The role of polar phytocomplexes on anticonvulsant effects of leaf extracts of Lippia alba (Mill.) N.E. Brown chemotypes

Objectives The purpose of the present work was to characterize the pharmacological profile of different L. alba chemotypes and to correlate the obtained data to the presence of chemical constituents detected by phytochemical analysis. Methods Essential oils from each L. alba chemotype (LP1—LP7) were characterized by gas chromatography–mass spectrometry (GC‐MS) and extracted non‐volatile compounds were analysed by HPLC and GC‐MS. The anticonvulsant actions of the extracted compounds were studied in pentylenetetrazole‐induced clonic seizures in mice and their effect on motor coordination was studied using the rota‐rod test in rats. The synaptosomes and synaptic membranes of the rats were examined for the influence of LP3 chemotype extract on GABA uptake and binding experiments. Key findings Behavioural parameters encompassed by the pentylenetetrazole test indicated that 80% ethanolic extracts of LP1, LP3 and LP6 L. alba chemotypes were more effective as anticonvulsant agents. Neurochemical assays using synaptosomes and synaptic membranes showed that L. alba LP3 chemotype 80% ethanolic extract inhibited GABA uptake and GABA binding in a dose‐dependent manner. HPLC analysis showed that LP1, LP3 and LP6 80% ethanolic extracts presented a similar profile of constituents, differing from those seen in LP2, LP4, LP5 and LP7 80% ethanolic extracts, which exhibited no anticonvulsant effect. GC‐MS analysis indicated the occurrence of phenylpropanoids in methanolic fractions obtained from LP1, LP3 and LP6 80% ethanolic extracts and also the accumulation of inositol and flavonoids in hydroalcoholic fractions. Conclusions Our results suggest that the anticonvulsant properties shown by L. alba might be correlated to the presence of a complex of non‐volatile substances (phenylpropanoids, flavonoids and/or inositols), and also to the volatile terpenoids (β‐myrcene, citral, limonene and carvone), which have been previously validated as anticonvulsants.     

Evidence of a high proportion of premature unbalanced separation of sister chromatids in the first polar bodies of women of advanced age

BACKGROUND: Maternal ageing is the only aetiological factor unequivocally linked to aneuploidy. Two mechanisms seem to explain these abnormalities in oocytes: non-disjunction and premature unbalanced separation of sister chromatids (PSSC). Previous studies of unfertilized oocytes argue for a major role of PSSC in the aetiology of aneuploidy for women of advanced age, but in vitro ageing of the oocytes could influence the results. METHODS: Owing to the high prevalence of aneuploidy in women of advanced age, chromosomal screening of the first polar body just before ICSI was offered to women (from 38 years of age) included in an assisted reproduction programme. RESULTS: Among 141 oocytes from 29 women (mean age 40 years and 2 months), 43 (30.5%) were abnormal. Sixty-five abnormalities were found and PSSC was involved in 80% of cases. CONCLUSION: These results are in accordance with previous studies and confirm, in 'fresh' oocytes, the major role of PSSC in the aetiology of aneuploidy in women of advanced age.     

Polar spongioblastoma: A high‐grade glioma that does not contain the IDH1 mutation or 1p/19q LOH

We report a case of an unusual glioma termed “primitive polar spongioblastoma” that displayed characteristic palisading tumor cells at the light microscopic level. The patient was a 52‐year‐old woman who underwent subtotal removal for a left frontotemporal tumor. The palisading pattern was present throughout the tumor. Several glial markers were revealed by immunohistochemical examination, but no neuronal markers were observed. Genetic studies showed O‐6‐methylguanine‐DNA methyltransferase (MGMT) methylation, wild type IDH1, and the absence of 1p/19q loss of heterozygosity (LOH) in the tumor genes. Based on histological and genetic features, this tumor might not be suited to any of neuroepithelial tumor in the recent WHO classification. We consider that cases such as this should be temporarily set under a separate heading and be entrusted to future investigation after more cases have been accumulated.     

