Impact of preimplantation genetic testing for aneuploidies (PGT-A) on first trimester biochemical markers - PAPP-A (placenta-associated plasma protein) and free ß-hCG (human chorionic gonadotropin).

OBJECTIVE: The objective of the study was to study the effect of preimplantation genetic testing for aneuploidies (PGT-A) performed at blastocyst stage on the levels of first trimester biomarkers. METHODS: This is an observational, collaborative, retrospective study. Seven hundred and twenty-eight patients were included in the study. Patients were with singleton pregnancies resulting from either natural conception (NC), or assisted reproductive techniques (ARTs) with PGT-A and frozen embryo transfer (FET) (ART/PGT-A/FET) or after ART without PGT-A and fresh ET (ART/no PGT-A/fresh ET) or FET (ART/no PGT-A/FET), who had first trimester combined screening test between 11 and 14 gestational weeks. They were stratified into four groups: group A (ART/PGT-A/FET) - 143 patients; group B (ART/no PGT-A/FET) - 100 patients; group C (ART/no PGT-A/fresh ET) - 346 patients, and group D (NC) - 139 patients. RESULTS: Statistically significant differences among the examined groups were observed for maternal age, BMI, ethnicity, and parity. The median placenta-associated plasma protein (PAPP-A) was lowest in the group with ART/PGT-A/FET and the highest result was obtained in the group with ART/no PGT-A/FET. Statistically significant difference in the median PAPP-A levels was identified among the examined groups (p = .0186). When a subgroup analysis was performed, a statistically significant difference was observed in the median PAPP-A between ART/PGT-A/FET group versus ART/no PGT-A/FET group (p = .01) and NC versus ART/no PGT-A/FET (p = .01). A similar trend toward statistical significance was noted when comparing NC versus ART/no PGT-A/fresh ET (p = .06). Multivariate analysis elucidated that when age is present in the model, the effect of any method of conception or testing for aneuploidy disappears. The other factors (BMI, ethnicity, and parity) do not influence the levels of PAPP-A. The lowest median free human chorionic gonadotropin (ß-HCG) was recorded in the NC group and the highest result was identified in the group with IVF/PGT-A/FET. No statistically significant difference was observed in the median concentration levels of free ß-hCG among the compared groups (p = .5789) and when subgroup analysis was performed (p>.05). The normality of the distribution of variables was analyzed by the Kolmogorov-Smirnov test and the median PAPP-A and free ßhCG concentration difference by the Wilcoxon rank-sum test with nonparametric ANOVA. CONCLUSIONS: Testing for aneuploidy (PGT-A) and the decision to transfer either fresh or cryopreserved embryos (ET) appear not to affect the levels of first trimester biochemical markers. The findings of the present study should be a baseline for future studies and could be used to improve the antenatal screening counseling for women with ART pregnancies and PGT-A.

Retinal Sensitivity in Patients with Type I Diabetes without Retinopathy or with Minor Retinal Changes.

Objective: To evaluate the ability of short-wavelength automated perimetry (SWAP) for the detection of visual impairment in patients with type I diabetes without retinopathy or with minor retinal vascular changes. Design: Comparative cross-sectional study. Participants: 30 eyes of 30 healthy subjects and 73 eyes of 73 patients with type I diabetes mellitus were studied. Methods: Ophthalmic examination of diabetic patients showed no retinopathy or minimal changes (less than 5 microaneurisms in each eye) with no previous laser treatment. All patients were examined by means of the SWAP 24-2 strategy. Mean Deviation (MD) and Pattern Standard Deviation (PSD) were compared between both groups. Results: There were differences in the clusters of altered points between both groups (p=0.004). SWAP MD was lower in the diabetic group than in the controls (-2.89 dB vs. -0.20 dB, p<0.001). SWAP PSD also differed between both groups (2.50 dB in control group, 3.12 dB in the diabetic group, p=0.003). In the diabetic group, mean period from the onset of diabetes was 12.6±6.7 years and minimal vascular changes were observed in the retina of 18 eyes (24.7%), while 55 had no lesions (75.3%). No differences in SWAP changes were found between patients without and with minimal diabetic retinopathy. Conclusions: Retinal sensitivity assessed by SWAP is depressed in patients with type I diabetes regardless of the presence of retinal vascular changes.

e-Learning strategies in occupational legal medicine based on problem solving through "CASUS" system.

