Investigation of liver-targeted peripheral focused ultrasound stimulation (pFUS) and its effect on glucose homeostasis and insulin resistance in type 2 diabetes mellitus: a proof of concept, phase 1 trial.

BACKGROUND: Mechanical waves produced by ultrasound pulses have been shown to activate mechanosensitive ion channels and modulate peripheral nerves. However, while peripheral ultrasound neuromodulation has been demonstrated in vitro and in pre-clinical models, there have been few reports of clinical tests. AIM: We modified a diagnostic imaging system for ultrasound neuromodulation in human subjects. We report the first safety and feasibility outcomes in subjects with type 2 diabetes (T2D) mellitus and discuss these outcomes in relation to previous pre-clinical results. DESIGN: The study was performed as an open label feasibility study to assess the effects of hepatic ultrasound (targeted to the porta hepatis) on glucometabolic parameters in subjects with T2D. Stimulation (peripheral focused ultrasound stimulation treatment) was performed for 3 days (i.e. 15 min per day), preceded by a baseline examination and followed by a 2-week observation period. METHODS: Multiple metabolic assays were employed including measures of fasting glucose and insulin, insulin resistance and glucose metabolism. The safety and tolerability were also assessed by monitoring adverse events, changes in vital signs, electrocardiogram parameters and clinical laboratory measures. RESULTS AND CONCLUSION: We report post-pFUS trends in several outcomes that were consistent with previous pre-clinical findings. Fasting insulin was lowered, resulting in a reduction of HOMA-IR scores (P-value 0.01; corrected Wilcoxon signed-rank test). Additional safety and exploratory markers demonstrated no device-related adverse impact of pFUS. Our findings demonstrate that pFUS represents a promising new treatment modality that could be used as a non-pharmaceutical adjunct or even alternative to current drug treatments in diabetes.

Rapid Microbiology Screening in Pharmaceutical Workflows.

Recently advances in miniaturization and automation have been utilized to rapidly decrease the time to result for microbiology testing in the clinic. These advances have been made due to the limitations of conventional culture-based microbiology methods, including agar plate and microbroth dilution, which have long turnaround times and require physicians to treat patients empirically with antibiotics before test results are available. Currently, there exist similar limitations in pharmaceutical sterility and bioburden testing, where the long turnaround times associated with standard microbiology testing drive costly inefficiencies in workflows. These include the time lag associated with sterility screening within drug production lines and the warehousing cost and time delays within supply chains during product testing. Herein, we demonstrate a proof-of-concept combination of a rapid microfluidic assay and an efficient cell filtration process that enables a path toward integrating rapid tests directly into pharmaceutical microbiological screening workflows. We demonstrate separation and detection of Escherichia coli directly captured and analyzed from a mammalian (i.e., CHO) cell culture with a 3.0 h incubation. The demonstration is performed using a membrane filtration module that is compatible with sampling from bioreactors, enabling in-line sampling and process monitoring.

Mesoscale blood cell sedimentation for processing millilitre sample volumes.

We demonstrate the efficient separation of blood cells from millilitre volumes of whole blood in minutes using a simple gravity sedimentation device. Blood cell and plasma separation is often the initial step in clinical diagnostics, and reliable separation techniques have remained a major obstacle for the success of point-of-care or remote diagnostics. Unlike plasma collection devices that rely solely on microchannels that restrict sample volume and throughput, we demonstrate the use of a hybrid micro/mesoscale sedimentation chamber to enable >99% capture of cells from millilitre blood samples in less than two minutes.

Counting single molecules in sub-nanolitre droplets.

We demonstrate single biomolecule detection and quantification within sub-nanolitre droplets through application of Cylindrical Illumination Confocal Spectroscopy (CICS) and droplet confinement within a retractable microfluidic constriction.

Quantum dots in molecular detection of disease.

The unique photophysical properties of semiconductor quantum dots (QDs) have made them ideal for use as spectral labels and luminescent probes. In this review, applications are presented in which QDs function as active participants in nanoscale biosensor assemblies, where replacing traditional molecular fluorophores results in improved assay performance. Specific focus is on disease detection with applications including multiplexed target detection, mutation detection by coincidence analysis and QD-based FRET reporters for miRNA detection and DNA methylation analysis.

Coupling confocal fluorescence detection and recirculating microfluidic control for single particle analysis in discrete nanoliter volumes.

