Effectiveness of countermeasure for polypharmacy by multidisciplinary team review in patients with diabetes mellitus.

AIMS/INTRODUCTION: Polypharmacy in diabetes patients is related to worse clinical outcomes. The aim of this study was to evaluate the usefulness of our countermeasure for polypharmacy, which combines a pharmacist check followed by a multidisciplinary team review in diabetic patients with polypharmacy. METHODS: A single-center, retrospective observational study was conducted at Gifu University Hospital. Study participants included diabetic patients taking six or more drugs on admission to the diabetes ward between July 2021 and June 2022. Drugs which were discontinued by the present countermeasure were examined, and the number of drugs being taken by each patient was compared between admission and discharge. RESULTS: 102 of 308 patients were taking six or more drugs on admission. The drugs being taken by these patients were evaluated by pharmacists using a checklist for polypharmacy. Eighty-four drugs which were evaluated as inappropriate or potentially inappropriate medications by pharmacists were discontinued following the multidisciplinary team review. The median and mean number of drugs taken by the 102 patients significantly decreased from 9.0 (IQR: 8-12) and 9.26 ± 2.64 on admission to 9.0 (IQR: 6-10) and 8.42 ± 2.95 on discharge (P = 0.0002). We followed up with these patients after discontinuation of the drugs and confirmed that their clinical status had not deteriorated. CONCLUSION: The present countermeasure for polypharmacy, which combines a pharmacist check based on a checklist for evaluating polypharmacy followed by a multidisciplinary team review, was useful for reducing the number of inappropriate or potentially inappropriate medications taken by diabetes patients with polypharmacy.

Physicochemical properties of wooden breast-extracted myosin and rheological properties of its heat-induced gel.

BACKGROUND: It is reported that broilers with 'wooden breast' have poor processing properties, such as low binding and water-holding capacities. However, the reason for the poor functional characteristics has not been clarified. In this study, myosin was extracted from a wooden breast. Its physicochemical properties were investigated to clarify the relationship between the structure and physicochemical properties of the heating gel of myosin obtained from the wooden breast. RESULTS: The turbidity of myosin solution extracted from wooden breast increased with increase in the heat treatment to a higher value than that from the normal breast meat myosin. The solubility of myosin collected from a wooden breast after heating decreased like normal breast muscle myosin. The surface hydrophobicity of myosin removed from wooden breast increased continually above 60 °C, unlike the change in surface hydrophobicity of normal breast myosin. The free thiol group of myosin extracted from the wooden breast was higher than normal breast myosin before and after heating. The apparent elasticity of heat-induced gels and chicken meat sausages was significantly lower in sausages and gel with wooden breast than normal ones (P < 0.05). The microstructure of the heated gel of normal myosin showed a fine network structure. In contrast, the heat-induced gel of wooden breast-extracted myosin showed a structure with loosely connected aggregates and many gaps. CONCLUSION: The coarseness of the internal gel structure of myosin extracted from wooden breast was shown to affect the apparent elasticity of the gel and sausages made from the chicken meat. © 2023 Society of Chemical Industry.

Immune Response in Chickens Vaccinated with Freeze-Thawed or Warmed Water-in-Oil Vaccine.

This study investigated whether freezing or warming water-in-oil (W/O) vaccines affected the immune responses of chickens. One of the conditions affecting the efficacy of commercially available animal vaccines is the storage temperature range. Previous studies have shown that the properties of some inactivated vaccines change owing to freezing, leading to reduced immune responsiveness after inoculation. In this study, we first determined the freezing temperatures of a commercial W/O vaccine using freezers maintained at -10, -13, -15, and -20°C. The results showed that the W/O vaccine froze from -10 to -12°C. Next, we evaluated the effect on antibody level transitions (sample-to-positive ratio) in 46-day-old broiler chickens vaccinated with the W/O vaccine that was maintained at -20°C, 5°C, and -10°C, in that order. In addition, the effect on antibody value transitions was evaluated in 45-day-old broiler chickens vaccinated with the W/O vaccines that were frozen and thawed between -20°C and 5°C repeatedly or warmed to 45°C. In these experiments, no remarkable effect of the freeze-thawing or warming treatments on antibody value transitions was observed. These results suggested that the efficacy of the W/O vaccine was not significantly affected when placed in a frozen environment or left in a room temperature environment of 42°C or lower for approximately 5 d. These data indicate the possibility of expanding the temperature range for handling W/O vaccines.

