Association between Circulating Levels of CXCL9 and CXCL10 and Physical Frailty in Older Adults.

INTRODUCTION: Dysregulation of pro-inflammatory chemokines is considered a potential mechanism for the development of age-related medical conditions such as frailty. However, evidence linking circulating chemokines with frailty remains lacking. MATERIALS AND METHODS: We performed a case-control study including 48 cases and 48 controls aged 65-90 years, using the National Center for Geriatrics and Gerontology outpatient registry data. Cases were outpatients with physical frailty and low habitual daily activity. Controls were robust outpatients who performed habitual daily activities. The Japanese version of the Cardiovascular Health Study criteria was used to diagnose physical frailty, and the modified Baecke questionnaire was used to evaluate habitual daily activities. Serum CXCL9 and CXCL10 levels were measured using enzyme-linked immunosorbent assay. RESULTS: The median age (interquartile range) in cases and controls was 78 (73-83) and 76 (72-80) years, with the proportions of men were 47.9% and 43.8%, respectively. In the logistic regression model with adjustment for age, sex, and other confounding factors, the multivariable odds ratios (95% confidence intervals) for the highest versus lowest tertile of CXCL9 and CXCL10 levels were 7.90 (1.61-49.80) and 1.61 (0.42-6.30), respectively. However, we did not observe a linear association between CXCL9 levels and physical frailty components. DISCUSSION/CONCLUSION: Our preliminary data exhibit that circulating CXCL9 levels were positively associated with the odds of physical frailty. However, these findings lack evidence of a dose-response relationship between CXCL9 levels and physical frailty components. Further research with a larger sample size is required to confirm these findings.

Dietary amino acid intake and sleep duration are additively involved in future cognitive decline in Japanese adults aged 60 years or over: a community-based longitudinal study.

BACKGROUND: Sleep duration and amino acid intake are independently associated with cognitive decline. This study aimed to determine the longitudinal association between sleep duration and cognitive impairment incidence and to examine the involvement of diet, particularly amino acid intake, in these associations in community dwellers. METHODS: In this longitudinal study in a community-based setting, we analyzed data from 623 adults aged 60-83 years without cognitive impairment at baseline. Sleep duration was assessed using a self-report questionnaire. Amino acid intake was assessed using 3-day dietary records. Cognitive impairment was defined as a Mini-Mental State Examination score ≤ 27. Participants were classified into short-, moderate-, and long-sleep groups according to baseline sleep duration (≤ 6, 7-8, and > 8 h, respectively). Using moderate sleep as a reference, odds ratios (ORs) and 95% confidence intervals (CIs) of short- and long-sleep for cognitive-impairment incidence were estimated using the generalized estimating equation. Participants were classified according to sex-stratified quartiles (Q) of 19 amino acid intake: Q1 and Q2-Q4 were low- and middle to high-intake groups, respectively. Using middle- to high-intake as a reference, ORs and 95% CIs of low intake for cognitive impairment incidence were estimated using the generalized estimating equation in each sleep-duration group. Follow-up period, sex, age, body mass index, depressive symptoms, education, smoking status, employment status, sleep aids use, physical activity, medical history, and Mini-Mental State Examination score at baseline were covariates. RESULTS: Mean follow-up period was 6.9 ± 2.1 years. Adjusted ORs (95% CIs) for cognitive impairment in short- and long-sleep groups were 0.81 (0.49-1.35, P = 0.423) and 1.41 (1.05-1.87, P = 0.020), respectively. Particularly in long sleepers (i.e., > 8 h), cognitive impairment was significantly associated with low cystine, proline, and serine intake [adjusted ORs (95% CIs) for cognitive impairment were 2.17 (1.15-4.11, P = 0.017), 1.86 (1.07-3.23, P = 0.027), and 2.21 (1.14-4.29, P = 0.019), respectively]. CONCLUSIONS: Community-dwelling adults aged ≥ 60 years who sleep longer are more likely to have cognitive decline, and attention should be paid to the low cystine, proline, and serine intake.

Severe neuroglycopenic symptoms due to nonketotic hypoglycemia in children with cardio-facio-cutaneous syndrome.

