RACE Procedure Research Articles

Abstract 3566: Mohs surgery trends in 104 cases of pediatric melanoma

Abstract Background: Cutaneous melanoma is rare in children with a global incidence of two to five cases per million people per year, accounting for roughly 1% of all pediatric malignancies.5 However, the incidence of pediatric melanoma in the United States is increasing.1 Mohs Micrographic Surgery (MMS) is a standard treatment for adult skin cancer due to its low recurrence rates and tissue preservation. However, it is less commonly used to treat melanoma in children because the required infrastructure is not widely accessible and many pediatric surgeons are not familiar with MMS.4 With more limited treatment options for pediatric melanoma, it is important to question the role of socioeconomic factors that may contribute to MMS treatment of pediatric patients with melanoma.2 Purpose: The purpose of the study is to investigate pediatric patients with melanoma and assess their tumor characteristics by comparing various skin cancer treatments (Mohs, other treatments [local excision], or no treatment).Methods: This is a retrospective cohort study of pediatric patients (age < 21 years) diagnosed with cutaneous Melanoma who received MMS between 2004 and 2020 in the National Cancer Database (NCDB). Descriptive statistics were collected on the following variables: the year of diagnosis, sex, race, tumor stage, tumor behavior, primary site, and surgical procedure. Overall survival was determined via the Kaplan-Meier test. Results: 104 cases of pediatric melanoma treated with MMS were identified. Per statistical analysis, it does not appear that surgical frequency has varied yearly from 2004 to 2020 (r² = 0). The median age of patients analyzed was 18 years old. Most patients were female (64.4%, N = 67), White (100%, N=104), non-Hispanic (100%, N=104), and had private insurance (87.3%, N = 90). 70.2% (N=73) of cases were defined as early-stage (stage 0 or I). 72.1% (N=75) of cases were identified as invasive. 45.2% (N=47) of MMS were performed with a margin of 1 cm or less, 19.2% (N=20) were performed with a margin of 1 cm or more, and the remaining had unspecified margins. The most common primary site for MMS was skin of the trunk (33.7%, N=35), followed by the upper limb and shoulder (16.3%, N=17), and the lower limb and hip (15.4%, N=16). All patients were alive at the last contact, with a median overall survival of 182 months (15.2 years). Discussion: MMS is used commonly in adults to treat a variety of skin tumors that can have aggressive traits such as melanoma. Although it offers low recurrence rates and tissue-sparing treatment in adult populations, MMS frequency has not changed within the pediatric population. MMS is mainly seen in privately insured, White patients with an early stage of melanoma; this is likely due to lower rates and delayed diagnosis in patients with darker skin that would otherwise present with metastatic disease. Given its clinical advantages and low rate of use, further research should address barriers that affect the availability of MMS in the pediatric population. Citation Format: Zachary G. Brandt, Xinxin Wu, Jared Kennard, Julia Griffin, Peter Silberstein. Mohs surgery trends in 104 cases of pediatric melanoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 3566.

Difference in presentation of pediatric and adult ependymoma: An NCDB analysis.

