Genetics of anophthalmia and microphthalmia. Part 1: Non-syndromic anophthalmia/microphthalmia.

Eye formation is the result of coordinated induction and differentiation processes during embryogenesis. Disruption of any one of these events has the potential to cause ocular growth and structural defects, such as anophthalmia and microphthalmia (A/M). A/M can be isolated or occur with systemic anomalies, when they may form part of a recognizable syndrome. Their etiology includes genetic and environmental factors; several hundred genes involved in ocular development have been identified in humans or animal models. In humans, around 30 genes have been repeatedly implicated in A/M families, although many other genes have been described in single cases or families, and some genetic syndromes include eye anomalies occasionally as part of a wider phenotype. As a result of this broad genetic heterogeneity, with one or two notable exceptions, each gene explains only a small percentage of cases. Given the overlapping phenotypes, these genes can be most efficiently tested on panels or by whole exome/genome sequencing for the purposes of molecular diagnosis. However, despite whole exome/genome testing more than half of patients currently remain without a molecular diagnosis. The proportion of undiagnosed cases is even higher in those individuals with unilateral or milder phenotypes. Furthermore, even when a strong gene candidate is available for a patient, issues of incomplete penetrance and germinal mosaicism make diagnosis and genetic counseling challenging. In this review, we present the main genes implicated in non-syndromic human A/M phenotypes and, for practical purposes, classify them according to the most frequent or predominant phenotype each is associated with. Our intention is that this will allow clinicians to rank and prioritize their molecular analyses and interpretations according to the phenotypes of their patients.

4q25 microdeletion encompassing PITX2: A patient presenting with tetralogy of Fallot and dental anomalies without ocular features.

Axenfeld-Rieger syndrome (ARS) is a heterogeneous clinical entity transmitted in an autosomal dominant manner. The main feature, Axenfeld-Rieger Anomaly (ARA), is a malformation of the anterior segment of the eye that can lead to glaucoma and impair vision. Extra-ocular defects have also been reported. Point mutations of FOXC1 and PITX2 are responsible for about 40% of the ARS cases. We describe the phenotype of a patient carrying a deletion encompassing the 4q25 locus containing PITX2 gene. This child presented with a congenital heart defect (Tetralogy of Fallot, TOF) and no signs of ARA. He is the first patient described with TOF and a complete deletion of PITX2 (arr[GRCh37]4q25(110843057-112077858)x1, involving PITX2, EGF, ELOVL6 and ENPEP) inherited from his ARS affected mother. In addition, to our knowledge, he is the first patient reported with no ocular phenotype associated with haploinsufficiency of PITX2. We compare the phenotype and genotype of this patient to those of five other patients carrying 4q25 deletions. Two of these patients were enrolled in the university hospital in Toulouse, while the other three were already documented in DECIPHER. This comparative study suggests both an incomplete penetrance of the ocular malformation pattern in patients carrying PITX2 deletions and a putative association between TOF and PITX2 haploinsufficiency.

Evolution of the corpus luteum volume determined ultrasonographically and its relation to the plasma progesterone concentration after artificial insemination in pregnant and non-pregnant dairy cows.

The aim of this study was to assess the relationship of the evolution of the corpus luteum (CL) volume that was determined ultrasonographically with the pregnancy status in lactating dairy cows during early pregnancy. Ultrasound examinations were carried out on 76 cows following artificial insemination (AI). Plasma concentrations of progesterone were determined from blood samples collected at each ultrasound examination. Conception was confirmed by ultrasonography on day 30 after AI. Around day 14 post-insemination (p.i.), the CL volume tended to decrease in pregnant and non-pregnant cows, and, after day 19 p.i., both groups differed significantly, indicating the luteal regression in non-pregnant cows. Reaching signification on day 20. The diminution in CL volume was also reflected in the plasma progesterone concentration. However, the patterns of CL volume, estimated by ultrasonography, differed more evidently and earlier between both groups (around 1 week p.i., at day 9 p.i. P < 0.05, whereas progesterone started to differ around 2 weeks p.i., at day 14 p.i, P < 0.05). These results indicate that the estimation of the CL volume by ultrasonography could be useful for assessing the presence of a functional CL.

