Seed vigor is a key factor affecting seed quality. The mechanical drying process exerts a significant influence on rice seed vigor. The initial moisture content (IMC) and drying temperature are considered the main factors affecting rice seed vigor through mechanical drying. This study aimed to determine the optimum drying temperature for rice seeds according to the IMC, and elucidate the mechanisms mediating the effects of drying temperature and IMC on seed vigor. Rice seeds with three different IMCs (20%, 25%, and 30%) were dried to the target moisture content (14%) at four different drying temperatures. The results showed that the drying temperature and IMC had significant effects on the drying performance and vigor of the rice seeds. The upper limits of drying temperature for rice seeds with 20%, 25%, and 30% IMCs were 45, 42, and 38 °C, respectively. The drying rate and seed temperature increased significantly with increasing drying temperature. The drying temperature, drying rate, and seed temperature showed extremely significant negative correlations with germination energy (GE), germination rate, germination index (GI), and vigor index (VI). A high IMC and drying temperature probably induced a massive accumulation of hydrogen peroxide (H₂O₂) and superoxide anions in the seeds, enhanced superoxide dismutase (SOD) and catalase (CAT) activity, and increased the abscisic acid (ABA) content. In the early stage of seed germination, the IMC and drying temperature regulated seed germination through the metabolism of H₂O₂, gibberellin acid (GA), ABA, and α-amylase. These results indicate that the metabolism of reactive oxygen species (ROS), antioxidant enzymes, GA, ABA, and α-amylase might be involved in the mediation of the effects of drying temperature on seed vigor. The results of this study provide a theoretical basis and technical guidance for the mechanical drying of rice seeds.
Abscisic Acid/metabolism* ; Antioxidants/pharmacology* ; Catalase/metabolism* ; Gene Expression Regulation, Plant/drug effects* ; Germination ; Gibberellins/metabolism* ; Hydrogen Peroxide/chemistry* ; Malondialdehyde/chemistry* ; Oryza/metabolism* ; Oxygen/chemistry* ; Plant Proteins/genetics* ; Reactive Oxygen Species ; Seeds/metabolism* ; Superoxide Dismutase/metabolism* ; Superoxides/chemistry* ; Temperature ; Weather ; alpha-Amylases/metabolism*

OBJECTIVETo explore the effect of Rhodiola on the expression of iNOS mRNA in severe acute pancreatitis (SAP) associated renal injury rats.

METHODSA total of 72 healthy rats were randomly divided into the sham-operated group (S), the SAP associated renal injury group (M), and the Rhodiola-treated group (RHO), 24 in each group. Rats in S and M groups were peritoneally injected with 10 mL/kg saline 3h before modeling, while rats in the RHO group were peritoneally injected with 10 mL/kg Rhodiola Injection 3 h before modeling. The peripheral ligament of pancreas was bluntly dissociated in rats of M and RHO groups. The head of pancreas was occlused by nontraumatic blood vessel forceps 3 h later to establish the model. Eight rats were randomly selected from each group at 12, 24, and 36 h after modeling to detect levels of serum amylase, creatinine, and blood urea nitrogen. Serum levels of interleukin 1β (IL-1β) and interleukin 10 (IL-10) were detected by enzyme-linked immunosorbent assay (ELISA). Pathological changes of the left kidney were observed under light microscope. The expression of inducible nitric oxide synthase (iNOS) mRNA in the right kidney was detected with real time polymerase chain reaction (RT-PCR).

RESULTSCompared with the S group, serum levels of amylase, creatinine (Cr), blood urea nitrogen (BUN), IL-1β, IL-10, and iNOS mRNA expression significantly increased in the M group (P < 0.01). The function of kidney and pancreas were obviously improved in the RHO group than in the M group. Levels of IL-1β and iNOS significantly decreased, but IL-10 levels significantly increased in the RHO group with statistical difference (P < 0.05).

