Safety and effectiveness of once-yearly zoledronic acid in Japanese osteoporosis patients: three-year post-marketing surveillance.

INTRODUCTION: Zoledronic acid (5 mg; ZOL), a once-yearly bisphosphonate, reduces osteoporotic fractures and increases bone mineral density (BMD). This 3-year post-marketing surveillance examined its real-world safety and effectiveness. MATERIALS AND METHODS: This prospective, observational study included patients who started ZOL for osteoporosis. Data were assessed at baseline, 12, 24, and 36 months for safety and effectiveness. Treatment persistence, potentially related factors, and persistence before and after the COVID-19 pandemic started were also investigated. RESULTS: The safety analysis and effectiveness analysis sets included 1406 and 1387 patients, respectively, with mean age of 76.5 years. Adverse reactions (ARs) occurred in 19.35% of patients, with an acute-phase reaction in 10.31, 1.01, and 0.55% after the first, second, and third ZOL infusions. Renal function-related ARs, hypocalcaemia, jaw osteonecrosis, and atypical femoral fracture occurred in 1.71, 0.43, 0.43, and 0.07% of patients, respectively. Three-year cumulative fracture incidences were 4.44% for vertebral, 5.64% for non-vertebral, and 9.56% for clinical fractures. BMD increased by 6.79, 3.14, and 1.78% at the lumbar spine, femoral neck, and total hip, respectively, after 3-year treatment. Bone turnover markers remained within reference ranges. Treatment persistence was 70.34% over 2 years and 51.71% over 3 years. Male, age ≥ 75 years, no previous medicines for osteoporosis, no concomitant medicines for osteoporosis, and inpatient at the first infusion were related to discontinuation. There was no significant difference in the persistence rate between before and after the COVID-19 pandemic (74.7% vs. 69.9%; p = 0.141). CONCLUSION: This 3-year post-marketing surveillance confirmed the real-world safety and effectiveness of ZOL.

Flip-back, an old trick to face highly contrasted relaxation times: application in the characterization of pharmaceutical mixtures by CPMAS NMR.

The (13)C-(1)H CPMAS with flip-back pulse NMR experiment is revisited in view of applications to pharmaceutical mixtures. The analysis of the kinetics of relaxation and CP transfer with and without the flip-back pulse shows that a significant gain in (13)C signal can be expected (thus in experimental time) from the flip-back pulse for protons with long T(1). The gain is of the order of T(1) of the protons expressed in seconds. The experiment is applied on samples with highly contrasted spin-lattice relaxation times T(1) for protons, situation encountered in pharmaceutical mixtures. The application of the flip-back increases significantly the relative signal intensity of the component with the longer T(1), making this component detectable even after using short recycle delays. Therefore, this CPMAS with flip-back experiment could be used routinely to get (13)C CPMAS NMR spectra of mixtures in constant experimental time and signal-to-noise ratio without the need for optimization of the recycle delays, and for whatever may be the degree of crystallinity of the active principal ingredient (API) and/or excipients.

Muraminomicins, new lipo-nucleoside antibiotics from Streptosporangium sp. SANK 60501-structure elucidations of muraminomicins and supply of the core component for derivatization.

We screened for bacterial phospho-N-acetylmuramyl-pentapeptide-translocase (MraY: EC 2.7.8.13) inhibitors with the aim of discovering novel antibiotics and observed inhibitory activity in the culture broth of an actinomycete, SANK 60501. The active compounds, muraminomicins A, B, C, D, E1, E2, F, G, H, and I exhibited strong inhibitory activity against MraY with IC50 values of 0.0105, 0.0068, 0.0104, 0.0099, 0.0115, 0.0109, 0.0089, 0.0134, 0.0186, and 0.0094 µg ml-1, respectively. Although muraminomicin F exhibited favorable antibacterial activity against drug-resistant Gram-positive bacteria, this activity was reduced with the addition of serum. To efficiently supply the core component for chemical modification studies, production was carried out in a controlled trial by adding myristic acid to the medium, and a purification method suitable for large-scale production was successfully developed.

Solution structure and model membrane interactions of temporins-SH, antimicrobial peptides from amphibian skin. A NMR spectroscopy and differential scanning calorimetry study.

