TZT-1027, an Antimicrotubule Agent, Attacks Tumor Vasculature and Induces Tumor Cell Death

TZT-1027, a dolastatin 10 derivative, is an antimicrotubule agent with potent antitumor activity both in vitro and in vivo. In this study, we performed biochemical and histopathological examinations, and evaluated TZT-1027-induced tumoral vascular collapse and tumor cell death in an advanced tumor model, murine colon 26 adenocarcinoma. In addition, we studied the effects of TZT-1027 on cultured human umbilical vein endothelial cells (HUVEC). Tolerable doses of TZT-1027 induced tumor-selective hemorrhage within 1 h. This hemorrhage occurred mainly in the peripheral area of the tumor mass. Measurements of tumoral hemoglobin content and dye permeation revealed that the hemorrhage occurred firstly and tumor blood flow stopped secondarily. The vascular damage was followed by continuous induction of apoptosis of the tumor cells, tumor tissue necrosis, and tumor regression. In cultured HUVEC, TZT-1027 induced marked cell contraction with membrane blebbing in 30 min. These cell changes were completely inhibited by K252a, a broad-spectrum inhibitor of protein kinases. These effects of TZT-1027 on both tumor vasculature and HUVEC were greater than those of vincristine. In conclusion, TZT-1027 quickly attacked the well-developed vascular system of advanced tumors by a putative protein kinase-dependent mechanism, and then blocked tumor blood flow. Therefore, TZT-1027 has both a conventional antitumor activity and a unique anti-tumoral vascular activity, making it a potentially powerful tool for clinical cancer therapy.     

Primary carcinoid tumor of the liver: report of a case with an emphasis on contrast-enhanced ultrasonographic findings

We report the case of a 82-year-old man with primary carcinoid tumor of the liver. Abdominal ultrasonography (US) showed 2 well-demarcated, round tumors in the right lobe (4 x 4 cm and 1 x 1 cm). The lesions were both markedly echogenic with many small cystic areas. Contrast-enhanced US showed the whole tumor to be quickly and highly enhanced, suggesting the highly vascular nature of the lesion, and the findings corresponded well with hepatic angiographic results. The patient underwent a US-guided needle biopsy, which yielded histological results consistent with a carcinoid tumor. Considering his age, we selected radiofrequency ablation (RFA) for treatment. After treatment, contrast-enhanced US demonstrated complete disappearance of blood flow signals, indicating satisfactory treatment. Our case showed that US and contrast-enhanced US findings can lead to a high suspicion of carcinoid tumor.     

Assessing nitrate metabolism in the intestinal tract by measuring breath nitric oxide and nitrous oxide, and its clinical significance

BACKGROUND: The toxicity of dietary nitrate (NO3-) is controversial. One reason is nitrate metabolism in the intestine is so complicated that it is far from fully understood. There is no study measuring breath nitric oxide (NO) and nitrous oxide (N2O) after ingesting vegetables and high-nitrate food at the same time. METHODS: Breath samples from 10 healthy young and 10 healthy old subjects were collected at 15-min intervals for 5 h after ingestion of 100 g of lettuce and during fasting (control). Breath NO and N2O were analyzed by a chemiluminescence and an IR-PAS analyzer respectively. RESULTS: N2O maximum concentration and excretions increased significantly after ingesting lettuce in each group [303 (30) vs. 750 (81) ppb, 771 (72) vs. 1668 (146) microg in young; 442 (52) vs. 1092 (109) ppb, 1088 (125) vs. 2100 (183) microg in old subjects; mean (SE), P<0.01], while NO did not. In addition, breath NO was strongly influenced by ambient NO, which varied greatly. N2O maximum level in old subjects after ingesting lettuce was higher than that of young subjects (750 vs. 1092 ppb, P<0.05), and significantly higher N2O concentration levels were seen at 30, 45, 60, and 105 min in old subjects. CONCLUSIONS: A large amount of N2O produced in the intestine and normal nitrate intake do not influence the breath NO concentration, probably due to its relatively small production. Higher maximum N2O concentration after ingesting lettuce in old subject is probably because more bacteria, which rapidly reduce dietary nitrate in the upper intestinal tract, inhabit the gut in old age. Our results suggested that breath N2O is a useful noninvasive maker to estimate dietary nitrate reduction in the intestinal tract.     

