A cytogenetic study of couples with recurrent spontaneous abortions and infertile patients with recurrent IVF/ICSI failure

PURPOSE:

This study was conducted to determine the frequency and contribution of chromosomal abnormalities in miscarriages and in couples with recurrent in vitro fertilization/intra cytoplasmic sperm injection (IVF/ICSI) failure.

MATERIALS and METHODS:

A total of 221 individuals; 79 with three or more recurrent spontaneous abortions and 142 with at least three IVF/ICSI failures. Chromosomal analysis from peripheral blood lymphocytes was performed according to standard cytogenetic methods using G-banding technique.

RESULTS:

Abnormal karyotype was found in 21 (9.50%) individuals. Of these 21 subjects, 4 (19.04%) exhibited sex chromosomal abnormalities and 17 (80.96%) had autosomal abnormalities. Male partners had significantly higher chromosomal abnormalities (5.88%) than of females (3.61%). These abnormalities were also higher in patients with recurrent spontaneous abortions than with IVF/ICSI failure (P < 0.05).

CONCLUSIONS:

These data may be indicative that chromosomal abnormalities are involved more in spontaneous abortions than in recurrent IVF/ICSI failure. Cytogenetic analysis could be valuable for these couples when clinical data fail to clarify the cause.     

Parental origin of chromatin in human monopronuclear zygotes revealed by asymmetric histone methylation patterns, differs between IVF and ICSI

In mouse zygotes, many post-translational histone modifications are asymmetrically present in male and female pronuclei. We investigated whether this principle could be used to determine the genetic composition of monopronuclear human zygotes in conventional IVF and ICSI. First we determined whether male female asymmetry is conserved from mouse to human by staining polypronuclear zygotes with antibodies against a subset of histone N-tail post-translational modifications. To analyze human monopronuclear zygotes, a modification, H3K9me3, was selected that is present in the maternal chromatin. After IVF a total of 45 monopronuclear zygotes were obtained. In 39 (87%) of zygotes a nonuniform staining pattern was observed, proof of a bi-parental origin and assumed to result into a diploid conception. Two zygotes showed no staining for the modification, indicating that the single pronucleus was of paternal origin. Four zygotes contained only maternally derived chromatin. ICSI-derived monopronuclear zygotes (n = 33) could also be divided into three groups based on the staining pattern of their chromatin: (1) of maternal origin (n = 15), (2) of paternal origin (n = 8) or (3) consisting of two chromatin domains as dominating in IVF (n = 10). Our data show that monopronuclear zygotes originating from IVF generally arise through fusion of parental chromatin after sperm penetration. Monopronuclear zygotes derived from ICSI in most cases contain uni-parental chromatin. The fact that chromatin was of paternal origin in 24% of ICSI and in 4% of the IVF zygotes confirms earlier results obtained by FISH on cleavage stages. Our findings are of clinical importance in IVF and ICSI practice.     

Clinical implications of sperm DNA damage in IVF and ICSI: updated systematic review and meta-analysis

