The Hydrolytic Peptides of Soybean Protein Induce Cell Cycle Arrest and Apoptosis on Human Oral Cancer Cell Line HSC-3.

Protein hydrolysates from various sources, including tuna cooking juice, soy protein isolate, sodium caseinate, wheat gluten and skin gelatin from porcine, tilapia, halibut and milkfish were analyzed to screen their antiproliferative activities against the human oral squamous carcinoma cell line, HSC-3. The soy protein isolate was selected for further investigations based on its hydrolysates with bromelain (SB) and thermolysin (ST), showing the greatest inhibition of cell growth. The SB and ST hydrolysates showed antiproliferative activities up to 35.45-76.39% against HSC-3 cells at 72 h, and their IC50 values were 0.74 and 0.60 mg/mL, respectively. SB and ST induced cell cycle arrest in the S phase through a pathway independent of p21 and p27 protein expression. Further, ST induced the apoptosis of HSC-3 cells by downregulating expression of Bcl-2, PARP, caspase 3 and caspase 9, but an upregulating expression of p53 and cleaved caspase 3. Unlike ST, SB may induce necrosis on HSC-3 cells. Thus, soybean hydrolysates may be a good source for providing antiproliferative peptides against HSC-3, while SB and ST may have the potential to be developed as functional foods.

A study to evaluate the potential of an in silico approach for predicting dipeptidyl peptidase-IV inhibitory activity in vitro of protein hydrolysates.

A total of 294 edible protein sequences and 5 commercial proteases listed in the BIOPEP database were analyzed in silico. The frequency (A), a parameter in silico described previously, was examined further to calculating the ratio of truncated peptides with Xaa-proline and/or Xaa-alanine to all peptide fragments in a protein hydrolyzed with a protease, using the BIOPEP database. Then the in vitro DPP-IV inhibitory activity was determined using the same 15 protein and protease combinations to evaluate their relationship. The result shows that A values considering the number of Xaa-proline+Xaa-alanine exhibited a strong correlation with in vitro DPP-IV inhibition rates by Pearson's correlation analysis (r=0.6993; P<0.05). Therefore, the in silico approach is effective to predict DPP-IV inhibitory activities in vitro of protein hydrolysates.

In silico, in vitro and in vivo analyses of dipeptidyl peptidase IV inhibitory activity and the antidiabetic effect of sodium caseinate hydrolysate.

The frequency (A), a novel in silico parameter, was developed by calculating the ratio of the number of truncated peptides with Xaa-proline and Xaa-alanine to all peptide fragments from a protein hydrolyzed with a specific protease. The highest in vitro DPP-IV inhibitory activity (72.7%) was observed in the hydrolysate of sodium caseinate by bromelain (Cas/BRO), and the constituent proteins of bovine casein also had relatively high A values (0.10-0.17) with BRO hydrolysis. 1CBR (the <1 kDa fraction of Cas/BRO) showed the greatest in vitro DPP-IV inhibitory activity of 77.5% and was used for in vivo test by high-fat diet-fed and low-dose streptozotocin-induced diabetic rats. The daily administration of 1CBR for 6 weeks was effective to improve glycaemic control in diabetic rats. The results indicate that the novel in silico method has the potential as a screening tool to predict dietary proteins to generate DPP-IV inhibitory and antidiabetic peptides.

Isolation of prolyl endopeptidase inhibitory peptides from a sodium caseinate hydrolysate.

Prolyl endopeptidase (PEP) has been associated with neurodegenerative disorders, and the PEP inhibitors can restore the memory loss caused by amnesic compounds. In this study, we investigated the PEP inhibitory activity of the enzymatic hydrolysates from various food protein sources, and isolated and identified the PEP inhibitory peptides. The hydrolysate obtained from sodium caseinate using bromelain (SC/BML) displayed the highest inhibitory activity of 86.8% at 5 mg mL(-1) in the present study, and its IC50 value against PEP was 0.77 mg mL(-1). The F-5 fraction by RP-HPLC (reversed-phase high performance liquid chromatography) from SC/BML showed the highest PEP inhibition rate of 88.4%, and 9 peptide sequences were identified. The synthetic peptides (1245.63-1787.94 Da) showed dose-dependent inhibition effects on PEP as competitive inhibitors with IC50 values between 29.8 and 650.5 µM. The results suggest that the peptides derived from sodium caseinate have the potential to be PEP inhibitors.

The Relationship between Stasis-Stagnation Constitution and Peripheral Arterial Disease in Patients with Type 2 Diabetes.