Diastolic Dysfunction in Liver Cirrhosis: Prognostic Predictor in Liver Transplantation?

BACKGROUND: Patients with liver cirrhosis may develop cirrhotic cardiomyopathy (CC), characterized by blunted contractile responsiveness to stress, diastolic dysfunction (DD), and electrophysiological abnormalities. It may adversely affect the long-term prognosis of these patients. METHODS: We conducted a retrospective analysis of patients undergoing liver transplantation (LT) for cirrhosis from January 2012 to June 2015. We analyzed demographic characteristics, the etiology of cirrhosis, Child-Pugh and Model for End-Stage Liver Disease (MELD) scores, the corrected QT (QTc) interval in the preoperative period, diastolic and systolic dysfunction, mortality and survival, and duration of mechanical ventilation and vasopressor support in the post-LT period. These variables were compared with diastolic dysfunction and prolongation of QTc, with the use of chi-square, Fisher, and Mann-Whitney U tests. RESULTS: The study included 106 patients, 80.2% male and overall average age 54.83 years. The median MELD score was 16, and Child-Pugh class C in 55.4%. Prolonged QTc interval before LT was present in 19% and DD in 35.8% of patients. QTc before LT or DD did not vary significantly with MELD or Child-Pugh score. CONCLUSIONS: The patients in the pre-LT period presented with a significant incidence of DD, which can predispose them to adverse cardiac events. The presence of DD correlates with mortality after LT in patients with hepatic cirrhosis.

Liver retransplantation in patients with acquired familial amyloid polyneuropathy: a Portuguese center experience.

In 1995 Furtado et al performed the first domino transplantation using a donor liver with familial amyloid polyneuropathy (FAP), thereby increasing the pool of donors. Our experience showed that the onset of FAP symptoms occurs earlier in some patients. Patients with FAP acquired by transplantation are candidates for liver retransplantation to minimize the progression of symptoms. Liver retransplantation is considered to be a high-risk procedure and has lower survival compared with the first transplantation. We evaluated the risk of liver retransplantation in patients with acquired FAP. We did a retrospective analysis of these patients based on the records of perioperative data. From 1995 to 2004 we carried out 81 domino transplantations, of which 10 were submitted to liver retransplantation because of acquired FAP. The better outcomes in this group lead us to think that the liver retransplantation in patients with acquired FAP is not associated with the same risks of liver retransplantation in candidates with graft failure.

Evaluation of operative risk in de novo familial amyloid polyneuropathy retransplantation.

Familial amyloid polyneuropathy (FAP) is the most common hereditary amyloidosis, characterized by progressive peripheral sensory and motor neuropathy. The livers of patients with FAP are used in domino liver transplantation in selected cases to increase the number of grafts available. In our department 10 patients underwent liver retransplantation (ReLTx) in the absence of liver dysfunction by de novo FAP after domino liver transplantation. Our aim was to compare the differences in the consumption of blood products and intraoperative hemodynamic support among patients with FAP undergoing liver transplantation (LTx) and patients with de novo FAP undergoing ReLTx in the same time frame. The anesthetic records of all patients who underwent LTx for FAP and ReLTx for de novo FAP were analyzed, from January 2009 to May 2014. Patients were divided into 2 groups: group 1 patients with FAP, and group 2 patients with de novo FAP. Statistical differences in the value of preoperative creatinine were found. Hemoglobin levels, preoperative international normalized ratio (INR), use of blood products, aminergic support, and surgical time showed no statistical difference. Major bleeding rates would be expected in patients undergoing ReLTx. Changes in renal function, chronic immunosuppressive therapy, and age may contribute to the increase in intraoperative complications. We did not find statistically significant differences, leading us to the conclusion that de novo FAP does not seem to be a predictor of perioperative risk.

