Recommended Resting-State fMRI Acquisition and Preprocessing Steps for Preoperative Mapping of Language and Motor and Visual Areas in Adult and Pediatric Patients with Brain Tumors and Epilepsy.

Resting-state (rs) fMRI has been shown to be useful for preoperative mapping of functional areas in patients with brain tumors and epilepsy. However, its lack of standardization limits its widespread use and hinders multicenter collaboration. The American Society of Functional Neuroradiology, American Society of Pediatric Neuroradiology, and the American Society of Neuroradiology Functional and Diffusion MR Imaging Study Group recommend specific rs-fMRI acquisition approaches and preprocessing steps that will further support rs-fMRI for future clinical use. A task force with expertise in fMRI from multiple institutions provided recommendations on the rs-fMRI steps needed for mapping of language, motor, and visual areas in adult and pediatric patients with brain tumor and epilepsy. These were based on an extensive literature review and expert consensus.Following rs-fMRI acquisition parameters are recommended: minimum 6-minute acquisition time; scan with eyes open with fixation; obtain rs-fMRI before both task-based fMRI and contrast administration; temporal resolution of ≤2 seconds; scanner field strength of 3T or higher. The following rs-fMRI preprocessing steps and parameters are recommended: motion correction (seed-based correlation analysis [SBC], independent component analysis [ICA]); despiking (SBC); volume censoring (SBC, ICA); nuisance regression of CSF and white matter signals (SBC); head motion regression (SBC, ICA); bandpass filtering (SBC, ICA); and spatial smoothing with a kernel size that is twice the effective voxel size (SBC, ICA).The consensus recommendations put forth for rs-fMRI acquisition and preprocessing steps will aid in standardization of practice and guide rs-fMRI program development across institutions. Standardized rs-fMRI protocols and processing pipelines are essential for multicenter trials and to implement rs-fMRI as part of standard clinical practice.

Glutamine Imaging: A New Avenue for Glioma Management.

The glutamine pathway is emerging as an important marker of cancer prognosis and a target for new treatments. In gliomas, the most common type of brain tumors, metabolic reprogramming leads to abnormal consumption of glutamine as an energy source, and increased glutamine concentrations are associated with treatment resistance and proliferation. A key challenge in the development of glutamine-based biomarkers and therapies is the limited number of in vivo tools to noninvasively assess local glutamine metabolism and monitor its changes. In this review, we describe the importance of glutamine metabolism in gliomas and review the current landscape of translational and emerging imaging techniques to measure glutamine in the brain. These techniques include MRS, PET, SPECT, and preclinical methods such as fluorescence and mass spectrometry imaging. Finally, we discuss the roadblocks that must be overcome before incorporating glutamine into a personalized approach for glioma management.

Ambulatory photodynamic therapy: a new concept in delivering photodynamic therapy.

BACKGROUND: Photodynamic therapy (PDT) has been shown to be effective in treating Bowen's disease, superficial basal cell carcinoma and actinic keratosis. OBJECTIVES: To investigate the feasibility of delivering PDT using a portable light-emitting diode device. METHODS: A prototype diode array, comprising 37 AlGaInP diodes cast in epoxy with a diffuser, and driven by a battery pack, was designed and constructed. A pilot study was carried out in five patients with histologically proven Bowen's disease who were referred for PDT with 5-aminolaevulinic acid. They were all treated in the hospital-based dermatology PDT suite such that each received the same level of supervision as patients receiving PDT with nonambulatory light sources. Patients recorded pain levels. In accordance with our usual practice, patients received two treatments at a 4-week interval. RESULTS: Four of five patients were clear at follow-up (range 6-13 months, median 9). Pain was classified as none or mild in 80% of treatments and moderate in the remainder. CONCLUSIONS: There are many potential benefits of ambulatory PDT, including the possibility of a much higher patient throughput, and allowing effective treatment at home. This pilot study provides early promising data of the safety and efficacy of this approach.