e14055 Background: Ependymomas originate from ependymal cells and account for 6.2% of primary brain tumors in the pediatric population. Adult and pediatric ependymomas are identical in histology, but present in different sites, with intracranial ependymomas seen in children and spinal ependymomas more frequent in adults. The unique presentation of ependymoma in children warrants exploration separate from adults; however, there is currently limited data providing a baseline understanding of pediatric ependymoma and those that it impacts. The National Cancer Database (NCDB) was analyzed to identify common presentations and demographic characteristics of ependymoma in children. Methods: Patients aged 18 and younger with a diagnosis of Ependymoma (ICD 9391-9394) were identified utilizing the National Cancer Database (NCDB) from 2004 – 2020 for a retrospective cohort analysis (N = 2,098). Demographic factors such as age, sex, race, ethnicity, primary site, insurance status, and surgical procedure were analyzed by descriptive statistics. Additionally, identical NCDB data was identified for patients over 18 years old for use as a comparison factor (N = 8,227). Results: A total of 2,098 pediatric patients were identified with an average age of diagnosis of 7.0 years (SD = 5.609, range = ( < 1) - 18 years) with a higher percentage being male (55.9%) than female (44.1%). The vast majority of cases of pediatric ependymoma originate in the brain (91.2%), followed by occurrences on the spinal cord and cranial nerves (8.8%). This differs from ependymoma in adults where only 44.8% of cases occur in the brain and 55.2% occur in the spinal cord. Of the 2,098 pediatric patients identified, they were histologically identified as either Ependymoma, NOS (51.5%), or Anaplastic Ependymoma (47.5%). Comparatively, Anaplastic Ependymoma makes up only 9.7% of ependymomas in adult populations. Additionally, the majority of patients were Non-Hispanic (76.3%) and white (76.9%). Most patients were privately insured (58.0%) followed by those insured by Medicaid (33.1%). Pediatric ependymoma is primarily treated with surgical resection (95.7%). Conclusions: Compared to adult occurrences, pediatric cases of ependymoma are not only unique in the site that they affect but also in histology, with a higher proportion of anaplastic ependymoma. This NCDB analysis is the first to specifically analyze pediatric ependymoma and fills a gap in our understanding of how this cancer presents in children. The majority of ependymoma patients are non-Hispanic and White with a primary site occurring in the brain rather than in the spinal cord, consistent with literature. These findings emphasize the importance of a deeper understanding of the nature of this disorder. Future research should further explore the impact of demographic and socioeconomic factors on the treatments utilized, and overall survival of pediatric patients with ependymoma.

The End of Batson? Rulemaking, Race, and Criminal Procedure Reform

The End of Batson? Rulemaking, Race, and Criminal Procedure Reform

Vascular Invasion Predicts Recurrence in Stage IA2-IB Lung Adenocarcinoma but not Squamous Cell Carcinoma

BackgroundLymphovascular invasion (LVI) is an adverse prognostic feature in resected stage I non-small cell lung cancer (NSCLC); however, it is unclear if the prognostic significance applies to both lung adenocarcinoma (LUAD) and squamous cell carcinoma (LUSC). Materials and MethodsA retrospective review of H&E-stained slides from surgically resected AJCC 8th ed. stage IA2-IB LUAD (n = 344) and LUSC (n = 102) from two institutions was performed. LVI was defined as either lymphatic (LI) or vascular (VI) invasion. Outcomes were assessed by 5-year recurrence-free survival (RFS) estimates using the Kaplan-Meier method. ResultsThe cohorts of LUAD and LUSC showed no significant differences in 5-year RFS (81% each), stage, age, race, or surgical procedure. The presence of LVI, VI, and LI was predictive of 5-year RFS for LUAD (LVI + 71% vs. LVI - 92%, P < 0.001; VI + 64% vs. VI - 90%, P < 0.001; LI + 75% vs. LI - 84%, P = 0.030) but not LUSC (LVI + 84% vs. LVI - 79%, P = 0.740; VI + 83% vs. VI- 80%, P = 0.852; LI + 84% vs. LI - 81%, P = 0.757). Among LUAD with LVI, VI was a stronger predictor of 5-year RFS than the remaining subset of VI-LI + tumors (64% vs. 87%, P = 004). Subset analysis of LI among LUAD stratified by VI showed no significant prognostic advantage to adding LI for risk stratification (VI-LI + 87% vs. VI-LI - 92%, P = 0.347 & VI+LI + 62% vs. VI + LI- 66%, P = 0.422). VI was present in 36% of LUAD. ConclusionVascular invasion is a strong predictor of recurrence in stage IA2-IB LUAD but not in LUSC. Adjuvant therapy trials should be directed at this subgroup.

Frequency and Results of Pregnancy Screening in Critically Ill Adolescents in Multiple Health Care Organizations