Mitochondrial characteristics of chicken breast muscle affected by wooden breast.

The growth rate of broiler chickens has increased by 400% over the past 50 years, and breast yields continue to increase. This has led to an increase in thoracic muscle abnormalities in broilers, with wooden breast becoming a major issue worldwide. The etiology and the mechanism underlying the etiology of wooden breasts have not yet been elucidated; however, it occurs due to oxidative stress. Reactive oxygen species, which cause oxidative stress, are mainly produced in mitochondria. Thus, in this study, we investigated the relationship between the severity of wooden breast in broilers and the characteristics of mitochondria as the source of reactive oxygen species. Sampling of the pectoralis major muscle at the ventral cranial position was conducted in 50-day-old broilers. The severity of wooden breast was classified into three groups based on the muscle fiber roundness and wing-wing contact test, with highest severity in severe wooden breast and lowest severity in normal breast. Nicotinamide adenine dinucleotide tetrazolium reductase staining revealed an increase in darkly stained muscle fibers, indicating high severity of wooden breast. The mitochondria were swollen in severe wooden breast cases, with highest swelling in severe wooden breast and lowest swelling in normal breast. The expression levels of the mitochondrial antioxidant enzyme genes superoxide dismutase 1 and superoxide dismutase 2 were significantly lower in wooden breast-severe tissue than in normal tissue. These results suggest that when the levels of reactive oxygen species in muscle fibers, which should be constant, are increased, mitochondrial homeostasis is not maintained and the damage levels increase in various membranes of the cell, leading to the disruption of normal physiological functions.

Novel antibody assessment method for microbial compositional alteration in the oral cavity.

Recently, it has been demonstrated that dysbiosis, an alteration in commensal microflora composition, is intimately involved in the onset of a variety of diseases. It is becoming increasingly evident that the composition of commensal microflora in the oral cavity is closely connected to oral diseases, such as periodontal disease, and systemic diseases, such as inflammatory bowel disease. Next-generation sequencing techniques are used as a method to examine changes in bacterial flora, but additional analytical methods to assess bacterial flora are needed to understand bacterial activity in more detail. In addition, the oral environment is unique because of the role of secretory antibodies contained in saliva in the formation of bacterial flora. The present study aimed to develop a new method for evaluating the compositional change of microbiota using flow cytometry (FCM) with specific antibodies against the bacterial surface antigen, as well as salivary antibodies. Using specific antibodies against Streptococcus mutans, a causative agent of dental caries, and human IgA, bacterial samples from human saliva were analyzed via FCM. The results showed that different profiles could be obtained depending on the oral hygiene status of the subjects. These results suggest that changes in the amount and type of antibodies that bind to oral bacteria may be an indicator for evaluating abnormalities in the oral flora. Therefore, the protocol established in this report could be applied as an evaluation method for alterations in the oral microbiota.

Comparison of single trough-based area under the concentration-time curve versus trough concentration for the incidence of vancomycin-associated nephrotoxicity.

INTRODUCTION: While the revised 2020 consensus guideline recommends the use of area under the concentration-time curve (AUC)-guided vancomycin monitoring, collecting multiple vancomycin serum samples to calculate the AUC may cause clinical complications. The aim of the present retrospective study was to evaluate whether AUC-guided vancomycin monitoring, in which AUC was calculated based on a single trough concentration, is a better predictor of nephrotoxicity than trough-guided monitoring in patients receiving vancomycin therapy. METHODS: A single-center, retrospective cohort study was conducted at the 614-bed Gifu University Hospital in Japan. Patients who received intravenous vancomycin for a documented or suspected infection and had their serum vancomycin trough concentration monitored between October 1, 2016 and September 30, 2020 were enrolled in the present study. RESULTS: Multivariate Cox proportional hazard analysis indicated that AUC (>600 µg•h/mL) was a significant risk factor for the incidence of acute kidney injury (AKI), while trough concentration (≥15 µg/mL) was not. Moreover, the AUC (>600 µg•h/mL) showed higher specificity and similar sensitivity to the trough concentration (≥15 µg/mL). Kaplan-Meier plots of the cumulative incidence of the AKI-free rate in patients indicated that the onset of AKI was significantly longer in patients with AUC ≤600 µg•h/mL than in patients with AUC >600 µg•h/mL (HR, 16.1; 95% CI, 6.3-41.2; p < 0.001). CONCLUSION: AUC based on a single trough concentration was a better predictor of nephrotoxicity than trough concentration.

