Single embryo transfer with comprehensive chromosome screening results in improved ongoing pregnancy rates and decreased miscarriage rates.

BACKGROUND: Single embryo transfer (SET) provides the most certain means to reduce the risk of multiple gestation. Regrettably, prospective trials of SET have demonstrated reductions in per-cycle delivery rates. A validated method of comprehensive chromosome screening (CCS) has the potential to optimize SET by transferring only euploid embryos. This retrospective study evaluates the efficacy of SET with CCS in an infertile population. METHODS: Overall and age-controlled ongoing pregnancy rates (OPR) were compared between women undergoing SET following CCS (CCS-SET, n= 140) and those undergoing SET without aneuploidy screening (control SET, n= 182). All transfers were at the blastocyst stage, with CCS performed after trophectoderm biopsy of expanded blastocysts and analysis with rapid PCR allowing for fresh transfer. RESULTS: In the CCS-SET and control SET groups, an OPR of 55.0 and 41.8%, respectively, was obtained. The OPR was lower for the control group (P< 0.01) despite a younger age than the CCS group (37.3 ± 3.4 versus 34.2 ± 3.9 years; P< 0.001). Birthweight and gestational age at delivery were equivalent. The proportion of clinical pregnancies resulting in miscarriage was higher in the control group (24.8 versus 10.5%, P< 0.01), with more patients requiring surgical interventions for aneuploid pregnancies. There was one monozygotic twin delivery in the CCS group and none in the control group. CONCLUSIONS: Compared with traditional blastocyst SET, SET after trophectoderm biopsy and rapid PCR-based CCS increases OPR and reduces the miscarriage rate. The enhanced selection empowered by CCS with SET may provide a practical way to eliminate multi-zygotic multiple gestation without compromising clinical outcomes per cycle.

Esophageal carcinoma: current controversial topics.

Worldwide, esophageal carcinoma is a common gastrointestinal cancer with a high mortality. The incidence of adenocarcinoma of the esophagus is increasing in the western world, but squamous cell carcinoma remains dominant in the underdeveloped parts of the world. Both types of esophageal carcinoma remain equally virulent. Currently, there are no optimal preventative screening programs available and most patients present with advanced or metastatic disease. Although many options are available for improving diagnostic accuracy, a single method has not displayed significant advantages over the others. In addition, selecting a superior treatment regimen has not surfaced. Preferred resection techniques exist, but one method has not illustrated improvements in survival over the others. A lack of improved survival rates with single modality therapies has led to a multi modality approach. However, developments in neoadjuvant and adjuvant therapies have led to mixed conclusions. Collectively, past studies have not shown an optimal neoadjuvant or adjuvant regimen in terms of survival benefit. This review highlights existing staging modalities and treatment regimens for esophageal carcinoma, in an effort to illustrate the controversial nature surrounding its management.

Social, neuroradiologic, medical, and neuropsychologic correlates of sexually aberrant behavior after traumatic brain injury: a controlled study.

OBJECTIVE: To identify social, neuroradiological, medical, and neuropsychological correlates of sexually aberrant behavior (SAB) after traumatic brain injury (TBI). DESIGN: A controlled study using a retrospective file review. SETTING: A brain injury unit providing inpatient and outpatient rehabilitation services. PARTICIPANTS: A sample of males (n = 25) exhibiting SABs and a control group (n = 25) matched for gender, severity of injury, age at injury, and time after injury. MAIN OUTCOME MEASURES: A protocol that recorded data on demographic, injury, radiological, medical, and neuropsychological variables. RESULTS: The SAB group had a significantly higher incidence of postinjury psychosocial disturbance in areas of nonsexual crime and failure to return to work than the matched TBI group. There were no significant differences between the two groups in the incidence of premorbid psychosocial disturbance or postinjury radiological, medical, or neuropsychological variables. CONCLUSIONS: The study results caution against simplistic explanations of SAB as the product of damage to the frontal-lobe systems or premorbid psychosocial disturbance. Furthermore, the results suggest that a wide-ranging assessment of people with TBI who exhibit SABs is required, because results of neuropsychological examination alone cannot be considered conclusive. Future research into the etiology of SABs could examine additional factors such as lack of insight, lack of empathy, and premorbid history of family dysfunction.

Human mdm2 mediates multiple mono-ubiquitination of p53 by a mechanism requiring enzyme isomerization.

