The detection and analysis of circulating tumor cells (CTCs) may enable a broad range of cancer-related applications, including the identification of acquired drug resistance during treatments. ...However, the non-scalable fabrication, prolonged sample processing times, and the lack of automation, associated with most of the technologies developed to isolate these rare cells, have impeded their transition into the clinical practice. This work describes a novel membrane-based microfiltration device comprised of a fully automated sample processing unit and a machine-vision-enabled imaging system that allows the efficient isolation and rapid analysis of CTCs from blood. The device performance was characterized using four prostate cancer cell lines, including PC-3, VCaP, DU-145, and LNCaP, obtaining high assay reproducibility and capture efficiencies greater than 93% after processing 7.5 mL blood samples spiked with 100 cancer cells. Cancer cells remained viable after filtration due to the minimal shear stress exerted over cells during the procedure, while the identification of cancer cells by immunostaining was not affected by the number of non-specific events captured on the membrane. We were also able to identify the androgen receptor (AR) point mutation T878A from 7.5 mL blood samples spiked with 50 LNCaP cells using RT-PCR and Sanger sequencing. Finally, CTCs were detected in 8 out of 8 samples from patients diagnosed with metastatic prostate cancer (mean ± SEM = 21 ± 2.957 CTCs/mL, median = 21 CTCs/mL), demonstrating the potential clinical utility of this device.
The detection and analysis of circulating tumor cells (CTCs) may enable a broad range of cancer-related applications, including the identification of acquired drug resistance during treatments. ...However, the non-scalable fabrication, prolonged sample processing times, and the lack of automation, associated with most of the technologies developed to isolate these rare cells, have impeded their transition into the clinical practice. This work describes a novel membrane-based microfiltration device comprised of a fully automated sample processing unit and a machine-vision-enabled imaging system that allows the efficient isolation and rapid analysis of CTCs from blood. The device performance was characterized using four prostate cancer cell lines, including PC-3, VCaP, DU-145, and LNCaP, obtaining high assay reproducibility and capture efficiencies greater than 93% after processing 7.5 mL blood samples from healthy donors, spiked with 100 cancer cells. Cancer cells remained viable after filtration due to the minimal shear stress exerted over cells during the procedure, while the identification of cancer cells by immunostaining was not affected by the number of non-specific events captured on the membrane. We were also able to identify the androgen receptor (AR) point mutation T878A from 7.5 mL blood samples spiked with 50 LNCaP cells using RT-PCR and Sanger sequencing. Finally, CTCs were detected in 8 of 8 samples from patients diagnosed with metastatic prostate cancer (mean \(\pm\) SEM = 21 \(\pm\) 2.957 CTCs/mL, median = 21 CTC/mL), thereby validating the potential clinical utility of the device.
Small bowel obstruction (SBO) is one of the most common causes of hospitalization and internal hernias represent a very rare etiology (0.2-0.9%). The most common manifestations include abdominal ...pain, nausea, and vomiting. SBO can progress to strangulated intestinal obstruction (SIO), a serious condition that requires early diagnosis and emergency surgery. We report the case of a SIO with septic shock in a 73-year-old female with comorbidities and a history of previous surgeries, which required emergency surgery with intestinal resection and discharge of the patient without complications.
An appendicovesical fistula is defined as an abnormal communication between the appendix and the urinary bladder, with only a few cases reported in the literature. It is very challenging to make an ...early diagnosis, due to the inability of conventional imaging modalities to detect this unique pathology. The symptoms are often mild, and there are not any specific signs or symptoms that might suggest this type of anomalous communication. We report a case of a 27-year-old male patient who presented difficulty for initiating urination, dysuria, and persistent urinary tract infections. An abdominal x-ray showed a large calculus inside the bladder. A cystoscopy was performed, where the tip of the appendix was seen protruding inside the bladder with a large fecalith adhered to the bladder wall. An appendectomy and partial cystectomy with primary repair were auspiciously achieved. A review of the literature is also presented.
OBJECTIVESeveral studies suggest that pirfenidone may have a potential off-label use for wound healing. However, the effectiveness of this medication in patients with burns remains uncertain. ...Accordingly, investigators sought to assess wound re-epithelialization in patients with second-degree burns after adding pirfenidone to usual care.
DESIGN AND SETTINGSingle-center pilot, proof-of-concept, single-blind randomized controlled trial.
PATIENTS AND INTERVENTIONEight patients with second-degree burns were treated with occlusive hydrocolloid dressings and were randomly allocated to receive either no additional treatment or pirfenidone.
OUTCOME MEASURESThe primary outcome of the study was to evaluate wound healing between groups based on the thickness of the re-epithelialized epidermis at day 7. Secondary outcomes were to qualitatively assess the development of fibrotic tissue in the dermis, anomalies in the basal membrane, and the development of collagen fibers by histologic analysis. Liver and renal functions were measured daily to assess the overall safety of oral pirfenidone.
MAIN RESULTSPatients treated with pirfenidone showed a remarkable improvement in wound re-epithelialization at day 7 (148.98 ± 13.64 vs 119.27 ± 15.55 μm; P = .029; 95% confidence interval, 4.14-55.29). Histologic evaluations showed less wound fibrosis in the pirfenidone group.
CONCLUSIONSA decrease in wound healing time by enhancing wound re-epithelialization was observed with pirfenidone. Larger clinical trials are needed to reach more reliable conclusions.
Glutathione S-transferases (GSTs) are a group of phase II detoxification enzymes, which catalyze the conjugation of glutathione (GSH) with carcinogens, among other xenobiotics. The GSTM3 gene is part ...of the GSTs gene family, and its polymorphism A/B has been associated with risk and protective effects of several cancers. This genetic variant is a deletion of 3 bp (AGG) in intron 6. Previous association studies have performed genotyping using techniques such as polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). In this study, we took advantage of the TaqMan® probes features and developed a reliable, faster, more simple and economic method to identify the 3-bp deletion. Our allelic discrimination method was able to distinguish between homozygous A/A , heterozygous A/B and homozygous B/B samples, as shown by TaqMan® based real-time PCR. Results were validated by Sanger Sequencing. In conclusion, we developed a specific and rapid method to detect the 3-bp deletion from the GSTM3 A/B polymorphism.
Currently, ultrasound-assisted liposuction has been widely used under empirical criteria. To date, no exhaustive description of the histological effects that are exerted during Vibration ...Amplification of Sound Energy at Resonance (VASER) application has been reported. Here, we describe the histological alterations in the skin of lipectomy flaps after using the VASER technology at different power settings.Four lipectomy flaps were resected and treated with VASER at 50% or 100% power for 1 minute in duplicate. Therefore, each experimental condition was tested on 8 tissue samples. We described the histopathological alterations in the epidermis, dermis, and hypodermis. The observations were performed by 2 independent morphologists unrelated to the lipectomy and treatment procedures. In the histopathological analysis, we did not find significant differences in the histopathological alterations of the epidermis, dermis, and hypodermis using both power settings. However, the dermis of the flaps subjected to 100% power showed considerable collagen fibers degradation and more adipocytes with ruptured membranes in comparison with the flaps treated with 50% power. Although we did not find significant differences between the 2 power conditions tested in this study, more histological alterations in the dermis were observed with the maximum power setting even at a “safe” distance reported in the conventional technique. However, this study was the first approach to determine the histological effects of VASER application, therefore, these results must be corroborated in further studies with a greater number of lipectomies, as well as more specific histopathological analyses. Finally, the data shown in this study reflect the importance of optimizing VASER procedures to increase the safety and efficacy of this technique.
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