Effects of a Cloth Panel Containing a Specific Ore Powder on Patients with Chamaecyparis obtusa (Cypress) Pollen Allergy.

Pollen allergy to Japanese cedar and cypress is a serious illness that impairs daily life and sleep, especially during pollen season. We have reported that placing a cloth panel containing a specific natural ore powder (CCSNOP) in a room may alleviate the symptoms of hay fever and may also benefit the immune system. This ore is from the Aso mountain range, a volcano on Kyushu Island in the southwestern part of Japan. The purpose of this study was to verify the effect of CCSNOP on cypress pollen. Thirty-one double-blind tests, which investigated cedar pollen allergies, were conducted from February to March 2018 and have already been reported. After this, in early April, 10 of these cases were recruited and all had CCSNOP installed in their bedrooms. Before that, various symptoms and changes in medication were recorded in a "Symptom Diary" and included a mood survey by a questionnaire, stress test using saliva amylase, changes in cypress-specific immunoglobulins IgE and IgG4 by blood sampling, and eosinophil changes. In addition, changes in 29 types of cytokines were investigated. Exposure to CCSNOP relieved symptoms and subjects decreased their intake of medication. There was no change in mood or stress, but eosinophil levels tended to decrease. Although there were no statistical changes in cypress-specific IgE or IgG4, an increase in the former and a decrease in the latter were observed in some individuals during the period of pollen dispersal. Furthermore, levels of GM-CSF and IL8 decreased significantly after use of CCSNOP. The CCSNOP was shown to be effective against cypress pollen allergy, and future investigations will be necessary to observe the long-term effects of CCSNOP.

Biological Effects of Cloth Containing Specific Ore Powder in Patients with Pollen Allergy.

OBJECTIVE: The custom-homebuilding company, Cosmic Garden Co. Ltd., located in Okayama City, Japan was established in 1997 and uses specific natural ore powder (SNOP) in wall materials and surveys customers in order to improve allergic symptoms. METHODS: To investigate the biological effects of SNOP, patients with a pollen allergy were recruited to stay in a room surrounded by cloth containing SNOP (CCSNOP), and their symptoms and various biological parameters were compared with those of individuals staying in a room surrounded by control non-woven cloth (NWC). Each stay lasted 60 min. Before and immediately after the stay, a questionnaire regarding allergic symptoms, as well as POMS (Profile of Mood Status) and blood sampling, was performed. Post-stay minus pre-stay values were calculated and compared between CCSNOP and NWC groups. RESULTS: Results indicated that some symptoms, such as nasal obstruction and lacrimation, improved, and POMS evaluation showed that patients were calmer following a stay in CCSNOP. Relative eosinophils, non-specific Ig E, epidermal growth factor, monocyte chemotactic protein-1, and tumor necrosis factor-α increased following a stay in CCSNOP. CONCLUSION: This ore powder improved allergic symptoms, and long-term monitoring involving 1 to 2 months may be necessary to fully explore the biological and physical effects of SNOP on allergic patients.

Prompt analgesic effect of antihistaminic diphenhydramine ointment on bone-joint-muscle pain as assessed by skin impedance.

Pain is sensed, transmitted, and modified via a variety of mediators and their receptors. Histamine is a well-known mediator of pain. In addition to their antagonistic effects against histamine, classical antihistaminics possess, to various degrees, antimuscarinergic, antiserotonergic, antiadrenergic, local anesthetic, membrane-stabilizing and other pharmacologic actions. Although there have been many attempts to use classical antihistaminics as analgesics and/or analgesic adjuvants, the appearance of non-steroidal anti-inflammatory drugs discouraged such efforts. Here, we compared the analgesic effect of an ointment containing 1% diphenhydramine (a typical first-generation antihistaminic drug) with that of indomethacin (a typical non-steroidal anti-inflammatory drug) in elderly patients with osteoarthritis and/or osteoporosis who complained of bone-joint-muscle pain. Analgesic effects were evaluated by measuring skin impedance and by subjective pain assessments (using a visual recording system) before and after ointment application. Diphenhydramine ointment exerted a prompt and marked analgesic effect that lasted for several hours, as assessed by either skin impedance or subjective pain evaluation. In contrast, the analgesic effect of indomethacin ointment was marginal, and significant only an hour or more later than that of diphenhydramine. These results suggest that diphenhydramine ointment may be useful for the relief of the bone-joint-muscle pains that are common in elderly subjects.