Decreasing the abundance of tetracycline-resistant Escherichia coli in pig feces during nursery using flavophospholipol as a pig feed additive.

Tetracyclines (TCs) are widely used for livestock, and the high prevalence of TC-resistant Escherichia coli in livestock has become a serious concern worldwide. In Japan, the National Action Plan on Antimicrobial Resistance in 2016 aimed to reduce the TC resistance rate in E. coli derived from livestock. Flavophospholipol (FPL), used as a feed additive, has an inhibitory effect on the spread of plasmid-mediated antimicrobial resistance. The number of TC-resistant E. coli was determined in pigs administered TCs and/or FPL to clarify the effect of FPL on reducing the number of TC-resistant E. coli in pigs. TC-resistant E. coli and their plasmids were then analyzed. The pigs were divided into four groups: control, doxycycline (DOXY; a TC), FPL, and a DOXY-FPL combination. Their feces were collected from the nursing period to the day before being transported to the slaughterhouse, followed by estimation of TC-resistant E. coli (colony-forming units [CFU]/g). The number of TC-resistant E. coli increased with the use of DOXY, suggesting that DOXY administration provides a selective pressure for TC-resistant E. coli. Supplementation with FPL as a feed additive significantly suppressed the increase in the number of TC-resistant E. coli, especially during the DOXY administration period. Transfer and growth inhibition analyses were performed for TC-resistant isolates. FPL inhibited the conjugational transfer and growth of a few TC-resistant E. coli isolates. These results suggest that FPL is effective against the spread of TC-resistant E. coli.

Accumulation of lipofuscin in broiler chicken with wooden breast.

Lipofuscin is one of the indicators of oxidative stress. To elucidate the role of oxidative stress in the development of wooden breast, this study investigates lipofuscin accumulation in various parts of wooden breast muscles. Sampling was performed using 46-day-old broiler chickens housed at a commercial Japanese poultry slaughterhouse. Fourteen wooden breast fillets and 13 normal breast fillets were collected in the deboning line. The samples used to measure shear force, 2-thiobarbituric acid reactive substance (TBARS), and for histological analysis were taken from the six portions of breast muscle fillets. In muscles affected by wooden breast, vacuolated muscle fibers were observed, and connective tissues appearing like perimysium were expanded with fibrosis. TBARS value and accumulation of lipofuscin were significantly higher in the wooden breast than in the normal breasts. A lot of lipofuscin granules were localized in the cytoplasm of collapsed muscle fibers of the wooden breast. The cranial portion of the wooden breast showed the highest shear force. The cranial position had a large amount of connective tissue and lipofuscin granules. The results of the present study strongly suggest that high oxidative stress, especially with a significant accumulation of lipofuscin, is associated with the development of wooden breasts.

Decreased colistin resistance and mcr-1 prevalence in pig-derived Escherichia coli in Japan after banning colistin as a feed additive.

BACKGROUND: Antimicrobial resistance to colistin, a widely used feed additive for farm animals across the world, has raised public health concern in recent years. Since July 2018, its use as feed additive has been banned in Japan to reduce the spread of plasmid-based mobilized colistin resistance (mcr) genes and the subsequent development of colistin-resistant bacteria. Evaluating the effects of these measures is required. METHODS: We evaluated the effects of colistin use, as a feed additive, on colistin resistance in pigs (n=5) from birth to finishing in the farm. Moreover, to evaluate changes in colistin resistance and mcr gene prevalence in response to colistin withdrawal, E. coli samples derived from pig faeces sourced from the fields of three geographically distinct farms were characterized before and after the withdrawal of colistin as a feed additive. RESULTS: Colistin-resistant Escherichia coli in pigs (n=5) increased during the colistin administration period and decreased immediately after its end. In three fields, the colistin resistance rate and prevalence of mcr-1 decreased immediately and significantly after the ban. However, colistin-resistant and mcr-1-positive E. coli were still detected in all three farm fields 12 months after the ban on colistin use. CONCLUSION: Agricultural colistin use caused selective pressure that contributed to widespread mcr dissemination in Japan. Colistin resistance and the presence of mcr genes should be continuously monitored in food-producing animals.