Objectives. In traditional Chinese medicine, Yu-Zhi (YZ, indicating stasis and stagnation) constitution describes a body that tends to express abnormal circulatory conditions. This study identified the linkage between YZ constitution and peripheral arterial disease (PAD) in patients with type 2 diabetes. Methods. Patients over 20 years of age who had had type 2 diabetes for 5 years or longer were recruited. PAD was diagnosed if the ankle-brachial index score was ≤0.9 in either leg. Level of YZ constitution was accessed by an YZ Constitution Questionnaire. Results. A total of 712 patients (354 men and 358 women) with a mean age of 61.5 ± 10.6 years and diabetes duration of 13.1 ± 6.7 years were recruited. The prevalence of PAD among our patients was 7.2%. Multivariate logistic regression revealed significant correlations between PAD and, respectively, YZ score, age, diabetes duration, current smoking, and hs-CRP. Conclusion. In addition to traditional risk factors, YZ constitution was statistically associated with PAD in patients with type 2 diabetes. This result invites further research into the effectiveness of traditional Chinese medicine to treat YZ constitution.

Oral hypoglycaemic agents and the development of non-fatal cardiovascular events in patients with type 2 diabetes mellitus.

AIMS: This study aimed to assess the risk of non-fatal cardiovascular events among patients with type 2 diabetes mellitus (T2DM) who are taking metformin, glimepiride or glyburide. MATERIALS AND METHODS: Using the National Health Insurance Research database in Taiwan, this retrospective cohort study identified 1159 patients with newly diagnosed T2DM from 1998 to 2007, 30 years and older and without a history of cardiovascular disease at baseline. Patients with cancer, liver cirrhosis or chronic kidney disease were excluded. On the basis of prescription, patients were grouped into three medication subcohorts: metformin (N = 595), glimepiride (N = 234) or glyburide (N = 330) monotherapy for 100% of the follow-up period without any oral anti-diabetic agents added or changed, by the end of 2009. Incidence and hazard ratios of non-fatal cardiovascular events including coronary artery disease, peripheral artery disease, stroke and heart failure among these three subcohorts were compared. RESULTS: The overall incidence of non-fatal cardiovascular events was the highest for patients taking glyburide (169.1 per 1000 person-years), followed by for those taking glimepiride and metformin (95.2 and 49.1 per 1000 person-years, respectively). Compared with the adjusted hazard ratio for patients taking glyburide, the adjusted hazard ratio for those taking glimepiride was 0.52 (95% CI 0.40-0.69) and for those taking metformin was 0.31 (95% CI 0.24-0.40). CONCLUSIONS: T2DM patients taking metformin and glimepiride are at lower risk of non-fatal cardiovascular events than those taking glyburide.

Chylous ascites as a manifestation of thyrotoxic cardiomyopathy in a patient with untreated Graves' disease.

BACKGROUND: Thyrotoxicosis is an uncommon cause of heart failure, and patients with heart failure rarely present with chylous ascites. In this report, we describe a patient with uncontrolled Graves' disease with thyrotoxicosis, heart failure, and chylous ascites. SUMMARY: A 39-year-old woman with no previous cardiac disease presented with dyspnea, orthopnea, palpitations, exophthalmos, goiter, distended abdomen, and pedal edema. The thyroid function tests demonstrated hyperthyroid Graves' disease (serum-free triiodothyronine level, 7.12 pg/mL [reference range, 2.0-4.0]; free thyroxine level, 4.33 ng/dL [reference range, 0.54-1.40]; thyroid-stimulating hormone level, <0.015 microU/mL [reference range, 0.34-5.60]; and thyrotropin receptor antibodies, 84.5% [reference value, <15%]). The chest radiograph showed moderate cardiomegaly and bilateral pleural effusions, electrocardiogram revealed atrial fibrillation, and the abdominal sonography found ascites. Chylous ascites was diagnosed by paracentesis and analysis of the ascitic fluid (triglyceride level, 347 mg/dL). Laboratory and imaging studies demonstrated no apparent hepatic dysfunction, abnormal tumor, lymphadenopathy, or lymphatic drainage deficit. With aggressive treatment of the heart failure and hyperthyroid state, her dyspnea, pleural effusion, chylous ascites, and edema resolved completely within a few days. CONCLUSIONS: Chylous ascites may develop as a result of heart failure secondary to thyrotoxic cardiomyopathy and resolve promptly if treated appropriately.

Permanent central diabetes insipidus after mild traumatic brain injury.