Cardio-facio-cutaneous syndrome (CFC) (OMIM 115150) is a congenital disease caused by constitutive activation of the Raf/MEK/ERK signaling cascade. Unlike aspects of morphological anomalies, metabolic functions related to the disease have garnered little attention. We present severe neuroglycopenic symptoms due to nonketotic hypoglycemia in two children with CFC (Case 1, a 4-year-old male with c.389A > G heterozygous variant in MAP2K1; Case 2, a 3-year-old male with c.770A > G heterozygous variant in BRAF). Case 1 exhibited a nonketotic hypoglycemic coma and clustered left-hemispheric convulsions despite receiving infusion therapy, leading to severe sequelae with choreoathetosis. Brain magnetic resonance imaging of Case 1 showed T2-elongation with restricted diffusion on the bilateral basal ganglia and thalamus, with the dominance of the right putamen. Case 2 presented a prolonged generalized seizure as an initial clinical symptom but fully recovered. The presence of growth hormone and cortisol deficiency was ruled out in both cases. Blood spots acylcarnitine profiles excluded the co-occurrence of mitochondrial HMG-CoA synthase deficiency and HMG-CoA lyase deficiency. These cases demonstrate the potential vulnerability to nonketotic hypoglycemia, especially during lipid shortages. As children with CFC frequently have difficulties feeding, we suggest great attention should be paid to the potential risk of severe nonketotic hypoglycemia.

Diet-induced inflammation is associated with sarcopenia and muscle strength in older adults who visit a frailty clinic.

BACKGROUND: Diet-induced inflammation may be associated with sarcopenia; however, few reports have examined this relationship. AIM: To examine the association between the dietary inflammatory index (DII) and sarcopenia in older adults who visited a frailty clinic in Japan. METHODS: This cross-sectional study used outpatient data from the Frailty Registry Study. The DII is an index of diet-induced inflammation, and a dietary assessment was performed using a brief self-administered diet history questionnaire to calculate the DII score. We classified DII scores by quartiles (Q1-Q4), and sarcopenia was diagnosed according to the Asian Working Group for Sarcopenia 2019 consensus. Logistic regression analyses for sarcopenia were performed. Age, sex, comorbidities, and physical activity were entered as confounding factors (Model 1) and Models 2, 3, and 4 with BMI, protein intake, and energy intake added to Model 1. RESULTS: We included 304 patients in the analysis (mean age, 77.6 ± 6.3 years; female, 67.4%). The prevalence of sarcopenia was 14.5%. Logistic regression analyses showed that DII scores were significantly associated with sarcopenia in Model 1 and 2 (Model 1, reference: Q1, Q4: OR 3.10, P = 0.020; Model 2, Q4: OR 3.40, P = 0,022) but not in Model 3 and 4. DISCUSSION: Diet-induced inflammation is associated with a higher likelihood of sarcopenia; however, this association disappeared after confounding for protein and energy intake. CONCLUSIONS: The results demonstrated that dietary protein and energy parameters were the main drivers for muscle health in medical patients.

[Cross-sectional association between the amino acid intake and episodic memory in middle- and old-aged community dwellers].

AIM: The specific amino acid intake has been suggested to be positively associated with the cognitive function. However, few reports have investigated the association between the amino acid intake and episodic memory (EM). Therefore, we investigated this association. METHODS: Data were obtained from the fourth survey (2004-2006) of the National Institute for Longevity Sciences - Longitudinal Study of Aging. We analyzed 2,082 participants 40-85 years old (50.1% male). The dietary intake was assessed by the three-day dietary records, and participants were classified into sex- and age-specific tertiles of protein and amino acid intakes. EM was assessed using the Logical Memory II of the Wechsler Memory Scale. The association of protein and amino acid intakes with EM was analyzed using the general linear model. Covariates were sex, age, body mass index, education, depressive symptoms, smoking status, employment status, living alone, and medical history in model 1. The energy intake was added to model 1 in model 2. The protein intake was added to model 2 in model 3. RESULTS: The mean (standard deviation) age was 59.4 (12.3) years old. After adjusting for the energy intake, the EM tended to be higher with a higher protein intake (p=0.053 for group differences and p=0.015 for trends). Furthermore, after adjusting for energy and protein intake, EM was significantly higher with higher intakes of isoleucine, leucine, lysine, methionine, phenylalanine, tyrosine, tryptophan, valine, and histidine (p< 0.05, both for group differences and trends). CONCLUSION: Our findings suggest a positive association between EM and the intake of essential and semi-essential amino acids, independent of the protein and energy intake.