张力带固定与下极切除治疗髌骨下极骨折

目的：对比张力带固定与下极切除治疗髌骨下极骨折的疗效。方法：手术治疗髌骨下极新鲜骨折８９例,其中张力带固定４９例,下极切除４０例,术后平均随访１１个月,并根据陆氏标准评价疗效。结果;张力带固定组４５例达到优良标准,下极切除组２８例达到优良标准,张力带固定组疗效显著优于下极切除组。     

Prevention of Age-Related Aneuploidies by Polar Body Testing of Oocytes

Purpose: We previously demonstrated that aneuploidy-free oocytes may be preselected by testing the first and second polar bodies removed from oocytes following their maturation and fertilization. The present paper describes the results of the application of the method in 659 in vitro fertilization cycles from patients of advanced maternal age. Methods: Using micromanipulation techniques, 3943 oocytes were tested by polar body sampling and fluorescent in situ hybridization analysis using specific probes for chromosomes 13, 18, and 21. Results: Fluorescent in situ hybridization results were available for 3217 (81.6%) of 3943 oocytes studied, of which 1388 (43.1%) had aneuploidies; 35.7% of the aneuploidies were of first meiotic division origin, and 26.1% of second meiotic division origin. Most errors in the first meiotic division were represented by chromatid malsegregation. The transfer of embryos deriving from 1558 of 1829 aneuploidy-free oocytes in 614 treatment cycles resulted in 131 clinical pregnancies and 88 healthy children born after confirmation of the polar body diagnosis. Conclusions: Polar body testing of oocytes provides an accurate and reliable approach for prevention of age-related aneuploidies in in vitro fertilization patients of advanced maternal age.     

Second polar body extrusion is highly predictive for oocyte fertilization as soon as 3 hr after intracytoplasmic sperm injection (ICSI)

Objective Our objective was to evaluate the time course and the predictive value of the extrusion of the second polar body after intracytoplasmic injection (ICSI) related to the fertilization rate, embryo cleavage and quality.Setting The setting was the in vitro fertilization program of a university hospital.Patients Twenty-one patients were treated with intracytoplasmic single sperm injection either for fertilization failure in IVF, low fertilization in IVF (<5%), or severe male factors.Design One hundred thirty-five of 205 metaphase 2 oocytes treated with intracytoplasmic single sperm injection were observed 1, 2, and 3 hr after the assisted fertilization procedure. Extrusion of the second polar body was recorded. For each of these oocytes, fertilization was noted 18 hr after ICSI and cleavage and embryo quality were assessed 24 hr later. The 70 remaining oocytes were used to assess a possible negative effect of repeated exposure to light microscopy.Results The extrusion of the second polar body 3 hr after injection was an observation with a sensitivity of 0.87, a specificity of 0.58, and a high positive predictive value (0.90) toward oocyte fertilization. Twenty-nine and four-tenths percent of the oocytes extruded a second polar body within the first hour, 56.6% within the first 2 hr, and 78.3% had a second polar body 3 hr after injection. This time course was related neither to the speed of embryo cleavage nor to the embryo quality. Fertilization, cleavage, and embryo quality were not affected by repeated observation as deduced from comparison with the control group and confirmed by a high pregnancy (62% per oocyte retrieval) and implantation rate (22% per replaced embryo).Conclusion Oocytes can be checked, in all safety, 3 hr after a single sperm injection for the presence of a second polar to predict oocyte fertilization with a high certainty.     

