Comparison of the prognosis of bisphosphonate-related osteonecrosis of the jaw caused by oral and intravenous bisphosphonates.

Bisphosphonates (BPs) have been used in medical practice for the treatment of osteoporosis, bone metastasis, and multiple myeloma. Although many studies have been published, the treatment and prognosis of bisphosphonate-related osteonecrosis of the jaw (BRONJ) remain unclear. This study included 59 patients with BRONJ: 29 had taken oral BPs and 30 had taken intravenous (IV) BPs. All received conservative treatments. When separated sequestra were seen, a sequestrectomy was performed. Segmental mandibular resection was performed when pathological fractures were diagnosed. The outcomes of treatments were compared between groups. For patients treated with oral rinses or mandibular resection, the number in whom clinical healing was observed did not differ between the oral BP and IV BP groups. With regard to sequestrectomy, 94% of patients in the oral BP group showed improvement with this treatment compared to 50% in the IV BP group. The number of patients in whom clinical healing of BRONJ was achieved was statistically better in the oral BP group than in the IV BP group after 6 months of treatment (P<0.001). The results showed that >90% of patients treated with oral BPs could be cured. However, 50% of patients treated with IV BPs did not show an improvement. Additional research is needed to further increase the therapeutic efficacy for the resolution of BRONJ.

Anti-U1RNP antibody and aseptic meningitis in connective tissue diseases.

OBJECTIVE: To investigate the relationship between aseptic meningitis and anti-U1RNP antibody in patients diagnosed with CTD. METHODS: Fourteen patients with aseptic meningitis were selected from among patients with CTDs who had visited our hospital. We analyzed their medical records to clarify the clinical and immunological features of aseptic meningitis. RESULTS: A total of 14 patients with aseptic meningitis were subsequently diagnosed as having either SLE (seven cases), MCTD (four), UCTD (one), overlap syndrome (one), or Sjögren's syndrome (one). Eight of the 14 patients had received NSAIDs, such as sulindac, naproxen, or loxoprofen, before the onset of aseptic meningitis. CRP levels were increased (mean +/- SD: 7.1 +/- 7.1 mg/dL) and CRP levels (10.4 +/- 7.7) in the drug-induced group were significantly increased (p < 0.01). The anti-U1RNP antibody was found in 13 of the 14 patients. There were no significant differences in cerebrospinal fluid findings between the drug-induced group and the non-drug-induced group. CONCLUSIONS: SLE or MCTD patients with aseptic meningitis tend to have anti-U1RNP antibody.

[Effectiveness of carboplatin and etoposide selective intra-arterial chemotherapy for progressive soft-tissue sarcoma].

Four patients with advanced soft tissue sarcomas were treated by selective intra-arterial injection of carboplatin (450 mg/m2/2 hr) and etoposide (200 mg/m2/2 hr). After 2-4 courses of treatment, tumor size was remarkably reduced. Three of the 4 patients became candidates for limb-salvage surgery with a safety margin. During the treatment, patients showed no significant complications except for slight nausea and myelosuppression. This protocol might be a recommendable treatment for advanced soft tissue sarcomas.

Heterozygous ras mutations are preserved in serially passaged human tumor xenografts and established cell lines.

We examined c-K-ras gene point mutations in human tumor xenografts and established cell lines as markers of genetic stability. Our previous study demonstrated the stability of c-K-ras gene mutations in human primary neoplasms and their tumor xenografts through serial passages in mice. In this study, we established 27 human cell lines derived from various human tumor xenografts in nude mice. Point mutation of the c-K-ras gene at codon 12 was found in 29.6% (8/27) of the cell lines, as well as in 29.6% (8/27) of the xenografts. The eight ras-mutated cell lines were derived from corresponding tumor xenografts carrying the ras mutation. Heterozygous ras gene mutation was confirmed in seven of the eight ras-mutated cell lines, as well as their corresponding xenografts. The incidence, type and heterozygosity of the c-K-ras gene mutation showed no discrepancies between the original xenografts and the established cell lines. From these findings, we concluded that point mutation of the c-K-ras gene was very stable in human tumor xenografts and established cell lines derived from the xenografts.

Cloning and sequencing of a cDNA encoding chicken mannan-binding lectin (MBL) and comparison with mammalian analogues.

The serum lectin, mannan-binding lectin (MBL) (also denoted mannan-binding protein or mannose-binding protein, MBP) has been identified in mammals (humans, monkey, cow, rabbit, mouse and rat). Upon binding to carbohydrates on the surface of microorganisms, MBL mediates activation of the complement system, leading to killing of the microorganism. MBL thus exerts a role in the innate immune defence. We have described the isolation and partial characterization of an analogous protein in chicken serum. Oligonucleotides based on the N-terminal sequence of this protein were used in a reverse transcription-polymerase chain reaction (RT-PCR) with chicken liver RNA as template. The PCR product was sequenced and found to encode part of the NH2 terminus of chicken MBL. A perfect match probe was synthesized and used to screen a chicken liver cDNA library. The isolated clones carried a cDNA insert of 1692 bp with an open reading frame of 714 bp encoding a mature protein of 238 amino acids including a signal peptide of five amino acids. The deduced amino acid sequence agrees with those determined by conventional amino acid sequence analysis of the peptides except for four residues. We have compared the deduced primary structure of chicken MBL with the mammalian analogues. The phylogenetic analysis indicates that the gene duplication leading to two different MBL forms in mammals occurred after the split from birds and reptiles. This concurs with the finding of only one form of MBL in chickens.