Comparison of the effects of elcatonin and risedronate on back and knee pain by electroalgometry using fall of skin impedance and quality of life assessment using SF-36.

Back and knee pain is a widespread health problem and a serious threat to the quality of life (QOL) in middle-aged and older adults, as it frequently accompanies osteoporosis and osteoarthritis. In order to compare the effects of elcatonin and risedronate on such pain, 20 units of elcatonin was intramuscularly injected to 18 patients, and 5 mg of risedronate was orally administered daily to 20 others with similar backgrounds. Exercise-induced pain was analyzed by measuring the fall of skin impedance by electroalgometry (EAM), and subjective pain was recorded by a visual rating system (VRS) on a scale of 0 (no pain) to 100 (unbearable pain). In patients treated with elcatonin, the mean EAM-estimated pain was significantly reduced after 4, 5 and 6 months of treatment, and the VRS score after 3, 5 and 6 months, indicating a significant analgesic effect. In the risedronate group, however, improvement was less remarkable. Two-way analysis of variance using pain as a dependent variable and treatment group and time as independent variables revealed a significantly greater effect of elcatonin over risedronate on both the EAM and VRS scores, and the influence of treatment time on pain was indistinguishable between the two treatment groups. Effect of exercise load on pain was less on knee load than knee and spine load and spine load, but indistinguishable between the two groups. Changes in QOL were evaluated by the SF-36 system. Norm-based scoring showed significant improvements in 3 of 4 categories for elcatonin and in 2 of 4 for risedronate, suggesting comparable effects on the physical aspects of QOL, whereas responses to emotionally and socially directed questions indicated significant improvements in all 4 categories for risedronate, but none for elcatonin, suggesting a more physical than emotional component in elcatonin effects compared to risedronate.

Vitamin D insufficiency defined by serum 25-hydroxyvitamin D and parathyroid hormone before and after oral vitamin D3 load in Japanese subjects.

Vitamin D insufficiency is a risk for both skeletal and nonskeletal health. However, some ambiguity remains about threshold serum 25(OH)D for vitamin D insufficiency. To determine the threshold serum 25(OH)D to maintain normal calcium availability without elevation in serum parathyroid hormone (PTH) among Japanese subjects with various calcium intakes, we conducted a multicenter prospective open-labeled study. We recruited 107 ambulatory subjects without disorders affecting vitamin D metabolism to whom oral vitamin D3 800 IU/day for 4 weeks or 1,200 IU/day for 8 weeks was given. Serum 25(OH)D, PTH, calcium, phosphate, and magnesium were measured before and after vitamin D3 supplementation. Calcium intake was assessed by questionnaires. When all the data were combined, serum 25(OH)D was negatively correlated with PTH. The cubic spline curve between serum 25(OH)D and PTH indicated PTH reached its plateau between 35 and 40 pg/ml at 25(OH)D between 25 and 30 ng/ml. Vitamin D3 supplementation increased serum 25(OH)D and decreased PTH. Change in PTH correlated positively with baseline serum 25(OH)D. From the regression analyses, baseline serum 25(OH)D above 28 ng/ml corresponded to the threshold level without reduction in PTH after vitamin D3 supplementation. In multivariate regression analyses, age but not calcium intake was a significant determinant of PTH. We concluded that a serum 25(OH)D level of 28 ng/ml was identified as a threshold for vitamin D insufficiency necessary to stabilize PTH to optimal levels.

Analgesic effect of raloxifene on back and knee pain in postmenopausal women with osteoporosis and/or osteoarthritis.

