Amjad Hossain

Spontaneously developed tail swellings (SDTS) influence the accuracy of the hypo-osmotic swelling test (HOS-test) in determining membrane integrity and viability of human spermatozoa

PurposeTo investigate the prevalence of spontaneously developed tail swellings (SDTS) in human sperm samples that are commonly encountered in the laboratory, and their influence on the hypo-osmotic swelling test (HOS-test).MethodsEjaculated, epididymal, and testicular sperm were evaluated for SDTS. Further, HOS-test scores were compared with those of vital stains using column washed sperm maintained in the laboratory.ResultsSDTS, at <10%, was present in all types of sperm samples. The highest and lowest occurrences of SDTS were found in cryopreserved sperm, and column-washed sperm respectively. SDTS can inflate the HOS-test score, and so lower the accuracy of the HOS-test. However, the HOS-test efficiency can be improved by assessing SDTS in the sample.ConclusionHOS-test and vital stain cannot be used interchangeably in all circumstances for sperm viability determination. The accuracy of the HOS-test can be enhanced by incorporating SDTS as a correction factor.

Sole Use of Sucrose in Human Sperm Cryopreservation

Glycerol alone or in combination with other additives is one of the most widely used and successful cryoprotectants for human sperm. The glycerol method requires rigorous post thaw sample washing for use in ART and this may lead to low sperm yield from oligospermic samples. In this study the feasibility of the use of sucrose in sperm cryopreservation was explored. Sucrose as cryoprotectant was combined with direct plunging of sample into liquid nitrogen (vitrification) as a freezing method. Sucrose treated sperm from normozoospermic and severly oligozoospermic samples underwent rapid freeze and thaw. Motility and viability were evaluated before freezing (after sucrose equilibration) as well as post freezing (after thaw). The 100 mM concentration of sucrose showed better cryoprotectant features compared to that of higher concentrations (200–1000 mM). Sucrose (100 mM)treated sperm maintained low but acceptable motility (30%) and satisfactory viability (60%) after freezing and thawing. The cryoprotectant capacity of sucrose for normozoospermic and oligozoospermic samples were identical. The sucrose method utilizes rapid freezing of a micro volume of sample and thus quickly freezes, thaws, and maximizes recovery of the sperm from the sample.

Kininogen present in rat reproductive tissues is apparently synthesized by the liver, not by the reproductive system

OBJECTIVE The purpose of this study was to determine the source(s) of the reproductive tract kininogen and to assess whether kininogen transcription is influenced by reproductive conditions. STUDY DESIGN Rats in various reproductive states (immature, mature, ovulatory, luteal phase, pregnancy, parturition, postpartum) were used to obtain reproductive tissues (follicles, corpora lutea, oviduct, uterus, testes) and liver. Complementary deoxyribonucleic acid probes for rat prekininogens were used to quantify kininogen messenger ribonucleic acid synthesis. RESULTS The T-prekininogen complementary deoxyribonucleic acid probe detected a single 1.6 kb message, whereas the k-prekininogen complementary deoxyribonucleic acid probe identified two messages, an abundantly expressed 1.6 kb band and a 2.2 kb band. The source of all the three prekininogen messages appears to be the liver. Naturally occurring reproductive conditions such as ovulation, implantation, and parturition, did not turn on prekininogen message transcription in the rat gonad or genital tract. Only decidualization of the uterus was associated with the induction of kininogen transcription in the liver. CONCLUSION There appears to be little, if any, contribution of local gene expression to the kininogen present in the reproductive tissues. Apparently, the reproductive tract increases uptake of kininogen from plasma as needed.

Preimplantation genetic diagnosis: a systematic review of litigation in the face of new technology.