Using Ga-67 scintigraphy in prostatic abscess.

The use of Ga-67 scintigraphy (Ga) in prostate inflammatory diseases may be restricted by the difficulty in distinguishing between the accumulation of Ga-67-citrate (Ga-citrate) in the lesion and feces. The diagnosis of prostatic abscess has been mainly made by other radiologic methods without scintigraphic studies and no finding of Ga has been reported. This patient demonstrated that coordinating the findings of Ga-citrate accumulation can be helpful in making a prompt diagnosis of a possibly fatal prostatic abscess, especially in those patients with poorly defined clinical symptoms and high risk factors.

Immunohistochemical demonstration of p53 protein in colorectal adenomas and adenocarcinomas. Reliable application of the heat-induced antigen retrieval method to formalin-fixed, paraffin-embedded material.

Ninety-nine polypoid neoplasms and eight advanced adenocarcinomas of the colon were studied immunohistochemically for p53 protein expression. For reproducible antigen retrieval, formalin-fixed, paraffin-embedded archival sections were heated at 90 degrees C for 120 min in 0.01 mol/L phosphate-buffered saline, pH 7.2, prior to immunostaining. Proliferating cell nuclear antigen served as a positive control marker for effective antigen retrieval. The 99 polyps were categorized into 24 high-grade adenomas, 60 non-invasive cancer-in-adenomas (CIA), and 15 CIA with stromal invasion. All the polyps contained portions of low-grade adenoma. Positive nuclear staining of p53 protein was observed in none of the non-neoplastic mucosa, nine (9%) of 99 low-grade adenomas, 17 (71%) of 24 high-grade adenomas, 46 (77%) of 60 non-invasive CIA, 10 (67%) of 15 invasive CIA, and five (63%) of eight advanced carcinomas. When the antigen retrieval treatment was omitted, the positivity rates were 0, 2, 17, 35, 40, and 63%, respectively. When the antigen-retrieved staining pattern was classified into (i) 'sparse' (< 25% of the nuclei of neoplastic glands labeled), 'scattered' (25-75%) and 'dense' (> 75%); or (ii) 'focal' (the positively labeled glands occupying < 25% of the tumor area), 'intermediate' (25-75%) and 'diffuse' (> 75%), the sparse and focal patterns predominated in high-grade adenomas and non-invasive CIA with low-grade atypia, while the dense and diffuse patterns predominated in invasive CIA and all the advanced carcinomas revealed the dense and diffuse patterns. Non-invasive CIA with high-grade atypia belonged to an intermediate type between the two groups.(ABSTRACT TRUNCATED AT 250 WORDS)

Heat-induced antigen retrieval of proliferating cell nuclear antigen and p53 protein in formalin-fixed, paraffin-embedded sections.

Immunohistochemical demonstration of proliferating cell nuclear antigen (PCNA) and p53 protein is important, particularly for the surgical diagnosis of neoplastic disorders. An effective, simple and reproducible method was established for observing the expression of these intranuclear antigens in routinely processed, formalin-fixed paraffin-embedded sections. Dramatic improvement of the antigenicity was obtained when the deparaffinized sections were heated in a hot water bath at 90 degrees C for 120 min in 0.01 mol/L citrate buffer, pH 6.0, for PCNA and in 0.01 mol/L phosphate-buffered saline, pH 7.2, for p53 protein. These reliable pretreatments are useful for the detailed comparative analysis of the expression of PCNA and p53 protein and fine histologic architecture and for retrospective study using a large number of archival specimens.

A side reaction in solid phase synthesis. Insertion of glycine residues into peptide chains via Nim----N alpha transfer.

In solid-phase peptide synthesis using N alpha-Boc-Nim-tosyl-histidine (Boc-His(Tos)), byproducts having extra Gly residues in the peptide chain were observed at a high rate. When a Boc-amino acid such as Asn was incorporated after assembly of Boc-His(Tos), the Nim-tosyl group was partially or fully cleaved by an activating agent, 1-hydroxybenzotriazole. In the successive coupling reactions, Boc-Gly was incorporated into the free Nim ring as well as the alpha-amino function, and the Nim-Gly was then transferred to the alpha-amino group of Gly of the peptide chain after removal of these Boc groups to give extra Gly residues at the position of Gly. This was observed in only the coupling reaction with Boc-Gly and could be circumvented using a more stable Nim protecting group for His, such as a dinitrophenyl group.

Synthesis of RNA oligomer using 9-fluorenylmethoxycarbonyl (Fmoc) group for 5'-hydroxyl protection.

An approach using a new combination of protecting groups in RNA oligomer synthesis is proposed, in which 5'-hydroxyl group of ribose moiety is temporarily protected with the alkaline labile 9-fluorenylmethoxycarbonyl (Fmoc) group and the 2'-hydroxyl group is protected with the acid labile 1-ethoxyethyl (EE) group. The adoption of this method presented great selectivity in removing the 5'-hydroxyl protecting group and facilitated the RNA oligomer synthesis. A RNA pentamer was synthesized by the phosphotriester method in solution.