The use of online teaching tools facilitate the incorporation of self-learning methods. With a view to encouraging convergence in teaching tools and methods in Occupational Legal Medicine, an initiative was set up within the classes of Legal and Forensic Medicine at Saragossa University, as part of the EU funded NetWoRM project, which has been led since 1999 by Ludwig-Maximilians-Universität in Munich (Germany). The interest of medical students in Occupational Legal Medicine has so far been low and in addition different aspects complicate the teaching of Occupational Legal Medicine at medical schools: One reason for the low interest is the limited availability of bedside teaching, one of the students' most favourite and effective way to learn. The reason for that is that most medical schools with occupational departments only have outpatient clinics. "Interesting" patients who be need for educational purposes are therefore only available for a limited part of the day. However, in order to recognize and prevent occupational disorders each medical student and physician needs profound clinical knowledge in Occupational Legal Medicine. This project has proven to be highly efficient in permitting the creation and validation of teaching tools which cover and improve the traditional training of the Occupational Legal Medicine programme imparted in the degree of Medicine.

A preliminary study on the incidence of heteroplasmy in mitochondrial DNA from vitreous humour.

Vitreous humour is routinely sampled in Forensic Medicine as several post-mortem analyses can be performed. However, it is not used for DNA analyses probably due to its scarce cellularity. In these samples, in which the study of nuclear DNA is difficult, the analysis of mtDNA is an alternative approach. The aim of this study was to investigate the utility of vitreous humour for forensic identification purposes. Samples were collected during vitrectomy from retinopathy patients, in collection bags with saline solution. Blood samples were also obtained in order to contrast results. Before DNA organic extraction, several centrifugation steps were needed to concentrate the vitreous humour samples. Unlike blood, direct amplification of 400-bp fragments of the hipervariable regions I and II (HVI and HVII) was not successful, possibly due to damage to the DNA strand caused by the surgery conditions (UV radiation, oxidative stress). Therefore, amplification of two overlapping fragments for each control region was performed in vitreous humour. In order to eliminate undesired products, all samples were purified by an enzymatic method. Thereafter, mtDNA fragments were sequenced using dye terminators in a MegaBACE 500 capillary sequencer. Sequences of HVI and HVII of approximately 400 bp were obtained from all samples. The sequences obtained from each patient matched almost perfectly those from blood. In summary, herein we describe for the first time a methodology suitable for the mtDNA analysis of vitreous humour samples.

Aggressions against healthcare workers: an approach to the situation in Spain and the victims psychological effects.

Aggression against healthcare workers is a problem of important consequences which is becoming a focus of research. However, its possible effects on psychological health have not been studied sufficiently in spite of the fact that they may be of importance even in the absence of physical aggression [Winstnaley S, Whittington R. Aggression towards health care staff in a UK general hospital: variation among professions. J Clin Nurs 2004;13:3-10,[1]].

[Retinal toxicity following chloroquine therapy]. / Toxicidad retiniana secundaria a tratamiento con cloroquina.

CASE REPORT: Two cases of bilateral ocular toxicity following chloroquine therapy in two women with systemic lupus erythematosus are presented. DISCUSSION: Retinal toxicity due to chloroquine is an uncommon iatrogenic disease. It is dose-dependent. New cases are still being described despite the application of ophthalmic screening techniques.

Functional and structural measurements in a multifactorial glaucoma risk model.

PURPOSE: To evaluate the relationship between a multifactorial probability risk model of developing glaucomatous visual field defects and the assessment of the retinal nerve fiber layer (RNFL) and short-wavelength automated perimetry (SWAP) in glaucoma suspects. METHODS: 157 eyes of 157 glaucoma suspects were included in the study. The risk of developing glaucomatous defects was assessed by applying a multifactorial model that included intraocular pressure, vertical cup-to-disk ratio, age and family history. Photographs of the RNFL and SWAP were performed. RESULTS: The SWAP was abnormal in 54 cases (34.3%); 21% of abnormal perimetries were obtained in the low-risk group, 37% in the moderate risk group and 56% in the high risk group. The nerve fiber layer evaluation in the different risk stages showed a pattern similar to the results obtained with SWAP. Multiple regression analyses, performed with the four variables included in the model with SWAP and RNFL evaluation, showed the relationship between the risk factors and the presence of glaucomatous damage--evaluated by RNFL and SWAP (p<0001). CONCLUSIONS: The probability model showed a good correlation between the risk scale and the RNFL and SWAP assessment.

