HBL-3 Cell Line,Derived from Precursor B-cell lymphoblastic Leukemia,Lacks Somatic Hypermutation of Immunoglobulin Heavy Chain Variable Region Genes

Somatic mutation (SM) analysis provides a useful tool for understanding the stages at which neoplastic differentiate from normal B-cells. B-cell precursor neoplasms are considered to be somatically premutational. However, the variable frequency of SM of the variable region (VH) genes has been described in cases of precursor B-cell acute lymphoblastic leukemia (PB-ALL). To better characterize PB-ALL based on the differentiation stage, we investigated the SM of the VH genes expressed by tumor cells of the surface immunoglobulin (sIg)^– HBL-3 cell line derived from childhood PB-ALL. In the HBL-3 cell line, the rearranged Ig heavy chain VH gene sequence showed no SM in the complementarity-determining regions of 1, 2, and 3, or in the framework regions of 1, 2, and 3 relative to the putative germline VH gene sequences. In addition, the VH segment of HBL-3 cells showed no intraclonal sequence heterogeneity, indicating ongoing SM. Our data demonstrated that HBL-3 cells express unmutated and developmentally regulated rearrangement of VH genes at the stage of B-cell precursor cells. HBL-3 cells, which are derived only from the sIg^– PB-ALL, showed that SM cannot be recognized in VH genes of tumor cells before the expression of sIg.     

Pregnancy by insemination of cryopreserved spermatozoa from a man with retrograde ejaculation: a case report

BACKGROUND: Owing to the prevalence of diabetes mellitus, spinal injuries and aggressive surgical treatment of cancer, the number of younger patients with retrograde ejaculation is increasing. Since medical treatment to restore antegrade ejaculation often fails, several options for accomplishing insemination by these patients, including the use of sperm-rich urine obtained after masturbation and in vitro fertilization with sperm retrieved from the seminal tract, have been reported. We used the least invasive and most inexpensive procedure in a patient/couple with this condition. CASE: A 23-year-old man suffered from retrograde ejaculation after a spinal injury. He could achieve erection and engage in sexual intercourse but seldom had an orgasm or the sensation of ejaculation. We obtained spermatozoa from urine produced after masturbation at home and froze them. We used these frozen-thawed spermatozoa for intrauterine insemination, leading to the term birth of a healthy infant. CONCLUSION: In selected patient/couples, frozen spermatozoa obtained from postmasturbation urine can be used successfully for intrauterine insemination. This minimally invasive and most inexpensive procedure should be tried before planning in vitro fertilization.     

Functional aortic stenosis diagnosed in fetal period

We report a case of fetal congenital heart disease prenatally diagnosed as critical aortic valve stenosis at 25 weeks of gestation. Fetal echocardiography demonstrated severe mitral regurgitation, aortic valve stenosis, hypocontractility of the left ventricle, and showed retrograde flow in the aortic arch like HLHS (hypoplastic left heart syndrome). However, soon after delivery, improvement in the baby's hemodynamics and myocardial contractility were recognized without any treatment.     

Placental site trophoblastic tumor of the uterine cervix occurring from undetermined antecedent pregnancy

A cervical polyp complicated by severe hemorrhage was removed from a 43-year-old Japanese woman (gravida 0), who had undergone tubectomy on the right side 10 years previously. The polyp was diagnosed by immunohistochemical studies as placental site trophoblastic tumor of the cervix, but no metastatic foci were found in any other uterine site. The tumor was further demonstrated by PCR polymorphisms to possess two genomic DNA of the patient and her husband. Serum beta-hCG and urinary hCG titers were both low, which rapidly fell to 0.8 mIU/mL after a total hysterectomy and remained 0.2 mIU/mL after dismission. She has been uneventful for 3 years.     