The combination of fatty liver and increased gamma-glutamyl transpeptidase levels as a risk factor for atherosclerotic plaque development in apparently healthy people

Background/aim: To elucidate how the combination of fatty liver and increased serum gamma-glutamyl transpeptidase (GGT) levels influences atherosclerotic plaque development in apparently healthy people. Materials and methods: The study population included people who had received an annual health checkup for more than 7 years and had no evidence of carotid plaque at baseline. We investigated the risk factors for carotid plaque occurrence using the Cox proportional hazards model. Results: A total of 107 people (76 men and 31 women; median age, 49 years) were enrolled. At baseline, fatty liver and a serum GGT level ≥50 U/L were observed in 13 and 38 people, respectively. During a median follow-up period of 13.3 years, carotid plaques appeared in 34 people. Multivariate analysis revealed that the combination of fatty liver and a serum GGT level ≥50 U/L was the only significant risk factor for carotid plaque occurrence (age- and sex-adjusted hazard ratio: 5.55; 95% confidence interval 1.70­18.14; P = 0.005). Conclusion: The combination of fatty liver and increased serum GGT levels raises the risk for atherosclerotic plaque development in apparently healthy people.

[A homebound status is related to a reduction of meal intake in older outpatients, independent depressive mood and their physical function].

AIM: We aimed to investigate the relationship between the frequency at which patients went out and the reduction of meal intake among older outpatients who did not require care. METHODS: The subjects were outpatients of ≥65 years of age who visited the department of geriatric medicine in our hospital for the first time. We analyzed 463 subjects (male, n=184; female, n=279), after excluding patients who had dementia, required care, lived in a nursing home, or had an acute disease.The outcome measure was the reduction of meal intake (a moderate or higher decrease in the patient's meal intake in the past 3 months). The independent measure was homebound status (going-out less than once a week). The covariates were sex, age, number of medications, and Kihon Checklist (categories of undernutrition, oral function, physical function, and mood). A logistic regression analysis was performed. RESULTS: The average age was 79.6±5.9 years in men, 79.9±6.1 years in women. Among the participants, 104 (22.5%) had a homebound status. In the logistic regression analyses, a homebound status was significantly associated with a reduction in meal intake, even after adjustment for potential confounding factors, including depressive mood and a low physical function (OR: 2.0; 95% CI: 1.1-3.6). CONCLUSIONS: A homebound status in older outpatients was related to a decline in their meal intake, independent of depressive mood and a low physical function. A reduction in meal intake leads to a lack of energy and results in malnutrition. Our results suggest that assessing the frequency at independently living older outpatients go out is important for the early prevention of malnutrition.

Isoniazid- and streptomycin-resistant miliary tuberculosis complicated by intracranial tuberculoma in a Japanese infant.

In Japan, the incidence of severe pediatric tuberculosis (TB) has decreased dramatically in recent years. However, children in Japan can still have considerable opportunities to contract TB infection from adult TB patients living nearby, and infants infected with TB may develop severe disseminated disease. A 3-month-old girl was admitted to our hospital with dyspnea and poor feeding. After admission, miliary TB and multiple brain tuberculomas were diagnosed. Anti-tuberculous therapy was initiated with streptomycin (SM), isoniazid (INH), rifampicin and pyrazinamide. Symptoms persisted after starting the initial treatment and mycobacterial cultures of gastric fluid remained positive. Drug sensitivity testing revealed the TB strain isolated on admission as completely resistant to INH and SM. Treatments with INH and SM were therefore stopped, and treatment with ethambutol and ethionamide was started in addition to rifampicin and pyrazinamide. After this change to the treatment regimen, symptoms and laboratory data gradually improved. The patient was treated with these four drugs for 18 months, and then pyrazinamide was stopped. After another 2 months, ethambutol was stopped. Treatment of tuberculosis was completed in 24 months. No adverse effects of these anti-TB drugs were observed. The patient achieved a full recovery without any sequelae. On the other hand, the infectious source for this patient remained unidentified, despite the extensive contact investigations. The incidence of drug-resistant TB is increasing in many areas of the world. Continuous monitoring for pediatric patients with drug-resistant TB is therefore needed.