Central retinal vein occlusion and HELLP syndrome.

PURPOSE: To present a rare case of central retinal vein occlusion in conjunction with the HELLP syndrome. METHODS: A 30-year-old woman presented in the 28th week of her second pregnancy with severe pre-eclampsia with HELLP syndrome; delivery by caesarean section was recommended. Ten days later, the patient complained of severely decreased visual acuity in her right eye. RESULTS: Ophthalmoscopy revealed a central retinal vein occlusion with venous engorgement and tortuosity, multiple flame hemorrhages, and disc and macular edema. Electroretinography revealed a reduction of b-wave/a-wave ratio. Fluorescein-angiography showed a blockage due to extensive retinal hemorrhages with late mild staining of the walls of veins. The patient presented a spontaneous improvement in visual acuity (0.8 two months after) and a complete resolution of ophthalmoscopic findings. CONCLUSION: Ophthalmic complications are possible during and soon after this syndrome. This is the first description of a patient suffering a central retinal vein occlusion during puerperium after the HELLP syndrome.

Population genetics of the STR loci HUMCSF1PO, HUMF13A01, HUMFES/FPS and D12S391 in Asturias (northern Spain).

In order to use genetic loci in forensic identity testing, some population data are needed. This paper presents a report of allele frequency data for the loci HUMCSF1PO, HUMF13A01, HUMFES/FPS and D12S391 in a population sample from Asturias (northern Spain). No deviation from the Hardy-Weinberg equilibrium was detected in any of the four markers investigated and there was no evidence of association between the alleles of these loci. Statistical analysis was also carried out to obtain some parameters of medico-legal interest and comparative studies were carried out with other populations studied to date for these loci. The Asturian sample does not differ substantially from other Caucasian and Spanish populations.

Anatomic results and complications in a long-term follow-up of pneumatic retinopexy cases.

BACKGROUND: The use of pneumatic retinopexy is controversial because 1) intraocular gas could increase the risk of proliferate vitreoretinopathy and 2) it has been reported to achieve low rates of success after a single procedure. METHODS: A total of 219 consecutive cases of SF6 pneumatic retinopexy were reviewed retrospectively. The follow-up period ranged from 2 to 5 years. RESULTS: A total of 179 (81.73%) of 219 eyes were treated successfully with a single procedure. The success rate increased with reoperations to 98.89%. Preoperative factors related to primary failure or redetachment were aphakia or pseudophakia (P = 0.0058) and breaks localized on horizontal meridia (P = 0.0305). Cryopexy was significantly associated with failure (P = 0.0007). A total of 31.05% of eyes showed early complications. The most frequent incidence was delay in subretinal fluid reabsorption (36.76%), followed by new breaks (26.47%). Late complications were observed in 6.84% of the eyes (six eyes with macular pucker, three with new retinal detachments, three with cataracts, and three with new breaks without detachment). CONCLUSIONS: Failures of pneumatic retinopexy were related to lens status, localization of the break, and cryopexy. The occurrence of early and late complications was similar to that in other procedures.

[Inhibitory effect of alpha tocopherol succinate on fibroblast wound healing]. / Efecto del succinato ácido de alfa-tocoferol sobre los mecanismos de cicatrización fibroblásticos.

PURPOSE: To evaluate and compare the effect of acid tocopherol succinate (ATS) on fibroblast migration and proliferation. METHODS: In vitro wound repair was determined in confluent fibroblast monolayer. Conjunctival fibroblasts were incubated with serum-deprived medium for 24 hours. After this time an artificial wound was made and the cells were incubated with fresh medium plus the doses of ATS to be tested. The cell free area was monitored at 0, 18, 24 and 48 hours. Groups of treatment: Group 1: ethanol 0.1%. Group 2: ATS 25 microM dissolved in ethanol (final concentration 0.1%) Group 3: ATS 50 microM in ethanol 0.1%. Each experiment was carried out in triplicate and repeated 4 times. RESULTS: There were no differences among the groups during the first 24 hours. ATS showed significantly larger cell-free area at 48 hours. There were no signs of cellular toxicity. CONCLUSIONS: 25 microM and 50 microM ATS inhibit fibroblast proliferation without cellular toxicity.