The Escherichia coli cytochrome c maturation (Ccm) apparatus can mature cytochromes with an extra cysteine within or adjacent to the CXXCH motif.

c-Type cytochromes are characterized by covalent attachment of haem to protein through thioether bonds between the vinyl groups of the haem and the thiols of a Cys-Xaa-Xaa-Cys-His motif. Proteins of this type play crucial roles in the biochemistry of the nitrogen cycle. Many Gram-negative bacteria use the Ccm (cytochrome c maturation) proteins for the post-translational haem attachment to their c-type cytochromes. The Ccm system can correctly mature c-type cytochromes with CCXXCH, CCXCH, CXCCH and CXXCHC motifs, even though these are not found naturally and the extra cysteine might, in principle, disrupt the biogenesis proteins. The non-occurrence of these motifs probably relates to the destructive chemistry that can occur if a free thiol reacts with haem iron to generate a radical.

What is the substrate specificity of the System I cytochrome c biogenesis apparatus?

c-Type cytochromes are characterized by covalent attachment of haem to protein through thioether bonds between the vinyl groups of the haem and the thiols of a CXXCH motif. Proteins of this type play crucial roles in the biochemistry of the nitrogen cycle. Many Gram-negative bacteria use the Ccm (cytochrome c maturation) proteins for the post-translational haem attachment to their c-type cytochromes; in the present paper, we discuss the substrate specificity of the Ccm apparatus. The main conclusion is that the feature recognized and required in the apocytochrome is simply the two cysteines and the histidine of the haem-binding motif.

Favorable early results of gastric banding for morbid obesity: the American experience.

BACKGROUND: In 2001 a new device for surgical weight loss was approved by the Food and Drug Administration (Lap-Band, Inamed Health). We describe initial results of laparoscopic gastric banding for morbid obesity in two American academic centers. METHODS: Prospective data was collected on consecutive morbidly obese patients undergoing laparoscopic adjustable gastric banding, and evaluated retrospectively. RESULTS: Four hundred forty-five consecutive patients underwent Lap-Band from May 2001 through December 2002. The 103 men and 341 women had an average age of 42.1 years (range 17-72 years) and an average body mass index (BMI) of 49.6 kg/m2 (range 35.2-92.2 kg/m2). One operation required conversion to laparotomy due to bleeding; the rest were completed laparoscopically. Mean length of stay was 1.1 days (range 1-10 days). There was one death. Additional complications included band slippage in 14 patients (3.1%), gastric obstruction without slip in 12 (2.7%), port migration in 2 (0.4%), tubing disconnections in 3 (0.7%), and port infection in 5 (1.1%). Two bands (0.4%) were removed due to intraabdominal abscess 2 months after placement. There was one band erosion (0.2%) and no clinically significant esophageal dilation. Ninety-nine patients have 1-year follow-up and have lost an average of 44.3% excess body weight. CONCLUSION: Laparoscopic gastric banding has much to offer the morbidly obese. We present data showing weight loss rivaling gastric bypass and acceptably low complications. These results parallel success with this device outside America.

Laparoscopic management of Meckel's diverticulum in adults.

BACKGROUND: Meckel's diverticulum is an uncommon entity. A high index of suspicion is necessary for opportune diagnosis and prompt treatment. Technetium (TC) 99m pertechnetate scintigraphy is a sensitive and specific test for Meckel's diverticulum. In adults, the scan contributes little to clinical decision making and often will not change the need for surgical intervention. We describe our experience with four patients. METHODS: Between August 2000 and August 2001, four patients were seen with Meckel's diverticula. Three were male and one was female. The mean age was 39 years (range, 18-64). Three patients presented with anemia and one with an acute abdomen. A 99mTc pertechnetate scan was performed at a cost of 900 dollars in the three anemic patients after other endoscopic and radiographic tests were nondiagnostic. Only one patient had a positive scan. All four patients underwent exploratory laparoscopy and small bowel resection. In one patient, a minilaparatomy had to be performed. RESULTS: All patients had a satisfactory outcome without complications. Three patients were discharged within 3 days of surgery. The remaining patient had a prolonged hospital stay because of ongoing chemotherapy for small cell lung cancer. In the three anemic patients who underwent enterectomy, ulcerated small bowel outside the diverticulum was found by the pathologist. CONCLUSION: Laparoscopy is safe, cost-effective, and efficient for the diagnosis and definitive management of Meckel's diverticulum. Technetium 99m pertechnetate scintigraphy scanning adds considerable time and expense to the care of the patient without significant benefits in adults. The practice of exploratory laparoscopy rather than scintigraphy is recommended.