OBJECTIVE To study legal cases against IVF facilities pertaining to preimplantation genetic diagnosis (PGD) misdiagnosis. DESIGN Systematic case law review. SETTING University medical center using US legal databases. PATIENT(S) The IVF recipients using PGD services. INTERVENTION(S) Lawsuits pertaining to PGD against IVF facilities. MAIN OUTCOME MEASURE(S) Lawsuits, court rulings, damage awards, and settlements pertaining to PGD after the birth of a child with a genetic defect. RESULT(S) Causes of action pertaining to PGD arise from negligence in performing the procedure as well as failure to properly inform patients of key information, such as inherent errors associated with the PGD process, a facilitys minimal experience in performing PGD, and the option of obtaining PGD. Courts have sympathized with the financial burden involved in caring for children with disabilities. Monetary damage awards are based on the costs of caring for children with debilitating defects, including lifetime medical and custodial care. CONCLUSION(S) Facilities offering PGD services expose themselves to a new realm of liability in which damage awards can easily exceed the limits of a facilitys insurance policy. Competent laboratory personnel and proper informed consent--with particular care to inform patients of the inherent inaccuracies of PGD--are crucial in helping deter liability.

Preconceptional Sex Selection: Past, Present, and Future

Predetermination of sex in human and in farm animals is reviewed. Preconceptional sex selection has generated great interest and controversy over the years. Medical and commercial benefits outweigh the ethical issues. Technology has not yet provided a routine method for separating the X- and Y-chromosome-bearing sperm. Flow cytometry is the only technique that produces a clinically significant enrichment of X- or Y-bearing spermatozoa However, concern has been raised about the methodological implications of the flow technique because of the use of DNA stains and UV light. Some other techniques, such as gradient columns, appear to produce a slight enrichment of one type of sperm over the other, but this level of enrichment appears unlikely to affect the sex ratio at birth. It thus remains speculative whether 100% pure preparation of X or Y sperm can be obtained unless a major improvement in methodology is achieved. Fluorescence in situ hybridization (FISH) and polymerase chain reaction (PCR) are currently the methods of choice for evaluating the validity of the sex selection procedure. In view of the extraordinary pace of the technological and scientific progress, it can be expected that the clinical and commercial application of the technology of preconceptional sex selection by X and Y sperm separation will be a reality in near future.

Food deprivation induces differential changes in contents and microstructures of digestive tract and appendages in bluegill fish, Lepomis macrochirus

Abstract 1. 1. The digestive tract was compared with the tract appendages (caeca) in bluegill fish, Lepomis macrochirus in their response to short and long term food deprivation. 2. 2. Fasting for 7 days resulted in 80% reduction of food content in the main tract, but only 40% reduction in appendages (caeca). 3. 3. The intestine exhibited two different patterns of food distribution under fed and food deprived conditions. 4. 4. The histopathological impact of starvation was more prominent on the intestine than on caeca. 5. 5. These results suggest that digestive tract and appendages concommitantly conserve food during food scarcity, but appendages may offer advantages in retaining food longer, and in their greater resistance to starvation-induced effects.

Acid phosphatase activity in the intestine and caeca of bluegill, exposed to methyl mercuric chloride

The toxic effects of mercury on fish can be measured accurately by studying the enzymes and other biochemical indicators. The gross organismic changes in fish due to mercury toxicity occur only after significant damage to the internal tissue or physiological-biochemical changes in the tissue. The intestine has contact with the external environment via food. Therefore, any significant alteration in the environment may induce some changes in the structure and function of the digestive tract. An attempt has been made to measure the changes in acid phosphatase activity in different regions of intestine and intestinal diverticulae of bluegill fish as these enzymes are highly sensitive to methyl mercury.