Genetic variation at six STR loci (HUMTH01, HUMTPOX, HUMCSF1PO, HUMF13A01, HUMFES/FPS, HUMVWFA31) in Aragon (north Spain).

The STR loci HUMTH01, HUMPTPOX, HUMCSF1PO, HUMF13A01, HUMFES/FPS and HUMVWFA31 are widely used in forensic casework analyses and population data are necessary to estimate the frequency of a DNA profile. This paper presents the results of a survey aimed to investigate the allele frequency distribution of these loci in an important Spanish population (Aragon, North Spain). Statistical analysis to determine whether allele frequencies were in Hardy-Weinberg Equilibrium was carried out and also to obtain some parameters of medico-legal interest. There was no evidence of association between the alleles of the loci. The Aragonese sample does not differ substantially from other Caucasian populations.

Quantification of intraoperative administration of mitomycin-C in filtering surgery with surgical sponge material.

PURPOSE: To determine the absorption and release of mitomycin-C 0.4 and 0.2 mg/mL from sponge-like specimens of Spongostan film (Ferrosan, Copenhagen, Denmark) and the scleral and conjunctival impregnation in an experimental model of filtering surgery. METHODS: The maximum amount of mitomycin per volume unit that Spongostan is able to absorb was determined physically as the difference between dry weight and soaked weight. Mitomycin-C activity in known volumes of Spongostan after mitomycin-C release in vitro also was determined at 0, 1, 10, and 30 seconds and 1, 3, 5, 10, 15, and 30 minutes. Antibiotic activity of the specimens was evaluated by means of bioassay. Millimeters of inhibition of bacterial growth were related to microg of mitomycin activity according to a reference curve obtained from known amounts of mitomycin-C. Finally, 10 eyes of 10 rabbits underwent filtering surgery with intraoperative application of mitomycin by means of the Spongostan film. The Spongostan implants then were removed and tested for mitomycin activity. Scleral and conjunctival specimens were obtained for bioassay. RESULTS: The maximum capacity of 25 mm2 x 0.5 mm thick Spongostan films saturated in 0.4 and 0.2 mg/mL solutions of mitomycin-C were 8.49 microg and 4.23 microg, respectively. Biologic activity (bioassay determination) was 8.24 microg and 4.19 microg of mitomycin-C, respectively. In vitro release of mitomycin was gradual until 30 minutes. In vivo mitomycin release from Spongostan after 5 minutes was 6.91 microg. Impregnation with the antimitotic was better in conjunctiva than sclera. CONCLUSION: Bioassay permits quantification of mitomycin-C activity. The release from sponge specimens is gradual, and impregnation was better in conjunctiva than sclera.

Fluorescence-based amplification of the STR loci D18S535, D1S1656 and D12S391 in a population sample from Aragon (north Spain).

Population data were generated for the STR loci D18S535, D1S1656 and D12S391 in a population sample of unrelated healthy individuals born and living in Aragon (North Spain). The three loci were amplified using a fluorescence-based PCR method and were typed automatically. No deviation from Hardy-Weinberg expectations were observed. The three loci proved to be highly discriminating and valuable polymorphisms for forensic analyses.

Short-wavelength automated perimetry and retinal nerve fiber layer evaluation in suspected cases of glaucoma.

OBJECTIVES: To determine if short-wavelength automated perimetry (SWAP) provides evidence that indicates early functional losses in ocular hypertensive subjects and to establish a direct comparison with early structural abnormalities in the retinal nerve fiber layer (RNFL). METHODS: A total of 160 eyes belonging to 83 patients with ocular hypertension (intraocular pressure >21 mm Hg and normal results on standard automated perimetry evaluation), on which a SWAP and RNFL study were performed, were examined. One hundred twenty-eight age-matched subjects without ocular hypertension were evaluated to establish the 95% and 99% confidence intervals at each of the 76 exploration points of the SWAP test. RESULTS: The RNFL study results were normal in 83 cases (51.8%) and pathologic in 77 cases (48.1%). The SWAP results were pathologic in 57 cases (35.6%). Significant differences (P<.001) were observed when comparing the distribution of normal and pathologic SWAP results among the types of defects in the RNFL (focal wedge, diffuse atrophy, and mixed atrophy). CONCLUSIONS: Short-wavelength automated perimetry is a useful test for the early detection of visual field losses. It is more sensitive than standard automated perimetry and provides a high association with RNFL assessment, which has proved capable of detecting signs of glaucomatous damage several years before the onset of the typical visual field defects.

