Directly selected cytomegalovirus-reactive donor T cells confer rapid and safe systemic reconstitution of virus-specific immunity following stem cell transplantation.

BACKGROUND: Adoptive transfer of virus-specific T cells may accelerate reconstitution of antigen-specific immunity and limit the morbidity and mortality of viral infections following allogeneic hematopoietic stem cell transplantation. The logistics of producing virus-specific T cells has, however, limited the application of cellular therapies, particularly following the introduction of more-recent regulatory stipulations. METHODS: We investigated the ability of cytomegalovirus-specific T cells, directly isolated from donor leucapheresates on the basis of interferon γ secretion, to restore antiviral immunity in a group of 25 patients following related-donor transplantation in a single-arm phase I-II study. Selected cells were administered early following transplantation, either after the detection of cytomegalovirus DNA by polymerase chain reaction-based surveillance or prophylactically between day 40 and day 50. RESULTS: Cell selection was successful in all cases, yielding a product biased towards CD4(+) over CD8(+) T cells. The target cell dose of 1 × 10(4) CD3(+) T cells/kg of recipient weight contained a median of 2840 cytomegalovirus-specific CD4(+) cells/kg and 630 cytomegalovirus-specific CD8(+) cells/kg, with a median purity of 43.9% interferon γ-secreting cells. Expansions of both CD4(+) and CD8(+) cytomegalovirus-specific T cells were observed in vivo within days of adoptive transfer. These cells were predominantly terminally differentiated effector-memory cells and showed the same T cell receptor variable ß chain (TCRBV) -restriction as the infused cells. They offered protection from reinfection in the majority of patients. CONCLUSIONS: These data indicate that application of cytomegalovirus-specific T cells generated by direct selection using γ-capture is both feasible and effective in a clinical environment. These simple in vitro methodologies should allow more widespread application of virus-specific T cell immunotherapies.

Methemoglobin suppression in a 0.3 Tesla magnet: an in vitro and in vivo study.

RATIONALE AND OBJECTIVES: The aim of this study was to design a pulse sequence that suppresses methemoglobin and is applicable in a 0.3-T permanent magnet. MATERIALS AND METHODS: Blood samples were collected from six healthy volunteers. Magnetic resonance imaging was performed with a 0.3-T scanner until the typical signal intensities of methemoglobin were obtained. Each blood sample was then scanned using repetition times of 300, 600, 900, 1200, 1500, and 1800 ms and a constant echo time of 20 ms. All other parameters (field of view, slice thickness, matrix, and number of signal averages) were kept constant for all six sequences. Signal intensities and repetition time data were used to calculate the T1 relaxation time of extracellular methemoglobin. T1 was determined by ordinary least square regression according to the equation S =k(1 - e(-repetition time/T1)), where S is signal intensity and k is the proportional constant. A short tau inversion recovery sequence with an inversion time calculated from the T1 value of methemoglobin was used on fat, water, and methemoglobin blood samples and in 11 patients diagnosed with subacute brain hemorrhages. The inversion time was calculated as ln2 T1 (tissue), where T1 is the relaxation time of extracellular methemoglobin to be suppressed. RESULTS: The T1 relaxation time of extracellular methemoglobin was determined to be 231.24 +/- 9.068 ms, and inversion time was calculated to be 160 +/- 6.67 msec. Application of an inversion time of 160 ms showed complete suppression on extracellular methemoglobin blood samples and all of the 11 subjects. CONCLUSION: A methemoglobin suppression technique using an inversion time of 160 +/- 6.67 ms is applicable in a 0.3-T permanent magnet.

Phase I study of high-stringency CD8 depletion of donor leukocyte infusions after allogeneic hematopoietic stem cell transplantation.