Abordaje histeroscópico de la trompa de Falopio (faloposcopia). Esterilización transcervical tubárica / Hysteroscopy of the fallopian tube (falloscopy). Transcervical tubal sterilization

El gran desarrollo de diferentes técnicas de reproducción asistida, así como la mejoría en los sistemas ópticos han permitido a muchos endoscopistas la visualización correcta de los ostium tubáricos, su cateterización y la visualización de la luz tubárica en toda su extensión. Muchos años han pasado desde los primeros intentos, obtenindo conocimientos suficientes como para eliminar oclusiones tubáricas, transferir gametos o embriones y colocar dispositivos intratubáricos como alternativa a los métodos quirúrgicos de esterilización femenina

The great improvement of many assisted reproduction techniques and the visual systems, have made possible the correct view of tubal ostia, the current catheterization and the evaluation of every tube segment. Many years have passed since initial attempts and a lot of information was obtained to make possible the elimination of tubal occlusions, the transfer of gametes and embryos, and the placement of intratubal devices as an alternative to surgical methods of female sterilization

Principios básicos en histeroscopia diagnóstica / Basic principles of diagnostic hysteroscopy

La histeroscopia diagnóstica es una técnica sencilla, segura y que proporciona una gran cantidad de información sobre la patología endouterina. No es extraño pues que su extensión se haya producido de una manera muy rápida, arrinconando a otras técnicas más antiguas y más agresivas, como el legrado

Diagnostic hysteroscopy is a simple and safety method, which give us a very important information about endouterine diseases. So, the fast developement of this technique has made that traditional curettage, has been forgotten

gfp-Tagged cells as a useful tool to study the survival of Escherichia coli in the presence of the river microbial community.

We have used an Escherichia coli strain DH5a containing pGreenTIR to study the survival of this bacterium in river water. As green fluorescence was maintained throughout survival both in dark and illuminated conditions, gfp-tagged E. coli cells were clearly distinguished from the microbial community of the river Butrón. gfp-tagged E. coli cells were monitored to estimate total density as well as the density of the culturable and viable (active electron transport system, CTC+) cells. Our results indicate that autochthonous bacteria and introduced E. coli are predated by flagellates. The autochthonous bacterial community behaves as predation-escaping prey, showing a tendency to cellular miniaturization and so maintaining the density of the population. In contrast, introduced E. coli behaves as predation-non-escaping prey, so E. coli was eliminated from the system. When comparing the elimination by predation of heat-treated and non-heated gfp-tagged E. coli cells we deduce that the flagellates do not discriminate between live and heat-treated cells. Finally, in the presence of the river microbial community, the E. coli cells appeared to be ingested before cellular deterioration could occur. Thus predation reduces the quantitative importance of the viable but nonculturable (VBNC) population of E. coli in the aquatic systems.

Participation of oxygen and role of exogenous and endogenous sensitizers in the photoinactivation of Escherichia coli by photosynthetically active radiation, UV-A and UV-B.

We studied the mechanisms by which photosynthetically active radiation (PAR) and ultraviolet (UV-A and UV-B) radiation damage Escherichia coli suspended in water. The roles played by oxygen and exogenous and endogenous sensitizers were analyzed by monitoring changes in the physiological state of irradiated cells. Impairment of the cellular functions was more severe in the case of UV radiations. Radiation caused cellular damage in the absence of oxygen. PAR, UV-A, and UV-B radiation induced photobiological and photodynamic reactions mediated by endogenous sensitizers, which significantly shortened the T90 (time needed to reduce a cellular parameter by 90%) based on the growth ability of the cells. In addition, when exogenous sensitizers were present, the photodynamic reactions also had a negative effect on the operation of the electron transport chains. The presence of oxygen might enhance photoinactivation, affecting both the growth ability and the electron transport chains. Endogenous sensitizers were responsible for the noxious action of oxygen. The presence of dissolved organic material played a protective role against the oxygen by absorbing the incident radiation, thereby reducing the energy that reached the endogenous sensitizers.

