Functional Analysis of PTH1R Variants Found in Primary Failure of Eruption.

Although many variants of the parathyroid hormone 1 receptor (PTH1R) gene are known to be associated with primary failure of eruption (PFE), the mechanisms underlying the link remains poorly understood. We here performed functional analyses of PTH1R variants reported in PFE patients-namely, 356C>T (P119L), 395C>T (P132L), 439C>T (R147C), and 1148G>A (R383Q)-using HeLa cells with a lentiviral vector-mediated genetic modification. Two particular variants, P119L and P132L, had severe reduction in a level of N-linked glycosylation when compared with wild-type PTH1R, whereas the other 2 showed modest alteration. PTH1R having P119L or P132L showed marked decrease in the affinity to PTH1-34, which likely led to severely impaired cAMP accumulation upon stimulation in cells expressing these mutants, highlighting the importance of these 2 amino acid residues for ligand-mediated proper functioning of PTH1R. To further gain insights into PTH1R functions, we established the induced pluripotent stem cell (iPSC) lines from a patient with PFE and the heterozygous P132L mutation. When differentiated into osteoblastic-lineage cells, PFE-iPSCs showed no abnormality in mineralization. The mRNA expression of RUNX2, SP7, and BGLAP, the osteoblastic differentiation-related genes, and that of PTH1R were augmented in both PFE-iPSC-derived cells and control iPSC-derived cells in the presence of bone morphogenetic protein 2. Also, active vitamin D3 induced the expression of RANKL, a major key factor for osteoclastogenesis, equally in osteoblastic cells derived from control and PFE-iPSCs. In sharp contrast, exposure to PTH1-34 resulted in no induction of RANKL mRNA expression in the cells expressing P132L variant PTH1R, consistent with the idea that a type of heterozygous PTH1R gene mutation would spoil PTH-dependent response in osteoblasts. Collectively, this study demonstrates a link between PFE-associated genetic alteration and causative functional impairment of PTH1R, as well as a utility of iPSC-based disease modeling for future elucidation of pathogenesis in genetic disorders, including PFE.

Influenza 2014-2015 among pregnant Japanese women: primiparous vs multiparous women.

This study was performed to determine whether multiparous pregnant women are prone to influenza. A questionnaire survey was conducted at 19 centres located throughout Japan, targeting all 6,694 postpartum women within 7 days after birth before leaving the hospital. All women gave birth during the study period between March 1, 2015, and July 31, 2015. Data regarding vaccination and influenza infection in or after October 2014, age, previous experience of childbirth, and number and ages of cohabitants were collected. Seventy-eight percent (n = 51,97) of women given questionnaires responded. Of these, 2,661 (51 %) and 364 (7.0 %) women reported having been vaccinated and having contracted influenza respectively. Multiparous women had a higher risk of influenza regardless of vaccination status (8.9 % [121/1362] vs 5.7 % [74/1299], relative risk [95 % confidence interval], 1.80 [1.36 to 2.38] for vaccinated and 9.3 % [112/1198] vs 4.3 % [57/1328], 2.18 [1.60 to 2.97] for unvaccinated women) compared to primiparous women. The risk of influenza increased with increasing number of cohabitants: 4.8 % (100/2089), 7.5 %, (121/1618), 9.0 %, (71/785), and 10.4 % (58/557) for women with 1, 2, 3, and ≥4 cohabitants respectively. Family size is a risk factor for influenza infection in pregnancy.

Ultrasound-guided surgery of particulate cancellous bone and marrow from the iliac crest for cleft palate.

Minimally invasive procedures for treatment of disease have become increasingly popular and require specialized instruments and precise imaging guidance. We here propose a technique of minimally invasive surgery with ultrasound echo-guided procedures as a less traumatic and invasive procedure to get particulate cancellous bone and marrow from the iliac crest for cleft palate. This technique has been used successfully at our institutions. Our experience suggests that it can provide reliable ultrasound echo imaging-guided surgery.

Interleukin-1 beta in unstimulated whole saliva is a potential biomarker for oral squamous cell carcinoma.