The mdm2 gene product is an important regulator of p53 function and stability. mdm2 is an E3 ubiquitin ligase for p53 and the RING finger domain of mdm2 is critical for ligase activity. Ubiquitin (Ub) conjugation is a general targeting modification and poly-ubiquitin chains specifically target proteins to the proteasome for degradation. In this report, we show that the multistep cascade of mdm2-mediated p53 ubiquitination can be reduced to three purified recombinant proteins: ubiquitin-conjugated E2, mdm2, and p53. This simplification allows enzymatic analysis of the isolated ligase reaction. The simplified reaction recapitulates the ubiquitination of p53 observed with individual components and the p53-Ub((n)) is qualitatively similar to p53-Ub((n)) detected in lactacystin-treated cells. Surprisingly, we find that p53 is modified with multiple mono-ubiquitin moieties as opposed to a poly-ubiquitin chain. Finally, kinetic analysis indicates the transfer reaction proceeds either through a modified Ping Pong mechanism involving requisite enzyme isomerization steps, or through a Rapid Equilibrium Random Bi Bi mechanism involving very large anti-cooperative interactions between the two substrate binding pockets on the enzyme, mediated through allosteric changes in enzyme structure.

Increased nucleotide excision repair in cisplatin-resistant ovarian cancer cells: role of ERCC1-XPF.

Increased platinum-DNA adduct removal has been shown by several DNA repair assays to be associated with cisplatin resistance in the A2780/C-series human ovarian cancer model system. In the present study, we provide further evidence that the resistance phenotype of these cell lines is due, in part, to enhanced nucleotide excision repair (NER). Cisplatin resistance was found to be associated with increased UV resistance. Northern blot analysis revealed that increased expression of ERCC1 was also associated with cisplatin resistance in this panel. Several other NER genes were found to be constitutively overexpressed in the most resistant cell line, C200, as compared with the parental A2780 cells. A plasmid substrate containing a site-specific cisplatin adduct was used to measure the nucleotide excision activity of cell extracts prepared from cisplatin-sensitive and -resistant cells. Using this in vitro assay, extracts prepared from C200 cells exhibited approximately 3-fold more activity than extracts prepared from A2780 cells, similar to the difference in UV sensitivity. Complementation of A2780 extracts with ERCC1-XPF protein resulted in approximately 2-fold increased activity, but had little effect on excision in C200 extracts. Overall, these results support a role for the ERCC1-XPF endonuclease as a determinant of increased NER in this cisplatin resistance model.

Changing management trends in penetrating colon trauma.

PURPOSE: Recent prospective studies have recommended primary repair for all penetrating colon injuries. We evaluated our management trends given these recommendations and assessed our results of primary repair. METHODS: A retrospective review was conducted of 145 patients with penetrating colon injuries received between January 1, 1991, and December 31, 1997. The patients were characterized according to demographics and severity of injury. Morbidity was defined as failure of a primary repair, abscess, fistula, wound dehiscence, fasciitis, sepsis, organ failure, or coagulopathy. The periods 1991 to 1993 (early period) and 1994 to 1997 (late period) were chosen for comparison. RESULTS: Primary repairs were performed in 53 of 75 patients (71 percent) during the early period and in 61 of 70 patients (87 percent) during the late period (P = 0.03). No significant differences in demographics or injury severity were found to account for the increased rate of primary repairs. The number of suture repairs was nearly equal in both periods (59 vs. 61 percent). The number of resections and anastomoses for destructive colon injuries was significantly higher in the late period (26 percent) compared with the early period (12 percent; P = 0.05). Morbidity was equal (24 percent) in the two periods. There were no failures of resections and anastomoses and one failure of suture repair. CONCLUSIONS: Increased primary repair occurred because of more liberal use of resection and anastomosis for destructive injuries. Suture repair was performed for the amenable colonic injury throughout the study period. Risk factors for failure of resection and anastomosis cannot be defined from our study. Further investigation is needed to determine if resection and anastomosis is safe for the most severely injured patients.

The effect of surgical office-based thyroid ultrasound on clinical decision making.

An important diagnostic tool for the evaluation of thyroid disease, thyroid ultrasound has recently become available for use in surgical offices. The purpose of this report is to determine the lesional sensitivity of office-based thyroid ultrasound and its impact on clinical decision making. Surgical office-based thyroid ultrasound was performed on 49 consecutive patients who presented with thyroid disease. Indications for sonography included a solitary palpable nodule (n = 32), multiple palpable nodules (n = 3), diffuse enlargement (n = 5), or other hormonal or radiologic abnormalities (n = 9). Thyroid ultrasound demonstrated 104 lesions compared with 38 lesions found on physical examination (P < 0.0001). In the subpopulation who underwent scintigraphy (n = 10), 24 nodules were identified by ultrasound and only 10 nodules were identified by scan (P < 0.01). Overall, office-based thyroid ultrasound impacted the clinical management of 40 patients (80%): in 16 patients, thyroid ultrasound was the only modality that demonstrated a multinodular condition, thus contributing to a decision to avoid surgery; 19 patients had ultrasound-guided fine-needle aspiration of vaguely palpable or nonpalpable lesions; and 5 patients underwent ultrasound-guided cyst aspiration and follow-up. Office-based thyroid ultrasound performed by surgeons is a highly accurate imaging modality that identified significantly more lesions than physical examination or scintigraphy. Clinical management was affected through the identification of a multinodular process or through facilitation of accurate image-guided biopsy.