Transient receptor potential 1 regulates capacitative Ca(2+) entry and Ca(2+) release from endoplasmic reticulum in B lymphocytes

Capacitative Ca(2+) entry (CCE) activated by release/depletion of Ca(2+) from internal stores represents a major Ca(2+) influx mechanism in lymphocytes and other nonexcitable cells. Despite the importance of CCE in antigen-mediated lymphocyte activation, molecular components constituting this mechanism remain elusive. Here we demonstrate that genetic disruption of transient receptor potential (TRP)1 significantly attenuates both Ca(2+) release-activated Ca(2+) currents and inositol 1,4,5-trisphosphate (IP(3))-mediated Ca(2+) release from endoplasmic reticulum (ER) in DT40 B cells. As a consequence, B cell antigen receptor-mediated Ca(2+) oscillations and NF-AT activation are reduced in TRP1-deficient cells. Thus, our results suggest that CCE channels, whose formation involves TRP1 as an important component, modulate IP(3) receptor function, thereby enhancing functional coupling between the ER and plasma membrane in transduction of intracellular Ca(2+) signaling in B lymphocytes.     

A case of acute rejection with adenovirus infection after ABO-incompatible kidney transplantation

Abstract:  We report clinical and histopathologic findings of a case of acute rejection with adenovirus infection after kidney transplantation. A 63-yr-old woman with end-stage renal disease caused by lupus nephritis received an ABO-incompatible living kidney transplantation from her husband. On the 7th post-operative day (POD), she had fever, hematuria, and bladder irritation. Although she was treated with an antibiotic, the symptoms were not improved. We diagnosed adenovirus infection as positive with the urine shell vial method and blood PCR analysis. Cyclophosphamide was interrupted and immunoglobulin therapy was performed. However, urine output decreased and serum creatinine levels increased. An episode biopsy was performed on POD 20. We diagnosed acute antibody-mediated rejection. She was treated with plasma exchange for acute rejection and antiviral drug (rivabirin) for active adenovirus infection. However, the renal graft dysfunction was deemed irreversible and the renal graft was removed on POD 34. The graftectomy specimen showed acute rejection and acute tubular necrosis with adenovirus infection.     

Removal of blood group A/B antigen in organs by ex vivo and in vivo administration of endo-beta-galactosidase (ABase) for ABO-incompatible transplantation

BACKGROUND: ABO incompatibility in organ transplantation is still a high risk factor for antibody-mediated rejection, despite the progress in effective treatments. We have explored the possibility of using the enzyme to remove the blood type A/B antigen in organs. METHODS: Recombinant endo-beta-galactosidase (ABase), which releases A/B antigen, was produced in E. coli BL-21. Human A/B red blood cells (RBC) were digested with ABase, and subjected to flow cytometric analysis after incubation with human sera. Purified recombinant ABase was intravenously administered to a baboon. Biopsies were taken from kidney and liver before and 1, 4 and 24 h after in vivo administration. Excised baboon kidneys were perfused with cold UW solution+/-purified recombinant ABase and preserved at 4 degrees C. Biopsies were taken before and 1 and 4 h after ex vivo perfusion. The change in A/B antigen expression was analyzed by immunohistochemical study. RESULTS: ABase removed 82% of A antigen and 95% of B antigen in human A/B red blood cells, and suppressed anti-A/B antibody binding and complement activation effectively. ABase was also found to remain active at 4 degrees C. In vivo infusion of ABase into a blood type A baboon demonstrated a marked reduction of A antigen expression in the glomeruli of kidney (85% at 1 h, 9% at 4 h and 13% at 24 h) and the sinusoids of liver (47% at 1 h, 1% at 4 h and 3% at 24 h) without serious adverse effects. After ex vivo perfusion and cold storage of excised baboon kidney (blood type B) with ABase, the expression levels of B antigen in glomeruli were reduced to 49% at 1 h and 6% at 4 h. CONCLUSIONS: This alternative approach might be useful for minimizing antibody removal and anti-B cell immunosuppression as an adjuvant therapy in ABO-incompatible kidney, liver and possibly heart transplantation.     

