to medical treatment, 6 needed percutaneous drainage, and 3 required surgery. Morbidity was 12.5% and mortality 0%. Average hospital stay was 7.68 days. CONCLUSION: our results are similar to those in other published series. The addition of two new groups to N Gbesso s classification provides better therapeutic orientation. We believe that early percutaneous drainage for collected abscesses bigger than 5 cm may improve symptoms and shorten hospital stay.

1,4- and 1,3-dipolar reactivity of alpha-alkoxycarbonylcycloimmonium N-aminides with dipolarophiles: synthesis of new imidazo[2,1-f][1,2,4]triazinium inner salts.

2-Alkoxycarbonylazolium N-aminides are interesting species, as they have the potential to act as efficient 1,4-dipole equivalents when they react with heterocumulenes, such as iso(thio)cyanates and carbodiimides. These reactions give heterobetaines containing the imidazo[2,1-f][1,2,4]triazinium system, in a formal [4 + 2] cyclocondensation process. These N-aminides, however, can also behave as 1,3-dipoles when they react with isocyanates to afford a cycloadduct that, depending on the position of alkoxycarbonyl group, can undergo a reversion process or a ring expansion to the more stable heterobetaine system.