Novel Y283C mutation of the A subunit for coagulation factor XIII: molecular modelling predicts its impaired protein folding and dimer formation

In an Italian patient with severe factor XIII deficiency, a novel mutation, Y283C (TAT to TGT), was identified heterozygously by nucleotide sequencing analysis in exon VII of the gene for the A subunit. The presence of this mutation was confirmed using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis in the proband and his brother. Molecular modelling predicts that the mutant molecule would be misfolded. It is probable that the impaired folding of the mutant Y283C A subunit led to its instability, which is at least in part responsible for the factor XIII deficiency of this patient.     

Completely resected stage IIIA non-small cell lung cancer: the significance of primary tumor location and N2 station

BACKGROUND: The number of N2 stations (single vs multiple N2 stations) is an important prognostic factor in patients with completely resected stage IIIA-N2 non-small cell lung cancer. However, the significance of both the N2 station(s) actually involved and the primary tumor location remains unclear. METHODS: The database was built with the use of a questionnaire survey on the survival of patients with pathologic stage IIIA-N2 non-small cell lung cancer completely resected between January 1992 and December 1993. The survey was performed by the Japan Clinical Oncology Group as of July 1999. The data include information on the survival and N2 stations of 402 patients. RESULTS: A frequently metastasized single N2 station was the lower pretracheal station in primary tumors in the right upper lobe, the subaortic station in the left upper lobe, and the subcarinal station in the right middle or lower lobe and the left lower lobe. In multiple N2 stations, the frequency of metastasis of the N2 station observed in a single N2 station was as high as 72% to 89%, and one or two other frequently metastasized stations were added to each group. Regarding the survival of patients with a primary tumor in each lobe except for the left lower lobe, a single N2 station resulted in a significantly better survival than did multiple N2 stations. Furthermore, the overall survivals classified according to each primary site showed a significant difference among the four primary sites (P =.04). CONCLUSIONS: The primary tumors in each lobe showed a prevalence of N2 station(s). The number of N2 stations is a good prognosticator except in patients with a primary tumor in the left lower lobe. In addition, the site of a primary tumor itself is also considered to influence the survival of the patients.     

Establishment of gene-vaccinated skin grafting against Toxoplasma gondii infection in mice

Vaccine effects of in vivo gene-vaccinated skin graft were evaluated against Toxoplasma gondii (T. gondii) infection. By using a gene gun, cDNA coding T. gondii SAG1 molecule was intracutaneously vaccinated into C57BL/6 (B6; a susceptible strain), BALB/c (a resistant strain) and (C57BL/6 x BALB/c) F1 (CBF1) mice, and the gene-vaccinated skin of these strains was transplanted to CBF1 mice. Regarding the antibody production against SAG1, CBF1-recipient mice transplanted with the SAG1 gene-vaccinated B6 skin were high responders, whereas CBF1 mice skin grafted with vaccinated skin of both BALB/c and CBF1 mice were low responders. The donor-derived LC/DC migrated to the draining lymph nodes of the recipients from the skin graft within 3 days. The vaccine effect against T. gondii challenge infection was obtained in CBF1 mice which received the skin graft of the SAG1 gene-vaccinated BALB/c mice.     

Brown-Séquard syndrome and cervical CSF leakage due to a knife injury: a case report

We report a case of Brown-Séquard syndrome and cervical CSF leakage caused by a knife injury. A 34-year-old man was involved in a fight and was stabbed on his occiput and back with a knife. Neurological examination on admission showed right hemiparesis, right hemihypesthesia and left hemihypalgesia, indicating Brown-Séquard syndrome. Furthermore, cerebrospinal fluid was leaking from the occipital stab wound. Head CT scan showed massive accumulation air in the subarachnoid space. Cervical MRI showed that the injury tract reached to the space between the occipital bone and the atlas. One week after admission, suboccipital craniectomy and duraplasty were performed because of continuous CSF leakage. Although, the CSF leakage recurred due to the wound infection, it disappeared naturally as the patient's general condition improved. Follow-up MRI studies demonstrated the cervical spinal lesion as hyperintensity on T2WI, which localized at the right side of the spinal cord. The patient's hemiparesis gradually improved and he underwent rehabilitation. Spinal cord injury due to a stab wound by a knife is rare in Japan. In this case, we suppose that the mechanism of spinal cord injury was due to direct injury by a knife avoiding the lateral corticospinal tract because his right hemiparesis obviously improved.     

Development of Aberrant Crypt Foci Involves a Fission Mechanism as Revealed by Isolation of Aberrant Crypts

Morphological analysis of isolated colonic crypts in rats, postnatally, indicated that the crypts reproduce themselves by a fission mechanism, the division beginning at the crypt base and proceeding upwards until there are two separate crypts. Occasionally, before the separation is complete, a second fission process starts on one or both sides of a bifurcating crypt and a triple-branched or quadruple-branched crypt results. Analysis of isolated aberrant crypt foci (ACF) in rats treated with 1,2-dimethylhydrazine revealed that the development of ACF consisting of multiple crypts is also due to a fission mechanism. Initially, an indentation appears at the base of a single ACF crypt, with subsequent formation of a bifurcation and eventual crypt division.     