In vivo insulin action in adrenodemedullated rats after voluntary running

The purpose of this study was to estimate the influence of epinephrine on in vivo insulin sensitivity and responsiveness after voluntary running. Wistar rats that had previously undergone adrenodemedullation or sham-operation were kept in a sedentary state or trained over a 4 week period. An euglycemic insulin clamp study was performed on adrenodemedullated sedentary rats (ADMX), adrenodemedullated voluntary running rats (ADMX-T), sham-operated voluntary running rats (SHAM-T), and control rats (C) at 18 h after the last bout of exercise. The insulin infusion rate was 3.0, 6.0, and 303.0 mU/(kg min), respectively. The blood glucose concentration was maintained constant at basal levels. Metabolic clearance rate of glucose (MCR) was calculated as an index of whole-body insulin action. In the presence of physiological hyperinsulinemia (an insulin infusion rate of 6.0 mU/(kg min)), MCR (ml/(kg min)) was significantly higher in ADMX-T rats (31.2 +/- 2.0) than in ADMX rats (19.8 +/- 0.8, P<0.001) and SHAM-T rats (23.8 +/- 0.8, P<0.05). Also, the MCR values of SHAM-T and ADMX rats were significantly (P<0.001, and P<0.05, respectively) greater than that of C rats (12.7 +/- 0.4). At maximal hyperinsulinemia (an insulin infusion rate of 303.0 mU/(kg min)), there was no difference of MCR between ADMX-T rats (49.8 +/- 4.3) and C rats (38.2 +/- 2.2). The GDR values of SHAM-T rats (43.5 +/- 3.7) and ADMX rats (43.5 +/- 2.1) were also not different from that of C rats. These results provide indirect evidence that epinephrine is one of factors that suppresses increased insulin sensitivity after physical training, although it seems to have no significant influence on insulin responsiveness.     

A neuronal mechanism of propofol-induced central respiratory depression in newborn rats

The neural mechanisms of propofol-induced central respiratory depression remain poorly understood. In the present study, we studied these mechanisms and the involvement of gamma-aminobutyric acid (GABA)A receptors in propofol-induced central respiratory depression. The brainstem and the cervical spinal cord of 1- to 4-day-old rats were isolated, and preparations were maintained in vitro with oxygenated artificial cerebrospinal fluid. Rhythmic inspiratory burst activity was recorded from the C4 spinal ventral root. The activity of respiratory neurons in the ventrolateral medulla was recorded using a perforated patch-clamp technique. We found that bath-applied propofol decreased C4 inspiratory burst rate, which could be reversed by the administration of a GABAA antagonist, bicuculline. Propofol caused resting membrane potentials to hyperpolarize and suppressed the firing of action potentials in preinspiratory and expiratory neurons. In contrast, propofol had little effect on resting membrane potentials and action potential firing in inspiratory neurons. Our findings suggest that the depressive effects of propofol are, at least in part, mediated by the agonistic action of propofol on GABAA receptors. It is likely that the GABAA receptor-mediated hyperpolarization of preinspiratory neurons serves as the neuronal basis of propofol-induced respiratory depression in the newborn rat.     

Renal hypothermia using ice slush for retroperitoneal laparoscopic partial nephrectomy

Renal hypothermia using ice slush for retroperitoneal laparoscopic partial nephrectomy was performed in 2 patients. A cylindrical device was secured in the enlarged port site. Through the device, ice slush was introduced and placed around the kidney. The nadir renal temperature under renal ischemia in the 2 patients was 18.4 degrees C and 25.8 degrees C.     

Alendronate inhibits urinary calcium microlith formation in a three-dimensional culture model