Double-subtraction maximum intensity projection MR angiography for detecting the artery of Adamkiewicz and differentiating it from the drainage vein

PURPOSE: To evaluate the efficacy of double-subtraction magnetic resonance angiography (MRA) (subtraction of the subtracted venous phase image from the subtracted arterial dominant phase image) for depicting the artery of Adamkiewicz and differentiating it from the drainage vein. MATERIALS AND METHODS: A total of 170 patients (123 men, 47 women; aged 17-84 years, mean = 67 years), with a thoracoabdominal vascular lesion underwent MRA for detection of the artery of Adamkiewicz. MRA was performed as a five-phase dynamic-enhanced three-dimensional (3D) fast spoiled gradient recalled acquisition in steady state (GRASS) sequence on a 1.5-T system, with double-dose bolus contrast and saline injection. The levels at which the artery of Adamkiewicz and drainage vein originated were determined. Signal intensities of the two vessels were measured with source images to assess the signal-to-noise ratio (SNR), contrast-to-noise ratio (CNR), and necessity of the double-subtraction technique. RESULTS: The artery of Adamkiewicz was detected in 140 patients (82.4%). Branching occurred at levels T8-T12 on the left and T8-T11 on the right. An additional anterior radiculomedullary artery was detected in 18 patients. The drainage vein was detected in 133 patients (78.2%). It merged at the T9-L2 level on both sides. In six of the 133 patients (4.5%), the drainage vein branched upwardly. Neither SNR nor CNR differed significantly between the artery of Adamkiewicz and the drainage vein in the arterial phase; but on the subtraction image, signal intensity of the artery was higher than that of the drainage vein (P < 0.05). CONCLUSION: Double-subtraction MRA is useful for detecting the artery of Adamkiewicz when it is necessary to differentiate it from the drainage vein.     

Expression of Parafibromin in Distant Metastatic Parathyroid Tumors in Patients with Advanced Secondary Hyperparathyroidism Due to Chronic Kidney Disease

Background Recently, somatic inactivating mutations in HRPT2 have been reported in the majority of sporadic parathyroid carcinoma in primary hyperparathyroidism (HPT). Parafibromin is a tumor suppressor protein encoded by HRPT2, and loss of nuclear expression of parafibromin was found in approximately 70% of the carcinoma. In secondary HPT due to chronic kidney disease (CKD), parathyroid carcinoma is very rare and whether HRPT2 plays a role in the carcinogenesis in these cases is not clear. We evaluated the expression of parafibromin in hemodialysis patients with distant metastatic parathyroid tumors. Methods Between June 1973 and December 2006, 2,142 patients underwent parathyroidectomy (PTx) for secondary HPT in our department. We encountered five (0.23%) patients with distant metastatic parathyroid tumors. We evaluated the immunohistochemistry for parafibromin in eight primary parathyroid glands removed from the neck at the initial operation and/or at reoperation and seven distant metastatic tumors resected at reoperation. Results In only one lung metastatic parathyroid tumor, negative staining for parafibromin was detected. In the other three lung, two regional node, and one chest wall metastatic parathyroid tumor, parafibromin was strongly stained in the nuclei of the parathyroid cells. Among eight primary glands, except for one with weakly positive staining, the expression of parafibromin was detected diffusely and strongly. Conclusion We conclude that the inactivating mutations and/or allelic loss of the HRPT2 gene may not play a major role in parathyroid carcinogenesis in secondary HPT due to CKD, but in these cases cancer development may be associated with a heterogeneous genetic disorder.