The Clock-Drawing Test as a Useful Screening Assessment of Preoperative Cognitive Impairment with Readmission After Transcatheter Aortic Valve Implantation.

Background: Transcatheter aortic valve implantation (TAVI) has recently become more common as a treatment for severe, symptomatic aortic stenosis (AS). Cognitive impairment (CI) is strongly associated with the prognosis of TAVI patients. However, some cognitive assessments currently in use are difficult to perform routinely in the clinical setting. To easier CI evaluation, we investigated whether CI using the clock-drawing test (CDT), one part of the Mini-Cog, affects the postoperative prognosis of TAVI patients with AS. Methods: The present study enrolled 52 patients (median age, 85 years; 28.8% male) who underwent TAVI and were discharged between 2019 and 2021. The outcome was readmission for all causes within one year of discharge and patients were grouped according to whether they were readmitted or not. Cognitive function was assessed using the Mini-Cog which combines verbal playback and CDT. Results: Of the 52, 11 patients (21.2%) comprised readmission group, including 4 (36.4%) each for fracture and infection, and 1 (9.1%) each for heart failure, subdural hematoma, and pneumothorax. Median Mini-Cog score was lower in the readmission group than in the non-readmission group (4 vs. 5; P < 0.05). The frequency of Mini-Cog score < 3 (indicative of CI) and CDT failure were significantly higher in the readmission group than in the non-readmission group, respectively (46% vs. 7%, P < 0.01) (46% vs. 12%, P < 0.05). Both of Mini-Cog score < 3 and CDT failure were independently associated with readmission. The areas under the curve showed CDT was an indicator of readmission with similar accuracy to the Mini-Cog score < 3. Kaplan-Meier curves showed significant differences in readmission after 1 year between the 2 Mini-Cog groups with scores of < 3 or ≥ 3 points and CDT failure and success. Conclusion: The CDT may be a very easy and simple screening assessment of preoperative CI with readmission within one year after TAVI.

Validation of a dietary balance score in middle-aged and older community-dwelling Japanese.

This study assessed the validity of dietary balance scores (DBSs) by investigating the association between DBSs and nutrient adequacy (NA) in two Japanese populations. The participants were 65 community-dwelling Japanese from Tokushima Prefecture and 2,330 community-dwelling Japanese from Aichi Prefecture. Based on food frequency questionnaires or 3-day dietary records, we obtained 18 food groups. The NA score integrates nine beneficial nutrients and two nutrients that should be limited. We calculated four different DBSs: DBS1 consisted of five food groups (score range:0?20), DBS2 consisted of nine food groups (score range:0?36), DBS3 consisted of eight food groups (score range:0?32), and DBS4 consisted of 10 food groups (score range:0?40). Both the Spearman rank correlation coefficient with NA and the area under the receiver operating characteristic curve (AUC-ROC) for the nine beneficial nutrients were then estimated to test the performance of each DBS in predicting nutrient intake. The results showed that DBS1 and DBS4 were positively correlated with NA, while the AUC-ROC showed that DBS4 could moderately discriminate individuals with adequate intake levels of all nine nutrients. These findings suggest DBSs (especially DBS4) are useful in assessing dietary balance in middle-aged and older community-dwelling Japanese. J. Med. Invest. 70 : 377-387, August, 2023.

A novel alkaline esterase from Sporosarcina sp. nov. strain eSP04 catalyzing the hydrolysis of a wide variety of aryl-carboxylic acid esters.