To assess the effect of raloxifene on bone and joint pain, 24 postmenopausal women with back or knee pain or both were randomly divided into two groups, based on the chronological sequence of consultation, to be treated with 60 mg raloxifene and 1 microg alfacalcidol (RA)/day (group RA) or 1 microg alfacalcidol alone (A)/day (group A), respectively, for 6 months. Pain following knee loading (KL) by standing up from a chair and bending the knee by squatting, knee and spine loading (KSL) by walking horizontally and ascending and descending stairs, and spine loading (SL) by lying down supine on a bed and leaving the bed to stand was evaluated by electroalgometry (EAM), based on measurement of the fall of skin impedance, and a visual rating scale (VRS), recording subjective pain on a scale of 0-100 between no pain and unbearable pain. The two groups showed no significant difference as to age, indices of mineral metabolism, back and knee pain, and bone status. RA gave a significantly greater analgesic effect than A by both EAM (P = 0.0158) and VRS (P = 0.0268) on overall comparison of the mean response to all modalities of exercise loading. Paired comparison between pretreatment and posttreatment indicated a significant effect of RA by both EAM (P = 0.0045) and VRS (P = 0.0017), but not that of A. The analgesic effect was more clearly noted on combined knee-spine loading (KSL) and spine loading (SL) than simple knee loading (KL). Monthly comparison of the analgesic effect indicated a significantly better analgesic effect in the fifth month by VRS. RA effect greater than A was more evident by EAM than VRS and during months 3-6 than during 1-2 months, suggesting a slowly progressive effect of RA. Pain evaluation by EAM and VRS mostly gave parallel results, except for a few occasions such as knee loading and spine loading by sitting up and leaving a bed, when EAM detected a positive effect but VRS failed to do so. RA appeared to be more effective on bone and joint pain than A in postmenopausal women according to both EAM and VRS measurements.

Effects of a Cloth Panel Containing a Specific Ore Powder on Patients with Japanese Cedar Pollen Allergy During the Pollen Dispersal Season.

Pollen allergy remains a big problem in contemporary societies. We have shown in previous studies that a cloth containing a special natural ore powder (CCSNOP) is effective in relieving symptoms in patients with pollen allergies. However, in that study, subjects were exposed to CCSNOP for only one hour. In the present study, CCSNOP or control (non-woven cloth; NWC) panels were placed in the bedrooms of pollen allergy patients for two weeks during the pollen dispersal season in 2018, and the effects were investigated. Twenty-one subjects were exposed to CCSNOP panels and 10 subjects were exposed to NWC panels. Our investigations showed that use of CCSNOP resulted in relief of symptoms and reduced use of therapeutics. Moreover, the ratio of eosinophil count decrease during exposure was higher in the CCSNOP group. Furthermore, a formula for measuring various cytokines and other parameters was established and clearly showed a distinction between the CCSNOP and NWC groups. In this formula, Granulocyte Macrophage colony-stimulating Factor (GM-SCF), Interleukin (IL)-12p40, Immunoglobulin (Ig) G4 and eosinophil count were extracted. These results indicated that CCNSNOP has a beneficial effect on pollen allergy patients. Future studies shall engage in long-term monitoring of pollen allergy patients who will utilize this mineral powder for at least one year.

Fluid secretion caused by aerolysin-like hemolysin of Aeromonas sobria in the intestines is due to stimulation of production of prostaglandin E2 via cyclooxygenase 2 by intestinal cells.

To clarify the mechanisms of diarrheal disease induced by Aeromonas sobria, we examined whether prostaglandin E2 (PGE2) was involved in the intestinal secretory action of A. sobria hemolysin by use of a mouse intestinal loop model. The amount of PGE2 in jejunal fluid and the fluid accumulation ratio were directly related to the dose of hemolysin. The increase over time in the level of PGE2 was similar to that of the accumulated fluid. In addition, hemolysin-induced fluid secretion and PGE2 synthesis were inhibited by the selective cyclooxygenase 2 (COX-2) inhibitor NS-398 but not the COX-1 inhibitor SC-560. Western blot analysis revealed that hemolysin increased the COX-2 protein levels but reduced the COX-1 protein levels in mouse intestinal mucosa in vivo. These results suggest that PGE2 functions as an important mediator of diarrhea caused by hemolysin and that PGE2 is produced primarily through a COX-2-dependent mechanism. Subsequently, we examined the relationship between PGE2, cyclic AMP (cAMP), and cystic fibrosis transmembrane conductance regulator (CFTR) Cl- channels in mouse intestinal mucosa exposed to hemolysin. Hemolysin increased the levels of cAMP in the intestinal mucosa. NS-398 inhibited the increase in cAMP production, but SC-560 did not. In addition, H-89, a cAMP-dependent protein kinase A (PKA) inhibitor, and glibenclamide, a CFTR inhibitor, inhibited fluid accumulation. Taken together, these results indicate that hemolysin activates PGE2 production via COX-2 and that PGE2 stimulates cAMP production. cAMP then activates PKA, which in turn stimulates CFTR Cl- channels and finally leads to fluid accumulation in the intestines.

