Molecular characterization of severe α-Thalassemias causing hydrops fetalis in Taiwan

α-Thalassemia hydrops fetalis is a common disorder in Taiwan. The condition causes perinatal death and many maternal obstetrical complications. In order to determine the molecular defects of this condition in Chinese, 87 unrelated families with this disorder were collected in the past 4 years. The molecular defects were studied by Southern blotting and DNA hybridization with φζ1-globin gene and LO (a 0.4 kb BamHI/EcoRI fragment in the 5′ flanking region of the ζ2-globin gene) probes. Eighty-one (93.1%) fetuses had homozygous Southeast Asian deletion (– –^SEA/– –^SEA). Five (5.7%) fetuses were compound heterozygotes for the Southeast Asian deletion and Thailand deletion (– –^SEA/– –^THAI). The remaining fetus was a compound heterozygote for the Southeast Asian deletion and an uncharacterized nondeletional defect (– –^SEA/(αα)Th). The molecular defects of α-thalassemia hydrops fetalis in Chinese are heterogeneous. This fact has important implications for genetic counseling and prenatal diagnosis.     

A simplified technique for embryo biopsy: Use of the same micropipette for zona drilling and blastomere aspiration

Purpose: Using different micropipettes for zona drilling and blastomere aspiration for embryo biopsy is prevalent at centers of preimplantation genetic diagnosis. The purpose of our study was to simplify the technique by using only one micropipette. Methods: In this animal model, ICR mouse embryos at the four-cell stage (n=446) were randomly allocated into two groups: a biopsied group (n=224) for blastomere aspiration and a control group (n=222) without micromanipulation. We used a drilling/biopsy micropipette to drill a hole in the zona by expulsion of acidified Tyrode’s solution and to aspirate the blastomere by gentle suction with the same micropipette and pull it out of the zona. One blastomere was biopsied from each embryo. Results: In all, 222 (99.1%) intact blastomeres were successfully biopsied from 224 embryos. Only two blastomeres were damaged during aspiration. The capacity for blastocyst development (92.4 vs 93.7%) was not different between the two groups, but the percentages of embryos hatching (51.8 vs 18.0%) and hatched (29.9 vs 8.1%) were significantly higher in the biopsied group than in the control group. Conclusions: This simplified technique of embryo biopsy is safe and highly efficient for obtaining blastomeres for preimplantation genetic diagnosis and may also facilitate hatching of the blastocysts.     

Andrology: Intracytoplasmic sperm injection (ICSI) for severe semen abnormalities: dissecting the tail of spermatozoa at the tip

Recently, several investigators have emphasized that damagingthe membrane of spermatozoa by compressing the mid-piece orcutting the mid-portion of the tail prior to injection yieldsbetter results than using motile spermatozoa in intracytoplasmicsperm injection (ICSI). Here we report our experience usinga modified immobilization technique of dissecting the tail ofthe spermatozoon at the tip in 78 cycles on 60 patients. In55 treatment cycles purely using this modified technique, 468mature oocytes were injected. A total of 35 oocytes (7.5%) wereinjured. Of the intact oocytes, 282 (65.1%) were normally fertilizedand 266 (94.3%) subsequently cleaved. A single pronucleus wasobserved in 16 (3.7%) oocytes, and three pronuclei were notedin 11 (2.5%) oocytes. Embryo transfers were performed in 54cycles, and 18 women (32.7%) achieved clinical pregnancies.In 23 cycles, we compared the effects of these three immobilizationtechniques on the sibling oocytes obtained from the same patientregarding normal fertilization, abnormal fertilization, andembryo cleavage and quality. The results were comparable amongthem. Seven pregnancies (30.4%) were achieved in this series.Dissecting a sperm tail at the tip is easily and quickly performedand achieves permanent immobilization. Compression of the mid-pieceis also easy, but usually takes several actions to achieve immobilization.Cutting the tail at the mid-portion requires more skill. Therefore,dissecting the tail of the spermatozoon at the tip may providean alternative method to immobilize the spermatozoon permanentlyprior to ICSI.     

Central cyanosis due to severe pulmonary hypertension combined with pericarditis as the initial manifestation of systemic lupus erythematosus

Systemic lupus erythematosus (SLE) is a multisystemic autoimmune disorder. The initial manifestation complicating any organ system either singly or in combination is protean. Herein we report a 26-year-old female patient with fresh SLE whose initial manifestation was central cyanosis caused by severe pulmonary hypertension and acute pericarditis. The symptoms were relieved dramatically after treatment with steroid and bosentan. Accurate and timely diagnosis in SLE-associated pulmonary hypertension may be life saving.     