Temporin-SHa and temporin-SHc are 13 residue long antimicrobial peptides from frog skin that have similar sequences but differ markedly in their membrane-damaging properties. Temporin-SHa contains a single basic lysine residue and has a unique antimicrobial spectrum of action among temporins, being very potent against Gram-positive and Gram-negative bacteria, yeasts, fungi, and protozoa. Temporin-SHc, which contains a single basic histidine residue, is inactive against Gram-negative bacteria, has a reduced efficacy against Gram-positive bacteria, but is still active against yeasts and fungi. Temporin-SHb, with no basic residue, has no antimicrobial activity. The three-dimensional structures of the peptides bound to SDS micelles were analyzed by CD and NMR spectroscopy combined with restrained molecular dynamics calculations. The peptides adopt well-defined amphipathic alpha-helical structures extending from residue 3 to residue 12, when bound to SDS micelles. The structures are stabilized by extensive interactions between aliphatic and aromatic side chains on the nonpolar face. Relaxation enhancements caused by paramagnetic probes showed that the peptides adopt nearly parallel orientations to the micelle surface and do not deeply penetrate into the micelle. The interaction of the peptides with model membranes was investigated by differential scanning calorimetry on anionic and zwitterionic multilamellar vesicles and membrane-permeabilization assays on calcein-loaded large unilamellar vesicles. Calorimetric data indicated that both temporin-SHa and -SHc reside at the hydrocarbon core-water interface of the anionic lipid bilayer but interact with anionic bilayers in a very different manner. This suggests that the charge-induced activity of temporins-SH for bacterial cells is due to changes in the membrane-disturbing mechanism of the bound peptides.

[A case of follicular lymphoma derived from the conjunctiva].

BACKGROUND: Follicular lymphoma, derived from ocular adnexa as defined by World Health Organization Classification of neoplastic diseases of the hematopoietic and lymphoid tissues, is quite rare in Japan. CASE: A 65-year-old woman visited our clinic for treatment of a conjunctival tumor in the left eye. FINDINGS: Incisional biopsy revealed that the lesion was a lymphoma with well-defined follicles, and it was L 26(CD20) and BCL-2 positive immunohistochemically. Flow cytometric analysis showed that more than 70% of the cells were double positive for CD10 and CD20. Southern blot hybridization demonstrated gene rearrangement bands in the immunoglobulin gene JH region. The chromosome had a translocation t(14, 18) (q32, q21). Systemic evaluation detected no other lymphoma. CONCLUSION: The lesion was a follicular lymphoma derived from conjunctival tissue. Additional examinations such as flow cytometric analysis, gene analysis, and chromosome examination were useful for differential diagnosis of marginal zone B cell lymphoma of mucosa associated lymphoid tissue type and benign reactive lymphoid hyperplasia.

The prevalence of obesity and its relationship with lifestyle factors in Jamaica.

We surveyed the prevalence of obesity in the general population in Jamaica, and examined the relationship between it and lifestyle. The survey population consisted of 1,935 inhabitants in Jamaica, whose body weight, height, marital status, educational history, employment status and other obesity-associated lifestyle factors were surveyed. Six major findings emerged. The first finding is that the proportion of obesity in women was very high, and there was a big gender difference. Secondly, a lower prevalence towards obesity was associated with cohabitation of the subjects in both genders, and higher educational levels in female subjects. Thirdly, the proportion of the subjects who considered their weight to be quite acceptable was higher in the obese/overweight groups in both genders. Fourthly, exercise frequency showed a negative correlation with the body mass index (BMI) in men, and the frequency of exercising was apparently lower in women than in men. Fifthly, as for dietary factors, in both genders vegetables showed a negative correlation with the BMI. Sixthly, non-smokers were also associated with a lower obesity prevalence in men. In conclusions, these findings suggest that social and lifestyle factors such as the educational level, marital status and dietary habits of the general population influence Jamaican obesity.

The participation of nitric oxide in peritoneal exudate cell cytotoxicity of mice by Fusobacterium nucleatum.