Influence of boron supplementation on vertebral and femoral bone mass in rats on strenuous treadmill exercise. A morphometric, densitometric, and histomorphometric study.

We studied the effect of boron supplement on experimental osteopenia caused by strenuous exercise in 93- d-old female Wistar rats. A control group of 15 rats was not manipulated. The exercise group of 30 rats was divided into 2 groups of 15 rats each, one that was fed a diet supplemented with 50 mg/kg of boron in the form of Na(2)B(4)O(7), and other that, did not receive a boron supplement. The length and weight were determined in the femur and fifth lumbar vertebra and the bone mineral content and density were assessed through densitometry, and trabecular bone volume, trabecular number, trabecular thickness, and trabecular separation with histomorphometry. The femur length and weight, and vertebra weight, and femur and vertebra bone mineral content and density were significantly lower and the trabecular separation was higher in the exercise group than in the others (p < 0.005 in all). The femur weight, bone mineral content and density, trabecular bone volume and trabecular thickness, were significantly higher in the exercise plus boron group (p < 0.005 to 0.0001). It was concluded that boron preserves bone mass in rats that have been exposed to intense exercise.

Effect of uterine horn ligation on bone mass: an experimental study in rats.

OBJECTIVE: After tubal ligation, normal bone mass in the presence of gonadal deficit has been reported. These incongruent results motivated us to examine the topic. STUDY DESIGN: Bone mass was assessed by densitometry and ultrasonography 60 days after surgery on 100-day-old female Wistar rats. Fifteen Wistar rats with uterine horn ligation (TL) were compared with 15 unoperated, 15 with a sham uterine horn ligation (Sham-TL), and 15 ovariectomized (OVX), using ANOVA and a correlation test to determine the relations between results. RESULTS: Femoral and vertebral bone mass were significantly lower in the OVX y TL groups than in unoperated and controls groups (P<0.0001). CONCLUSIONS: Our study revealed significantly lower axial and peripheral bone mass in rats with uterine horn ligation.

Behavior of phalangeal bone ultrasound in normal women with relation to gonadal status and body mass index.

The behavior of phalangeal bone ultrasound was studied, measured by amplitude-dependent speed of sound (Ad-SOS) in meters per second, in 324 normal women (mean age 48.9 +/- 13.7 years) classified by gonadal status (premenopausal, perimenopausal and postmenopausal) and body mass index (BMI, thin, normal, overweight and obese). Ad-SOS differed significantly with gonadal status and BMI (p<0.0001 for all). In the overall group of women, Ad-SOS correlated negatively with age (r=-0.84, p<0.0001), weight (r=-0.16, p<0.005), BMI (r=-0.27, p<0.0001), and tartrate-resistant acid phosphatase concentration (TRAP) (r=-0.35, p<0.0001). The negative correlation remained significant in the groups separated by gonadal status, but to a lesser extent. After adjusting for confounding variables such as age and weight, Ad-SOS was dependent on age (but not on weight or BMI) in the overall group of women and in the gonadal status groups. In conclusion, Ad-SOS values differed significantly with gonadal status and BMI, and correlated negatively with TRAP. The plot of Ad-SOS against age differed significantly with gonadal status as well as BMI.

Do ultrasound measurements reflect bone microarchitecture rather than bone mass?: An in vitro study of the rat femur with the use of ultrasound, densitometry, and histomorphometry.