PM and D1S80 loci gene frequencies in the Zaragoza population of northern Spain.

LDLR, GYPA, HBGG, D7S8, GC (PM loci) and D1S80 are widely used in forensic casework analyses and population data are required to estimate the frequency of a DNA profile. This paper presents the results of a survey aimed at investigating the allele and genotype frequency distribution of these loci in an important Spanish population (Zaragoza, North Spain). Statistical analysis to determine whether allele frequencies were in Hardy-Weinberg equilibrium was carried out as well as to obtain some parameters of medicolegal interest. There was no evidence of association between the alleles of the loci. The Zaragoza sample does not differ substantially from other Caucasian populations.

Choroidal metastasis of mixed carcinoma of the parotid gland.

BACKGROUND: A case of mixed carcinoma of the parotid gland (an epidermoid carcinoma located in a pleomorphic adenoma) metastatic to the choroid is presented. The histopathology of the tumor is discussed. METHODS: A 65-year-old man was admitted complaining of blurred vision in his right eye for 1 day. He underwent parotidectomy for mixed carcinoma of the parotid gland (an epidermoid carcinoma located in a pleomorphic adenoma that was completely excised) 6 months before. Funduscopic examination showed a nasal retinal detachment, with gray-whitish, minimally elevated nodular choroidal lesions. Fluorescein angiography and contact B-scan ultrasonography confirmed the presence of an underlying mass. RESULTS: The right eye was enucleated and an epidermoid infiltrating carcinoma was identified. CONCLUSIONS: Metastatic tumors are the most common intraocular malignancies, and the choroid is by far the most common location for intraocular metastases. There are few cases reported of parotid tumors metastatic to the orbit. To the best of our knowledge, no histological examination of an ocular metastatic mixed carcinoma of the parotid gland has yet been reported.

Distribution of types for six PCR-based loci; LDLR, GYPA, HBGG, D7S8, GC and HLA-DQA1 in central Pyrenees and Teruel (Spain).

The PCR-based DNA loci LDLR, GYPA, HBGG, D7S8, GC and HLA DQA1 are widely used in forensic casework analyses. Population data on the distribution of allele frequencies are desired to estimate the rarity of a DNA profile. We studied the allele distributions at these forensically important DNA markers in two Spanish populations (Central Pyrenees and Teruel). Results were in agreement with Hardy-Weinberg expectations. Furthermore, there was little evidence for departures from expectation of independence between loci within the two sample populations. Tests for homogeneity were carried out between the two Spanish populations and a U.S. Caucasian population.

Frequencies of the five PCR-based genetic markers LDLR, GYPA, HBGG, D7S8 and GC in the population of Asturias (North Spain).

Allele and genotype frequencies of the loci LDLR, GYPA, HBGG, D7S8 and GC (PM loci) were investigated in a population sample of 215 unrelated individuals from Asturias (North Spain). Multiplex amplification and simultaneous typing of the five loci was carried out using the polymarker PCR amplification and typing kit. All loci met Hardy-Weinberg expectations. The Asturian sample does not differ significantly from other Caucasians, but significant differences were observed between this population and SW Hispanic, Afro-american and Korean populations.

Bilateral optic neuritis in occupational exposure to trichloroethylene.

Central nervous system depression is the predominant response to acute trichloroethylene exposure. Cranial or peripheral nerves are also affected. The paper is a case report of a bilateral retrobulbar optic neuritis caused by occupational trichloroethylene poisoning. The urinary concentration of trichloroacetic acid one week after the onset of symptoms was positive. Visual fields and visual evoked potentials recovered eight months later. Other neurological diseases, such as multiple sclerosis were excluded.