PurposeAdolescent females may undergo pregnancy screening while receiving critical care services, but the frequency and results are unknown. The objectives of this study are to evaluate patient characteristics, pregnancy screening frequency, and rate of positive pregnancy screens in adolescent females of childbearing age who require critical care services. We hypothesize that when adolescent pregnancy screening is performed in the critical care setting, it occurs in a higher frequency in older subjects. MethodsThis is a multicenter retrospective observational cohort study utilizing TriNetX, an electronic health record database. The following electronic health record data were collected and evaluated in adolescent females aged 12–18 years and billed for critical care services: age, race, ethnicity, diagnostic codes, selected radiology and surgical procedure codes, number of deaths, pregnancy screening laboratory codes, and pregnancy screening results. ResultsA total of 5,241 subjects (2,242 [42.8%] subjects for whom pregnancy screen was noted and 2,999 [57.2%] subjects for whom it was not noted) were included in this study. Subjects aged 15–18 years (odds ratio = 1.56, 95% confidence interval = 1.38–1.77, p value < .0001) and had Hispanic or Latina ethnicity (odds ratio = 1.46, 95% confidence interval = 1.28–1.66, p value < .0001) had a higher association with pregnancy screening. A positive pregnancy screen was identified in 18 (0.8%) subjects. DiscussionIn our study, positive pregnancy screens were infrequent, not all subjects were screened, and there was an association between pregnancy screening and ethnicity. Because of the potential for screening bias, this study suggests that clinicians should strongly consider routine pregnancy screening for all females of childbearing age and that hospital policies should require this type of screening.

A Virtual Hybrid Care Hotel Model Supports the Recovery of Post-procedural Patients with Mild to Severe Systemic Diseases.

Patients with mild to severe chronic systemic disease undergoing low-risk procedures are often hospitalized for observation. The Care Hotel is a novel virtual medicine hybrid model of care that offers patients a comfortable, out of hospital environment where they can receive both in-person and virtual care after a surgery or procedure. This study aimed to analyze if virtual hybrid post-procedure care in a hotel could be both conducted on and accepted by patients, even those with moderate to severe chronic diseases. This retrospective cohort study was conducted between July 23, 2020 and June 4, 2021 at Mayo Clinic in Florida, a 306-bed community academic hospital. We collected the sex, age, race, ethnicity, acceptance rate, ASA score, and primary procedure of patients using the Care Hotel. Out of 392 patients, 272 (69.4%) opted for care in the program. Median patient age was 61.5 years, 59.56% were males, and 86.40% were white. We found that 50.37% had an ASA score of 2 and 43.4% had an ASA score of 3. Ten different surgical specialties were able to utilize the Care Hotel for care in 47 different procedure types. Urology had the most patients (n=70, 25.7%). Post-electrophysiologic procedures were the most common procedures (n=39, 14.3%). Our virtual hybrid Care Hotel program was widely accepted by patients and could care for a multitude of post-operative procedures. Additionally, this novel program can care for patients with both mild and severe systemic diseases.

Surgical anatomy of the upper esophagus related to robot-assisted cervical esophagectomy.

Robot-assisted cervical esophagectomy (RACE) enables radical surgery for tumors of the middle and upper esophagus, avoiding a transthoracic approach. However, the cervical access, narrow working space, and complex topographic anatomy make this procedure particularly demanding. Our study offers a stepwise description of appropriate dissection planes and anatomical landmarks to facilitate RACE. Macroscopic dissections were performed on formaldehyde-fixed body donors (three females, three males), according to the surgical steps during RACE. The topographic anatomy and surgically relevant structures related to the cervical access route to the esophagus were described and illustrated, along with the complete mobilization of the cervical and upper thoracic segment. The carotid sheath, intercarotid fascia, and visceral fascia were identified as helpful landmarks, used as optimal dissection planes to approach the cervical esophagus and preserve the structures at risk (trachea, recurrent laryngeal nerves, thoracic duct, sympathetic trunk). While ventral dissection involved detachment of the esophagus from the tracheal cartilage and membranous part, the dorsal dissection plane comprised the prevertebral compartment harboring the thoracic duct and right intercosto-bronchial artery. On the left side, the esophagus was attached to the aortic arch by the aorto-esophageal ligament; on the right side, the esophagus was bordered by the azygos vein, right vagus nerve, and cardiac nerves. The stepwise, illustrated topographic anatomy addressed specific surgical demands and perspectives related to the left cervical approach and dissection of the esophagus, providing an anatomical basis to facilitate and safely implement the RACE procedure.

A retrospective study of treatment of genu valgum/varum with guided growth: Risk factors for a lower rate of angular correction.

