Citrobacter Infection and Wnt Signaling

Gut flora generally contributes to a healthy environment, but both commensal and pathogenic bacteria that influence the innate and adaptive immune responses can cause acute and/or chronic mucosal inflammation. Citrobacter rodentium is a member of the family of enteropathogens that provide an excellent in vivo model to investigate host?Cpathogen interactions in real time. It is the etiologic agent of transmissible murine colonic hyperplasia, and inflammation following C. rodentium infection is dependent upon the genetic background. Ongoing and completed studies in this model have established that Wnt/??-catenin, Notch, and phosphatidylinositol 3-kinase pathways regulate colonic crypt hyperplasia, whereas epithelial?Cstromal cross talk, mediated by MEK/ERK/nuclear factor ??B signaling, regulates inflammation and/or colitis in susceptible strains. The C. rodentium-induced hyperplastic state also increases the susceptibility to either mutagenic insult or in mice heterozygous for the Apc gene. The ability to modulate the host response to C. rodentium infection therefore provides an opportunity to delineate the mechanisms that determine mucosal hyperplasia, intestinal inflammation, and/or neoplasia as disease outcomes.     

Immunoglobulin-like domain containing receptor 1 mediates fat-stimulated cholecystokinin secretion

Cholecystokinin (CCK) is a satiety hormone produced by discrete enteroendocrine cells scattered among absorptive cells of the small intestine. CCK is released into blood following a meal; however, the mechanisms inducing hormone secretion are largely unknown. Ingested fat is the major stimulant of CCK secretion. We recently identified a novel member of the lipoprotein remnant receptor family known as immunoglobulin-like domain containing receptor 1 (ILDR1) in intestinal CCK cells and postulated that this receptor conveyed the signal for fat-stimulated CCK secretion. In the intestine, ILDR1 is expressed exclusively in CCK cells. Orogastric administration of fatty acids elevated blood levels of CCK in wild-type mice but not Ildr1-deficient mice, although the CCK secretory response to trypsin inhibitor was retained. The uptake of fluorescently labeled lipoproteins in ILDR1-transfected CHO cells and release of CCK from isolated intestinal cells required a unique combination of fatty acid plus HDL. CCK secretion secondary to ILDR1 activation was associated with increased [Ca²⁺]_i, consistent with regulated hormone release. These findings demonstrate that ILDR1 regulates CCK release through a mechanism dependent on fatty acids and lipoproteins and that absorbed fatty acids regulate gastrointestinal hormone secretion.     

An evaluation of brachytherapy and external beam radiation used with wide-margin surgical resection in the treatment of extra-abdominal desmoid tumors

Surgical resection has had control rates of 53% to 77% in the treatment of extra-abdominal desmoid tumors. Surgical excision combined with external beam radiation therapy (EBRT) has had local control rates of up to 83% in some series. The purpose of this study was to evaluate the effectiveness of resection combined with radiotherapy (brachytherapy, EBRT, or both) in the treatment of extra-abdominal desmoid tumors. We retrospectively reviewed the charts of 24 consecutive patients (27 histologically confirmed extra-abdominal desmoid tumors). Patients were included in the study if they had a lesion that was potentially resectable with a wide margin, allowing for limb salvage, and if they did not have a contraindication to radiotherapy. Limb functioning was assessed with the Musculoskeletal Tumor Society (MSTS) scoring system. Seventeen patients (7 men, 10 women) with 19 tumors met the inclusion criteria. Mean age at diagnosis was 23.4 years. Follow-up (mean, 4.28 years) involved serial clinical examinations and magnetic resonance imaging of tumor sites. After surgery, the tumors were treated with brachytherapy (n = 6), EBRT (n = 10), or both (n = 3). Two of the 17 tumors in patients with negative margins of resection recurred locally (local control rate, 88.2%). Mean MSTS score was 29/30 (96.7%). The role of surgery, radiotherapy, chemotherapy, hormone therapy, and other treatments for extra-abdominal desmoid tumors is not well defined. When wide-margin resection and radiotherapy can be performed with limb preservation surgery, local control and complication rates compare favorably with those of other reported methods of treatment. Given the results and limitations of our study, we cannot make a definitive recommendation as to which modality--brachytherapy or EBRT--should be used in the treatment of extra-abdominal desmoid tumors.     

Long-term culture of Japanese human embryonic stem cells in feeder-free conditions

Human pluripotent embryonic stem cells (hESCs) have great promise for research into human developmental biology, development of cell therapies for the treatment of diseases, toxicology, and drug discovery. Traditionally, undifferentiated hESCs are maintained on mouse embryonic fibroblasts (MEFs), which impede the clinical applications of hESCs. Here we have examined the long-term stability of the Japanese hESC line (KhES-1) in feeder-free culture. KhES-1 cells were cultured with MEF conditioned medium (CM) and different doses of basic fibroblast growth factor (bFGF) in six-well-plates of which the surface was coated with Matrigel. KhES-1 cells were maintained for at least 40 passages. In this culture system, the cells maintained stable proliferation rates and steadily expressed Oct-4, Nanog, and alkaline phosphatase. In addition, KhES-1 cells maintained without direct feeder contact formed embryonic bodies with expression of markers from the three germ layers. Here we demonstrated that Japanese human embryonic stem cells KhES-1 were cultured long term in a feeder-free method, while retaining pluripotency in vitro.     