AIM: The patients in the permanent diabetes insipidus (DI) group are more likely to have more severe TBI, which is defined by a post-resuscitational and pre-sedational Glasgow Coma Scale (GCS) score of 8/15 or less. This study presents a case of permanent, central DI following mild traumatic brain injury with post-resuscitation GCS 13/15. CASE REPORT: A 17-year-old boy suffered from mild brain injury and experienced permanent DI without any anatomical changes on image in the early stage of traumatic brain injury. However, 1 year later, magnetic resonance imaging (MRI) of the brain in this patient has revealed some sequel of contusion. Moreover, the patient still has DI after treatment with diamino-8-D-arginine vasopressin (DDAVP). CONCLUSION: This patient had a rare clinical presentation of permanent, central DI, following a mild traumatic brain injury. Identification of head trauma as the aetiology of hypopituitarism may be overlooked if there is a long delay in onset after trauma. Since anterior hypopituitarism can develop decades after the episode of head trauma, monitoring for endocrine dysfunction during follow-up of these patients is important.

p53 codon 72 proline/arginine polymorphism and autoimmune thyroid diseases.

p53 protein participates in the processes of apoptosis, which is involved in a number of immunological reactions. In order to test whether the p53 gene could be used as a genetic marker for the prediction of the development of autoimmune thyroid diseases (AITD), we screened, by using polymerase chain reaction (PCR) analysis, for the C (CCC)/G (CGC) polymorphism at the p53 codon 72 (Pro 72/Arg 72) to determine the genotypes of 107 Hashimoto's thyroiditis (HT) and 90 Graves' disease (GD) patients, and 105 normal controls. The data demonstrated that, for the genotype analysis, HT patients featured an enhanced numerical ratio for the Arg/Arg homozygous genotype (33.7%) and a diminished ratio for the Arg/Pro heterozygous genotype (41.1%) at the p53 codon 72 than was the case for normal controls (Arg/Arg: 17.1% and Arg/Pro: 61.9%; P=0.005). The odds ratio for the risk of the Arg/Arg genotype's appearance, compared with that of the Arg/Pro and Pro/Pro genotypes combined, for the HT patient group was 2.450 (95% confidence interval: 1.274-4.716). With respect to allelic analysis, we did not observe significant difference in the frequency of appearance of the Arg allelic variant and the Pro allelic variant for the p53 codon 72 when comparing the HT patient group with the control group (P=0.208). On the other hand, GD patients presented no significant difference in distribution for both genotype and allelic frequencies (P=0.344 and 0.245, respectively) when compared with normal controls. In conclusion, HT patients feature a greater ratio of arginine homozygosity at p53 codon 72 than in the case for normal subjects. The p53 codon 72 proline/arginine polymorphism may be a genetic marker to predict the increased susceptibility of development of HT.

Comparing dual phase Tl-201 thyroid scan and fine-needle aspiration cytology to detect follicular carcinoma.

UNLABELLED: It remains difficult to pre-operatively differentiate follicular carcinoma and adenoma even under intraoperative investigations including histopathologic diagnosis of frozen sections. METHODS: We retrospectively compared dual phase thallium (Tl-201) thyroid scan and fine-needle aspiration (FNA) cytology to detect follicular carcinoma in 12 patients with cold thyroid nodules. Fine-needle aspiration cytology results were assessed and categorized by experienced pathologists into benign, suspicious, and malignant. Dual phase Tl-201 thyroid scan including an early (10-min) image and a delayed (3-hour) image were acquired after 2 mCi (74 MBq) of Tl-201 was injected intravenously. Dual phase Tl-201 thyroid scan findings were visually interpreted as positive and negative results. RESULTS: Based on FNA cytologic interpretation criteria, three nodules were diagnosed as benign, five nodules were diagnosed as suspicious, and four nodules were diagnosed as malignant. However, based on dual phase Tl-201 thyroid scan interpretation criteria, all of the 12 nodules could accurately diagnose as malignant. CONCLUSIONS: We conclude that dual phase Tl-201 thyroid scan was more accurate in diagnosing follicular carcinoma when comparing with FNA cytology.

Tc-99m-tetrofosmin thyroid scan in patients with low I-131 thyroid uptake.

UNLABELLED: Low 24-hour thyroid uptake of I-131 (I-131 uptake) is a common finding that influences evaluation of the thyroid gland. METHODS: We studied 20 female patients aged 16-82 years with low I-131 uptake. The following causes of reduced I-131 uptake were defined as (A) six cases with subacute thyroiditis, (B) seven cases with T4 suppression therapy, and (C) seven cases with iodinated pharmaceuticals use. Thirty-minute Tc-99m tetrofosmin thyroid scan was performed within 24 hours of the I-131 thyroid study. RESULTS: The results were correlated with neck ultrasound, serum TSH and biopsy histopathological findings. Tc-99m tetrofosmin thyroid scans revealed 10 normal thyroids, five diffuse goiters, four multinodular goitres, and one solitary thyroid nodule. CONCLUSIONS: Our results suggest that Tc-99m tetrofosmin thyroid scans may provide additional information to diagnose thyroid pathology in patients with low 24-hour I-131 uptake.