The rate of angular correction (ROAC) is very unpredictable and may be affected by various factors in the treatment of genu valgum and varum by means of guided growth. The purpose of this study was to assess the ROAC in cases from our institution and to identify risk factors associated with the occurrence of lower ROAC.We retrospectively reviewed the chart records of 68 patients undergoing guided growth with figure-eight plate for the correction of genu valgum and varum. Based on the data from these patients, the annual increment of physeal growth was calculated and compared with data from the Anderson chart. The associations between patient characteristics and ROAC were evaluated with the use of univariate logistic regression.The mean rate of femoral angular correction was 10.29 degrees/year, while the mean rate of tibial angular correction was 7.92 degrees/year. In a univariate logistic regression analysis, the variables associated with a higher risk of lower ROAC included non-idiopathic coronal deformity of the knee (odds ratio = 13.58, p < 0.001) and body weight at or above the 95th percentile for children (odds ratio = 2.69, p = 0.020).Obesity and non-idiopathic coronal deformity of the knee are risk factors for lower ROAC. It is still uncertain whether severity of deformity, race, and operative procedure have a substantial effect on the rate of correction.Level III evidence.

Robot-assisted cervical esophagectomy (RACE procedure) using a single port combined with a transhiatal approach in a rendezvous technique: a case series.

Difficulties in thoracic access and the risk of pulmonary complications are major problems in esophageal surgery. Transhiatal techniques have been described to avoid the thoracic approach, but their oncological radicality continues to be questioned. A combination of a cervical and transhiatal approach, however, appears promising. We describe the technique of a robot-assisted cervical esophagectomy (RACE procedure), combined with a transhiatal approach in a rendezvous technique. The da Vinci Xi® robotic system was docked in a single port technique via a cervical approach. The upper third of the esophagus and the surrounding lymphatic tissue was dissected thoracically. Subsequently, the system was docked abdominally to allow us to completely dissect the esophagus in the rendezvous procedure. The patients (n = 4) suffered no trauma or injury to surrounding structures during the procedure, and sensitive structures were preserved. Almost no robot arm collision occurred, and the arms did not contact the patients' head or shoulders. No patient converted to conventional robotic-assisted transthoracic esophagectomy. Complications included anastomotic leakage (n = 1), transient palsy of the recurrent laryngeal nerve (n = 1), and pneumonia (n = 1). The cervical approach to esophagectomy allows comfortable preparation and facilitates transhiatal access, while the rendezvous procedure enables easy identification of the cranial dissection plane. The degrees of freedom of movement of the robotic instruments allow for precise and controlled preparation, and the latest technology minimizes the risk of robot arm collision in single-excision surgery. This combined, robot-assisted approach appears to be a promising procedure for esophagectomy.

Pediatric perioperative measures of sleep, pain, anxiety and anesthesia emergence: A healing touch proof of concept randomized clinical trial

Background and purposeThe purpose of this study was to determine the impact of healing touch (HT) on sleep, anxiety, anesthesia emergence and pain. MethodsHT, sham HT, control with an aide (CP) and control groups without the presence of an aide (CNP), underwent polysomnography (PSG) preoperatively. The Yale Preoperative Anxiety Scale (YPAS) score was obtained preoperatively before medications were given and in the preoperative surgery area. Sedation score, anesthesia emergence score and vital signs were recorded. Pain scores were determined by the Observation Pain Assessment Scale (OPAS) postoperatively and at time of discharge. Preoperative laboratory blood was drawn for C-reactive protein (CRP), glucose, cortisol and vitamin D25 levels as indicators of stress and anxiety, and a HT satisfaction survey was given. ResultsThirty-nine patients consented to participate and were randomly assigned to HT (9), HT sham (12), CP (7) and CNP (11) groups. Mean patient age was 13.0 years, and no significant group differences were found for age, sex, race or patient procedure, categorized as laser, burn reconstruction and plastic surgery reconstruction. Additionally, no significant group differences were detected for any of the PSG parameters, YPAS scores, OPAS scores, medications, anesthesia emergence score, bloodwork or satisfaction survey score. CRP, glucose and cortisol levels were higher in the CNP group, suggesting that pediatric patients undergoing elective surgeries may benefit from more pre-operative support, possibly by HT. ConclusionsAlthough no tracked parameters showed statistically significant findings, anecdotal HT benefits included enhanced relaxation and sounder sleep.