The objective of this study was to evaluate cytokines in unstimulated whole saliva (UWS) of oral squamous cell carcinoma (OSCC) patients as compared to those with pre- and post-operation for evaluation as markers of OSCC. Sixteen OSCC patients were included in this study. Cytokine concentrations in resting saliva were measured using a Bio-Plex suspension array system. Only interleukin-1 (IL-1) beta showed significantly different cytokine concentration in saliva between pre- and post-operation. IL-1 beta was released from cultured OSCC cells confirmed by ELISA and immunohistochemistry. From this study, IL-1 beta in UWS may be useful for detection of early stage OSCC. More studies are needed to accept the utility of IL-1 beta in UWS for predicting, diagnosis and evaluation of OSCC.

Magnesium intake decreases Type 2 diabetes risk through the improvement of insulin resistance and inflammation: the Hisayama Study.

AIMS: Early studies have shown that magnesium intake decreases the risk of Type 2 diabetes, but the results are still inconsistent. We prospectively examined the association between magnesium intake and incidence of Type 2 diabetes in a general Japanese population. METHODS: A total of 1999 subjects without diabetes aged 40-79 years who underwent a 75-g oral glucose tolerance test were followed up prospectively for a mean of 15.6 years. RESULTS: During the follow-up, 417 subjects developed Type 2 diabetes. The age- and sex-adjusted incidence of Type 2 diabetes significantly decreased with increasing magnesium intake quartile levels (≤ 148.5, 148.6-171.5, 171.6-195.5 and ≥ 195.6 mg/day, P for trend = 0.01). In multivariate analyses, after adjusting for comprehensive risk factors and other dietary factors, the hazard ratio of Type 2 diabetes was 0.67 (95% CI 0.49-0.92; P = 0.01) in the third quartile and 0.63 (95% CI 0.44-0.90; P = 0.01) in the highest quartile compared with the first quartile. In addition, the risk of Type 2 diabetes was 14% lower (P = 0.04) for a 1-sd increment of log-transformed magnesium intake in the multivariate-adjusted model. In stratified analysis, there were statistically significant interactions between magnesium intake and levels of homeostasis model assessment of insulin resistance, high-sensitivity C-reactive protein or alcohol intake on the risk of Type 2 diabetes (all P < 0.05). CONCLUSIONS: Our findings suggest that increased magnesium intake was a significant protective factor for the incidence of Type 2 diabetes in the general Japanese population, especially among subjects with insulin resistance, low-grade inflammation and a drinking habit.

Antitumor efficacy of oncolytic herpes simplex virus adsorbed onto antigen-specific lymphocytes.

Although several studies have reported that locally administering oncolytic viruses effectively targets malignancies, the efficacy of systemically administered oncolytic viruses is restricted. Recently, however, it was reported that systemic administration of oncolytic vesicular stomatitis virus adsorbed onto antigen-specific lymphocytes was effective against malignancies. We hypothesized that intravenously administering such virus might have significant potential in treatment of the malignant tumors. We adsorbed oncolytic herpes simplex virus-1 mutant R3616 onto lymphocytes harvested from mice with acquired antitumor immunity. We administered adsorbed R3616 to peritoneally disseminated tumors and analyzed the efficacy of this treatment. Mice administered adsorbed R3616 survived significantly longer than mice administered R3616 adsorbed onto non-specific lymphocytes, or mice administered either virus or tumor antigen-specific lymphocytes alone. In this context, herpes oncolytic virus is a promising treatment not only for primary lesions, but also for multiple metastasizing lesions. This treatment strategy may become one of the most effective methods for systemic virus delivery.

Zygoma implant-supported prosthetic rehabilitation of a patient with a maxillary defect.

This clinical report describes the successful management of a patient who underwent extensive resection of a maxillary cancer, by introduction of a maxillary obturator prosthesis using zygoma implants. The patient was a 57-year-old man with cancer of the upper anterior gingiva. The maxillary bone in the affected region had been extensively excised by radical surgery. Owing to loss of teeth retaining the denture, the existing prosthesis was unstable, and the patient experienced severe speech and mastication disorders. Four zygoma implants (two on each side), and two conventional dental implants (one each at both maxillary tuberosities) were used as denture retainers. The obturator prosthesis was stabilized by the implants, and the patient's oral function improved. High-level compatibility between the implant and surrounding tissue was obtained by mucosal regeneration around the implant. The results suggest that the combination of zygoma and conventional dental implants improves postoperative oral function by facilitating retention of the obturator prostheses.