A novel esterase showing activity specific for esters of aryl-carboxylic acids was discovered in Sporosarcina sp. nov., which was identified by the 16S rDNA sequencing method in addition to morphological and physiological analyses. The aryl-carboxylesterase (named EstAC) was purified 780-fold from crude cell extracts by a 5-step procedure. EstAC was characterized as a monomeric protein with a molecular weight of 43,000, an optimum pH of around 9.0, and an optimum temperature of 40 °C. The pH optimum and the effects of inhibitors together with an internal amino acid sequence suggested that EstAC is a member of family VIII esterases. EstAC was found to be highly active on a wide variety of substrates such as alkyl benzoates, alkyl phenylacetates, ethyl α- or ß-substituted phenylpropionates, dialkyl terephthalates, dimethyl isophthalate, and ethylene glycol dibenzoate. However, monomethyl terephthalate was not hydrolyzed. It was suggested that EstAC had 4-hydroxybenzoyl and cinnamoyl esterase activities as well.

Heparin-induced thrombocytopenia in essential thrombocytosis.

We report a 67-year-old woman with essential thrombocytosis who developed cerebral infarction and heparin-induced thrombocytopenia during treatment for the cerebral infarction. She developed additional cerebral infarcts, acute femoral artery occlusion, and thrombophlebitis of her lower extremities. She was successfully treated with argatroban. This is the first report of a patient with essential thrombocytosis who developed heparin-induced thrombocytopenia and serious conditions, which included multiple thromboembolisms and coagulation disorders mimicking disseminated intravascular coagulation.

Breakfast Protein Quality and Muscle Strength in Japanese Older Adults: A Community-Based Longitudinal Study.

OBJECTIVES: The amount of breakfast protein intake is important for maintaining muscle strength. However, the effect of breakfast protein quality (ie, bioavailability) remains unclear. We investigated the association between breakfast protein quality and the incidence of muscle weakness. DESIGN: Longitudinal study. SETTING AND PARTICIPANTS: Healthy older adults age 60-83 years without stroke, arthritis, Parkinson disease, or muscle weakness at baseline (maximum follow-up period and participations were 9.2 years and 5 times, respectively). METHODS: Weakness was defined by the Asian Working Group for Sarcopenia 2019 criteria, using grip strength. Breakfast protein quality was evaluated using the protein digestibility-corrected amino acid score (PDCAAS), where higher scores represent higher quality, calculated from 3-day dietary records. Participants were classified according to sex-stratified tertiles of breakfast PDCAAS (ie, low to high groups). The association between PDCAAS and incident weakness was analyzed using the generalized estimating equation, after adjusting for sex, age, follow-up time, grip strength, body mass index, physical activity, cognition, education, smoking, economics, medical history, lunch and dinner PDCAASs, and energy and protein intake during 3 regular meals at baseline. RESULTS: Overall, 14.4% of the initial sample was excluded owing to a diagnosis of weakness-related diseases, and 58.3% (n = 701) had at least 1 follow-up measurement for inclusion in the analysis. The mean ± SD follow-up period was 6.9 ± 2.1 years; the cumulative number of participants was 3019, and 282 developed weakness. Using the low PDCAAS group as the reference, the adjusted odds ratios (95% CIs) for incident weakness in the middle and high PDCAAS groups were 0.71 (0.43-1.18) and 0.50 (0.29-0.86), respectively. CONCLUSIONS AND IMPLICATIONS: Higher breakfast protein quality was associated with a reduction in incident weakness in older adults, independent of protein intake. These findings may highlight the role of protein quality for muscle health in older adults.

Establishment of diagnosis by bisulfite-treated methylation-specific PCR method and analysis of clinical characteristics of pseudohypoparathyroidism type 1b.