High hemoglobin A1c associated with increased adverse limb events in peripheral arterial disease patients undergoing revascularization

ObjectiveDiabetes and peripheral arterial disease (PAD) are independently associated with increased risk of amputation. However, the effect of poor glycemic control on adverse limb events has not been studied. We examined the effects of poor glycemic control (high hemoglobin A1c level) on the risk of amputation and modified major adverse limb events (mMALEs) after lower extremity revascularization. MethodsPatients undergoing PAD revascularization who had hemoglobin A1c (HbA1c) levels available within 6 months were identified in the Veterans Affairs database of 2003 to 2014 (N = 26,799). The diagnosis of preoperative diabetes mellitus (PreopDM) was defined using diabetes diagnosis codes and evidence of treatment. Amputation and mMALE risk was compared for HbA1c levels using Kaplan-Meier analysis. Cox proportional hazards models were created to assess the effect of high HbA1c levels on amputation and mMALE (adjusted for age, gender, race, socioeconomic status, comorbidities, cholesterol levels, creatinine concentration, suprainguinal or infrainguinal procedure, open or endovascular procedure, severity of PAD, year of cohort entry, and medications) for all patients and stratified by PreopDM. ResultsHigh HbA1c levels were present in 33.2% of the cohort, whereas 59.9% had PreopDM. Amputations occurred in 4359 (16.3%) patients, and 10,580 (39.5%) had mMALE. Kaplan-Meier curves showed the worst outcomes in patient with PreopDM and high HbA1c levels. In the Cox model, incremental HbA1c levels of 6.1% to 7.0%, 7.1% to 8.0%, and >8% were associated with 26% (hazard ratio [HR], 1.26; 95% confidence interval [CI], 1.15-1.39), 53% (HR, 1.53; 95% CI, 1.37-1.7), and 105% (HR, 2.05; 95% CI, 1.87-2.26) higher risk of amputation, respectively. Similarly, the risk of mMALE also increased by 5% (HR, 1.05; 95% CI, 0.99-1.11), 21% (HR, 1.21; 95% CI, 1.13-1.29), and 33% (HR, 1.33, 95% CI, 1.25-1.42) with worsening HbA1c levels of 6.1% to 7.0%, 7.1% to 8.0%, and >8%, respectively (vs HbA1c ≤6.0%). In stratified analysis by established PreopDM, the relative risk of amputation or mMALE was much higher with poor glycemic control (HbA1c >7.0%) in patients without PreopDM. ConclusionsPAD patients with worse perioperative glycemic control have a significantly higher risk of amputation and mMALE. Incremental increases in HbA1c levels are associated with higher hazards of adverse limb outcomes independent of PreopDM status. Poor glycemic control (HbA1c >7.0%) in patients without a PreopDM diagnosis carries twice the relative risk of amputation and mMALE than in those with good glycemic control. These results suggest that screening of diabetic status and better management of glycemic control could be a target for improvement of perioperative and long-term outcomes in PAD patients.

3' Rapid Amplification of cDNA Ends (3' RACE) Using Arabidopsis Samples.

Production of functional eukaryotic RNA is a very elaborate process that involves a complex interplay between transcription and various RNA processing activities, including splicing, 5' capping, and 3' cleavage and polyadenylation (Bentley, 2014). Accurate mapping of RNA ends provides a valuable tool to assess transcriptional and post-transcriptional events giving rise to different gene transcripts. The abundance of such transcripts most likely depends on exogenous and developmental cues, or mutations. In the reference plant Arabidopsis, perturbation of the HUA-PEP post-transcriptional regulatory factors (Rodríguez-Cazorla et al., 2015) leads to the accumulation of aberrant transcripts of the key floral homeotic gene AGAMOUS (AG) (Yanofsky et al., 1990) that retain intronic sequence. It was determined by 3' RACE reactions that such erroneous transcripts correspond to premature processing and polyadenylation events taking place at the AG intron region. Here we describe a protocol that is suitable for analysis of relatively abundant transcripts and also for detecting aberrant RNA species that are likely prone to rapid turnover. Likewise, the method, here adapted to Arabidopsis reproductive tissues, can be applied to characterize RNA species from other organs (leaf, root) and/or other plant species. We provide a detailed protocol of our 3' RACE procedure comprising four major parts: Total RNA extraction, RNA amount determination and quality control, the RACE procedure itself, and isolation of the resulting RACE products for cloning and sequencing.