The clinical effect of sperm DNA damage in assisted reproduction has been a controversial topic during recent decades, leading to a variety of clinical practice recommendations. While the latest European Society of Human Reproduction and Embryology (ESHRE) position report concluded that DNA damage negatively affects assisted reproduction outcomes, the Practice Committee of the American Society for Reproductive Medicine (ASRM) does not recommend the routine testing of DNA damage for in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI). Herein, our aim was to perform a systematic review and meta-analysis of studies investigating whether sperm DNA damage affects clinical outcomes in IVF and ICSI, in order to contribute objectively to a consistent clinical recommendation. A comprehensive systematic search was conducted according to PRISMA guidelines from the earliest available online indexing year until March 2020, using the MEDLINE-PubMed and EMBASE databases. We included studies analysing IVF and/or ICSI treatments performed in infertile couples in which sperm DNA damage was well defined and assessed. Studies also had to include information about pregnancy, implantation or live birth rates as primary outcomes. The NHLBI-NIH quality assessment tool was used to assess the quality of each study. Meta-analyses were conducted using the Mantel–Haenszel method with random-effects models to evaluate the Risk Ratio (RR) between high-DNA-damage and control groups, taking into account the 95% confidence intervals. Heterogeneity among studies was evaluated using the I² statistic. We also conducted sensitivity analyses and post-hoc subgroup analyses according to different DNA fragmentation assessment techniques. We identified 78 articles that met our inclusion and quality criteria and were included in the qualitative analysis, representing a total of 25639 IVF/ICSI cycles. Of these, 32 articles had sufficient data to be included in the meta-analysis, comprising 12380 IVF/ICSI cycles. Meta-analysis revealed that, considering IVF and ICSI results together, implantation rate (RR = 0.74; 95% CI = 0.61–0.91; I² = 69) and pregnancy rate (RR = 0.83; 0.73–0.94; I² = 58) are negatively influenced by sperm DNA damage, although after adjustment for publication bias the relationship for pregnancy rate was no longer significant. The results showed a non-significant but detrimental tendency (RR = 0.78; 0.58–1.06; I² = 72) on live birth rate. Meta-analysis also showed that IVF outcomes are negatively influenced by sperm DNA damage, with a statistically significant impact on implantation (RR = 0.68; 0.52–0.89; I² = 50) and pregnancy rates (RR = 0.72; 0.55–0.95; I² = 72), although the latter was no longer significant after correction for publication bias. While it did not quite meet our threshold for significance, a negative trend was also observed for live birth rate (RR = 0.48; 0.22–1.02; I² = 79). In the case of ICSI, non-significant trends were observed for implantation (RR = 0.79; 0.60–1.04; I² = 72) or pregnancy rates (RR = 0.89; 0.78–1.02; I² = 44), and live birth rate (RR = 0.92; 0.67–1.27; I² = 70). The current review provides the largest evidence to date supporting a negative association between sperm DNA damage and conventional IVF treatments, significantly reducing implantation and pregnancy rates. The routine use of sperm DNA testing is therefore justified, since it may help improve the outcomes of IVF treatments and/or allow a given couple to be advised on the most suitable treatment. Further well-designed controlled studies on a larger number of patients are required to allow us to reach more precise conclusions, especially in the case of ICSI treatments.     

Roles of kisspeptin in IVF/ICSI-treated infertile women and in human granulosa cells

Kisspeptin, a crucial central regulator of reproduction, has been used as a trigger in in vitro fertilization (IVF) treatment. This study aimed to investigate the roles of kisspeptin in IVF treatment in infertile females (n = 30); and in steroidogenesis in human granulosa-like tumor cell line (KGN). In the human study, blood was collected at three time points including (1) the beginning of gonadotropin stimulation (Phase I), (2) around eight days after gonadotropin stimulation (Phase II), and (3) on the day of ovum pick-up (Phase III). Follicular fluid (FF) was collected at Phase III. Serum human chorionic gonadotropin (hCG) was measured 15 days after embryo transfer and fetal heart beats were determined around 42 days of menstrual cycle to classify the subjects into successful and unsuccessful groups. FF kisspeptin levels were higher in successful compared with unsuccessful subjects (P < 0.01). Kisspeptin levels were significantly higher in FF than in serum in successful subjects (P < 0.05) but were comparable in unsuccessful subjects. Serum kisspeptin was comparable among three phases in the successful group but its levels in Phase III were significantly lower compared with Phase I in the unsuccessful group (P < 0.01). Serum kisspeptin in Phase II/III had positive correlations with serum E2 in Phases II and III and the outcomes of IVF/intracytoplasmic sperm injection (ICSI) treatment including serum hCG levels. For the cell experiment (n = 3), kisspeptin treatment in the presence of FSH together with IGF-1 enhanced CYP19A1 (aromatase) mRNA expression compared with control. FSH alone increased aromatase concentrations in the supernatant compared with control and kisspeptin at the dose of 10^-2 mmol/L with FSH enhanced aromatase concentrations in the supernatant compared with FSH alone (P < 0.001 all). In conclusion, kisspeptin enhanced aromatase expression and secretion and was associated with positive outcomes of IVF/ICSI treatment. Further studies regarding supplementation of kisspeptin could reveal its beneficial effects on IVF/ICSI treatment.     