Decreased cisplatin damage-dependent DNA synthesis in cellular extracts of mismatch repair deficient cells.

The proficiency of both nucleotide excision repair (NER) and DNA mismatch repair (MMR) influences cellular sensitivity to cisplatin (cis-diamminedichloroplatinum). To gain further insight into how MMR may influence platinum drug sensitivity, the effect of loss of MMR on repair synthesis was measured in vitro by a commonly used method that relies on whole-cell extracts to drive [alpha-32P]dATP incorporation into cisplatin-damaged plasmid DNA. Extracts evaluated include those from cells with or without functional hMLH1 (HCT116+ch2 versus HCT116+ch3, respectively) and hMSH2 (HEC59 versus HEC59+ch2, respectively). Loss of MMR in the HCT116 system was associated with a 2.8-fold reduction in cisplatin damage-specific DNA synthesis, whereas it was associated with a 3.0-fold reduction in the HEC59 system, suggesting that a decrease in the ability to repair cisplatin-damaged DNA accompanies loss of MMR. An in vitro DNA excision assay that utilized a substrate containing a site-specific cisplatin adduct was performed. Using this highly NER-specific assay, no significant difference was apparent between the extracts derived from NER-proficient versus -deficient cells. These and other data lead us to suggest that the increase in apparent repair synthesis in platinum-damaged plasmids by extracts from MMR-proficient versus -deficient cellular extracts may reflect a distinct and possibly adverse DNA synthetic process rather than productive NER.

Sentinel lymph node biopsy for melanoma.

BACKGROUND: The most powerful predictor of survival for patients with melanoma is the status of the regional lymph nodes. Sentinel lymph node biopsy may provide improved staging accuracy without the morbidity of elective lymph node dissection (ELND). METHODS: Sixty-eight patients with intermediate thickness melanoma underwent gamma probe guided sentinel node biopsy without ELND and were followed up over a mean of 22 months. RESULTS: A sentinel node was found in all patients. Six patients (9%) had positive sentinel nodes; all underwent complete lymphadenectomy. Two patients (3%) with negative sentinel nodes developed nodal recurrence; 1 of these patients was found to have microscopic disease on reexamination of the sentinel node. Two patients (3%) developed systemic disease. CONCLUSION: Gamma probe guided sentinel node biopsy can be performed with a high rate of technical success. It provides accurate pathological staging with a low incidence of nodal basin failure.

Recent insights into platinum drug resistance in cancer.

Cisplatin and its analogs have become important components of chemotherapeutic regimens for the treatment of solid tumors, however, their overall effectiveness is limited by the emergence of drug-resistant tumor cells. Resistance to the platinum drugs is multifactorial consisting of mechanisms that prevent the formation of lethal platinum-DNA adducts and mechanisms that operate downstream of the drug/target interaction to promote cell survival. Continued progress in the study of the drug resistance phenotype as well as the development of new platinum analogs may eventually lead to improved therapies and increased survival rates.

No evidence of hemispheric facilitation following the induction of negative and positive affect.

A dichotomy is thought to exist between the hemispheres whereby the right hemisphere is specialized for the processing of negative emotions and the left hemisphere is specialized for positive emotions. Van Strien and Morpurgo (Neuropsychologia, 1992, 30, 845-848) demonstrated that the activation of negative and positive emotional states resulted in the allocation of attentional resources to the contralateral hemispace. In the present experiment we sought to replicate this effect and improve upon certain methodological features of the experiment. The effect of positive and negative emotions on performance in the left and right visual fields was investigated in 30 dextral students. Positive and negative emotional states were generated by presenting subjects with an emotive word prior to each trial and subsequently requiring them to use that word within a sentence. Performance within the left and right fields was measured using a gap detection task which was neutral in relation to functional asymmetry. No evidence of right or left visual field facilitation was found for the positive or negative conditions, respectively. These results are not interpreted as a refutation of hemispheric specialization for emotional valence. Instead, they are seen to highlight the frailty of hemispheric facilitation effects.

Analysis of breast-tissue cathepsin-d isoforms from patients with breast-cancer, benign breast disease and from normal controls.