RATIONALE AND OBJECTIVES: In an experimental study in 40 rat femurs, the authors correlated the amplitude-dependent speed of bone ultrasound (Ad-SOS) with the bone mineral content and density and with the bone trabecular connectivity: trabecular bone volume, trabecular number, trabecular thickness, and trabecular separation to evaluate and compare the usefulness of the Ad-SOS to determine bone quantity and/or quality. METHODS: Bone mineral content and density were determined with dual-energy x-ray absorptiometry. Trabecular connectivity was determined with histomorphometric techniques. RESULTS: There was a strong correlation between the Ad-SOS and the other parameters studied, with a particularly high positive correlation with trabecular bone volume and trabecular thickness, and an inverse correlation with trabecular separation. The correlation was weaker with the bone mineral content and bone mineral density and with the trabecular number. For the trabecular separation, the correlation was significant in all cases, but it was negative. CONCLUSIONS: Bone ultrasound, in this case Ad-SOS, defines the quality of the bone in terms of trabecular architecture rather than bone density; however, this conclusion is valid only for the rat femur model that the authors used.

Effects of sodium bicarbonate supplementation on axial and peripheral bone mass in rats on strenuous treadmill training exercise.

We observed the effects of sodium bicarbonate supplement on bone mass in rats on strenuous treadmill training. Sixty female Wistar rats (93-days-old; mean initial weight 261 +/- 16 g) were studied. One group of 15 rats was killed at the beginning of the experiments (basal control group), while another group of 15 rats was not manipulated (Exer-NaB-). Another group of 15 rats was exercised but did not receive sodium bicarbonate (Exer+NaB-), while the final group of 15 rats exercised and received sodium bicarbonate (Exer+NaB+) at a dose of 0.05 mg/kg/day, administered by esophageal catheter on exercise days. These rats were killed at the end of 11 weeks. Femoral and vertebral length, weight, and bone mineral content (BMC) and density (BMD) were measured. According to ANOVA with the Tukey-Kramer test, femur length and weight, vertebral weight, femur BMC and BMD, vertebral BMC and BMD and the ratio between femur and vertebral BMC and final body weight, and plasma bicarbonate were lower in the basal control and Exer+NaB- groups than in the two other groups (P < 0.005-0.0001). Overall, there was a positive correlation between femur and vertebral BMC and femur BMC and length (P < 0.0001 for all). Only in the Exer+NaB- group was there a positive association between plasma bicarbonate levels and femur length (r = 0.78; P < 0.0005). Our study demonstrates the adverse effects of strenuous exercise on bone, and the usefulness of sodium bicarbonate supplements in preventing and minimized these effects.

Valoración de beta-HCG, estradiol y progesterona en mujeres portadoras de DIU / Evaluation of beta-human chorionic gonadotrophin, estradiol and progesterone in women with intrauterine devices

Objetivo: Se pretende investigar una posible alteración del ciclo ovárico en mujeres portadoras de dispositivo intrauterino (DIU), al mismo tiempo que se intenta averiguar mediante el estudio de la beta-HCG si estos dispositivos son abortivos.Material y método: Se estudian tres grupos. El primero compuesto por 103 mujeres portadoras de DIU, el segundo por 300 mujeres que utilizaban un método de barrera, y el tercero por 300 mujeres que deseaban gestación y no utilizaban método anticonceptivo. En todos ellos se determinaron los valores de progesterona y estradiol en la fase lútea, así como la beta-HCG sérica.Resultados: En el grupo I no se detectó ninguna beta-HCG positiva, frente a un caso en el grupo II y 57 en el grupo III. No se encontraron diferencias significativas respecto a la determinación del estradiol y de la progesterona.Conclusiones: De los hallazgos de nuestro estudio no podemos concluir que la acción anticonceptiva del DIU se produzca durante el ciclo ovárico, no existiendo tampoco fecundación, por lo que se descarta también la acción abortiva. (AU)

The effect of supplemental copper on osteopenia induced by ovariectomy in rats.