CONCLUSIONRhodiola had better protective effect on SAP associated renal injury, which might be achieved through inhibiting the expression of IL-1β, stimulating the expression of IL-10, down-regulating iNOS mRNA expression, reducing the generation of oxygen free radicals and NO damage to cells, and improving hypoxia tolerance capabilities of the kidney.

Amylases ; Animals ; Blood Urea Nitrogen ; Creatinine ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Interleukin-1beta ; Kidney ; Nitric Oxide Synthase Type II ; genetics ; metabolism ; Pancreas ; Pancreatitis ; drug therapy ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley ; Rhodiola

Salivary function in mammals may be defective for various reasons, such as aging, Sjogren's syndrome or radiation therapy in head and neck cancer patients. Recently, tissue-specific stem cell therapy has attracted public attention as a next-generation therapeutic reagent. In the present study, we isolated tissue-specific stem cells from the human submandibular salivary gland (hSGSCs). To efficiently isolate and amplify hSGSCs in large amounts, we developed a culture system (lasting 4-5 weeks) without any selection. After five passages, we obtained adherent cells that expressed mesenchymal stem cell surface antigen markers, such as CD44, CD49f, CD90 and CD105, but not the hematopoietic stem cell markers, CD34 and CD45, and that were able to undergo adipogenic, osteogenic and chondrogenic differentiation. In addition, hSGSCs were differentiated into amylase-expressing cells by using a two-step differentiation method. Transplantation of hSGSCs to radiation-damaged rat salivary glands rescued hyposalivation and body weight loss, restored acinar and duct cell structure, and decreased the amount of apoptotic cells. These data suggest that the isolated hSGSCs, which may have characteristics of mesenchymal-like stem cells, could be used as a cell therapy agent for the damaged salivary gland.
Amylases/genetics/metabolism ; Animals ; Antigens, CD/genetics/metabolism ; Apoptosis ; Cell Differentiation ; Humans ; Male ; Mesenchymal Stromal Cells/*cytology/metabolism ; Radiation Injuries, Experimental ; Rats ; Rats, Wistar ; *Regeneration ; Salivary Glands/cytology/injuries/physiology/*surgery ; *Salivation ; *Stem Cell Transplantation

OBJECTIVETo investigate the effect of modified Xiaochaihu Decoction (, MXD) on transforming growth factor-β1/Sma- and Mad-related proteins (TGF-β1/Smads) signaling pathway in rats with chronic pancreatitis (CP) induced by dibutyltin dichloride.

METHODSThirty healthy male Wistar rats were randomly divided into the normal control group, CP group and CP+MXD-treated group. CP was induced by injection of dibutyltin dichloride (DBTC, 7 mg/kg of body weight) into the right caudal vein, and the control rats were treated with vehicle. MXD was given daily by gavage at a dose of 10 g/kg of body weight, starting from the day after CP induction. After 28-day treatment, the n-benzoyl-tyrosyl para-aminobenzoic acid (NBT-PABA) test was carried out to evaluate exocrine pancreatic function. Then, rats were sacrificed, and pancreatic tissues were harvested for histological evaluation. In addition, the mRNA expression of TGF-β1, TGF-β1 type II receptor (TGFβRII), Smad3 and Smad7 was determined in pancreatic tissues by using real-time polymerase chain reaction.

RESULTSTreatment of CP with MXD improved the PABA recovery, decreased the histological lesion, and reduced the mRNA expression of TGF-β1, TGFβRII and Smad3 (P<0.05). However, MXD had no effect on Smad7 mRNA level.

CONCLUSIONSMXD could protect the pancreas against chronic injury and improve pancreatic exocrine function in DBTC induced rat CP model. Its mechanism may involve inhibition of the TGF-β1/Smads signaling pathway.