Totally laparoscopic management of gallstone ileus.

Gallstone ileus is an uncommon disease of elderly patients who present with bowel obstruction. Mortality and severe complications are common, even in modern series, due to the comorbidities in the affected patient population. A number of less invasive ways to treat this disease are described. We report on a case where enterolithotomy was performed laparoscopically. The patient is a 60-year-old diabetic woman who presented with a bowel obstruction and pneumobilia on abdominal radiographs. She underwent exploratory laparoscopy using three reusable ports, an enterolithotomy, and her remaining bowel was examined. The benefits to a minimal access approach to this rare disorder are discussed. The ability to suture laparoscopically is emphasized.

Laparoscopic choledochojejunostomy and gastrojejunostomy in a porcine model.

BACKGROUND: Surgical extirpation remains the only known curative treatment for cancer of the pancreas. Because of locally advanced or metastatic tumor, up to 80% of patients are unresectable at the time of initial diagnosis [13]. Other investigators previously have suggested that laparoscopy before laparotomy aids in the diagnosis of unresectable pancreatic cancer in a fair number of patients even after negative computed tomography scans [3, 17]. Many surgeons are reluctant to incorporate laparoscopy into the workup of patients with cancer of the pancreas because of the frequent need for surgical bypass in the management of either biliary tract obstruction or gastric outlet obstruction [9, 13]. Previous studies have demonstrated the feasibility of laparoscopic cholecystojejunostomy combined with gastrojejunostomy in a porcine model, as well as the individual accomplishment of laparoscopic choledochojejunostomy. The purpose of this study was to document the feasibility of performing laparoscopic choledochojejunostomy with gastrojejunostomy. METHODS: Under general anesthesia, seven pigs underwent laparoscopic choledochojejunostomy and gastrojejunostomy using an intracorporeal hand-sutured technique. RESULTS: The mean operating time ranged from 150 to 450 min. All the animals recovered completely from the operation and had patent anastomoses at the time of necropsy. One pig died of gastric bleeding on postoperative day 13, and two animals had intraabdominal fluid collections discovered at the time of necropsy. CONCLUSIONS: These results suggest that synchronous laparoscopic bypass of biliary and gastric outlet obstruction is feasible, and can be performed in a manner similar to that used in open operations. We believe this lends support to the argument promoting laparoscopy in the evaluation of pancreatic cancer.

Effects of heat shock protein 70 (Hsp70) on arsenite-induced genotoxicity.

Arsenic, a human carcinogen, is genotoxic, although its mechanism(s) of action for tumorigenesis is not well understood. Among the toxicity-related properties of this chemical are its clastogenic and aneugenic activities, as well as its capacity for inducing stress-response in the form of elevated heat shock protein (HSP) expression. In the present study, we evaluated the effects of Hsp70 expression on arsenite (As)-induced structural and numerical chromosome anomalies in human cells. Human MCF-7 Tet-off cells stably transfected with a pTRE/Hsp70-1 transgene construct were used to regulate Hsp70 levels prior to in vitro As exposures. Separate cultures of relatively high vs. low Hsp70-expressing cells were established. A cytokinesis block micronucleus assay with kinetochore immunostaining was used to detect micronuclei (MN) derived from chromosome breakage (K-MN) or loss (K+MN). These studies demonstrated significant increases in micronucleus frequencies in response to As following either a long exposure (5 or 10 microM for 46 hr), or short exposure (10 or 40 microM for 8 hr) protocol. Overall, the long protocol was more efficient in producing K+MN and cells with multiple MN. Overexpressing Hsp70 resulted in significant reductions in the percent of cells positive for MN for both the long and short As exposure protocols. Both K+ and K- types of As-induced MN were lower in cells with elevated Hsp70 as compared to cells without overexpression of Hsp70. We conclude that the dose and duration of As exposure influence the type as well as amount of chromosomal alteration produced and that inducible Hsp70 protects against both the clastogenic and aneugenic effects of this chemical.