A phase I dose-escalation clinical trial of intraoperative direct intratumoral injection of HF10 oncolytic virus in non-resectable patients with advanced pancreatic cancer.

In 2005, we initiated a clinical trial that examined the efficacy of the oncolytic virus HF10 to treat pancreatic cancer. Pancreatic cancer continues to have a high mortality rate, despite multimodal treatments for patients, and new therapeutic methods are greatly needed. The current mainstream methods for cancer treatment include biological therapeutics such as trastuzumab (Herceptin) for breast cancer or erlotinib (Tarceva) for non-small cell lung cancer. Oncolytic virus therapy is a new and promising treatment strategy for cancer. Oncolytic viruses are novel biological therapeutics for advanced cancer that appear to have a wide spectrum of anticancer activity with minimal human toxicity. To examine the efficacy of oncolytic virus therapy for pancreatic cancer, we initiated pilot studies by injecting six patients with non-resectable pancreatic cancer with three doses of HF10. All patients were monitored for 30 days for local and systemic adverse effects and were not administered any other therapeutics during this period. There were no adverse side-effects, and we observed some therapeutic potential based on tumor marker levels, survival, pathological findings and diagnostic radiography. The tumors were classified as stable disease in three patients, partial response in one patient and progressive disease in two patients.

Friend leukaemia insertion (Fli)-1 is a prediction marker candidate for radiotherapy resistant oral squamous cell carcinoma.

Radiotherapy is commonly used to treat oral squamous cell carcinoma (OSCC), but its therapeutic effects are unpredictable. To determine which genes correlate with radiation resistance in oral cancer, the authors evaluated radiation sensitivity using a standard colony formation assay with a gene microarray system for seven OSCC cell lines. They found significant associations between dozens of gene-expression levels and radiation resistance of OSCC cell lines. Following analysis of the different radiosensitive cancer cell lines, the friend leukaemia insertion (Fli)-1 gene was selected as a prediction marker gene for OSCC radiotherapy resistance. Fli-1 expression was associated with radiation resistance in OSCC patients. These data help to predict the effects radiation therapy has on OSCC, in turn contributing to the development of alternative radiation therapies.

Analysis of bone volume using computer simulation system for secondary bone graft in alveolar cleft.

The purpose of this study was to measure the bone volume necessary for secondary bone grafting in the alveolar cleft using surgical simulation software based on three-dimensional computed tomography (CT) scan data, to compare this measurement with the actual volume of the bone graft, and to evaluate consistency. The subjects were 13 patients with cleft lip and palate who underwent CT using a cone-beam CT unit (CB-CT) 1 month before surgery, followed by bone grafting with particulate cancellous bone and marrow (PCBM) to close the cleft. The bone volume necessary for grafting was measured based on the CB-CT scan data. Correlation analysis, a test of the population mean between two samples, and Wilcoxon's signed rank test were conducted between these measurements and the actual bone volume (PCBM volume) used for grafting. SPSS was used for statistical analysis, and the level of significance was set below the 5% level. The results showed a significant correlation, with no significant differences between the two in all tests. These results suggest that measuring and preoperatively calculating the bone volume necessary for bone grafting with surgical simulation software using CB-CT scan data is beneficial.

Mandibular reconstruction using 2-step bone transport in an irradiated cancer patient with type 2 diabetes mellitus.

This report describes mandibular reconstruction using a 2-step bone transport technique in a cancer patient with type 2 diabetes mellitus (DM) who received radiotherapy before ablative surgery. The bone, measuring 90 mm in length with the mandibular curvature and good alveolar form, was regenerated using a plate-guided method. Three implants were placed 21 months after completion of the first distraction and 14 months after the second distraction. These implants integrated successfully and have remained stable more than 2 years after loading. The results suggest that mandibular reconstruction using bone transport is possible after segmental osteotomy in irradiated cancer patients with DM, although a long treatment period is required.

Analyses of speech intelligibility in patients after glossectomy and reconstruction with fasciocutaneous/myocutaneous flaps.