Rotationally oscillating drill (Ros-Drill) for mouse ICSI without using mercury

Intracytoplasmic sperm injection (ICSI) is an important assisted reproductive technology (ART). Due to deployment difficulties and low efficiency of the earlier (conventional) version of ICSI, especially in the mouse, a piezo-assisted ICSI technique had evolved as a popular ART methodology in recent years. An important and remaining problem with this technique, however, is that it requires small amounts of mercury to stabilize the pipette tip when piezoelectric force pulses are applied. To eliminate this problem we developed and tested a completely different and mercury-free technology, called the "Ros-Drill" (rotationally oscillating drill). The technique uses microprocessor-controlled rotational oscillations on a spiked micropipette without mercury or piezo. Preliminary experimental results show that this new microinjection technology gives high survival rate (>70% of the injected oocytes) and fertilization rate (>80% of the survived oocytes), and blastocyst formation rates in early trials (approximately 50% of the survived oocytes). Blastocysts created by Ros-Drill ICSI were transferred into the uteruses of pseudopregnant surrogate mothers and healthy pups were born and weaned. The Ros-Drill ICSI technique is automated and therefore; it requires a very short preliminary training for the specialists, as evidenced in many successful biological trials. These advantages of Ros-Drill ICSI over conventional and piezo-assisted ICSI are clearly demonstrated and it appears to have resolved an important problem in reproductive biology.     

Partitioning of Soluble Calcium in the Water Hyacinth (Eichhornia crassipes (Martius) Solms) in Lake Victoria,Uganda

Abstract: An investigation of soluble calcium partitioning in organs of the water hyacinth growing along the shores of L. Victoria, bordering Jinja town was carried out. The levels of Na⁺, K⁺, Mg²⁺ and Ca²⁺ in the lake water were also estimated. The pH and electrical conductivity of lake water were determined. There was a differential accumulation of Ca and oxalate between component organs of the water hyacinth. The concentration of oxalate in the various organs was more than twice that of Ca. Soluble Ca decreased with age in all organs except the leaf, while oxalate increased with age in all organs except the rhizome. There were site variations in the partitioning of both Ca and oxalate in the water hyacinth.     

胞浆内精子注射技术生产小鼠

以piezo操作系统为技术支撑 ,在掌握小鼠卵母细胞胞浆内精子注射技术 (ICSI)的基础上 ,进行了ICSI技术生产试管小鼠的尝试。来自成年昆明 (KM)小鼠附睾尾的新鲜精子 ,剪切去尾后 ,直接将精子头注射到B6D2F1小鼠卵母细胞质中 ,注射后 1h ,83.3%的卵母细胞存活。6h时 ,84.0 %的成活卵子成功受精 ,形成原核 ,排出PB2 体外培养的ICSI胚胎 ,卵裂率 (98%vs 94.7% )和 4-细胞期胚胎比率 (89.5%vs 92.1% )均与培养的体内受精卵没有差异 (P >0.05 ) ;但是 ,桑椹胚(63.8%vs84.2% )和囊胚发育率 (25.7%vs68.4% )极显著地 (P <0.01)低于对照组。120枚原核期胚胎移植给 7只假孕受体后 ,4只受孕小鼠共产出 28只ICSI小鼠 (23.3% )。健康成年的 25只ICSI小鼠都没有明显的生理和行为异常。随机选择其中的 20只小鼠 ,分别进行ICSI小鼠间、ICSI与KM小鼠间共 12组的交配 ,结果所有雌鼠妊娠产仔。在成功建立小鼠ICSI技术的基础上,成功获得了我国的首例ICSI小鼠,并且证明这些ICSI小鼠都具有正常的繁殖后代的能力。     

Founding Pioneers of IVF: Independent innovative researchers generating livebirths within 4 years of the first birth

In this 40th anniversary year of the first IVF live birth, it is pertinent to look at all those teams endeavouring to generate live births from this unique technology and who succeeded within 4 years of the first. There were 9 teams who achieved this and a further 3 who were successful soon after, by the end of 1982. This historical review is compiled by 2 authors who were actively engaged in the field of IVF at the time of the first birth and who have remained active in Reproductive Medicine throughout their professional lives. They bring intimate and relevant knowledge of those pioneer researchers from the early years who can be classified as the "Founding Pioneers" of IVF.     