Assessment of the Relationship of Sperm Morphology with Seminal and Other Clinical Conditions of Semen Donors

The occurrence of abnormal forms of spermatozoa in human semen is quite common. According to WHO, semen is considered normal even if it contains 50% morphologically abnormal spermatozoa. This study assessed whether the sperm morphology maintains any relation with the relevant clinical conditions of the semen donor. One hundred samples representing normal and different types of male factor etiologies underwent semen and morphological analysis. Clinical information such as race, age, weight, profession, medication, medical history, and smoking habit of the semen contributors were recorded. The influence of seminal and clinical features on sperm morphology was evaluated with multiple regression analysis. Head abnormalities were more common than tail abnormalities. Acrosomal defects and coiled tails were the most prevalent head and tail abnormalities, respectively. Regression analysis failed to confirm any strong association between sperm morphology and other seminal parameters. Accessory gland-related seminal parameters such as viscosity, volume, pH, and liquefaction showed the least association with the morphological variability. Sperm morphology also showed poor correlation with race, age, weight, smoking habit, and work environment.

Human sperm bioassay has potential in evaluating the quality of cumulus-oocyte complexes

Human sperm bioassay is routinely used as a quality control check for the culture media. This is one of the three bioassays chosen by the College of American Pathologists (CAP) for interlaboratory proficiency testing to assess the standards of in vitro fertilization (IVF) and andrology laboratories. This study utilized sperm bioassay to assess the quality of cumulus-oocyte complexes (COCs) retrieved in IVF procedures COCs, harvested from the female partner of IVF couples, undergoing identical ovarian stimulation protocols, were individually inseminated with the sperm of the corresponding male partner. Sperm motility in sperm-COC cocultures were compared. Cocultures were established by inseminating the 103 COCs, retrieved from 18 IVF couples with 1 x 10(5) to 2 x 10(5) sperm of the corresponding male partners of the couples. In all 18 cases, the sperm were prepared identically using the Percoll wash method. The cocultures were maintained for 48 h but the oocytes were removed immediately after the fertilization check (approximately 16 h). The motility of sperm in the cocultures and in the insemination stocks were noted and 17 of 18 sperm stocks used for insemination had similar high preinsemination motility (90.2 +/- 5.0%). At 48 h the sperm motility had significantly decreased in the cocultures compared to the insemination stocks; 52.7 +/- 19.9% versus 67.2 +/- 10.4%. There was no difference in the motility among the small, medium, and large COCs (56.4 +/- 24.6%, 52.5 +/- 17.9%, and 50.8 +/- 20.9%, respectively). In 45% of IVF cases, the motility in cocultures varied widely, falling below as well as above that of their corresponding insemination stocks. Furthermore, the sperm motility varied among the cocultures in both pregnant and nonpregnant patients but the extent of variation appears to be greater in the latter. The inter-COC coculture sperm motility variation most likely is due to the differences in the quality of cumulus-oocyte complexes.

Fractions of Percoll eliminated sperm : Increased sperm yield in Percoll wash

Sperm preparation by Percoll is one of the most widely used techniques, but it has the disadvantage of producing a low final yield of motile sperm. This study compared the quality of Percoll-eliminated sperm with that of Percoll recovered sperm to determine if the profile of the Percoll preparation could be improved. Semen specimens were fractionated by discontinuous Percoll gradient column. This resulted in 4 sperm fractions: sperm retained in the semen (SRS), sperm clumped at the interface of 40 and 80% Percoll (SCI), sperm scattered in the column (SSC), and sperm-forming pellet, called Percoll-recovered sperm (PRS). The sperm fractions were evaluated for count, motility, viability, vitality, and morphology. The SRS exhibited the lowest motility, viability, and vitality (40, 68, and 64%, respectively), which were significantly different from those of SCI, SSC, and PRS. The motility, viability, and vitality differences between SSC and PRS did not reach a statistically significant level. Good motility (grades a and b) was found in SSC (73%) when compared with that of PRS (82%). The lower part of SSC and PRS exhibited identical sperm morphology. The Percoll sperm recovery was 18%, but if SSC is combined with PRS, the total recovery becomes 40%. These data suggest that the SSC fraction, particularly the lower part, which maintains the physical contact with the PRS, can be pooled together to increase the final sperm yield without compromising the quality of the sperm to be used for insemination.