Identification and characterization of cold-responsive microRNAs in tea plant (Camellia sinensis) and their targets using high-throughput sequencing and degradome analysis.

BackgroundMicroRNAs (miRNAs) are approximately 19 ~ 21 nucleotide noncoding RNAs produced by Dicer-catalyzed excision from stem-loop precursors. Many plant miRNAs have critical functions in development, nutrient homeostasis, abiotic stress responses, and pathogen responses via interaction with specific target mRNAs. Camellia sinensis is one of the most important commercial beverage crops in the world. However, miRNAs associated with cold stress tolerance in C. sinensis remains unexplored. The use of high-throughput sequencing can provide a much deeper understanding of miRNAs. To obtain more insight into the function of miRNAs in cold stress tolerance, Illumina sequencing of C. sinensis sRNA was conducted.ResultSolexa sequencing technology was used for high-throughput sequencing of the small RNA library from the cold treatment of tea leaves. To align the sequencing data with known plant miRNAs, we characterized 106 conserved C. sinensis miRNAs. In addition, 215 potential candidate miRNAs were found, among, which 98 candidates with star sequences were chosen as novel miRNAs. Both congruously and differentially regulated miRNAs were obtained, and cultivar-specific miRNAs were identified by microarray-based hybridization in response to cold stress. The results were also confirmed by quantitative real-time polymerase chain reaction. To confirm the targets of miRNAs, two degradome libraries from two treatments were constructed. According to degradome sequencing, 455 and 591 genes were identified as cleavage targets of miRNAs from cold treatments and control libraries, respectively, and 283 targets were present in both libraries. Functional analysis of these miRNA targets indicated their involvement in important activities, such as development, regulation of transcription, and stress response.ConclusionsWe discovered 31 up-regulated miRNAs and 43 down-regulated miRNAs in ‘Yingshuang’, and 46 up-regulated miRNA and 45 down-regulated miRNAs in ‘Baiye 1’ in response to cold stress, respectively. A total of 763 related target genes were detected by degradome sequencing. The RLM-5′RACE procedure was successfully used to map the cleavage sites in six target genes of C. sinensis. These findings reveal important information about the regulatory mechanism of miRNAs in C. sinensis, and promote the understanding of miRNA functions during the cold response. The miRNA genotype-specific expression model might explain the distinct cold sensitivities between tea lines.Electronic supplementary materialThe online version of this article (doi:10.1186/s12870-014-0271-x) contains supplementary material, which is available to authorized users.

Identification of underserved areas for urologic cancer care

The delivery of urologic oncology care is susceptible to regional variation. In the current study, the authors sought to define patterns of care for patients undergoing genitourinary cancer surgery to identify underserved areas for urologic cancer care in Washington State. The authors accessed the Washington State Comprehensive Hospital Abstract Reporting System from 2003 through 2007. They identified patients undergoing radical prostatectomy, radical cystectomy (RC), partial nephrectomy (PN), radical nephrectomy, and transurethral resection of the prostate (TURP). TURP was included for comparison as a reference procedure indicative of access to urologic care. Hospital service areas (HSAs) are where the majority of local patients are hospitalized; hospital referral regions (HRR) are where most patients receive tertiary care. The authors created multivariate hierarchical logistic regression models to examine patient and HSA characteristics associated with the receipt of urologic oncology care out of the HRR for each procedure. Greater than one-half of patients went out of their HRR in 7 HSAs (11%) for radical prostatectomy, 3 HSAs (5%) for radical nephrectomy, 10 HSAs (15%) for PN, and 14 HSAs (22%) for RC. No HSAs had high export rates for TURP. Few patient factors were found to be associated with surgical care out of the HRR. High-export HSAs for PN and RC exhibited lower socioeconomic characteristics than low-export HSAs, adjusting for HSA population, race, and HSA procedure rates for PN and RC. Patients living in areas with lower socioeconomic status have a greater need to travel for complex urologic surgery. Consideration of geographic delineation in the delivery of urologic oncology care may aid in regional quality improvement initiatives.