Totally laparoscopic feeding jejunostomy.

BACKGROUND: A feeding jejunostomy should be used for nutritional support in a small subset of patients. Minimal-access approaches for the placement of jejunal tubes have been described, but they often require special equipment not common to all operating theaters. We describe a technique of totally laparoscopic jejunostomy tube (LJT) placement using equipment found in most operating theaters. METHODS: Thirty-five patients underwent LJT over a 12-month period. Indications included gastroparesis, anorexia nervosa, oral cancer, cerebral palsy, and Huntington's chorea. The technique involved three incisions for trocars (one for a 10-mm camera and two for 5-mm working ports) and one small incision for the tube. A 16-Fr T-tube was passed transabdominally under direct vision, and a jejunotomy was made approximately 20 cm distal to the ligament of Trietz. Each limb of the T-tube was passed into the lumen of the bowel, and a purse-string suture was placed around the enterotomy and tied intracorporeally. After insertion, the serosa surrounding the insertion site is tacked to the anterior abdominal wall in four places with a reusable stainless steel suture passer. To test whether the tube was watertight, we injected methylene blue solution into the tube. RESULTS: All of the patients tolerated the procedure well. There were no operative deaths. Five LJTs were electively removed in the office. One patient was reoperated on 10 days postoperatively because of intractable pain, but the source of pain was not found and the LJT was intact. CONCLUSIONS: LJT may be placed safely using the described technique. No significant morbidity or mortality occurred in our series. The results of this study have prompted us to consider LJT for any patient requiring access to the jejunum for feeding.

The poor quality of information about laparoscopy on the World Wide Web as indexed by popular search engines.

BACKGROUND: This study was undertaken to determine the quality of information on the Internet regarding laparoscopy. METHODS: Four popular World Wide Web search engines were used with the key word "laparoscopy." Advertisements, patient- or physician-directed information, and controversial material were noted. RESULTS: A total of 14,030 Web pages were found, but only 104 were unique Web sites. The majority of the sites were duplicate pages, subpages within a main Web page, or dead links. Twenty-eight of the 104 pages had a medical product for sale, 26 were patient-directed, 23 were written by a physician or group of physicians, and six represented corporations. The remaining 21 were "miscellaneous." The 46 pages containing educational material were critically reviewed. At least one of the senior authors found that 32 of the pages contained controversial or misleading statements. All of the three senior authors (LKN, NAO, GAF) independently agreed that 17 of the 46 pages contained controversial information. CONCLUSION: The World Wide Web is not a reliable source for patient or physician information about laparoscopy. Authenticating medical information on the World Wide Web is a difficult task, and no government or surgical society has taken the lead in regulating what is presented as fact on the World Wide Web.

Primary skeletal muscle lymphoma presenting as refractory cellulitis.

The right torso of a 55-year-old woman showed diffuse skin and soft-tissue changes suggestive of cellulitis. However, several clinical and radiologic features, including the subacute and non-toxic nature of the illness and the patient's lack of response to antibiotic therapy, indicated a noninfectious etiology. Malignancy was suggested by striking changes seen on computed tomographic scanning--including extensive infiltration and enlargement of the musculature of the right shoulder girdle, the intercostal musculature, the latissimus dorsi, and the rhomboids; focal enlargement of the right paraspinal muscles; and enlargement of the psoas and the iliacus muscles and of the musculature around the hip joint. The mediastinal, hilar, and paraaortic regions showed no adenopathy. A large hypodense lesion of approximately 4.5 cm, which was seen in the caudate lobe of the liver, raised the concern of a metastatic focus of malignancy. Because of these findings, an immediate muscle biopsy was performed. Results showed a non-Hodgkin's lymphoma with a B-cell phenotype. Although primary skeletal muscle lymphoma is very uncommon in patients without human immunodeficiency virus infection, clinical presentation of refractory cellulitis, as seen in the current case, is extremely rare.

