CD138-negative clonogenic cells are plasma cells but not B cells in some multiple myeloma patients.

Clonogenic multiple myeloma (MM) cells reportedly lacked expression of plasma cell marker CD138. It was also shown that CD19(+) clonotypic B cells can serve as MM progenitor cells in some patients. However, it is unclear whether CD138-negative clonogenic MM plasma cells are identical to clonotypic CD19(+) B cells. We found that in vitro MM colony-forming cells were enriched in CD138(-)CD19(-)CD38(++) plasma cells, while CD19(+) B cells never formed MM colonies in 16 samples examined in this study. We next used the SCID-rab model, which enables engraftment of human MM in vivo. CD138(-)CD19(-)CD38(++) plasma cells engrafted in this model rapidly propagated MM in 3 out of 9 cases, while no engraftment of CD19(+) B cells was detected. In 4 out of 9 cases, CD138(+) plasma cells propagated MM, although more slowly than CD138(-) cells. Finally, we transplanted CD19(+) B cells from 13 MM patients into NOD/SCID IL2Rγc(-/-) mice, but MM did not develop. These results suggest that at least in some MM patients CD138-negative clonogenic cells are plasma cells rather than B cells, and that MM plasma cells including CD138(-) and CD138(+) cells have the potential to propagate MM clones in vivo in the absence of CD19(+) B cells.

Immunoglobulin prophylaxis against cytomegalovirus infection in patients at high risk of infection following allogeneic hematopoietic cell transplantation.

Reports on the efficacy of intravenous immunoglobulin (IVIG) prophylaxis against cytomegalovirus (CMV) infection after allogeneic hematopoietic cell transplantation (HCT) have often sparked controversy. In addition, we are not aware of any study that has examined whether prophylaxis with IVIG affects the incidence of CMV infection in high-risk patients--those who are elderly or have received human leukocyte antigen (HLA) mismatched HCT. In the present open-label, phase II study, we addressed this question. We enrolled 106 patients in the study. The cumulative incidences of CMV infection at 100 days after HCT were similar in the intervention and the control groups (68% and 64%, P=.89; 89% and 87%, P=.79, respectively, for patients 55 years or older and those who received HLA-mismatched HCT). In those who received HLA-mismatched HCT, 1-year overall survival after HCT was 46% in the intervention group and 40% in the control group (P=.31); for age≥55 years, the corresponding values were 46% and 40% (P=.27). Our data showed that prophylaxis with regular polyvalent IVIG did not affect the incidence of CMV infections or survival among older patients or those who receive HLA-mismatched HCT.

Three novel repetitive units of reelin.

Reelin is a protein involved in the organization of the developing brain of vertebrates. Reelin is divided into three parts, a F-spondin-like domain, a region including the epitopes of the monoclonal and polyclonal anti-Reelin antibodies, and eight-fold Reelin repeats. Reelin repeats consists of a subrepeat A, an EGF-like domain, and a subrepeat B. We found that the anti-Reelin antibodies' recognition region consists of three repetitive units. The correspondence of the repetitive units with the antigenicity is discussed.

Simultaneous determination of D-lactic acid and 3-hydroxybutyric acid in rat plasma using a column-switching HPLC with fluorescent derivatization with 4-nitro-7-piperazino-2,1,3-benzoxadiazole (NBD-PZ).

A highly sensitive method for the determination of D-lactic acid and 3-hydroxybutyric acid (3-HB) in rat plasma was developed using high-performance liquid chromatography with octadecylsilica (ODS) connected to a chiral column. At first, (D + L)-lactic acid and 3-HB in the plasma were derivatized with a fluorescent reagent, 4-nitro-7-piperazino-2,1,3-benzoxadiazole (NBD-PZ), separated on the ODS column and determined fluorimetrically at 547 nm with 491 nm of excitation wavelength. During the separation step on the ODS, the peak fraction of (D + L)-lactate derivative was introduced directly to a phenylcarbamoylated beta-cyclodextrin chiral column by changing the flow of the eluent via a six-port valve. Then, D-lactate derivative was separated enantiomerically from the L-lactate derivative, and the enantiomeric ratio was determined from the chromatogram. Intra- and inter-day accuracy values for the determination of D-lactic acid in 10 microL of rat plasma were 97.8-109.2 and 98.4-109.9%, and those for 3-HB were 99.8-108.4 and 99.8-103.8%, respectively. The intra- and inter-day precision values were within 4.6 and 5.1% for D-lactic acid, and 2.7 and 2.4% for 3-HB, respectively. The detection limits for D-lactic acid and 3-HB were approximately 2.0 and 0.04 microM, respectively (signal-to-noise ratio 3). The proposed method was applied to the plasma of diabetic rats induced by intraperitoneal administration of streptozotocin, and the significant increases of both D-lactic acid and 3-HB concentrations were observed in the diabetic rats as compared to the normal rats.