Preoperative Enoxaparin Is Safe to Use in Major Gynecologic Surgery for Prophylaxis of Venous Thromboembolism

To evaluate the safety of preoperative enoxaparin in patients undergoing major gynecologic oncology surgery. We identified a retrospective cohort group of patients undergoing major gynecologic oncology surgery from June 2002 to June 2004. Exclusion criteria included laparoscopic surgery, inferior vena cava filter, history of venous thromboembolism, and current anticoagulation for prior venous thromboembolism. All patients received prophylaxis with sequential pneumatic compression devices and early ambulation. We identified patients who received (preoperative and postoperative) enoxaparin (20-40 mg) and compared them to patients who received no additional prophylaxis other than pneumatic compression alone. Patient outcomes including estimated blood loss, blood transfusions, operative time, and length of hospital stay were collected. Statistical analysis was performed using the χ Wilcoxon rank sum tests. This study was approved by the institutional review board. We identified 122 patients who met our study criteria; there were 63 patients who received preoperative enoxaparin and 59 patients who received no additional prophylaxis. Both groups were similar in age, body mass index, race, comorbidities, cancer diagnosis, and surgical procedure. There was no significant difference between the enoxaparin group and the sequential pneumatic compression devices-only group regarding transfusion rates (29% and 27%; P = 0.86), operating time (150 and 140 minutes; P = 0.16), blood loss greater than 500 cc (35% and 37%; P = 0.79), and length of stay (5 vs 6 days). The use of preoperative enoxaparin is not associated with increased blood loss, transfusion requirements, operative time, or hospital stay among patients having major gynecologic surgery.

Morphological “primary homology” and expression of AG -subfamily MADS-box genes in pines, podocarps, and yews

The morphological variation among reproductive organs of extant gymnosperms is remarkable, especially among conifers. Several hypotheses concerning morphological homology between various conifer reproductive organs have been put forward, in particular in relation to the pine ovuliferous scale. Here, we use the expression patterns of orthologs of the ABC-model MADS-box gene AGAMOUS (AG) for testing morphological homology hypotheses related to organs of the conifer female cone. To this end, we first developed a tailored 3'RACE procedure that allows reliable amplification of partial sequences highly similar to gymnosperm-derived members of the AG-subfamily of MADS-box genes. Expression patterns of two novel conifer AG orthologs cloned with this procedure-namely PodAG and TgAG, obtained from the podocarp Podocarpus reichei and the yew Taxus globosa, respectively-are then further characterized in the morphologically divergent female cones of these species. The expression patterns of PodAG and TgAG are compared with those of DAL2, a previously discovered Picea abies (Pinaceae) AG ortholog. By treating the expression patterns of DAL2, PodAG, and TgAG as character states mapped onto currently accepted cladogram topologies, we suggest that the epimatium-that is, the podocarp female cone organ previously postulated as a "modified" ovuliferous scale-and the canonical Pinaceae ovuliferous scale can be legitimally conceptualized as "primary homologs." Character state mapping for TgAG suggests in turn that the aril of Taxaceae should be considered as a different type of organ. This work demonstrates how the interaction between developmental-genetic data and formal cladistic theory could fruitfully contribute to gymnosperm systematics.

Late steps of ribosome assembly in E. coli are sensitive to a severe heat stress but are assisted by the HSP70 chaperone machine

The late stages of 30S and 50S ribosomal subunits biogenesis have been studied in a wild-type (wt) strain of Escherichia coli (MC4100) subjected to a severe heat stress (45–46°C). The 32S and 45S ribosomal particles (precursors to 50S subunits) and 21S ribosomal particles (precursors to 30S subunits) accumulate under these conditions. They are authentic precursors, not degraded or dead-end particles. The 21S particles are shown, by way of a modified 3′5′ RACE procedure, to contain 16S rRNA unprocessed, or processed at its 5′ end, and not at the 3′ end. This implies that maturation of 16S rRNA is ordered and starts at its 5′-terminus, and that the 3′-terminus is trimmed at a later step. This observation is not limited to heat stress conditions, but it also can be verified in bacteria growing at a normal temperature (30°C), supporting the idea that this is the general pathway. Assembly defects at very high temperature are partially compensated by plasmid-driven overexpression of the DnaK/DnaJ chaperones. The ribosome assembly pattern in wt bacteria under a severe heat stress is therefore reminiscent of that observed at lower temperatures in E. coli mutants lacking the chaperones DnaK or DnaJ.