Previously we reported that mice infected recurrently with live Fusobacterium nucleatum(Fn) synthesize a significant amount of NO between 12 hr and 24 hr after Fn injection. Fn is a gram-negative rod periodontal pathogen. NO could not be induced by heat-killed Fn or in untreated mice. This NO, derived from the iNOS after infection of live Fn, was not involved in the Fn reduction because Fn clearance occurs within 6 hr. We investigated in this study whether this NO was involved in cytotoxicity in peritoneal exudate cells (PEC) in vivo. The mice were divided into two groups: those treated with live Fn (immune) and those left untreated (normal). PEC number, NO production, detection of apoptosis or death cells, and lactate dehydrogenase (LDH) release activity after injection of live Fn were compared in these groups. In the immune group, the increase of the total cell numbers caused by an increase in neutrophils, a significant NO production only after injection of live Fn at 24 hr and identification of iNOS positive macrophages were confirmed. The apoptotic rate was very low and did not increase at 24 hr in vivo. Therefore, apoptosis was seldom relevant to the NO. In the immune group, LDH activity was remarkable high at 24 hr, and dead cells and macrophages phagocytizing cell fragments increased at the same time. Pretreatment of L NMMA, an inhibitor of iNOS, suppressed LDH activity and cell death. Therefore, the NO derived from the iNOS is involved in the cytotoxicity. These results suggest that NO may contribute to the inflammatory response during Fn infection in periodontitis.

The reduction of Fusobacterium nucleatum in mice is irrelevant to the nitric oxide induced by iNOS.

Previously we reported that mice infected recurrently with live Fusobacterim nucleatum (Fn) synthesize a significant amount of NO between 12 hr and 24 hr after the Fn injection. We now investigated whether the NO has the capability of killing Fn, a gram-negative rod periodontal pathogen. The mice were divided into three groups: treated with live bacteria (LB), treated with heat-killed bacteria (HKB) and untreated: normal (N). The Fn reduction, NO production and cell number after Fn injection were then compared in these mice. In the LB group, no Fn was detected at 6 hr, whereas it was still detected in the HKB and N groups at 24 hr as assessed by both colony counts and PCR assays. A significant amount of NO was synthesized in the LB group at 24 hr after the Fn injection. Fn is not killed by SNAP-generated NO in vitro. An increase in the total cell number was accompanied by an increase of the neutrophil numbers in the LB group. Intracellular O2(-) generation (including ONOO(-)) was visualized using dihydrorhodamine (DHR)-123. The peak of O2(-) generation by PEC was shown to be at 3 hr in all 3 groups. The number of O2(-) positive cells in the LB group at 3 hr was remarkably high, and most of them were likely to be neutrophils. The Fn reduction would be performed cooperatively via oxygen dependent and oxygen independent mechanisms. Thus reactive oxygen species (ROS) included in the oxygen dependent mechanism appear to be important for Fn reduction. However the significant amounts of NO derived from the iNOS synthesized in the LB group between 12 hr and 24 hr after injection of LFn were not involved in the Fn reduction.

Intracellular production and extracellular release of oxygen radicals by PMNs and oxidative stress on PMNs during phagocytosis of periodontopathic bacteria.

In this study we investigated intracellular and extracellular oxygen radical production by polymorphonuclear leukocytes (PMNs) during the phagocytosis of periodontopathic bacteria. In in vitro assays, bacteria of the species Porphyromonas gingivalis, Actinobacillus actinomycetemcomitans, and Fusobacterium nucleatum were phagocytosed at 37 degrees C for 4 h by purified peripheral human PMNs from healthy subjects (n = 6). Superoxide production during phagocytosis was determined by flow cytometry and with a fluorescence/luminescence microplate reader. After phagocytosis, oxidative stress was determined by flow cytometry. Both the intracellular and extracellular oxygen radical production by PMNs phagocytosing F. nucleatum was significantly greater than that of PMNs phagocytosing P. gingivalis and A. actinomycetemcomitans ( P < 0.01 by the Mann-Whitney test). Moreover, after 4 h of incubation, the oxidative stress of PMNs phagocytosing F. nucleatum was significantly greater than that of PMNs phagocytosing P. gingivalis and A. actinomycetemcomitans. We conclude that a high level of superoxide production by PMNs may damage not only periodontopathic bacteria but also PMNs themselves, and may be correlated with the destruction of periodontal tissue.

Usefulness of measuring plasma cytokines in oral and maxillofacial surgery.

We determined the intra- and postoperative plasma concentrations of cytokines (tumor necrosis factor [TNF]-Alpha, interleukin [IL]-1Beta, IL-6, and IL-8) in oral surgery patients with procedures ranging in duration from 20 to 375 min and investigated their relationship to the intensity of the surgical stress over time. No significant differences from baseline levels were observed in the levels of TNF-Alpha, IL-1Beta, and IL-8. By contrast, increased IL-6 levels were noted only on the first postoperative day, when they reached 1500% of the baseline level, after which they decreased to preoperative levels by the third postoperative day and tended to reflect the intensity of surgical stress.