The recent proliferation of platforms designed to handle arrays of nano- and picolitre volumes is in response to the need to perform biological assays on discrete entities, such as single cells. However, a critical challenge associated with this trend for in vitro compartmentalization is the need for highly sensitive, yet low-volume detection platforms. In this paper, we coupled confocal fluorescence detection with recirculating microfluidic control to perform single particle DNA assays within five nL chambers. The performance of this low-volume assay was shown to match that of traditional single molecule detection platforms. However, volume requirements per measurement were nearly 3 orders of magnitude less than conventional systems, enabling future integration with lab-on-a-chip systems that require discrete or digitalized sample processing.

Advances in skeletal tissue engineering with hydrogels.

OBJECTIVES: Tissue engineering has the potential to make a significant impact on improving tissue repair in the craniofacial system. The general strategy for tissue engineering includes seeding cells on a biomaterial scaffold. The number of scaffold and cell choices for tissue engineering systems is continually increasing and will be reviewed. DESIGN: Multilayered hydrogel systems were developed to coculture different cell types and develop osteochondral tissues for applications including the temporomandibular joint. EXPERIMENTAL VARIABLE: Hydrogels are one form of scaffold that can be applied to cartilage and bone repair using fully differentiated cells, adult and embryonic stem cells. OUTCOME MEASURE: Case studies represent an overview of our laboratory's investigations. RESULTS: Bilayered scaffolds to promote tissue development and the formation of more complex osteochondral tissues were developed and proved to be effective. CONCLUSION: Tissue engineering provides a venue to investigate tissue development of mutant or diseased cells and potential therapeutics.

Hyperthermic isolated limb perfusion for extremity sarcomas.

BACKGROUND: The treatment options available for extremity sarcomas are amputation or limb-sparing surgery with radiation, which may incur significant morbidity and body disfigurement. Hyperthermic isolated limb perfusion (HILP) may be an attractive option in extremity sarcomas for unresectable lesions to preserve limb function and maintain quality of life. METHODS: We report the outcomes of 5 patients who underwent HILP for unresectable primary or recurrent extremity sarcomas from 1994 to 2000 at our institution. RESULTS: All patients had initial complete clinical responses to HILP, and the limb was salvaged in 4 of the 5 patients. Complications included chronic lymphedema, neuropathic pain, and prolonged wound healing. CONCLUSIONS: HILP with melphalan is a safe and effective treatment option for selected patients with locally advanced and unresectable extremity sarcomas. The response rates are high, with limb salvage occurring in most patients. Further studies of larger groups of patients are warranted.

Long-term results after a low anterior resection with mucosectomy and colo-anal sleeve anastomosis for a diffuse cavernous haemangioma of the rectum.

Diffuse cavernous hemangioma of the rectum is an unusual lesion. We reporting the case of an 18-year-old man with a rectal cavernous hemangioma in whom recurrent rectal bleeding and marked anemia were thought to be caused by his co-existing internal hemorrhoids. This resulted in a 2-year delay in reaching the correct diagnosis. Digital rectal examination revealed a walnut-sized, wide-based, elastic, soft mass 3 cm proximal to the anal verge. Colonoscopy revealed a bluish, submucosal lesion with superficial capillary dilatation at the same site. Arteriography demonstrated vascular tumours in the territory of the right hypogastric artery and the superior rectal artery. In 1972, Parks and co-workers described resection and colo-anal sleeve anastomosis as an alternative operation in the treatment of this rare malformation. We will describe the clinical presentation, diagnosis, and long-term results in a patient with this condition managed with this surgical technique. The patient has done well without any recurrence of rectal bleeding for over 10 years since his operation. Resection with a colo-anal sleeve anastomosis offers major advantages such as a lower risk of intraoperative bleeding, no risk of damaging the pelvic nerves, sparing of continence and avoidance of a permanent colostomy. It should therefore be considered the treatment of choice for this uncommon condition.

[Acute cholecystitis in patients over 80 years of age: indication for immediate surgical treatment]. / La colecistite acuta nei pazienti ultraottantenni: un'indicazione al trattamento chirurgico immediato.

The Authors analyse the results of very early surgical treatment in 43 patients over 80 years of age. The severity of morphological changes in the gallbladder and complicated course of the disease are the main factors causing an unfavorable effect on the results of a later treatment. Echography allows, in a high percentage of cases, to confirm the clinical doubt of acute cholecystitis. Cholecystectomy was carried out in all the patients within the first 6 hours (very early surgery-VES). The overall mortality rate was 7%; the morbility rate was 18%.

Malignant schwannoma of the rectum: a clinical and pathological contribution.