Understanding fertilization through intracytoplasmic sperm injection (ICSI)

Since the establishment of in vitro fertilization, it became evident that almost half of the couples failed to achieve fertilization and this phenomenon was attributed to a male gamete dysfunction. The adoption of assisted fertilization techniques particularly ICSI has been able to alleviate male factor infertility by granting the consistent ability of a viable spermatozoon to activate an oocyte. Single sperm injection, by pinpointing the beginning of fertilization, has been an invaluable tool in clarifying the different aspects of early fertilization and syngamy. However, even with ICSI some couples fail to fertilize due to ooplasmic dysmaturity in relation to the achieved nuclear maturation marked by the extrusion of the first polar body. More uncommon are cases where the spermatozoa partially or completely lack the specific oocyte activating factor. In this work, we review the most relevant aspects of fertilization and its failure through assisted reproductive technologies. Attempts at diagnosing and treating clinical fertilization failure are described.     

Changes in termite (Isoptera) communities due to the clearance and cultivation of tropical forest in Uganda

Changes in termite communities following clearance and cultivation of part of Mabira Forest Reserve were studied. The study sites included natural forest, a cleared but not cultivated site and six sites cultivated for different periods, ranging from 1 to 20 years.
A total of 25 species, including Odontotermes amanicus (Sjostedt) and Microtermes luteus (Harris), both not previously identified in Uganda, was recorded in the natural forest. Forest clearance resulted in drastic reduction of the number of species to about 40% of what existed in the natural forest, while cultivation led to further elimination, leaving only four and two species, respectively, in the 1- and 20-year-old banana groves.
The study showed that termite species diversity declines when the natural forest is cleared and cultivated. It is suggested that reduction in termite diversity alters ecological processes and may result in diminishing productivity with time.     

Metal elements associate with in vitro fertilization (IVF) outcomes in 195 couples

PurposeEnvironmental factors may affecting reproductive function reduction and embryonic development. Couples who are exposed to heavy metals for a long time may affect the outcome of in vitro fertilization (IVF). To evaluate the effect of elements on IVF outcomes, a total of 195 couples undergoing IVF were included in this study.MethodsElements including V, Cr, Mn, Ni, Cu, Zn, As, Mo, Cd, Ba, Hg, Tl, Pb were measured in serum and follicular fluid (FF) samples of female and semen samples of male by Inductively Coupled Plasma Mass Spectrometry (ICP-MS). Multiple linear regression were applied to evaluate the association between metal elements and semen quality parameters and the number of oocytes in MII stage. Poisson regression and the robust variance estimation of the generalized estimation equation were used to evaluate the association between elements and IVF outcomes.ResultsThe statistical results showed that Cr had a significant negative correlation with total sperm concentration (TSC) and total motile sperm count (TMC), the correlation coefficients were -0.52 (-0.27∼1.43) and -0.4(-1.24, 0.45), respectively. At the same time, Ba was significantly correlated with TSC and TMC, the correlation coefficients were 0.1(-0.15∼0.34) and 0.12(-0.13, 0.36), respectively. Cr, Ba and Pb in follicular fluid (FF) had a significant positive correlation with the number of oocytes in MII stage. The correlation coefficients were 3.15 (0.79, 5.52), 1.54 (-0.27, 3.36), 12.27 (7.49, 17.04). The Tl level of FF was significantly associated with the high probability of blastocyst formation and high-quality blastocysts (RR: 2.83, 95 % CI: 0.92∼7.95; RR: 3.12, 95 % CI: 0.64, 12.84). The Hg level (RR: 3.98, 95 % CI: 0.78∼14.77) and the Ba level in serum (RR: 12.75 95 % CI: 1.31∼89.71) were significantly correlated with high-quality blastocysts. The levels of Ni, Cu, Mo in seminal plasma of men were significantly correlated with blastocyst formation and high-quality blastocysts (RR values were all greater than 1.5). In addition, the level of Ba was significantly correlated with the high probability of blastocyst formation (RR: 1.7, 95 % CI: 1.14∼2.52).ConclusionOur results reveal that Cr, Ba and Pb may affect TSC, TMC and MII oocytes. Moreover, Ba, Cr, As, Hg and Tl in serum and Mo in seminal plasma were related to fertilization results, good embryos, blastocyst formation, high-quality embryos, and pregnancy and live birth rates. Tl in FF may related to the quality of embryonic development, Ba was an important risk factor which closely related to the outcomes of IVF in both male and female. Through our detection and statistical analysis of clinical samples, it is shown that although not all elements will affect the outcome of IVF the key elements we have selected need to arouse our attention, which benifit to the diagnosis and prevention of clinical infertility.     