Effect of wooden breast on postmortem changes in chicken meat.

Wooden breast is a common problem in the poultry industry, occurring when broiler breast meat becomes rubber-like and extremely hard. Unclear points remain regarding the mechanical strength changes caused by post-mortem biochemical changes in wooden breasts. This study aimed to investigate this knowledge gap. We found endogenous protease activity to be high in wooden breasts and observed a 30 kDa fragment of troponin T (an indicator of postmortem tenderness) from day 1 postmortem. The amount of intramuscular connective tissue in wooden breasts was greater than that of normal breast meat, particularly in the perimysium. The intramuscular connective tissue structure and quantity significantly affect the mechanical strength of meat. It became clear that the wooden breasts are much more mechanically stronger than normal breasts at postmortem day 5 because the large amount of intramuscular connective tissue in the wooden breasts has hardly changed even 5 days postmortem.

Rapid growth rate results in remarkably hardened breast in broilers during the middle stage of rearing: A biochemical and histopathological study.

The high incidence of meat of impaired quality poses a serious problem in the poultry industry. In recent years, the incidence of the pectoralis major muscle that appeared pale colored, remarkably hardened, and exudative, called "wooden breast" or "woody breast" has increased in slaughter houses. In the present study, 19-day-old Ross 308 broiler chickens affected (n = 10) and unaffected (n = 10) with remarkably hardened breast were selected from a commercial broiler farm, and reared to 55 days of age under a controlled environment. Among the affected birds, 5 of 10 birds appeared exhausted with markedly suppressed weight gain and 4 of 10 birds died during the rearing period. In contrast, all unaffected birds survived and most gained weight. Four of 10 unaffected birds lost the ability of back-to-back wing contact by the late stage of rearing. The biochemical analysis of blood plasma samples of 20-day-old birds revealed that creatine kinase and L-aspartate aminotransferase values in most affected birds were higher than those in unaffected birds; however, these values in unaffected birds increased rapidly with lost wing contactability and increasing age. Postmortem examinations revealed that the mean diameter of myofibers in affected birds was smaller than that in unaffected birds. Moreover, symptoms of degenerative and regenerative muscles were observed in most birds in both groups. Among them, a decrease in, or defect of, the characteristic polygonal shape of myofibers was the most common change within the pectoralis major muscles in both groups. The present study demonstrated that broilers affected with remarkably hardened breast during the middle stage of rearing would have suppressed physical status and weight gain, or would die. It was suggested that rapid growth in broilers might be a cause of remarkably hardened breast.

Morphometric analysis of growing tenocytes in the superficial digital flexor tendon of piglets.

The fine structures of different tendons in various animals at different ages have been studied extensively to reveal their arrangement and growth patterns. However, knowledge of the microstructures of the growing tenocytes in the tendons of piglets is still lacking. Thus, we performed the first morphometric analysis to describe the characteristics of tenocytes in the metacarpal superficial digital flexor tendon of 0-, 10- and 20-day-old piglets. In the present study, hydrochloric acid/collagenase digestion was applied to remove the interstitial connective tissue to obtain clear visualization of intact tenocytes and their cytoplasmic processes (Cp). Then, the morphometry of the tenocytes was investigated by optical and electron microscopy. The mean ± SE values of the fascicle area, number of tenocytes/fascicle, cell density, number of Cp/tenocyte, length of Cp, and thickness of Cp were compared among the three age groups. Significant differences (judged at P<0.05) were found in almost all morphometric aspects among the age groups, except for the number of Cp/cell (P=0.545) and thickness of the Cp (P=0.105). A decrease of cell density corresponded with an increase in the length of the Cp, which were extended to connect either with the Cp of the other tenocytes or the surrounding endotendineum. Moreover, an increase of the fascicle area reflected the increase in tendon diameter. The revealed morphometric characteristics are thus the outcome of tendon growth.