OBJECTIVE: The effect of a copper supplement on preventing bone mass loss induced by ovariectomy in rats was investigated. DESIGN: Three groups of fifteen 100-day-old female Wistar rats, each with a mean initial weight of approximately 260 g per animal, were selected for a 30-day experiment. One group of 15 ovariectomized rats was fed a diet supplemented with 15 mg of copper per kilogram of feed. The other two groups: 15 ovariectomized and 15 Sham- ovariectomized rats did not receive the supplement. Morphometric (weight and length) and densitometric studies with dual-energy x-ray absorptiometry were performed on the whole femur and the fifth lumbar vertebra of each animal at the end of the 30-day period. RESULTS: The ovariectomized rat group fed a diet supplemented with copper did not show the bone mass loss at the axial (fifth lumbar vertebra) or peripheral (femur) level that was evidenced in the ovariectomized group. CONCLUSIONS: The results of the measurement of axial and peripheral bones show that a supplement of copper may have a potential therapeutic application in the treatment and prevention of involutional osteoporosis.

Gynecological factors and body mass index as determinants of bone mass in normal postmenopausal women. A study with peripheral quantitative computed tomography (pQCT).

OBJECTIVE: This paper studied the influence of several gynecological factors (years since menopause (YSM), age at menarche and gynecological age or reproductive life) simultaneously with anthropometric factors as determinants of bone mass in 189 healthy postmenopausal women. METHODS: Bone mass was determined by peripheral quantitative computed tomography. RESULTS: An overall evaluation showed that YSM correlated negatively with trabecular and cortical bone density (BMDTrab and BMDCorti) (P<0.05 in both cases). Age at menarche correlated negatively with BMDCorti (P<0.05) and gynecological age correlated positively with BMDTrab (P<0.05). Classifying the women according to their body mass index (BMI), the YSM correlation persisted in those subjects whose BMI was >25 kg/m(2), and in age at menarche and gynecological age of women whose BMI was <25 kg/m(2) (P<0.05). After separating women according to their age at menarche, their gynecological age and BMI, the only significant difference that persisted was in BMDTrab which was lower in the group with gynecological age <33 years, with a BMI <25 kg/m(2) (P=0.020). Parity and smoking had no impact on our results. By multiple regression, with BMD as the dependent variable and the gynecological factors as independent variables, we only observed significance between YSM and BMDCorti (P<0.005). The same was observed after separating women according to their BMI in the >25 kg/m(2) group (P<0.05). CONCLUSIONS: Our data stress the importance of YSM on BMDTrab and BMDCorti, of age at menarche on BMDCorti and of gynecological age on BMDTrab. However, YSM is the gynecological factor that mainly determines BMD. The differences observed between measurements taken with pQCT and other methods commonly used to estimate bone mass indicate that results obtained with one technique cannot be extrapolated to other methods.

Calcitonin versus clodronate in the prevention of ovariectomy-induced osteopenia in rats.

The effects of salmon calcitonin and clodronate were compared in ovariectomised rats. Sixty female Wistar rats ( 260 g in weight) were fed the same diet and had the same living conditions. The rats were divided into the following groups: 15 rats with sham ovariectomy and no drug treatment (Sham-OVX); 45 rats with bilateral ovariectomy subdivided into 15 rats not receiving drug treatment (OVX group), 15 rats treated with subcutaneous salmon calcitonin, 2 U/kg/day every 2 days (OVX + CT group) and 15 rats treated with subcutaneous clodronate, 5 mg/kg/day every 2 days (OVX + Cl group). Sixty days after surgery, the rats were sacrificed and their femurs and fifth lumbar vertebrae were dissected and cleaned of soft tissue. Femur length, vertebral height, and bone mineral content and bone mineral density of the femur and fifth lumbar vertebra by dual-energy X-ray absorptiometry were measured. Calcitonin had a significant and stronger effect in preventing ovariectomy-induced osteopenia in the femur (OVX + CT vs OVX groups, p < 0.0001); both calcitonin and clodronate had a significant effect on the fifth lumbar vertebra, which was greater in the calcitonin group (OVX + CT vs OVX + Cl groups, p<0.005). These findings indicate that calcitonin has a protective effect on both the axial (trabecular bone) and peripheral (cortical bone) skeletons, but clodronate only has a protective effect on the axial skeleton.