Role of sample adequacy in fine needle aspiration biopsy of palpable breast lesions.

BACKGROUND: The role of fine needle aspiration biopsy (FNAB) in breast lesions remains uncertain because its accuracy has been questioned. We hypothesized that this is related to failure to define standards for adequacy of sample preparation. METHODS: We reviewed cytology and pathology reports from 351 patients undergoing FNAB at a 350-bed community, university-affiliated teaching hospital over a 2-year period. Patients were included if they had an open biopsy within 2 years of an FNAB of the same lesion. Cytology reports were reviewed by a cytopathologist unaware of the clinical setting or subsequent histology; surgeons unaware of the cytology reports reviewed histology reports. Cytological diagnoses were benign, likely benign, probable cancer, and definite cancer. Samples were characterized as satisfactory, less than optimal (few mammary epithelial clusters), or inadequate (no mammary epithelial clusters.) We assessed the proportion of inadequate samples, the accuracy of FNAB, and the influence of sample adequacy upon FNAB sensitivity. RESULTS: Ninety-nine (28%) FNABs were inadequate, 77 (22%) less than optimal, and 175 (50%) satisfactory. Ninety-five patients (27%) had a final diagnosis of malignancy. FNABs classified as satisfactory or less than optimal and characterized as benign (n = 102) had a negative predictive value (NPV) of 0.91; those termed definite cancer (n = 43) had a positive predictive value (PPV) of 0.98. Only 10% of all cancers were identified in the 28% of FNABs that were classified as inadequate (P <0.01). CONCLUSIONS: We concluded that too few FNABs are performed in a fashion that permits definitive cytological interpretation. Inadequate FNABs are less likely to detect malignancy. After excluding inadequate preparations, FNABs interpreted as definite cancer and as benign are highly accurate in identifying patients with and without cancer.     

Recovery of Functional Memory T Cells in Lung Transplant Recipients Following Induction Therapy with Alemtuzumab

Profound T-cell depletion with the monoclonal antibody alemtuzumab facilitates reduced maintenance immunosuppression in abdominal and lung transplantation. While the phenotype of the post-depletional T cells has been characterized, little is known about their function. In the present study, global and CMV-specific T-cell function was assessed longitudinally in 23 lung transplant (LTx) recipients using T-cell assays (ImmuKnow and T Cell Memory, Cylex, Columbia, MD) during the first year posttransplant after induction therapy. Recovery of mitogen responses were seen at 2 weeks posttransplantation (65%PHA; 58% Con A), despite the low number of circulating T cells (<2%). These responses declined at 4-5 months (24%PHA; 54% Con A) and were partially reconstituted by 9 months (46% PHA; 73% Con A). CMV-specific responses recovered in 80% of R+ patients as early as 2 weeks posttransplant (n = 5) and 72% of patients had a memory response by 3 months (n = 11). In contrast, only 2 of 5 patients who did not exhibit memory responses pre-transplant (R-) developed transient CMV-specific T-cell responses. Our results show that profound depletion of T cells induced by alemtuzumab spares the functional subset of CMV-specific memory T cells. Conversely, CMV R- patients predepletion may require a prolonged period of prophylaxis.     

CT Scan Diagnosis of Bleeding Peptic Ulcer after Gastric Bypass

Investigation of the bypassed stomach in patients with suspected peptic ulcer disease presents a major challenge to bariatric surgeons. Various methods have been suggested for visualization of the duodenum and bypassed stomach. These include endoscopy via percutaneous gastrostomy access, retrograde endoscopy and virtual gastroscopy using CT scan. We present a case of peptic ulcer bleeding diagnosed with the help of conventional CT scan. To the best of our knowledge, this is the second such case reported in the literature and the first in the bariatric population.     

Cortical network for gaze control in humans revealed using multimodal MRI

Functional magnetic resonance imaging (fMRI) techniques allow definition of cortical nodes that are presumed to be components of large-scale distributed brain networks involved in cognitive processes. However, very few investigations examine whether such functionally defined areas are in fact structurally connected. Here, we used combined fMRI and diffusion MRI-based tractography to define the cortical network involved in saccadic eye movement control in humans. The results of this multimodal imaging approach demonstrate white matter pathways connecting the frontal eye fields and supplementary eye fields, consistent with the known connectivity of these regions in macaque monkeys. Importantly, however, these connections appeared to be more prominent in the right hemisphere of humans. In addition, there was evidence of a dorsal frontoparietal pathway connecting the frontal eye field and the inferior parietal lobe, also right hemisphere dominant, consistent with specialization of the right hemisphere for directed attention in humans. These findings demonstrate the utility and potential of using multimodal imaging techniques to define large-scale distributed brain networks, including those that demonstrate known hemispheric asymmetries in humans.     

Spilled Gallstones: the Source of an Enigma

Spillage of gallstones may occur in the course of laparoscopic cholecystectomy. The incidence of this mishap and its consequences are variable. Ignored by many surgeons, stone spillage may be the source of significant morbidity many years after surgery. In this report, we describe the clinical course of a patient who presented with upper abdominal pain and swelling. The past history was positive for laparoscopic cholecystectomy 15 years earlier. After excision, the swelling was found to be a pseudocyst formed around spilled gallstones during a previous cholecystectomy. Apart from postoperative wound infection, the patient recovered well and remains so. Here, we discuss the problem and provide suggestions for spillage prevention and stone retrieval once spillage occurs.