This study analyzed the results of speech intelligibility tests in patients with tongue cancer who had undergone resection with the aim of making surgical recommendations for flap design and inset, to improve speech function. A total of 126 patients, enrolled from 13 Japanese institutions, were classified into 3 groups according to the resected site: lateral, anterior, and combined. The lateral group was further divided into 3 subgroups and the anterior group into 2 subgroups according to the size of resection. The speech intelligibility score was analyzed based on articulatory site and mode: 5 articulatory sites (linguodentoalveolar, linguopalatal, linguovelar, and their intermediates); and 7 articulatory modes (plosives, fricatives, affricatives, grids, nasals, vowels and semivowels). Low speech intelligibility scores were recorded at sites where flaps contribute directly to the pronunciation in the lateral and combined groups and at the anterior part of the reconstructed tongue in the anterior group. Plosives and glides displayed low values in general. A radial forearm flap had higher function in the lateral group than other flaps. The type of flap had no effect in the anterior and combined groups. Surgical techniques and flap selection to improve functional status in each type of resection are discussed.

Physical activity level in healthy free-living Japanese estimated by doubly labelled water method and International Physical Activity Questionnaire.

OBJECTIVE: To measure total energy expenditure (TEE) for normal healthy Japanese by the doubly labelled water (DLW), and to compare the physical activity level (PAL) among categories classified by the categories used in daily reference intake (DRI), Japan and the International Physical Activity Questionnaire (IPAQ). SUBJECTS AND METHODS: A total of 150 healthy Japanese men and women aged 20- to 59-year-old living in four districts of Japan. TEE was measured by the DLW method, and the PAL was calculated from TEE divided by basal metabolic rate. Simultaneously with TEE measurement, the PAL was assessed employing the categories used in DRI, Japan and IPAQ. RESULTS: The average TEE and PAL were 10.78+/-1.67 MJ/day and 1.72+/-0.22 for males and 8.37+/-1.30 MJ/day and 1.72+/-0.27 for females, respectively. The subjects in the highly active categories assessed by both DRI and IPAQ showed significantly higher PAL compared with less active categories. However, PALs among light and moderate categories by DRI, and insufficient and sufficiently active by IPAQ were not significantly different. CONCLUSIONS: In developed countries, highly active subjects could be assessed by a simple questionnaire. However, the questionnaire should be improved to clarify the sedentary to moderately active subjects by assessing carefully very light to moderate physical activity.

Primary omental gamma/delta T-cell lymphoma involving the central nervous system.

Gamma/delta T-cell lymphoma (GDTL) is an uncommon lymphoma that was initially reported to involve only the liver and spleen. GDTL other than the hepatosplenic type is extremely rare. Frequent primary sites include skin and subcutaneous tissue, intestine, or nasal region. We report a case of GDTL of the omentum in a 54 year-old-man. The tumor cells are CD2-, CD3+, CD4-, CD5-, CD8+, CD56+, TIA-1+, granzyme B+/-. They expressed the identical phenotype of intestinal GDTL. The patient was treated with 2 courses of CHOP which comprised cyclophosphamide, doxorubicin, vincristine and prednisolone, and 3 courses of EPOCH which comprised etoposide, prednisolone, vincristine, cyclophosphamide and doxorubicin. However, he failed to obtain remission. During the fourth course of EPOCH, muscle weakness of the lower extremities developed and intracranial masses were observed by computed tomographic scan of the brain. Dissemination of lymphoma to the central nervous system was considered and it may be attributable to the expression of CD56 in this case. High dose methotrexate (HD-MTX) chemotherapy successfully eliminated the omental tumor and reduced the size of the intracranial masses, thus HD-MTX appears to be an effective treatment against GDTL.

Mineralized bone loss in partially edentulous trabeculae of ovariectomized rabbit mandibles.