Distribution of Acacia senegal (L.) Willd. (Fabaceae) in Uganda and its relationship to climatic factors

The aim of the study was to map the distribution of Acacia senegal and its infraspecific taxa in Uganda and predict its suitable range of occurrence based on climatic factors. The distribution of the infraspecific taxa was analysed for richness, diversity, turnover and reserve selection. Regression analysis was performed to understand the relationship between distribution of the species and climatic variables. Georeferenced species occurrence points were superimposed over temperature and vapour maps. Areas with highest taxa richness, diversity and turnover were mapped and found in the Lake Kyoga basin. The species shows preference for the northern region of the study area with dissimilar climatic pattern from the southern region and the superimposition supported findings. Variety kerensis exhibited narrower climatic, altitudinal and distribution range preference. Temperature seasonality, maximum temperature of warmest month, temperature annual range, mean temperatures of warmest and driest quarters had the highest coefficients of determination (r² > 0.7) hence most important in influencing species distribution. The most appropriate locations for in situ conservation and for germplasm collection to ensure maximum diversity is secured are found in Wabisi‐Wajala, Kiula, Kyalubanga, Bajo, Kasagala, Kabwika‐mujwalanganda, Maruzi, Moroto and Napak Central Forest Reserves. The study recommends ecological studies to understand status of A. senegal.     

Distribution and Races of Xanthomonas axonopodis pv. malvacearum on Cotton (Gossypium hirsutum) in Uganda

Surveys in 1995 and 1996 showed that bacterial blight caused by Xanthomonas axonopodis pv. malvacearum occurs throughout the main cotton growing areas of Uganda, causing seedling blight, angular leaf spot and bacterial boll rot. During the vegetative and early fruiting stages of crop growth, severe symptoms of `blackarm' spread from leaves to the stem, causing loss of fruiting branches. A set of Upland cotton cultivars ( Gossypium hirsutum ) were then used to determine the races of the blight bacterium present in Uganda. Many of the isolates induced moderate to severe symptoms on all the test hosts except 101–102B, indicating infection with race 10 or 18. The next most common isolate was race 7. Races 16 and 6 were also identified and 23% of isolates caused symptoms on all the differential cultivars including 101–102B, results indicating the presence of a race of the pathogen which may be the same as that identified in countries neighbouring Uganda and designated as race 20.     

Status of forests in Uganda

Trees, forests and woodlands cover about 14% of Uganda’s land surface. Over the last 30–40 years, growth in human population and corresponding increase in demand for forest products for domestic and industrial use, expansion of agricultural land, illegal settlements and weak forest management capacity have adversely affected the status of natural forests in Uganda, particularly the biodiversity. Until recently, little attention had been paid to development of commercial forests which should have provided alternative forest products and services to relieve the pressure on natural forests and conserve biodiversity. As a result, Uganda’s forests have been degraded, and in some cases, the biodiversity has been eroded. There is a need for regular data collection and monitoring of the status of the forests in terms of areal extent, distribution, plantation species introductions and biodiversity.     

Killing of a pearl-spotted owlet (Glaucidium perlatum) by male red colobus monkeys (Procolobus tephrosceles) in a forest fragment near Kibale National Park, Uganda

Adult male red colobus (Procolobus tephrosceles) were observed capturing and killing an owl (Glaucidium perlatum) in the Rurama forest fragment near Kibale National Park in western Uganda. The owl was not subsequently eaten by the colobus, their conspecifics, or the other primates present during the attack. Because the incident was preceded by an agonistic encounter with a raptor, the event is best interpreted as a misdirected antipredator behavior. Although antipredator behaviors are not unknown in red colobus, this is the first such incident directed against a raptor to be documented.     

In vitro production of embryos in South American camelids

Studies in reproductive biotechnology techniques have been minimal in South American camelids (SAC). Complex reproductive characteristics of these species contribute to slow progress. Nevertheless, some techniques, such as in vitro fertilization, intracytoplasmic sperm injection and nuclear transfer have been applied and have produced advances in knowledge on embryo environment and in vitro conditions necessary for development. Embryo production may have a high impact in both domestic and wild camelids population. Studies addressed to improve in vitro embryo production and oocyte collection could be a potential key to develop IVF and embryo production as a routine procedure in camelids.