Comparison of Intrathecal Chloroprocaine With Bupivacaine in Short Gynecological Procedures: A Randomized Double-Blind Study.

Background Neuraxial anesthesia, compared to general anesthesia, offers better patient comfort, early ambulation, and discharge with excellent post-operative pain relief for short gynecological procedures. Recently chloroprocaine, a short-acting local anesthetic agent became available for intrathecal use. This study aimed to compare intrathecal chloroprocaine with bupivacaine in short gynecological procedures. Methodology Consecutive patients undergoing short gynecological procedures, patients belonging to the American Society of Anesthesiology (ASA) I and II, between 18 and 60 years of age, and patients with a height between 150 cm and 180 cm were included in the study. Randomization was done using a computer-generated random number table. Patients were allocated to one of the two study groups. Group B received 4 mL of isobaric bupivacaine (0.25%) 10 mg intrathecal, and Group C received 4 mL of isobaric chloroprocaine (1%) 40 mg intrathecal. The primary outcome criteria were time to ambulation and discharge readiness. The secondary outcome criteria were onset, duration, and intensity of sensory and motor blockade, time to voiding, and any adverse effects. Results Patients receiving chloroprocaine had a significantly (p=0.001) faster time (158±31 min) to ambulation compared to bupivacaine (241±23 min). The regression of sensory blockade was substantially faster (p=0.001) with chloroprocaine (60±13 min) than with bupivacaine (94±24 min). Mean time to motor onset was significantly (p=0.05) faster in chloroprocaine (8±3 min) than bupivacaine (12±3 min) group. Significantly faster (p=0.001) recovery of motor blockade was observed with chloroprocaine (130±32 min) than bupivacaine (211±22 min). The time to first voiding was also significantly earlier with stable hemodynamics and no adverse effects in chloroprocaine group. Conclusion Intrathecal chloroprocaine may be an attractive alternative and is superior to isobaric bupivacaine as it provides early ambulation and discharge readiness for daycare anesthesia in short gynecological procedures.

Expanding primary cells from mucoepidermoid and other salivary gland neoplasms for genetic and chemosensitivity testing.

Restricted availability of cell and animal models is a rate-limiting step for investigation of salivary gland neoplasm pathophysiology and therapeutic response. Conditionally reprogrammed cell (CRC) technology enables establishment of primary epithelial cell cultures from patient material. This study tested a translational workflow for acquisition, expansion and testing of CRC-derived primary cultures of salivary gland neoplasms from patients presenting to an academic surgical practice. Results showed that cultured cells were sufficient for epithelial cell-specific transcriptome characterization to detect candidate therapeutic pathways and fusion genes, and for screening for cancer risk-associated single nucleotide polymorphisms (SNPs) and driver gene mutations through exome sequencing. Focused study of primary cultures of a low-grade mucoepidermoid carcinoma demonstrated amphiregulin-mechanistic target of rapamycin-protein kinase B (AKT; AKT1) pathway activation, identified through bioinformatics and subsequently confirmed as present in primary tissue and preserved through different secondary 2D and 3D culture media and xenografts. Candidate therapeutic testing showed that the allosteric AKT inhibitor MK2206 reproducibly inhibited cell survival across different culture formats. By contrast, the cells appeared resistant to the adenosine triphosphate competitive AKT inhibitor GSK690693. Procedures employed here illustrate an approach for reproducibly obtaining material for pathophysiological studies of salivary gland neoplasms, and other less common epithelial cancer types, that can be executed without compromising pathological examination of patient specimens. The approach permits combined genetic and cell-based physiological and therapeutic investigations in addition to more traditional pathologic studies, and can be used to build sustainable bio-banks for future inquiries.This article has an associated First Person interview with the first author of the paper.

HPV positive neuroendocrine cervical cancer cells are dependent on Myc but not E6/E7 viral oncogenes.

Using conditional cell reprogramming, we generated a stable cell culture of an extremely rare and aggressive neuroendocrine cervical cancer. The cultured cells contained HPV-16, formed colonies in soft agar and rapidly produced tumors in immunodeficient mice. The HPV-16 genome was integrated adjacent to the Myc gene, both of which were amplified 40-fold. Analysis of RNA transcripts detected fusion of the HPV/Myc genes, arising from apparent microhomologous recombination. Spectral karyotyping (SKY) and fluorescent-in-situ hybridization (FISH) demonstrated coordinate localization and translocation of the amplified Myc and HPV genes on chromosomes 8 and 21. Similar to the primary tumor, tumor cell cultures expressed very high levels of the Myc protein and, in contrast to all other HPV-positive cervical cancer cell lines, they harbored a gain-of-function mutation in p53 (R273C). Unexpectedly, viral oncogene knockdown had no effect on the growth of the cells, but it did inhibit the proliferation of a conventional HPV-16 positive cervical cancer cell line. Knockdown of Myc, but not the mutant p53, significantly inhibited tumor cell proliferation. On the basis of these data, we propose that the primary driver of transformation in this aggressive cervical cancer is not HPV oncogene expression but rather the overexpression of Myc.