OBJECTIVE: The purpose of this study was to evaluate trabeculae changes in partially edentulous bone in ovariectomized rabbits. BACKGROUND: Numerous clinical studies have suggested that the greater risk for oral bone loss in females may be correlated with osteoporosis after menopause. Knowledge of trabecular changes in partially edentulous bone in animals with loss of ovarian function may be beneficial in the diagnosis and treatment of partially edentulous patients of postmenopausal women. METHODS: Twelve adult female Japanese white rabbits were examined. The mandibular incisors were initially extracted to simulate the partially edentulous bone. Six animals were bilaterally ovariectomized and the other six sham-ovariectomized 12 weeks after tooth extraction. The partially edentulous parts of distal mandibular bodies were processed undecalcified 12 weeks after ovariectomized or sham-ovariectomized surgeries and examined by quantitative trabecular bone histomorphometry. RESULTS: In ovariectomized rabbits, there were significant increases in trabecular separation, osteoid volume, osteoid thickness, osteoid width, eroded surface, and mineral apposition rate, and a significant decrease in trabecular number. CONCLUSION: The results of sparser trabecular structure, more trabecular osteoid, and increased trabecular bone turnover demonstrate mineralized bone loss in partially edentulous trabeculae of ovariectomized rabbit mandibles and suggest that the same loss may occur in postmenopausal women.

Bone mineral density in mandibles of ovariectomized rabbits.

The purpose of this study was to evaluate the association between loss of ovarian function and bone mineral density changes in the mandibles of ovariectomized rabbits using peripheral quantitative computed tomography. Twenty-four adult female Japanese white rabbits were used in this experiment. The mandibular incisors were initially extracted to create implant bone. Twelve animals were bilaterally ovariectomized and the other twelve sham-ovariectomized 12 weeks after tooth extraction. All rabbits were sacrificed at 4 and 12 weeks after ovariectomy and sham-ovariectomy, with 6 rabbits in each of the four resultant groups. The edentulous parts of distal mandibular bodies were measured by peripheral quantitative computed tomography. There were significant decreases in total bone mineral density and trabecular bone mineral density in the mandibles of rabbits at 12 weeks post-ovariectomy. The results demonstrate the mineralized bone loss in the mandibles of ovariectomized rabbits.

Clinical pathway for impalpable or small lung lesions treated with coil marking and thoracoscopy.

OBJECTIVES: Advances in computed tomography are detecting increasingly impalpable or small pulmonary lesions. We propose a clinical pathway for managing such lesions. METHODS: We conducted a retrospective study in a community teaching hospital to describe the hospital schedules of 18 patients having 19 lesions 10 mm or less and ground glass attenuation. Under computed tomography, a coil (Complex Helical Fibered Platinum Coil-18) was placed at the proximal side of the lesion. Using thoracoscopy and radiographic fluorography, we conducted partial lung resection targeting the coil the next day, adding lobectomy, if required. RESULTS: Final diagnosis included primary and metastatic lung cancer (n = 14), atypical adenomatous hyperplasia (n = 1), and benignancy (n = 4). Patients were admitted 2* days before surgery (*Numbers are medians). On postoperative day 3, chest tubes were removed. Epidural analgesia was continued for 5 days. On postoperative day 7, patients were discharged. Their admission charge was a total of yen 979,610. CONCLUSIONS: The hospital course above may be applied to the clinical pathway for managing impalpable or small lung lesions.

Penetration of the duodenum by an ingested needle with migration to the pancreas: report of a case.

A case of a penetration of the duodenum by a needle with migration to the pancreas in a 50-year-old man is reported herein. The patient was referred to us with a chief complaint of diarrhea. An abdominal plain roentgenogram showed a needle in the upper abdominal area. An abdominal computed tomography scan and contrast X-ray revealed the foreign body to be located outside of the duodenum and in the head of the pancreas. An emergency operation was therefore performed on the first day and the needle in the head of the pancreas was thus extirpated safely. A perforation of the gastrointestinal tract by an ingested foreign body is difficult to accurately and quickly diagnose when no peritonitis or abscess formation is observed. Therefore, the use of contrast X-ray is considered to be useful in the diagnosis of such a perforation.

[Multiple myeloma with Sweet disease developing from monoclonal gammopathy of undetermined significance and Sjögren syndrome].

A 57-year-old woman was diagnosed as having monoclonal IgG kappa gammopathy of undetermined significance with Sjögren syndrome. Five years later, she was admitted with an increased level of serum IgG and diagnosed as having multiple myeloma. After admission, fever and painful erythema developed. Combined chemotherapy with adrenal cortical steroid diminished the skin lesions. Erythema recurred during treatment with granulocyte colony-stimulating factor for neutropenia due to chemotherapy. A biopsy specimen from the skin revealed dense neutrophilic infiltration in the dermis, and a diagnosis of Sweet disease was made.