Methylmercury inhibits cysteine uptake in cultured primary astrocytes, but not in neurons.

The maintenance of adequate intracellular glutathione (GSH) concentrations is dependent on the availability and transport of the rate-limiting substrate, cysteine. A suggested mechanism of methylmercury (MeHg) neurotoxicity in brain involves the formation of oxygen radicals, and a decrease in intracellular levels of GSH. Recently, we have characterized various cysteine transport systems (both Na(+)-dependent and -independent) in cerebrocortical astrocytes and hippocampal neurons. The present study was carried out to investigate the effect of MeHg on cysteine uptake in both astrocytes and neurons, and to determine whether cysteine transport is differentially affected in the two cell types by MeHg treatment. Sixty-minute pretreatment with MeHg caused significant concentration-dependent inhibition in cysteine uptake in astrocytes, but not in neurons. As most of the cysteine transport is Na(+)-dependent (80-90% of total), additional studies focused on MeHg's effect on the Na(+)-dependent cysteine transporters X(AG(-)) and ASC. An additive inhibitory effect on cysteine uptake was observed in astrocytes treated with MeHg (5 microM) plus sub-maximal inhibitory concentrations (0.1 and 0.5 mM) of threo-beta-hydroxy-aspartate (THA), a specific inhibitor of the Na(+)-dependent transporter, X(AG(-)), compared to astrocytes treated with MeHg (P<0.001) or THA alone (P<0.05). There was no additive effect of MeHg and maximal inhibitory concentrations of THA (1.0 and 5.0 mM) on astrocytic cysteine uptake inhibition. Additional studies examined the sensitivity of the Na(+)-dependent ASC transport system to MeHg treatment. Maximal inhibitory concentration of L-serine (10 mM) alone had a rather modest inhibitory effect on cysteine uptake, and when applied in the presence of MeHg there was no additive effect. These results suggest that the inhibition of cysteine uptake by MeHg in astrocytes occurs through specific inhibition of both the X(AG(-)) as well as the ASC transport system.

Lessons learned from laparoscopic gastric banding for morbid obesity.

BACKGROUND: Laparoscopic gastric banding is a minimally invasive bariatric operation that is increasing in popularity at many centers worldwide. Although this procedure is not yet approved in the United States, clinical trials are ongoing. METHODS: We report our results of a 3-year follow-up on 60 patients who underwent the laparoscopic gastric band procedure for the treatment of morbid obesity. The procedure was performed at the Wesley Obesity Clinic in Brisbane, Australia. RESULTS: At follow-up, 51 of the 60 patients (85%) still had the laparoscopic gastric band in place. All of the patients had a lower body weight after undergoing the procedure. The average weight loss was 39 kg (range 2 to 98 kg), representing a loss of 65% of average excess body weight. Twenty-five of 51 patients (49%) regained some weight after their initial loss, but the average amount was only 5 kg. The remaining 26 patients have remained at their lowest body weight recorded after the procedure or are continuing to lose weight. There was no operative mortality. Complications predominantly were caused by band slippage (21%), which has been nearly eliminated in recent practice (1 slip in the last 225 cases). Subsequent modifications in the technique to prevent band slippage included placing the band near the level of the esophagus, with minimal disruption of the posterior gastric attachments and diligent suturing of the band in place. CONCLUSIONS: We conclude that the laparoscopic gastric band is effective in short- and long-term weight loss. The high rate of reoperation for repositioning has been avoided in current practice.