Amylases ; blood ; Animals ; Base Sequence ; Blood Glucose ; metabolism ; Body Weight ; drug effects ; Chronic Disease ; DNA Primers ; Disease Progression ; Drugs, Chinese Herbal ; therapeutic use ; Lipase ; blood ; Male ; Pancreatitis ; drug therapy ; metabolism ; pathology ; RNA, Messenger ; genetics ; Rats ; Rats, Wistar ; Real-Time Polymerase Chain Reaction ; Signal Transduction ; drug effects ; Smad Proteins ; genetics ; metabolism ; Transforming Growth Factor beta1 ; metabolism

Saccharomycopsis fibuligera possesses high alpha-amylase and glucoamylase activities that enable it to utilize raw starch as a carbon source. A expression cassette containing the promoter sequence of 3-phosphogylycerate kinase gene (PGK1p), the alpha factor signal sequence from Saccharomyces cerevisiae and the alpha-amylase coding sequence of S. fibuligera was constructed. The alpha-amylase expression cassette was inserted in the ILV2 locus of industrial brewer's yeast strain YSF-5 encoding alpha-acetolactate synthase (AHAS) by homologous recombination. The transformed yeast strain was selected on the media with starch as the sole carbon source and verified by PCR. The transformant exhibited secretive alpha-amylase activity, low AHAS activity and reduced diacetyl production. Effects of temperature, pH, and metal ions on the activity of the alpha-amylase expressed by the transformant were examined. The fermentation performance of host strain YSF-5 and the transformant was also examined.
Beer ; microbiology ; Diacetyl ; metabolism ; Glycogen Synthase Kinase 3 ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; Recombination, Genetic ; Saccharomyces cerevisiae ; genetics ; metabolism ; Saccharomycopsis ; enzymology ; genetics ; alpha-Amylases ; biosynthesis ; genetics ; metabolism

Genetic modification of barley variety can be an efficient way to improve beer quality. The objective of this study was to understand the effect of trxS gene on hydrolases activities in transgenic and non-transgenic barley seeds. The results showed that alpha-amylase, free beta-amylase and limit dextrinase activity were increased in transgenic seeds in comparison with non-transgenic seeds. Sulfhydryl content of protein in transgenic seeds was also higher than that in non-transgenic seeds, suggesting that trxS gene could express in barley seeds, which opens a new way for breeding new barley varieties to improve beer quality.
Germination ; genetics ; Glucosyltransferases ; metabolism ; Hordeum ; enzymology ; genetics ; Plants, Genetically Modified ; enzymology ; genetics ; Seeds ; enzymology ; genetics ; Sulfhydryl Compounds ; metabolism ; Thioredoxins ; genetics ; alpha-Amylases ; metabolism ; beta-Amylase ; metabolism

Based on the constructed promoter probe vectors that could replicate both in E. coli and in a cold-adapted bacterium, several candidate promoters were isolated and their activities were evaluated by RT-PCR. The transcription initiation sites and core sequence of promoters were determined by primer extension analysis. A low-temperature expression vector was constructed by using the strongest promoter and a thermolabile alpha-amylase gene was successfully overproduced under control of this promoter at low temperature (7 degrees C), while the secreted alpha-amylase amounted up to 35% of the total extracellular proteins. The expression system is expected to be useful for the production of thermolabile exogenous proteins at low temperatures.
Acinetobacter ; genetics ; metabolism ; Adaptation, Physiological ; genetics ; Bacterial Proteins ; biosynthesis ; genetics ; Base Sequence ; Cold Temperature ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; genetics ; Molecular Sequence Data ; Promoter Regions, Genetic ; genetics ; Transformation, Genetic ; alpha-Amylases ; biosynthesis ; genetics