BACKGROUND: Donor leukocyte infusions (DLI) are given after hematopoietic stem-cell transplantation to eradicate persistent tumor or correct mixed chimerism (MC). The drawback of DLI is the risk of graft-versus-host disease (GVHD). In this phase I study, we examined the potential of highly extensive CD8 depletion of DLI as a means of improving its safety profile. METHODS: High-stringency immunomagnetic CD8 depletion of DLI was performed after steady state donor apheresis. Patients with persistent disease or MC received escalated dose CD8-depleted DLI at 3-month intervals starting from 6 months posttransplantation. The starting dose was 1 x 10(6) CD4 cells/kg in recipients of unrelated and 3 x 10(6) CD4 cells/kg in recipients of related donor transplantations. RESULTS: Twenty-eight patients received CD8-depleted DLI (n=16 unrelated or mismatched, n=12 human leukocyte antigen-identical sibling). Median CD8 depletion was more than 4 log. The median overall dose of CD4+ cells/kg given was 4 x 10(6) (range 1 x 10(6)-43 x 10(6)). Conversion from MC to full donor chimerism was observed in 8 of 16 evaluable patients, and disease responses occurred in 5 of 11 patients (complete response in four and partial response in one). Five of 28 patients developed severe acute pattern (grade II-IV) GVHD. Two patients died as a result of complications relating to GVHD. CONCLUSIONS: Graft-versus-tumor effects can be observed after high-stringency CD8-depleted DLI, although the major toxicity remains GVHD in this high-risk patient group. The safety and efficacy profile of this approach will require testing in a randomized controlled study.

Trismus and trigeminal neuralgia in one patient with colon cancer.

A 64-year-old man got trismus and trigeminal neuralgia under the diagnosis of colon cancer with mandibular metastasis after emergency appendectomy and elective hemicolectomy. The patient chose to forgo further surgery and was given only palliative chemotherapy and radiotherapy. He died six months after diagnosis. Metastatic tumors to the oral cavity are relatively uncommon. They are found most commonly in the mandible, and 70% of cases are adenocarcinoma-most commonly from breast and lung, followed by adrenals, kidneys, prostate, thyroid and colon. Mandibular mass is usually the first sign, then soft-tissue swelling, pain and paresthesias. Tissue proof is needed to confirm the diagnosis. The treatment depends on the nature of the primary, the degree of dissemination and the precise location. However, the prognosis is grim, with the mean survival after diagnosis being only about 6-7 months.

Simultaneous Klebsiella pneumoniae and amoebic liver abscess in an immunocompetent patient.

Simultaneous Klebsiella pneumoniae and amoebic liver abscess is rarely reported in immunocompetent patients. A 47-year-old man was hospitalized with abdominal pain, fever, chills, and hypotension. Physical examination revealed right upper quadrant tenderness. Abdominal computed tomography showed an area of low attenuation with some liquefaction in the liver. Echo-guided aspiration revealed 30 mL of pus, which grew Klebsiella pneumoniae, and the same organism was isolated from the blood. Cytology examination of the pus showed scattered amoeba. The patient gradually improved over 1 month on treatment with cefmetazole and metronidazole, along with repeated drainage of the abscess. His amoebic indirect hemagglutination titer was 1:128, but no parasite ova or amoeba were found in the stool. He had no evidence of immunocompromise. Parasitic diseases may be a predisposing factor for bacterial infections, including pyogenic liver abscess. The possible coexistence of amoebae and bacteria in a liver abscess should not be discounted.

Emphysematous pyelonephritis.

OBJECTIVE: The mortality rate among patients with emphysematous pyelonephritis (EPN) is high and the best therapy has long been debated. MATERIAL AND METHODS: Between January 1993 and December 2004, 26 patients diagnosed with EPN were treated at our hospital. Historical, demographic and laboratory data were obtained from medical records for analysis. RESULTS: The mean age of the patients was 58.7+/-12.7 years. All patients had diabetes, and women outnumbered men (23:3). The clinical findings were non-specific. A plain abdominal X-ray was diagnostic in only 9/26 patients (36%), ultrasonography was accurate in 13 (50%) and CT was diagnostic in all cases. Eighteen patients survived, including all five treated with open drainage and three out of four treated with percutaneous drainage (although one of these later required nephrectomy), and eight died. Diabetic ketoacidosis or non-ketotic hyperosmolar syndrome, obesity and smoking were significant predictors of death. CONCLUSIONS: Early diagnosis by means of CT and CT-guided percutaneous drainage (which is better than ultrasonography-guided drainage) or open drainage, along with antibiotic treatment, may be a reasonable alternative to nephrectomy for this high morbidity condition.