Pathogenesis of pseudohypoparathyroidism type 1b (PHP-1b) is related to the loss of methylation at the GNAS exon A/B region, which is combined with epigenetic defects at other differentially methylated GNAS regions in most sporadic cases. In this study, we established a method for evaluating the methylation status of a CpG island in exon A/B using a methylation-specific polymerase chain reaction (MSPCR). We designed two primer pairs specific for the methylated and unmethylated alleles and evaluated the methylation status of GNAS exon A/B in samples from PHP-1b patients and normal controls. We examined 20 Japanese patients and 20 normal controls, and one primer set was found to be appropriate for diagnosis. Furthermore, we evaluated the clinical data of patients. Weight and height of patients were not significantly different from the normal population. Short stature (adult height ≤ 2SD) was observed in one patient and short metacarpals in two. Obesity was observed in six patients, and no patient showed ectopic subcutaneous calcification. Seven patients showed subclinical hypothyroidism because of resistance to thyroid stimulating hormone, but no patient had an abnormally low free thyroxine level, and none received oral thyroid hormone replacement. For diagnosis of PHP-1b, only clinical data is not sufficient because a few PHP-1b patients show clinical features similar to PHP-1a, and hence, molecular biology techniques are required for correct diagnosis. We concluded that MSPCR is applicable for rapid molecular diagnosis of PHP-1b.

[A case of left posterior inferior temporal infarction with right homonymous upper quadrantanopsia and alexia with agraphia for Kanji].

BACKGROUND: Alexia with agraphia usually represents damage in angular gyrus. We report an unusual case of alexia with agraphia caused by a posterior inferior temporal lesion. CASE REPORT: A 82-year-old, right-handed man was admitted because of reading disorder. Visual acuity was 0.7 OD and 0.7 OS. Goldmann perimetry revealed right homonymous upper quadrantanopsia. Standard Language Test of Aphasia revealed mild anomia and severe alexia with agraphia for kanji. Magnetic resonance imaging demonstrated left temporal and posterior infarction in the posterior cerebral artery region. Single photon emission computed tomography revealed decreased blood flow in the left inferior temporal, parahippocampal, fusiform, lingual and inferior occipital gyri. There were no significant lesions nor any decreased of blood flow in the angular gyrus. CONCLUSION: It is suggested that reading and writing of ideograms such as Kanji is related not only to the angular gyrus but also to the posterior inferior temporal lobe and medial posterior lobe of the visual cortex.

Dietary diversity is associated with longitudinal changes in hippocampal volume among Japanese community dwellers.

BACKGROUND/OBJECTIVES: Dietary habits are known to affect health, including the rate of brain ageing and susceptibility to diseases. This study examines the longitudinal relationship between dietary diversity and hippocampal volume, which is a key structure of memory processing and is known to be impaired in dementia. SUBJECTS/METHODS: Subjects were aged 40-89 years (n = 1683, men: 50.6%) and participated in a 2-year follow-up study of the National Institute for Longevity Sciences-Longitudinal Study of Aging. Dietary intake was calculated from 3-day dietary records, and dietary diversity was determined using the Quantitative Index for Dietary Diversity at baseline. Longitudinal changes in hippocampal and total grey matter volumes were estimated by T1-weighted brain magnetic resonance imaging and FreeSurfer software. Estimated mean brain volume change in relation to dietary diversity score quintiles was assessed by the general linear model, adjusted for age, sex, education, smoking status, alcohol intake, physical activity, and comorbidities. RESULTS: The mean (± standard deviation) % decreases in hippocampal and total grey matter volume during the 2-year follow-up were 1.00% (±2.27%) and 0.78% (±1.83%), respectively. Multivariate-adjusted decreases in total grey matter volume were associated with dietary diversity score (p = 0.065, p for trend = 0.017), and the % decrease in hippocampal volume was more strongly associated with the dietary diversity score: the estimated mean (± standard error) values were 1.31% (±0.12%), 1.07% (±0.12%), 0.98% (±0.12%), 0.81% (±0.12%), and 0.85% (±0.12%), according to dietary diversity quintiles in ascending order (p = 0.030, p for trend = 0.003). CONCLUSIONS: Among community dwellers, increased dietary diversity may be a new nutritional strategy to prevent hippocampal atrophy.

A novel missense mutation in the AVPR2 gene of a Japanese infant with nephrogenic diabetes insipidus.