[ Nail calcium content in relation to age and bone mineral density].

The growth of nails, and their matrix components, are influenced by several physiological, pathological and environmental factors. Because of the slow rate of nail growth, the elemental composition of the nail is also expected to be affected by transient factors controlling serum components. The mineral components of nail clippings may therefore reflect the long-term patterns of mineral metabolism, such as the rise of creatinine concentration seen in nails in patients with renal failure with hypercreatinemia. Fingernail and toenail Ca concentrations decreased with age in both men and women, whereas Mg concentrations tended to increase. Postmenopausal women had lower finger nail Ca concentrations than premenopausal women. LBMD showed a significant positive correlation with finger nail Ca content. The measurement of finger nail Ca content may be useful as a predictor of osteoporosis.

Aeromonas sobria hemolysin causes diarrhea by increasing secretion of HCO3-.

Aeromonas sobria hemolysin (ASH) is one of the major virulence factors produced by A. sobria, a causative agent of diarrhea in humans. We investigated the effects of ASH on anion transport in human colonic epithelial cells. ASH increased short circuit currents across the intestinal epithelia, which were suppressed by anion channel antagonists, such as carbonic anhydrase inhibitors, and by the removal of external HCO3-. Iliac fluid accumulation was also inhibited by carbonic anhydrase inhibitors. The results suggest that ASH activates HCO3- secretion, whose level correlates with the severity of diarrhea.

Cleavage specificity of the serine protease of Aeromonas sobria, a member of the kexin family of subtilases.

Subtilisin-like proteases have been grouped into six families based on a sequence of the catalytic domain. One of the six is the kexin family, of which furin is a representative protease. All members of the kexin family, except one, are from eukaryotes. The one prokaryotic protease is a serine protease of Aeromonas sorbria (ASP). Here, we examined the substrate specificity of ASP based on the cleavage of short peptides. The results showed that ASP preferentially cleaves the peptide bond following two basic residues, one of which is Lys, but not the bond following a single basic residue. This indicates that the tertiary structure around the catalytic domain of ASP resembles, but is not identical to that of furin. Prekallikrein was cleaved into four fragments by ASP, indicating that the protein must be cleaved at specific sequences.

A pore-forming toxin produced by Aeromonas sobria activates cAMP-dependent Cl- secretory pathways to cause diarrhea.

Aeromonas sobria hemolysin (ASH) is one of the major virulence factors produced by A. sobria, a human pathogen that causes diarrhea. We investigated the effects of ASH on Cl(-) transport in human colonic epithelial cells. ASH increased short-circuit currents (Isc) and (125)I efflux from Caco-2 cells, indicating ASH activate Cl(-) secretion. Additions of inhibitors of cyclic AMP dependent Cl(-) channels, glybenclamide and NPPB suppressed the Isc and (125)I efflux increases induced by ASH. And ASH increased the intracellular cyclic AMP concentration. Moreover, ASH stimulated fluid accumulation in the iliac loop test, and glybenclamide and NPPB suppressed this fluid accumulation. Thus, cAMP-dependent Cl(-) secretory pathway could be related with diarrhea induced by A. sobria.

Properties of hemolysin and protease produced by Aeromonas trota.