Method for the determination of Aconitum alkaloids in dietary supplements and raw materials by reversed-phase liquid chromatography with ultraviolet detection and confirmation by tandem mass spectrometry: single-laboratory validation.

A study of single-laboratory validation (SLV) of a reversed-phase liquid chromatography (RP-LC) method was conducted for the determination of diester-diterpene Aconitum alkaloids, viz., aconitine, mesaconitine, and hypaconitine, in a variety of dietary supplements, including single- and multiple-ingredient dry powder extracts, pills, capsules, and raw materials. The Aconitum alkaloids in the samples were extracted by diethyl ether in the presence of ammonia. After cleanup with solid-phase extraction to remove the matrix interferences, the alkaloids were determined by RP-LC with UV detection at 235 nm, and the results were confirmed by tandem mass spectrometry. The linear responses for aconitine, mesaconitine, and hypaconitine based on the present LC system ranged from 0.5 to 200 microg/mL. Relative standard deviations of 2.0 to 6.9% were obtained from duplicate analysis of 6 test materials of different matrixes for the 3 Aconitum alkaloids performed by 2 analysts on 5 different days. The recoveries determined for supplements and raw materials spiked with 3 Aconitum alkaloids at levels of 2.5-10 microg/g were in the range of 86-99%. In view of the attainment of satisfactory results for accuracy, precision, and recovery in the SLV study, it is recommended that the method validation process proceed to a collaborative study.

Clinical features of human intestinal capillariasis in Taiwan.

Human intestinal capillariasis is a rare parasitosis that was first recognized in the Philippines in the 1960 s. Parasitosis is a life threatening disease and has been reported from Thailand, Japan, South of Taiwan (Kaoh-Siung), Korea, Iran, Egypt, Italy and Spain. Its clinical symptoms are characterized by chronic diarrhea, abdominal pain, borborygmus, marked weight loss, protein and electrolyte loss and cachexia. Capillariasis may be fatal if early treatment is not given. We reported 14 cases living in rural areas of Taiwan. Three cases had histories of travelling to Thailand. They might have been infected in Thailand while stayed there. Two cases had the diet of raw freshwater fish before. Three cases received emergency laparotomy due to peritonitis and two cases were found of enteritis cystica profunda. According to the route of transmission, freshwater and brackish-water fish may act as the intermediate host of the parasite. The most simple and convenient method of diagnosing capillariasis is stool examination. Two cases were diagnosed by histology. Mebendazole or albendezole 200 mg orally twice a day for 20-30 d is the treatment of choice. All the patients were cured, and relapses were not observed within 12 mo.

MR imaging of the musculoskeletal soft tissue mass: is heterogeneity a sign of malignancy?

BACKGROUND: Magnetic resonance (MR) is considered the imaging modality of choice to evaluate soft tissue lesions. Whether MR imaging can be used to differentiate benign from malignant soft tissue lesions is still controversial. To elucidate this controversy, MR images of 37 patients with soft tissue masses of the musculoskeletal system were reviewed at Christchurch Hospital, New Zealand. METHODS: There were 19 benign and 18 malignant lesions. MR images were evaluated with regard to lesion size, border definition, homogeneity, changes in pattern of homogeneity, signal characteristic (signal intensity on T1-weighted, T2-weighted), and demonstration of relation to neurovascular bundle and bone as well as edema in or around the lesion. RESULTS: Statistically significant imaging features favoring a diagnosis of malignancy included inhomogeneity at T2-weighted images (p = 0.002) and a change in pattern from homogeneity on T1-weighted images to inhomogeneity at T2-weighted images (p = 0.003). Malignant tumors also had neurovascular or bone involvement in 28% of cases, which was not seen in their benign counterparts. Size, border definition, and edema of surrounding tissues were generally not helpful in distinguishing benign from malignant soft tissue masses. CONCLUSIONS: The inhomogeneity of lesions on T2, the change from homogeneity on T1 to inhomogeneity on T2 sequence, and involvement of bone or neurovascular structures are features that may be helpful in differentiating benign from malignant soft tissue masses.