Treatment and prognosis of primary thymic carcinoma

Forty consecutive cases with invasive ductal carcinoma of the breast (samples of both the primary invasive area and the involved node were available for 19 cases, multiple sections of breast tissue including both the primary invasive area and forefront intraductal cancerous area were available for 15 cases; and multiple sections of breast cancerous tissue and the involved node were available for six cases) were examined by immunohistochemical analysis for evaluation of their proliferation rate. In this study, we selected the PCNA index as a marker of the proliferation rate of the tumor cells. We demonstrated that the PCNA index of the primary invasive area was significantly higher than that of the forefront intraductal area (P – 0.0017), and the PCNA index of the involved nodes was significantly higher than that of the primary invasive area (P = 0.0004). These preliminary findings suggest that the progression from the tumor cells of the intraductal cancerous area to the metastatic tumor cells must be accompanied by two phases of increase in the proliferation rate. © 1993 Wiley-Liss, Inc.     

A Case of Squamous Cell Lung Carcinoma with High Concentration of Parathyroid Hormone-related Peptide in Serum and Pleural Effusion Presenting Hypercalcemia

A 57-year-old man with lung squamous cell carcinoma revealed hypercalcemia, hypophosphoremia, elevation of nephrogenous cAMP and metabolic alkalosis. Serum parathyroid hormone (PTH) and l,25(OH)₂D₃ concentrations were not elevated. These findings were consistent with those in humoral hypercalcemia of malignancy (HHM), PTH-related peptide (PTHrP) concentrations were determined using N- and C-terminal specific radioimmunoassays (PTHrP-N, PTHrP-C), and elevation of both PTHrP-N and PTHrP-C concentrations in the serum was noted (PTHrP-N, 27 pmol/liter (norma<5); PTHrP-C, 1408 pmol/liter (norma<50)). High concentration of PTHrP (946 pmol/ liter for PTHrP-N and 5983 pmol/liter for PTHrP-C) was also found in the pleural fluid obtained at autopsy. Immunohistochemical study, using paraffin-embedded sections of the tumor tissue obtained at autopsy, revealed numerous PTHrP-positive cells and expression of PTHrP gene was confirmed by Northern blot analysis. These findings indicate that PTHrP, produced in the tumor tissue, was secreted into the blood stream, which caused HHM in the patient. Gel permeation chromatography of the serum and pleural fluid revealed several peaks of both PTHrP-N and PTHrP-C. Molecular forms of PTHrP-N were larger than those of PTHrP-C in the serum as well as pleural fluid. These findings indicate that multiple forms of PTHrP molecules are present in the serum and pleural fluid. Granulocytosis was also noted in the patient. However, granulocyte- and granulocyte macrophage-cotony stimulating factor were not detected in the serum, and the mechanism of the granulocytosis in the patient was unclear.     

Correlation of urinary 8-hydroxy-2'-deoxyguanosine (8-OHdG), a biomarker of oxidative DNA damage, and clinical features of hematological disorders: a pilot study

The 8-hydroxy-2'-deoxyguanosine (8-OHdG), presently the most popular marker for oxidative DNA damage, level has been reported to be elevated in patients with various malignancies. In the present study, urinary 8-OHdG was examined in 44 patients with hematological disorders (13 malignant lymphoma, 11 adult T cell leukemia/lymphoma (ATL), 10 acute leukemia, and 10 myelodysplastic syndrome (MDS)) by an enzyme-linked immunosorbent assay. The pre-therapy level of urinary 8-OHdG in ATL patients was significantly elevated compared with normal controls (25.3+/-12.9 vs. 11.9+/-7.3 ng/mg, P<0.05). Although patients with lymphoma, acute leukemia and MDS also showed higher urinary 8-OHdG levels than normal controls, the differences were not significant. However, two patients with refractory anemia with excess blasts in transformation (RAEB-t) having extreme monocytosis and neutrophilia showed exceptionally high urinary 8-OHdG levels (161.0 and 218.9 ng/mg). Urinary 8-OHdG excretion increased transiently with chemotherapy, and this fluctuation was significant irrespective of the disorder (P<0.05). Interestingly, lymphoma patients with high LDH, advanced stage, poor performance status or International Prognostic Index (IPI) of high/high-intermediate risk had significantly elevated urinary 8-OHdG levels (P<0.05-<0.001). These latter results suggest that urinary 8-OHdG may be a reliable prognostic marker in lymphoma patients and should encourage large scale and long term follow up studies.     

Utility of liposomes coated with polysaccharide bearing 1-amino-lactose as targeting chemotherapy for AH66 hepatoma cells

The cell recognition element is very important for drug delivery systems. We synthesized cholesteryl pullulan (CHP) bearing 1-aminolactose (1-AL) and introduced a saccharide, cholesteryl pullulan bearing 1-aminolactose (1-AL/CHP), to an outer layer of the conventional liposome as a cell recognition element. Lectin recognized the beta-galactose by aggregation of 1-AL/CHP coated liposome (1-AL/CHP liposome). The uptake of this liposome to AH66 rat hepatoma cells was greater than in liposomes without 1-aminolactose in vitro. Furthermore, 1-AL/CHP liposomal adriamycin showed a stronger antitumor effect in comparison with other types of liposomal adriamycin in vitro. When in vivo tumor-targeting efficacy was investigated in AH66 tumor transplanted mice using 3H-liposome, the tumor/serum radioactivity ratio in mice injected with 1-AL/CHP liposome was higher than that of mice injected with other liposomes. These observations suggest that 1-AL is effective as a cell recognition element. As a result, 1-AL/CHP liposome is considered to be a good carrier of anticancer drugs for the active targeting of tumor cells.