We examined the properties of exotoxins produced by Aeromonas trota (A. enteropelogenes), one of the diarrheagenic species of Aeromonadaceae. Nine of 19 A. trota isolates that grew on solid media containing erythrocytes showed hemolytic activity. However, the hemolytic activities of the culture supernatants of these hemolytic strains of A. trota were markedly lower than those of A. sobria when cultured in liquid medium, and the amount of hemolysin detected by immunoblotting using antiserum against the hemolysin produced by A. sobria was also low. A mouse intestine loop assay using living bacterial cells showed that A. trota 701 caused the significant accumulation of fluid, and antiserum against the hemolysin produced suppressed the enterotoxic action of A. trota 701. These results indicated that A. trota 701 was diarrheagenic and the hemolysin produced was the causative agent of the enterotoxic activity of A. trota. The hemolysin in A. sobria was previously shown to be secreted in a preform (inactive form) and be activated when the carboxy-terminal domain was cleaved off by proteases in the culture supernatant. Since mature hemolysin was detected in the culture supernatants of A. trota, we analyzed the extracellular protease produced by A. trota. Fifteen of 19 A. trota isolates that grew on solid media containing skim milk showed proteolytic activity. We subsequently found that most A. trota isolates possessed the serine protease gene, but not the metalloprotease gene. Therefore, we determined the nucleotide sequence of the serine protease gene and its chaperone A. trota gene. The results obtained revealed that the deduced amino acid sequences of serine protease and the chaperone were homologous to those of A. sobria with identities of 83.0% and 75.8%, respectively.

Maturation pathway of metalloprotease produced by Aeromonas sobria.

Previously, we cloned the metalloprotease gene of Aeromonas sobria (amp) and determined its nucleotide sequence (GenBank accession number DQ784565). The protease is composed of 591 amino acid residues. In this study, we purified the mature metalloprotease from the culture supernatant of A. sobria and determined the amino terminal sequence and molecular size of AMP. In addition, we examined the production of AMP diachronically and found that AMP emerges outside of the cell as an intermediate composed of mature and propeptide regions. Subsequently, we determined that the N-terminal amino acid sequence of the intermediate and found that the sequence is identical to that of the mature metalloprotease. This means that the intermediate is composed of a mature AMP region and a C-terminal propeptide. The cross culture experiment of mutants of metalloprotease and serine protease of A. sobria on skim milk agar medium indicates that the intermediate released outside of the cell is inactive and that serine protease produced by A. sobria accelerates the conversion of the intermediate from the inactive to the active form.

Fractional absorption of active absorbable algal calcium (AAACa) and calcium carbonate measured by a dual stable-isotope method.

With the use of stable isotopes, this study aimed to compare the bioavailability of active absorbable algal calcium (AAACa), obtained from oyster shell powder heated to a high temperature, with an additional heated seaweed component (Heated Algal Ingredient, HAI), with that of calcium carbonate. In 10 postmenopausal women volunteers aged 59 to 77 years (mean ± S.D., 67 ± 5.3), the fractional calcium absorption of AAACa and CaCO(3) was measured by a dual stable isotope method. (44)Ca-enriched CaCO(3) and AAACa were administered in all subjects one month apart. After a fixed-menu breakfast and pre-test urine collection (Urine 0), (42)Ca-enriched CaCl(2) was intravenously injected, followed by oral administration of (44)Ca-enriched CaCO(3) without carrier 15 minutes later, and complete urine collection for the next 24 hours (Urine 24). The fractional calcium absorption was calculated as the ratio of Augmentation of (44)Ca from Urine 0 to Urine 24/ augmentation of (42)Ca from Urine 0 to Urine 24. Differences and changes of (44)Ca and (42)Ca were corrected by comparing each with (43)Ca. Fractional absorption of AAACa (mean ± S.D., 23.1 ± 6.4), was distinctly and significantly higher than that of CaCO(3 )(14.7 ± 6.4; p = 0.0060 by paired t-test). The mean fractional absorption was approximately 1.57-times higher for AAACa than for CaCO(3). The serum 25(OH) vitamin D level was low (mean ± S.D., 14.2 ± 4.95 ng/ml), as is common in this age group in Japan. Among the parameters of the bone and mineral metabolism measured, none displayed a significant correlation with the fractional absorption of CaCO(3) and AAACa. Higher fractional absorption of AAACa compared with CaCO(3) supports previous reports on the more beneficial effect of AAACa than CaCO(3) for osteoporosis.

Comparison of the analgesic effects of bisphosphonates: etidronate, alendronate and risedronate by electroalgometry utilizing the fall of skin impedance.