Schwannomas of the rectum are uncommon and incompletely characterized tumours, and only a limited number of cases have been reported. On the basis of a case of rectal schwannoma and a review of the literature on this rare condition, we stress the importance of the clinical features, diagnostic difficulties and surgical indications for the various therapeutic approaches. The basis for radical operation, due to the tendency of such tumours to recur locally and the real possibility of malignant degeneration, is discussed. We also emphasize the difficulty of making a benign diagnosis with histological certainty.

[Systemic neoadjuvant chemotherapy in locally advanced gastric carcinoma: phase II study with 5-fluorouracil, epirubicin and etoposide]. / Chemioterapia neoadiuvante sistemica nel carcinoma gastrico localmente avanzato: studio in fase II con 5-fluorouracile, epirubicina ed etoposide.

Locally advanced gastric adenocarcinomas have a poor prognosis, particularly when the tumours are bulky, located in the cardia or when they present local/regional lymph node involvement. Neoadjuvant chemotherapy for locally advanced gastric cancer is an experimental treatment strategy that may increase resectability and improve survival in patients suffering from an almost uniformly fatal neoplasm. At our institution 11 patients younger than 70 years of age in good physical and mental condition with non-resectable adenocarcinomas of the stomach as determined by endoscopy, computed tomography scans and pathology examinations, were treated with combination chemotherapy [5-fluorouracil (375 mg/m2 i.v. for 5 days, epirubicin (60 mg/m2 i.v. on day 1), etoposide 80 mg/m2 on days 1, 2 and 3, leucovorin 100 mg/m2 for 5 days] every 4 weeks as neoadjuvant chemotherapy. The response to chemotherapy was evaluated after three courses. After three courses, we had one complete response, 8 partial responses or stable disease, and no response in two cases. One patient was still alive 36 months postoperatively. These preliminary results suggest that this protocol is an effective form of neoadjuvant chemotherapy for locally advanced gastric carcinoma.

[The echo-guided percutaneous treatment of dysontogenetic cysts of the liver]. / Il trattamento percutaneo ecoguidato delle cisti disontogenetiche del fegato.

Congenital cystic disease of the liver include two different entities, in the last years more frequently and easily recognized by ultrasonography or computed tomography: the simple, nonparasitic or biliary or solitary cysts of the liver and the polycystic liver disease. Our experience concern 153 patients affected by nonparasitic hepatic cysts and 17 patients affected by polycystic liver disease. 10/153 patients (6.5%) with symptomatic simple cysts have been treated by percutaneous US-guided aspiration and intracystic instillation of ethanol. All the patients were discharged asymptomatic after 24 hours with no complications; only in 2 patients a second treatment was necessary. Repeated follow-up examination showed non recurrence. The same procedure was also used for 3/17 symptomatic patients affected by polycystic liver disease in order to give temporary relief of the symptoms due to compression (pain, dyspnea, vomit, jaundice). A 45-years-old lady, with previous several surgical fenestrations, was treated for 10 years by 20 sessions of percutaneous aspiration and instillation of ethanol with no complications. We conclude that the percutaneous aspiration and intracystic instillation of ethanol would be recommended as primary treatment of patients affected by non parasitic cysts of the liver, because it offers good relief of the symptoms due to compression with good patients compliance and without complications.

Treatment of cutaneous and subcutaneous tumors with electrochemotherapy using intralesional bleomycin.

BACKGROUND: Electrochemotherapy (ECT) is performed by locally administering a chemotherapeutic agent in combination with electric pulses. Previous clinical studies have demonstrated the effectiveness of ECT. In these initial trials, the drug was administered intravenously, followed by administration of electric pulses directly to the tumor. This study was initiated to determine whether an intralesional injection of the drug in combination with electric pulses could provide an improved result. A group of 34 patients was studied. METHODS: The dose of intralesional bleomycin was based on tumor volume. This was followed 10 minutes later by 6 or 8 99-microsec pulses of electricity at an amplitude of 1.3 kV/cm. Both the bleomycin and the electric pulses were administered after 1% lidocaine with epinephrine solution was injected around the treatment site. RESULTS: All patients responded to the treatment. Responses were observed in 142 (99%) of 143 metastatic nodules or primary tumors within 12 weeks, with complete responses observed in 130 (91%) of the nodules. No complete responses were observed in nodules treated with bleomycin only or electric pulses only. Random biopsies confirmed the clinical findings. All patients tolerated the procedure well, and no significant side effects were noted. Muscle contraction was evident during administration of each electric pulse but promptly subsided after the pulse. CONCLUSIONS: ECT was shown to be an effective local treatment for cutaneous malignancies. The results suggest that ECT may have a tissue-sparing effect and result in minimal scarring. ECT may be a suitable alternative therapy for the treatment of basal cell carcinoma, local or regional recurrent melanoma, and other skin cancers.