Nitric oxide synthase expression in the central nervous system of Sepia officinalis: an in situ hybridization study

We recently reported the molecular cloning of nitric oxide synthase (NOS) mRNA from Sepia officinalis (SoNOS) using a strategy that involves hybridization of degenerate PCR primers to highly conserved NOS regions, combined with a RACE procedure. Here, in situ hybridization study has been performed on serial sections of the cuttlefish central nervous system to reveal localized specific staining of cell bodies in several lobes of the brain. Staining was found in many lower motor centres, including cells of the inferior and superior buccal lobes (feeding centres); in some higher motor centres (anterior basal and peduncle lobes); in learning centres (vertical, subvertical and superior frontal lobes); and in the visual system [medulla and deep retina (optic lobe)]. Positive staining was also found in the olfactory lobe. NOS-expressing cells have been detected also in the interbasal lobe. Double labelling experiments, performed on consecutive sections, showed that neurons containing NOS immunoreactivity were also positive in in situ hybridization staining. All these data support the presence of NOS in several systems in the cuttlefish brain.

A high-throughput method for monitoring changes in homeobox gene expression

Members of the homeobox gene superfamily of transcription factors are essential determinants of cellular identity during development. Their regulation and downstream gene targets are not well understood, however. This is due, in part, to the large number of genes that need to be analyzed simultaneously. A method has been developed for the rapid and simultaneous detection of changes in the expression of homeobox-containing genes, including members of the developmentally important Hox gene family. The method selectively amplifies and labels homeobox-containing mRNAs using a 3′-RACE procedure in combination with a degenerate forward primer that targets a highly conserved region of the homeobox found in all vertebrate Hox genes and many non- Hox homeobox-containing genes. The amplified sequences are identified by hybridization to a membrane-based array of covalently bound Hox and non- Hox homeobox gene sequences of interest. The method has been used here to demonstrate previously undetected changes in the expression of homeobox genes during retinoic acid-induced nerve cell differentiation in mouse pluripotent P19 cells.

Expression of genes encoding antimicrobial and bradykinin-related peptides in skin of the stream brown frog Rana sakuraii

Peptidomic analysis of an extract of the skin of the stream brown frog Rana sakuraii Matsui and Matsui, 1990 led to the isolation of a C-terminally α-amidated peptide (VR-23; VIGSILGALASGLPTLISWIKNR·NH 2) with broad-spectrum antimicrobial activity that shows structural similarity to the bee venom peptide, melittin together with two peptides belonging to the temporin family (temporin-1SKa; FLPVILPVIGKLLNGIL·NH 2 and temporin-1SKb; FLPVILPVIGKLLSGIL·NH 2), and peptides whose primary structures identified them as belonging to the brevinin-2 (2 peptides) and ranatuerin-2 (1 peptide) families. Using a forward primer that was designed from a conserved region of the 5′-untranslated regions of Rana temporaria preprotemporins in a 3′-RACE procedure, a cDNA clone encoding preprotemporin-1SKa was prepared from R. sakuraii skin total RNA. Further preprotemporin cDNAs encoding temporin-1SKc (AVDLAKIANIAN KVLSSL F·NH 2) and temporin-1SKd (FLPMLAKLLSGFL·NH 2) were obtained by RT-PCR. Unexpectedly, the 3′-RACE procedure using the same primer led to amplification of a cDNA encoding a preprobradykinin whose signal peptide region was identical to that of preprotemporin-1SKa except for the substitution Ser 18 → Asn. R. sakuraii bradykinin ([Arg 0,Leu 1,Thr 6,Trp 8] BK) was 28-fold less potent than mammalian BK in effecting B2 receptor-mediated relaxation of mouse trachea and the des[Arg 0] derivative was only a weak partial agonist. The evolutionary history of the Japanese brown frogs is incompletely understood but a comparison of the primary structures of the R. sakuraii dermal peptides with those of Tago's brown frog Rana tagoi provides evidence for a close phylogenetic relationship between these species .