The isoform composition of cathepsin D has been investigated by isoelectric focusing of breast tissue supernatant fluids from patients with breast cancer, benign breast disease and from normal controls. The results indicated the presence of several acid protease isoforms between pi values of 2 to 8. Three of these isoforms (with approximate pIs of 6.0, 6.4 and 7.0) were pepstatin-inhibitable but not inhibitable by a mixture of protease inhibitors for seryl, cysteinyl and metalloproteases. These three isoforms, and not the more acidic isoforms, contained a 31 kD protein band which was recognized by polyclonal antibodies against cathepsin D, suggesting that these isoforms are cathepsin D. Further evidence that these isoforms are cathepsin D came from studies in which the pepstatin-inhibitable protease activity and not the pepstatin-uninhibitable protease activity, bound to and was elutable from, an immunoaffinity resin made by coupling anticathepsin D polyclonal antibodies to agarose. The mean relative percentage of the total breast tissue protease activity associated with pepstatin-inhibitable activity (i.e. cathepsin D) was significantly increased (p<0.01) in five breast cancer isoform profiles (64+/-4%, mean+/-S.D.) when compared to five normal control breast profiles (32+/-5%). An analysis of the three cathepsin D isoforms between pIs 6 to 7 indicated a trend of increased relative amounts of the most acidic isoform in the breast cancer isoform profiles when compared to isoform profiles from benign breast disease and normal control breast tissues. Evidence that the more acidic cathepsin D isoforms are related to the more neutral isoforms by sialylation came from studies in which neuraminidase treatment of breast tissue supernatant fluids led to decreased amounts of the most acidic isoform with a concomitant increase in the more neutral isoforms. The apparent increased relative amounts of the most acidic cathepsin D isoform in malignant breast tissue, coupled with the neuraminidase treatment results, provide further evidence that malignant breast tissue cathepsin D is abnormally glycosylated.

Lumbar muscle usage in chronic low back pain. Magnetic resonance image evaluation.

Methods for detecting recruitment patterns of the lumbar muscles during exercise in patients with chronic low back pain are limited. This article discusses the use of magnetic resonance imaging with Roman chair extension exercise to examine lumbar muscle usage in five normal volunteers, five chronic low back pain patients without surgery, and five chronic low back pain patients with surgery. Changes in signal intensities of psoas, multifidus, and longissimus/iliocostalis with graded exercise were measured at three lumbar disc levels. At rest, there was a difference between multifidus and longissimus/iliocostalis signal intensity in chronic low back pain subjects without surgery (P = 0.0162) and in chronic low back pain subjects with surgery (P = 0.0036), but not in normal subjects. At peak exercise, there was a difference in signal intensities between multifidus and longissimus/iliocostalis in all groups (normal volunteers, P = 0.0069; chronic low back pain patients without surgery, P = 0.0125; chronic low back pain patients with surgery, P = 0.0060). The exercise response was attenuated in chronic low back pain patients with surgery. Thus, MRI demonstrates static and dynamic differences in lumbar paraspinal musculature in chronic low back pain subjects compared to normal subjects.

Osteomyelitis: characteristics and pitfalls of diagnosis with MR imaging.

Prospective and retrospective magnetic resonance (MR) imaging (0.35-T) interpretations were compared with final diagnoses in 110 patients suspected to have osteomyelitis. Diagnostic criteria of dark marrow on T1-weighted images and bright marrow on short-tau inversion-recovery images yielded a prospective sensitivity of 98% and a prospective specificity of 75%. Sixty percent of uncomplicated septic joint effusions demonstrated abnormal marrow signal intensity that was mistaken for osteomyelitis. Retrospective review revealed that overall specificity could be improved to 82% without loss of sensitivity if increased marrow signal intensity on T2-weighted images were included as an additional criterion. Specificity may be further increased by use of knowledge of morphologic patterns that distinguish various forms of osteomyelitis. Ten patients (9%) had potential pitfall diagnoses (eg, fracture, infarction, healed infection) that mimic osteomyelitis. MR imaging can be sensitive and specific for osteomyelitis if characteristic appearances and pitfall diagnoses are incorporated into the diagnostic criteria.

Addition of telomere-associated HeT DNA sequences "heals" broken chromosome ends in Drosophila.

Stocks of D. melanogaster X chromosomes carrying terminal deletions (RT chromosomes) have been maintained for several years. Some of the chromosomes are slowly losing DNA from the broken ends (as expected if replication is incomplete) and show no telomere-associated DNA added to the receding ends. Two stocks carry chromosomes that have become "healed" and are no longer losing DNA. In both stocks the broken chromosome end has acquired a segment of HeT DNA, a family of complex repeats found only at telomeres and in pericentric heterochromatin. Although the HeT family is complex, the HeT sequence joined to the broken chromosome end is the same in both stocks. In contrast, the two chromosomes are broken in different places and have no detectable sequence similarity at the junction with the new DNA. Sequence analysis suggests that the new telomere sequences have been added by a specific mechanism that does not involve homologous recombination.