Analysis of genetic alterations in salivary gland tumors by comparative genomic hybridization.

In order to define and map chromosomal copy number alterations in salivary gland tumors (SGTs), a comparative genomic hybridization (CGH) technique was applied to two pleomorphic adenomas (PAs), one adenoid cystic carcinoma (ACC), and one basal cell adenocarcinoma (BCAC). The PAs exhibited regional copy number losses at 5q12.4-q14.1, 9q12-q21.13, and 16q11.2, as well as a gain at 20p12.1; among these, the losses at the 9q12-q21.11 and 16q11.2 regions were common to both PAs. The ACC showed overrepresentations of the entire regions of chromosomes 16 and 20, a regional gain at 22q12.3-q13.1, and no losses. In the BCAC, regional gains at 9p21.1-pter, 18q21.1-q22.3, and 22q11.23-q13.31 as well as losses at 2q24.2 and 4q25-q27 were seen; the gain at 22q12.3-q13.1 was common in both the ACC and the BCAC. These CGH data indicate that different genetic alterations are present in the different types of SGTs, and that the alterations involve several chromosomes. The discovery of common alterations in the same and/or different types of tumors might be important in the understanding of the development and progression of the SGTs.

Marked therapeutic effect of dimyristoylphosphatidylcholine liposomes on carcinoma mice model in vivo.

Significantly prolonged survival rate was obtained for the first time using carcinoma mice models after the administration of single-component liposomes of dimyristoylphosphatidylcholine (DMPC) without any drug. An increase in lymphocytes under optical microscope was observed without any increase in the neutrophils count, suggesting that the DMPC liposomes might inhibit the tumor growth as well as increase in lymphocytes in vivo.

[Clinical effects of adjuvant therapy using Z-100 (Ancer 20 injection) for oral cancer--prevention of stomatitis and hematopoietic impairment].

A combination of radiotherapy and chemotherapy is a usual treatment method for malignant head and neck tumors, however chemoradiotherapy is associated with hematopoietic impairment and serious stomatitis in patients. The clinical effects and evaluation of hematopoietic activity (e.g., leukocyte count) and the degree of stomatitis under adjuvant therapy using Z-100 (Ancer 20 injection) for oral cancer were investigated for preoperative cancer therapy. In order to evaluate the clinical effects of Ancer 20 injection with regard to hematopoietic activity and the degree of stomatitis, a clinical study was performed for 18 patients with oral cancer in our department. The 18 patients, who had oral squamous cell carcinomas (5 of the tongue, 4 of the mandibular gingiva, 3 of the maxillary gingiva, 1 of the floor of the mouth, 3 of the buccal mucosa, and 2 others), were treated with this combination of adjuvant therapy with Ancer 20 injection, from March, 1991 to March, 1997. They were injected with Ancer 20 (twice a week, 40 micrograms) during the cancer treatment period. We investigated hematopoietic activity, (e.g., leukocyte and platelet counts) and the degree of stomatitis periodically, before and after the combined radiotherapy and chemotherapy treatment period. It was found that in the patients who were treated with Ancer 20 injection, the decrease in leukocyte and platelet counts was prevented and the condition of stomatitis was improved. These results suggest that Ancer 20 injection may generally improve various dysfunctions due to hematopoietic impairment by radiotherapy combined with chemotherapy, and improve immunological factors. We conclude that Ancer 20 injection is a useful adjuvant treatment for oral cancer.