Osteoporosis is associated with the pathogenesis of urinary stone formation. Urinary stones are similar to bone diseases such as osteoporosis and bone metabolism in terms of pathogenesis. Bisphosphonates are potent inhibitors of bone resorption, and are used in the management of bone disease. Furthermore, bisphosphonates have a strong affinity for calcium, and a reported inhibitory effect on calcium oxalate crystallization in vitro. Thus, bisphosphonates might also inhibit urinary stone formation. Madin-Darby canine kidney (MDCK) cells form calcium phosphate microliths at the basolateral side in vitro. We investigated the inhibitory effects of new generation bisphosphonates (alendronate and incadronate) on calcium phosphate microlith formation and on the expression of osteopontin, which is an important urinary stone matrix. MDCK cells formed two types of colonies in three-dimensional soft agar culture; dark colonies containing calcium phosphate microliths and clear colonies free from microliths. We applied purified alendronate and incadronate at concentrations of 10^–11, 10^–9, 10^–7 and 10^–5 M to MDCK cells cultured in three-dimensional soft agar and investigated the efficiency of colony formation and the dark colony ratio (number of dark colonies relative to the total number of colonies). The administration of 10^–9 and 10^–7 M alendronate decreased the dark colony ratio compared with controls, whereas incadronate did not significantly alter this colony ratio compared with controls. The expression of osteopontin in cultured cells was inhibited by the 10^–7 M alendronate administration. The present findings show that alendronate inhibits calcium stone formation, suggesting that it is effective in the prevention of urolithiasis.     

Thoracic CT findings of adult T-cell leukemia or lymphoma

OBJECTIVE: The aim of this study was to assess pulmonary CT findings in patients with adult T-cell leukemia or lymphoma. MATERIALS AND METHODS: We retrospectively reviewed CT scans of the lung in 87 patients with adult T-cell leukemia or lymphoma who had undergone chest CT between January 1996 and March 2002 at two institutions. The CT scans were interpreted by two chest radiologists working in consensus. Parenchymal abnormalities (ground-glass attenuation, consolidation, nodules, thickening of bronchovascular bundles, interlobular septal thickening, honeycombing, crazy-paving appearance, and bronchiectasis) were evaluated, as were enlarged lymph nodes, pleural effusion, and pleural thickening. In 46 patients who underwent surgical biopsy or autopsy, CT-pathologic correlation was performed with the actual specimens by a pathologist and two chest radiologists. RESULTS: On the CT scans, abnormal findings were seen in 60 patients (69.0%). CT findings consisted of ground-glass attenuation (n = 37), centrilobular nodules (n = 25), thickening of bronchovascular bundles (n = 22), and consolidation (n = 13). These abnormalities were predominantly seen in the peripheral lung parenchyma (n = 26). Pathologically, these findings corresponded with atypical lymphocyte infiltration along the interstitium and the alveolar spaces. Pleural effusion and enlarged lymph nodes were found in 22 and 27 patients, respectively. CONCLUSION: CT findings in patients with adult T-cell leukemia or lymphoma consisted mainly of ground-glass attenuation, centrilobular nodules, and thickening of the bronchovascular bundles in the peripheral lung. These findings, although nonspecific, are considered suggestive of thoracic involvement in patients with adult T-cell leukemia or lymphoma.     

Thoracic manifestation of myeloperoxidase-antineutrophil cytoplasmic antibody (MPO-ANCA)-related disease. CT findings in 51 patients

OBJECTIVE: To assess the pulmonary CT findings of patients with serum evidence of the myeloperoxidase-antineutrophil cytoplasmic antibody (MPO-ANCA). METHODS: The pulmonary CT scans of 62 patients with serum evidence of MPO-ANCA (51 with microscopic polyangiitis, 11 with Churg-Strauss syndrome) were retrospectively assessed with regard to parenchymal, pleural, and mediastinal abnormalities. RESULTS: On the CT scans, abnormal findings were seen in 51 of the patients (82%, n = 62). Of the patients, the CT findings consisted of ground-glass attenuation in 48 of the patients (94%, n = 51), consolidation in 40 (78%), and thickening of bronchovascular bundles in 26 (51%). Pathologically, these findings corresponded to alveolar hemorrhages, interstitial chronic inflammation in the alveolar septa, vasculitis, or fibrosis. These abnormalities were predominantly seen in peripheral lung parenchyma (n = 37). CONCLUSION: The CT findings in patients with MPO-ANCA consisted mainly of ground-glass attenuation and consolidation in the peripheral lung. These findings, although nonspecific, are considered as pulmonary involvement in patients with MPO-ANCA.