Multiple caspases are involved in beta-amyloid-induced neuronal apoptosis.

beta-amyloid peptide (Abeta) has been implicated in the pathogenesis of Alzheimer disease and has been reported to induce apoptotic death in cell culture. Cysteine proteases, a family of enzymes known as caspases, mediate cell death in many models of apoptosis. Multiple caspases have been implicated in Abeta toxicity; these reports are conflicting. We show that treatment of cerebellar granule cells (CGC) with Abeta25-35 causes apoptosis associated with increased activity of caspases-2, -3 and -6. Selective inhibition of each of these three caspases provides significant protection against Abeta-mediated apoptosis. In contrast, no change in caspase-1 activity was seen after Abeta25-35 application, nor was inhibition of caspase-1 neuroprotective. Similar to CGC, cortical neuronal cultures treated with Abeta25-35 demonstrate increased caspase-3 activity but not caspase-1 activity. Furthermore, significant neuroprotection is elicited by selective inhibition of caspase-3 in cortical neurons administered Abeta25-35, whereas selective caspase-1 inhibition has no effect. Taken together, these findings indicate that multiple executioner caspases may be involved in neuronal apoptosis induced by Abeta.

Methylmercury has a selective effect on mitochondria in cultured astrocytes in the presence of [U-(13)C]glutamate.

The effect of methylmercury on glutamate metabolism was studied by (13)C magnetic resonance spectroscopy. Cerebral cortical astrocytes were pretreated with methylmercury, either 1 microM for 24 h, or 10 microM for 30 min, and subsequently with 0.5 mM [U-(13)C]glutamate for 2 h. Labeled glutamate, glutamine, aspartate and glutathione were present in cell extracts, and glutamine, aspartate and lactate in the medium of all groups. HPLC analysis of these amino acids showed no changes in concentrations between groups. Surprisingly, the amounts of [U-(13)C]glutamate and unlabeled glucose taken up by the astrocytes were unchanged. Furthermore, the amounts of most metabolites synthesized from [U-(13)C]glutamate were also unchanged in all groups. However, formation of [U-(13)C]lactate was decreased in the 10 microM methylmercury group. This was not observed for labeled aspartate. It is noteworthy that both [U-(13)C]lactate and [U-(13)C]aspartate can only be derived from [U-(13)C]glutamate via mitochondrial metabolism. [U-(13)C]glutamate enters the tricarboxylic acid cycle (located in mitochondria) after conversion to 2-[U-13C]oxoglutarate and [U-(13)C]aspartate is formed from [U-(13)C]oxaloacetate, as is [U-(13)C]lactate. [U-(13)C]lactate can also be formed from [U-(13)C]malate. This differential effect on labeled aspartate and lactate indicates cellular compartmentation and thus selective vulnerability of mitochondria within the astrocytes to the effects of methylmercury. The decreased lactate production from glutamate might be detrimental to surrounding cells since lactate has been shown to be an important substrate for neurons.

Detection of early gene expression changes by differential display in the livers of mice exposed to dichloroacetic acid.

Dichloroacetic acid (DCA) is a major by-product of water disinfection by chlorination. Several studies have demonstrated the hepatocarcinogenicity of DCA in mice when administered in drinking water. The mechanism of DCA carcinogenicity is not clear and we speculate that changes in gene expression may be important. In order to analyze early changes in gene expression induced by DCA treatment we used the differential display method. Mice were treated with 2 g/l DCA in drinking water for 4 weeks. Total RNAs were obtained from livers of both control and treated mice for analysis. Of approximately 48 000 bands on the differential display gels representing an estimated 96% of RNA species, 381 showed differences in intensity. After cloning and confirmation by both reverse-northern and northern analyses, six differentially expressed genes were found. The expression of five of these genes was suppressed in the DCA-treated mice while one was induced. After sequencing, four genes were identified and two were matched to expressed sequence tags through the BLAST program. These genes are alpha-1 protease inhibitor, cytochrome b5, stearoyl-CoA desaturase and carboxylesterase. Stearoyl-CoA desaturase was induced approximately 3-fold in the livers of DCA-treated mice and the other three genes were suppressed approximately 3-fold. Stearoyl-CoA desaturase, cytochrome b5 and carboxylesterase are endoplasmic reticulum membrane-bound enzymes involved in fatty acid metabolism. The expression pattern of four of these genes was similar in DCA-induced hepatocellular carcinomas and the 4 week DCA-treated mouse livers. The expression of stearoyl-CoA desaturase and one of the unidentified genes returned to control levels in the carcinomas. Understanding the roles and interactions between these genes may shed light on the mechanism of DCA carcinogenesis.