Analgesic effects of etidronate, alendronate and risedronate were compared in patients with osteoporosis and/or osteoarthritis by measuring the fall of skin impedance along with conventional subjective pain-estimation by visual rating scale (VRS). One hundred ninety-nine postmenopausal women consulting the Osteoporosis and Osteoarthritis Clinic of Katsuragi Hospital complaining of back and/or knee pain were randomly divided into four groups; Group A (49 subjects) given 5 mg/day alendronate, Group E (50 subjects) 200 mg/day etidronate, Group R (50 subjects) 2.5 mg/day risedronate and Group P no bisphosphonate. None of the four groups showed significant deviation from others as to age and parameters of bone metabolism. Proportions of subjects with osteoporosis was 18-40%. Those with osteoarthritis of the spine and knee, higher than Grade II according to the Nathan and Lawrence-Kellgren scale, respectively, was 45 and 61%, respectively, without a significant difference among the four groups. Significant positive correlation was found between the fall of skin impedance and pain expressed in VRS. Attenuation of exercise-induced fall of skin impedance and also subjective pain expressed in VRS was greatest in Group E with a highly significant difference from Groups A (P = 0.0002 and P < 0.0001), R (P < 0.0001 and P = 0.0014) and P (P < 0.0001 and P < 0.0001). Neither A nor R showed significant difference from P as to the fall of skin impedance. Among the three bisphosphonates tested, etidronate appeared to be outstanding in analgesic effects.

Postural stabilizing effect of alfacalcidol and active absorbable algal calcium (AAA Ca) compared with calcium carbonate assessed by computerized posturography.

Sway and postural instability have drawn attention as a risk factor for osteoporotic fracture, in addition to low bone mineral density (BMD) and poor bone quality. In view of the fracture-reducing effect of alfacalcidol and active absorbable algal calcium (AAA Ca) not readily explained by rather mild increases of BMD, attempts were made to evaluate postural stabilizing effect of alfacalcidol, AAA Ca, and calcium carbonate (CaCO(3)) by computerized posturography. Track of the gravity center was analyzed to calculate parameters related to tract length, track range, and track density to express the degree of sway before and after supplementation in 126 subjects ranging in age between 20 and 81 years randomly divided into four groups. Supplementation with AAA Ca containing 900 mg elemental Ca (group A), no calcium (group B), CaCO(3) also containing 900 mg elemental Ca (group C), or alfacalcidol (group D) continued daily for 12 months. For each parameter, the ratio closed eye value/open eye value (Romberg ratio) was calculated to detect aggravation of sway by eye closure. Age, parameters of Ca and P, and proportions of subjects with fracture and those with low BMD showed no marked deviation among the groups. With eyes open, significant decreases of a track range parameter (REC) from group B was noted in groups A (P = 0.0397) and D (P = 0.0296), but not in group C according to multiple comparison by Scheffe, indicating superior postural stabilizing effect of A and D over C. In the first 2 months, a significant fall was already evident in REC from group B in group D (P = 0.0120) with eyes open. Paired comparison of sway parameters before and after supplementation revealed a significant increase of track density parameter (LNGA), indicating sway control efficiency and a significant decrease of REC in groups A and D compared to group B with eyes open. With eyes closed, only group A showed a significant improvement from group B (P = 0.0456; Fig. 1), with a significant shortening on paired After/Before comparison (P = 0.0142; Fig. 2). Computerized posturography appears to be useful in analyzing sway phenomena especially as to the effects of vitamin D and various Ca preparations.

Effect of age on body sway assessed by computerized posturography.

The swaying and postural instability frequently seen in elderly subjects had not been analyzed quantitatively in detail until the introduction of computerized posturography. In order to assess the changes of body sway with aging, we performed computerized posturography in 144 subjects (51 men and 93 women, between the ages of 22 and 88 years) without specific neurological or metabolic disorders. The total and timed track length of the center of gravity, reflecting the distance of sway, increased with advancing age, with a highly significant positive correlation, without marked sex differences. The total area covered by the track of the center of gravity (expressing the extent of sway) also showed a similar tendency. Track density per unit area, expressing the efficiency of postural control, in contrast, decreased with age, showing a significant negative correlation with age, but only when the subjects had their eyes open; this decrease did not occur when they had their eyes closed. The Romberg ratio, an index of exacerbation of sway on eye closure, showed little change with a tendency for slight alleviation of sway and improvement in the efficiency of its control. Computerized posturography appears to be a useful tool with which to analyze the mechanism of swaying associated with old age.