We describe an infant with nephrogenic diabetes insipidus (NDI) with a novel mutation in the arginine vasopressin receptor 2 (AVPR2) gene. A 1-month-old infant showed failure to thrive and hypernatremia. The water deprivation test revealed elevated serum osmolality and low urine osmolality. The patient showed a slight but not significant response to intramuscular injection of arginine vasopressin (AVP). DNA analysis revealed a novel missense mutation involving substitution of proline for leucine at position 173 (P173L), which was reported to be important for stabilizing the hydrogen bond between tyrosine at position 205 and leucine at position 169. This mutation was not detected in 116 ethnic-matched controls. This case, with clinical data including the water deprivation test and P173L mutation, will facilitate understanding the structure and function of the A VPR2.

Inhibition of γ-glutamyltransferase ameliorates ischaemia-reoxygenation tissue damage in rats with hepatic steatosis.

BACKGROUND AND PURPOSE: Hepatic steatosis may be associated with an increased γ-glutamyltransferase (γ-GT) levels. Ischaemia-reoxygenation (IR) injury causes several deleterious effects. We evaluated the protective effects of a selective inhibitor of γ-GT in experimentally induced IR injury in rats with obesity and steatosis. EXPERIMENTAL APPROACH: Otsuka Long-Evans Tokushima Fatty (OLETF) rats with hepatic steatosis were used in the current study. The portal vein and hepatic artery of left lateral and median lobes were clamped to induce ischaemia. Before clamping, 1 ml of saline (IR group) or 1-ml saline containing 1 mg·kg-1 body weight of GGsTop (γ-GT inhibitor; IR-GGsTop group) was injected into the liver via the inferior vena cava. Blood flow was restored after at 30 min of the start of ischaemia. Blood was collected before, at 30 min after ischaemia and at 2 h and 6 h after reoxygenation. All the animals were killed at 6 h and the livers were collected. KEY RESULTS: Treatment with GGsTop resulted in significant reduction of serum ALT, AST and γ-GT levels and hepatic γ-GT, malondialdehyde, 4-hydroxy-2-nonenal and HMGB1 at 6 h after reoxygenation. Inhibition of γ-GT retained normal hepatic glutathione levels. There was prominent hepatic necrosis in IR group, which is significantly reduced in IR-GGsTop group. CONCLUSION AND IMPLICATIONS: Treatment with GGsTop significantly increased hepatic glutathione content, reduced hepatic MDA, 4-HNE and HMGB1 levels and, remarkably, ameliorated hepatic necrosis after ischaemia-reoxygenation. The results indicated that GGsTop could be an appropriate therapeutic agent to reduce IR-induced liver injury in obesity and steatosis.

Recombinant thrombomodulin prevented hepatic ischemia-reperfusion injury by inhibiting high-mobility group box 1 in rats.

Recombinant thrombomodulin (rTM) is a novel anticoagulant and anti-inflammatory agent that inhibits secretion of high-mobility group box 1 (HMGB1) from liver. We evaluated the protective effects of rTM on hepatic ischemia-reperfusion injury in rats. Ischemia was induced by clamping the portal vein and hepatic artery of left lateral and median lobes of the liver. At 30 min before ischemia and at 6 h after reperfusion, 0.3 ml of saline (IR group) or 0.3 ml of saline containing 6 mg/kg body weight of rTM (IR-rTM group) was injected into the liver through inferior vena cava or caudate vein. Blood flow was restored at 60 min of ischemia. Blood was collected 30 min prior to induction of ischemia and before restoration of blood flow, and at 6, 12, and 24 h after reperfusion. All the animals were euthanized at 24 h after reperfusion and the livers were harvested and subjected to biochemical and pathological evaluations. Serum levels of ALT, AST, and HMGB1 were significantly lower after reperfusion in the IR-rTM group compared to IR group. Marked hepatic necrosis was present in the IR group, while necrosis was almost absent in IR-rTM group. Treatment with rTM significantly reduced the expression of TNF-α and formation of 4-hydroxynonenal in the IR-rTM group compared to IR group. The results of the present study indicate that rTM could be used as a potent therapeutic agent to prevent IR-induced hepatic injury and the related adverse events.