Fluorimetric determination of D- and L-lactate derivatized with 4-(N, N-dimethylaminosulfonyl)-7-piperazino-2,1,3-benzoxadiazole (DBD-PZ) by high-performance liquid chromatography.

A highly sensitive method, based on fluorescence derivatization with 4-N,N-dimethylaminosulfonyl-7-piperazino-2,1,3-benzoxadiazol e (DBD-PZ), was developed for the determination of D- and L-lactic acid. High-performance liquid chromatographic separation of the lactic acid derivative was achieved using an octadecylsilica column followed by enantiomeric separation on a phenylcarbamoylated beta-cyclodextrin chiral column. The separation factor for D, L-lactic acid derivatives was 1.30 using MeOH/H(2)O (80/20) as a mobile phase, and the detection limits were approximately 360 and 300 fmol on column for D- and L-lactic acid derivative, respectively. The proposed method was applied to determine D- and L-lactate in a wine and a lactic drink. Both D- and L-lactate could be determined simultaneously, with the precisions ranging from 3.50 to 5.78% for intra-day and 4.28-9.92% for inter-day determinations. The relative recovery was in the range of 91.6-100%.

Enantiomeric determination of D- and L-lactate in rat serum using high-performance liquid chromatography with a cellulose-type chiral stationary phase and fluorescence detection.

A method is described for the quantitative determination of d- and L-lactate in 10 microl of rat serum, which includes fluorescence derivatization of D- and L-lactate with 4-(N, N-dimethylaminosulfonyl)-7-piperazino-2,1,3- benzoxadiazole (DBD-PZ) followed by O-acetylation. The derivatives are separated by HPLC on an octadecylsilica, and, via column switching, on a cellulose-type chiral column. Levulinic acid was used as the internal standard. The enantiomers of lactate were separated with the separation factor (alpha) of 1.27 and the resolution (Rs) of 2.72, while the linearity for the detection was over the range of 10 nmol/ml to 20 micromol/ml (r = 0.999). Interday precision values for D-lactate in rat serum were 5.8, 5.3, and 4.1% for 10, 100, and 1000 nmol/ml, and accuracy values were 109.6, 98.2, and 103.1%, respectively (n = 5). The reduction of d-lactate concentration in rat serum by fasting was observed with the method.

[Recent progress on analytical chemistry and biochemistry of D-amino acids].

Recent findings that D-amino acids, especially D-aspartic acid and D-serine, exist in vivo in the mammalian tissues (brain and peripheries), prompted us now to investigate their biological and pathological roles in mammals. In this review, the overview of the progress of analytical chemistry and biochemistry of D-amino acids is described.

[Interstitial pneumonia complicated by rapidly progressive glomerulonephritis associated with anti-myeloperoxydase antibody].

A 78-year-old man was admitted to our hospital in October 1994 because of renal dysfunction. The level of anti-myeloperoxidase antibody in serum was 500 EU/ml, and examination of a specimen obtained by open renal biopsy revealed crescentic glomerulonephritis. A chest roentgenogram revealed no abnormality. Administration of prednisolone was started at 50 mg/day, and the dosage was then tapered. Renal function remained stable, but on interstitial shadow appeared on the chest roentgenogram. Pulmonary function tests showed a restrictive ventilatory abnormality and a low diffusing capacity. A thoracoscopic lung biopsy was done in April 1995, and microscopic examination of the specimen showed interstitial pneumonia with relatively young granulation. The dosage of prednisolone was increased to 50 mg/day, after which the interstitial shadow decreased and pulmonary function improved. The level of anti-myeloperoxydase antibody in serum was 16 EU/ml (weakly positive). Alveolar hemorrhage can occur in cases of rapidly progressive glomerulonephritis. In addition, interstitial pneumonia or pulmonary fibrosis can also complicate this condition. In the present case, glomerulonephritis associated with anti-myeloperoxydase antibody co-existed with interstitial pneumonia. This case is valuable because both renal biopsy and lung biopsy specimens were available.

[Isolated iliac artery aneurysms associated with disseminated intravascular coagulation (DIC)].