A pilot study utilizing intraoperative lymphoscintigraphy for identification of the sentinel lymph nodes in vulvar cancer.

OBJECTIVE: To identify sentinel lymph nodes using intraoperative lymphoscintigraphy. METHODS: Technetium-99-labeled sulfur colloid was injected at the site of primary vulvar carcinoma. An intraoperative gamma counter was used to identify one or more sentinel lymph nodes. RESULTS: Ten patients underwent bilateral inguinal and femoral lymphadenectomy. The clinical stages are as follows: T1 in 6, T2 in 2, and T3 in 2. A total of four groins (3 patients) were positive for metastases. In one patient only the sentinel node was positive for disease. In a second patient, two unilateral nodes were positive for disease and both were identified with the gamma counter as sentinel nodes. In the third patient, a single sentinel node was positive for malignancy in each groin. Multiple nonsentinel lymph nodes were positive in each groin in this patient. In no case was the sentinel node negative when other nonsentinel nodes were positive. CONCLUSION: Intraoperative lymphoscintigraphy quantitatively identifies one or more sentinel lymph nodes. Since sentinel lymph nodes can be localized transcutaneously, this technique may be useful for selective lymphadenectomy. Larger patient accrual is necessary to verify this technique.

The clinical relevance of sentinel lymph nodes identified with radiolymphoscintigraphy.

BACKGROUND: The purpose of this case report is to illustrate the utility of radio-guided mapping of sentinel lymph nodes (SLN's) as demonstrated by the technique's successful identification of nodes containing metastatic disease that would have been left behind if only the visual-oriented vital blue dye mapping technique had been used. METHOD: The patient underwent preoperative lymphoscintigraphy and intra-operative lymphatic mapping using vital blue dye and radiolymphoscintigraphy using the Neoprobe (handheld gamma probe). Nodes which were blue and/or "hot" (i.e., radioactive counts were three times the background count) were considered SLN's. RESULTS: Four SLN's were harvested, all of which were "hot" but only one of which was both "hot" and blue. Pathology revealed that the two SLN's positive for metastatic disease were not blue. CONCLUSION: While the blue dye lymphatic mapping technique provides the surgeon with a visual road map in the identification of SLN's, the Neoprobe increases the success rate of localization when compared to vital blue dye mapping due to the reliable migration of radiocolloid to the SLN's in the regional basin. Radiolymphoscintigraphy also increases the accuracy and efficiency of the SLN harvest by providing a directed dissection to the level of the nodes in the basin. The Neoprobe increases the yield of SLN's, some of which are clinically relevant since they contain metastatic disease.

Redefining cutaneous lymphatic flow: the necessity of preoperative lymphoscintigraphy in the management of malignant melanoma.

OBJECTIVE: The purpose of this study is to emphasize the instrumental role of preoperative lymphoscintigraphy in the surgical treatment of patients with malignant melanoma. SUMMARY BACKGROUND DATA: The efficacy of lymphoscintigraphy is reflected in its ability to reveal cutaneous lymphatic drainage to regional nodal basins that are at risk for melanoma metastases but not necessarily discernable to be at risk through standard historical anatomical guidelines or clinical experience. This preoperative lymphatic mapping technique has contributed greatly to the accuracy and efficiency of staging procedures including sentinel node biopsy and elective lymph node dissection. PATIENTS AND METHODS: After informed consent, a selected series of four patients with primary melanomas located in watershed areas of the body (left neck, right mid-abdomen, right scapula, left back) and two patients with extremity melanomas (right distal forearm and left ankle) underwent pre-operative lymphoscintigraphy to identify all basins for metastases. RESULTS: In all of the cases, lymphatic drainage occurred in an unusual and unexpected basin that could not have been predicted clinically and in three of the cases the resected basins contained positive sentinel nodes. If not for the preoperative lymphoscintigraphy, these nodal basins would not have been resected and metastatic disease would have been left behind. In addition, the staging of the melanoma patient would have been inaccurate. CONCLUSION: If the sentinel node biopsy of elective lymph node dissection (ELND) were based on clinical predictions only, nodes equally at risk for metastatic disease would not have been resected and in some cases, nodal basins not at risk for metastases would have been resected unnecessarily. Without lymphoscintigraphy, the validity and efficacy of the ELND or the sentinel node biopsy for nodal staging is greatly compromised. These six case studies illustrate the difficulty of predicating lymphatic drainage from primary sites located on the head and neck, truck and even the extremities and demonstrate the indispensability of preoperative lymphoscintigraphy in the management of malignant melanoma.