In vivo studies on the effect of UV-radiation on the eye lens in animals

Ultraviolet light is a non-ionizing radiation that induces photochemical reactions in the tissue. Its spectral A and B ranges are partially absorbed by the cornea and/or lens thus causing damage on the cellular, cell physiological and molecular level. UV-A does not seem to damage the cornea permanently and its effects in the lens have a very prolonged latency period. Typical reactions of the cornea are oedema, punctate keratitis (photoelectric keratitis) and neovascularization. In the lens all reactions that could be evidenced, were located in the epithelium and in the outer cortical fiber cells.In vivo UV-A induces swelling and slight vacuolation of the anterior suture system, but apart from these transient effects, only very limited permanent damage could be demonstrated. UV-B induces the formation of an anterior subcapsular cataract, starting also with vacuolation of the suture system. These morphological characteristics can be visualized at the slitlamp microscope. Histologically, sutural irregularities (UV-A) and epithelial hyperplasia with capsular multiplication (UV-B) as well as desintegration of the anterior suture system could be observed. Patho-physiologically, a reduction of lens fresh weight (UV-B) as well as changes of the equilibrium of reduced and oxidized glutathione (GSH/GSSG) could be demonstrated. On the protein-biochemical level, changes in the ratio of water-soluble versus water-insoluble protein could be evidenced, as well as effects on specific crystallin fractions, namely-crystallin. In addition, the appearance of a newly synthetized 31 kDa protein could be demonstrated in UV-B irradiated mice.     

HLA-DR-associated genetic control of the type of leprosy in a population from Surinam

The relationship between HLA phenotype and leprosy classification was studied in 73 unrelated patients and 92 healthy controls from a mixed Negroid-Caucasoid population originating from Surinam, South America. Heterogeneity in the distribution of HLA-DR (but not A, B, and C) was detected between tuberculoid (TT^* + BT^*) leprosy and lepromatous (BL^* + LL^*) leprosy patients (p = 0.024). This heterogeneity appeared to be caused almost exclusively by DR3. Most significantly, the frequency of DR3 was increased among polar tuberculoid (TT) leprosy patients as compared to the rest of the patients (p = 0.0003). Compared with healthy controls the frequency of DR3 was increased among TT patients p = 0.0006), unchanged in BT patients, and decreased among lepromatous (BL + LL) patients (p = 0.027). These data indicate that in this population an DR3-associated factor controls the type of the disease that develops after infection with Mycobacterium leprae.     

The chromosomal analysis of human oocytes. An overview of established procedures

The cytogenetic survey of mature human oocytes has been and remains a subject of great interest because of the prevalence of aneuploidy of maternal origin in abnormal human conceptuses, and the lack of understanding about the non-disjunction processes in human meiosis. The first attempts to analyse the chromosomal content of human female gametes were made in the early 1970s, and led to limited data because of the paucity of materials and the inadequacy of the procedure used. The years to follow brought a resurgence of interest in this field, because of the development of human IVF techniques which made oocytes unfertilized in vitro available for cytogenetic analysis. Numerous studies have since been performed. However, the difficulties in obtaining good chromosome preparations and of performing accurate chromosome identification have reduced the viability of these studies, resulting in large variations in the reported incidences of chromosomal abnormalities. The further introduction of new procedures for oocyte fixation and the screening of large oocyte samples have allowed more reliable data to be obtained and to identify premature chromatid separation as a major mechanism in aneuploidy occurrence. The last decade has been privileged to witness the adaptation of molecular cytogenetic techniques to human oocytes, and thus various powerful procedures have been tried not only on female gametes, but also on polar bodies, involving sequential and multicolour fluorescent in situ hybridization (FISH) labelling, comparative genomic hybridization (CGH), spectral karyotyping and alternative methods such as primed in situ labelling (PRINS) and peptide nucleic acid (PNA) techniques. A large body of data has been obtained, but these studies also display a great variability in the frequency of abnormalities, which may be essentially attributable to the technical limitations of these in situ methods when applied to human oocytes. However, molecular cytogenetic approaches have also evidenced the co-existence of both whole chromosome non-disjunction and chromatid separation in maternal aneuploidy. In addition, the extension of these techniques to oocyte polar body materials has provided additional data on the mechanism of meiotic malsegregation. Improvements of some of these techniques have already been reported. The further development of new approaches for the in situ analysis of human meiosis will increase the impact of cytogenetic investigation of human oocytes in the understanding of aneuploidy processes in humans.