Age,Environment, and Lymphocyte Immunization Influence the Spontaneous Resorption Rate in the CBA/J × DBA/2J Mouse Model

PROBLEM: This study was designed to examine the influence of age, environment, and lymphocyte immunotherapy on fetal resorption rates in the CBA/J × DJA/2J mouse model. METHODS: After weaning, CBA/J female mice were randomly allocated to two different environments: room A was a conventional housing facility, and room B was a specific pathogen-free room. They were further divided into two groups in each room according to age (> three months and ≤ three months), and mated with CBA/J, DBA/2J, or BALB/c males. The fetal resorption rates were observed. The immunization study was conducted using CBA/J females greater than three months old in room B. Preimmunization with various preparations and dosages of lymphoctyes was performed 1 wk before mating with DBA/2J males, and fetal resorption rates were measured to investigate the effect of immunotherapy. RESULTS: In room A, the fetal resorption rates (9.4 to 11.8%) were not significantly different among the various mating combinations using CBA/J females greater or less than three months of age. In room B, CBA/J females older than three months and mated with DBA/2J males had a higher rate of fetal resorption than those younger than three months (28.9 versus 14.2%) and had higher rate of fetal resorption than those housed and mated in room A (28.9 versus 10.9%). This fetal resorption rate was also higher than those seen with other mating combinations. Preimmunization with male BALB/c splenic lymphocytes (optimal dose 1 × 10⁷ cells) was effective in decreasing fetal resorption rate (7.9%), whereas administration of normal saline, or immunization with male CBA/J or DBA/2J lymphocytes was not (20.7 to 27.0%). CONCLUSION: Age and environment influence the spontaneous resorption rate in the CBA/J × DBA/2J mouse model. The high fetal resorption rate of older CBA/J females mated with DBA/2J males in a pathogen-free environment can be reduced by immunization with BALB/c splenic lymphocytes. These results suggested that a variety of mechanisms might initiate early pregnancy failure and that immunological modulation during implantation might be a nonspecific factor.     

Andrology: Co-culture with Vero cell monolayer maintains the motility of asthenozoospermic semen samples

The clinical effectiveness of co-culture with Vero (Green monkeykidney) cell monolayer in maintaining the motility and viabilityof fresh asthenozoospermic semen (18 samples) and frozen–thawedsemen with poor motility (motility fraction <50%) (15 samples)in a 24-h period was evaluated. Co-culture with Vero cell monolayerin human tubal fluid (HTF) medium for 24 h resulted in a statisticallybetter maintenance of motility percentage (P < 0.005), meanamplitude of lateral head displacement (ALH) (P < 0.005),and mean track speed (VCL) (P < 0.05) than culture in HTFmedium alone. However, these motility parameters (motility percentage,ALH, VCL) declined soon after removal of spermatozoa from themonolayer. Co-culture with Vero cell monolayer also maintainedthe viability percentage of these sperm samples (52% of theoriginal value) after the 24-h period compared with culturein HTF medium alone (22% of the original) (52% versus 22%, P< 0.05).It is concluded that Vero cell monolayer is effectivein the maintenance of motility and viability of asthenozoospermicsemen or frozen–thawed semen with poor motility.This co-culturesystem may be beneficial in enhancing the in-vitro performanceof asthenozoospermic semen samples in the practice of assistedreproductive technology.However, its safety needs further evaluation.     

Pre-therapy CT scan showing peritoneal thickening from metastatic renal pelvis carcinoma patients

We investigated clinical significance of peritoneal thickening from metastatic renal pelvis based on pretherapy computed tomography (CT) scan findings. The criteria for inclusion were as follows: (1) pathology and CT scan confirmed metastatic renal pelvis carcinoma and (2) peritoneal thickening based on pre-therapy CT scan findings. We investigated the route of spread, gastrointestinal (GI) complications, and response to chemotherapy. A total of 68 cases were enrolled in this study, including seven patients with liver metastases and three with abdominal wall invasion. GI complications included obstruction in ten patients and bleeding in three. Response to chemotherapy demonstrated by reduced peritoneal thickening was noted in 24 patients. In conclusion. peritoneal thickening with clinical suspicion of peritoneal involvement can get indirect evidence from route of spread (liver or abdominal wall), GI complications (obstruction or bleeding) or response to chemotherapy (obvious decrease peritoneal thickening) from metastatic renal pelvis carcinoma patients. Pretherapy CT scan with peritoneal thickening should be alert that tumor has spread to the peritoneum.     

Unrecognized paroxysmal ventricular standstill masquerading as epilepsy: a Stokes-Adams attack.

Recognition of cardiac syncope masquerading as epilepsy may be difficult in the Emergency Department. We report a middle-aged man with recent onset convulsions who posed a diagnostic puzzle before it was found that he had paroxysmal ventricular standstill with complete atrioventricular block: he made a complete recovery after temporary pacemaker insertion. The main lessons from this case were (1) a convulsive seizure of only seconds duration and with an abrupt return of consciousness suggests syncope not epilepsy, (2) repeated, convulsive syncopes without provocation suggest cardiac syncope, (3) a 12-lead ECG should be recorded as soon as possible after such a series of episodes and should not be discontinued until an event is captured, and (4) Emergency Department clinicians should be familiar with any automatic gain on their ECG machine, lest fast, atrial activity be mistaken for narrow complex tachycardia. In summary, a good clinical history is of prime importance in differentiating convulsive syncope from epilepsy, and a simple, non-invasive cardiovascular evaluation may help to diagnose the condition as cardiac syncope.