An 84-year-old man was referred to our hospital on December 3, 1993, because of excessive bleeding after tooth extraction. Coagulation studies revealed thrombocytopenia (2.2 x 10(4)/microliter), increased serum levels of FDP-E (3172 ng/ml) and D dimer (42.6 micrograms/ml), and increased plasma levels of thrombin-atithrombin III complex (48.0 ng/ml) and plasmin-alpha 2-plasmin inhibitor complex (6.8 micrograms/ml). Computed tomography showed bilateral common iliac artery aneurysms. A diagnosis of DIC due to bilateral common iliac artery aneurysms was made. Heparin (10,000 U/day) administered to treat DIC resulted in good control, and operative repair of the aneurysms was successful. This is the second reported case of isolated iliac artery aneurysms associated with DIC.

[Contamination of flexible fiberoptic bronchoscopes with Mycobacterium chelonae linked to an automated endoscope disinfection machine--on the relationship between the presence of the organism in the intestinal tract and contamination of disinfection machine, and a case of gallbladder and bile duct infection with M. chelonae].

In 1993, thirteen strains (8.7%) of M. chelonae were isolated from bronchoalveolar lavage fluid (BALF) obtained by bronchoscopy of 150 patients in Tachikawa-sogo (T) hospital, where the same automated disinfection machine was commonly used for cleaning, sterilization and disinfection of fiberbronchoscope and fibercolonoscope except 3 bronchoscopes disinfected by gas sterilization. Since January 1994, manual cleaning and sterilization has been applied for bronchoscope, and thereafter no strain of M. chelonae was isolated from BALF of 55 patients in the T hospital. While only one strain (3%) of M. chelonae was isolated from BALF of 33 patients in Ota (O) hospital, but many strains of M. chelonae were isolated from intestinal fluid obtained by fibercolonoscopy of the patients. Manual method of cleaning and disinfection was performed for both bronchoscopes and colonoscopes in the O hospital from the beginning. Based on these results, it was suggested that M. chelonae are commonly present in the colon (intestine) of normal persons. Thus colonoendscope may be often contaminated with the organism and subsequently the automated disinfecting machine may also be contaminated with the organism which is resist and against usual disinfection procedure, and resulted in bronchoscope contamination. If the presence of M. chelonae in intestinal tract is not rare, bile duct may be naturally infected with the organism. A case of cholecystitis and cholangitis caused by M. chelonae, which has not been reported previously, was found in the T hospital.

An unusual foreign body in an unerupted supernumerary tooth.

A case of an unusual foreign body (a tip fragment from a plastic chopstick) embedded in one of two unerupted supernumerary teeth in the premaxillary region of a 12-year-old Japanese boy is presented.

[The clinical effect of cisplatin and 5-FU infusion in oral cancer].

The combination chemotherapy of cisplatin and 5-fluorouracil (5-FU) is more effective than monotherapy of either drug in the head and neck cancer. In 1982, Kish reported that CF-therapy, the combination of cisplatin and 5-FU, had a high curative rate (about 82%). CF-therapy was utilized in 9 patients with carcinoma of the oral region from March, 1987 to September, 1990 in our hospital. The patients' ages ranged from 23 to 74 years old with an average of 55. All tumor were squamous cell carcinoma. There were 2 cases in stage II, 4 cases in stage III and 3 cases in stage IV. Two patients treated with only CF-therapy and the other patients treated with radiotherapy and operation in addition to CF-therapy. Our regimen is cisplatin 80-100 mg i.v. or i.a. in day 1 and 5-FU 250 mg i.a. or 5-FU 750 mg i.v. in day 2-6. The therapeutic results showed a 78% response rate to the therapy with 4 cases of CR (complete response), 3 cases of PR (partial response), 1 case of MR (minor response) and 1 case of PD (progressive disease). Side effects of cisplatin plus 5-FU (eg. nausea and vomiting) were so severe that the treatment had to the stopped in many patients. The method we have reported here was evaluated, and it is concluded that this CF-therapy is very effective as a first treatment of oral cancer.

Intermaxillary fixation using screws. Report of a technique.

A technique of intermaxillary fixation using screws anchored in the maxilla and mandible has been described. AO (Synthes) screws with a diameter of 3.5 mm and 12 mm to 16 mm in length were inserted at the antero-lateral surface of the maxilla and the buccal surface of the mandible. This technique is particularly suitable for mandibular fractures in denture wearing patients.