PURPOSE: The sarco/endoplasmic reticulum Ca(2+)-ATPase (SERCA), encoded by ATP2A2, is an essential component for G-protein coupled receptor (GPCR)-dependent Ca(2+) signaling. However, whether the changes in Ca(2+) signaling and Ca(2+) signaling proteins in parotid acinar cells are affected by a partial loss of SERCA2 are not known. MATERIALS AND METHODS: In SERCA2(+/-) mouse parotid gland acinar cells, Ca(2+) signaling, expression levels of Ca(2+) signaling proteins, and amylase secretion were investigated. RESULTS: SERCA2(+/-) mice showed decreased SERCA2 expression and an upregulation of the plasma membrane Ca(2+) ATPase. A partial loss of SERCA2 changed the expression level of 1, 4, 5-tris-inositolphosphate receptors (IP(3)Rs), but the localization and activities of IP3Rs were not altered. In SERCA2(+/-) mice, muscarinic stimulation resulted in greater amylase release, and the expression of synaptotagmin was increased compared to wild type mice. CONCLUSION: These results suggest that a partial loss of SERCA2 affects the expression and activity of Ca(2+) signaling proteins in the parotid gland acini, however, overall Ca(2+) signaling is unchanged.
Amylases/metabolism ; Animals ; Blotting, Western ; Calcium/metabolism ; Calcium Signaling/drug effects/genetics/*physiology ; Carbachol/pharmacology ; Immunohistochemistry ; Inositol 1,4,5-Trisphosphate Receptors/metabolism ; Mice ; Mice, Knockout ; Parotid Gland/*metabolism ; Sarcoplasmic Reticulum Calcium-Transporting ATPases/genetics/*metabolism ; Signal Transduction/drug effects/genetics/physiology

BACKGROUNDIt is accepted that inflammatory cytokines play a key role in the development of acute pancreatitis, so blocking the initiation of inflammatory reactions may alleviate pathological changes of acute pancreatitis. We studied the regulatory effect of arsenic trioxide (As(2)O(3)) on apoptosis and oncosis of pancreatic acinar cells in vitro and in vivo and its therapeutic effect on acute pancreatitis.

METHODSPancreatic acinar cells were isolated by collagenase digestion method. Apoptosis and oncosis of isolated pancreatic acinar cells were detected with Hoechst 33258 + PI or Annexin V + PI double fluorescent staining. Amylase and lactate dehydrogenase release were measured. Acute pancreatitis was induced in Wistar rats by intraperitoneal injections of caerulein, and apoptosis was detected with terminal dUTP nick-end labeling method. Tumor necorsis factor alpha (TNF-alpha) mRNA, myeloperoxidase, nuclear factor-kappaB and histological grading of pancreatic damage were measured.

RESULTSThere was an increased apoptosis but a decreased oncosis of pancreatic acinar cell after the treatment with As(2)O(3). The levels of lactate dehydrogenase and amylase release were markedly decreased in As(2)O(3) treated group. Myeloperoxidase content, TNF-alpha mRNA level, nuclear factor-kappaB activation and pathological score in As(2)O(3) treated group were significantly lower than in the untreated group.

CONCLUSIONSAs(2)O(3) can induce apoptosis and reduce oncosis of pancreatic acinar cell, thus resulting in reduced release of endocellular enzyme of acinar cells, reduced inflammatory cell infiltration and decreased the production of inflammatory cytokines, so that the outcome of alleviated pathological changes was finally achieved.

Amylases ; metabolism ; Animals ; Apoptosis ; drug effects ; Arsenicals ; pharmacology ; Flow Cytometry ; In Situ Nick-End Labeling ; Inflammation ; drug therapy ; genetics ; metabolism ; L-Lactate Dehydrogenase ; metabolism ; Male ; NF-kappa B ; metabolism ; Oxides ; pharmacology ; Pancreas ; drug effects ; metabolism ; pathology ; Pancreatitis ; drug therapy ; genetics ; metabolism ; Peroxidase ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Wistar ; Signal Transduction ; drug effects ; Tumor Necrosis Factor-alpha ; genetics

The effect of dual promoters on recombinant protein production from bacterial phage based Bacillus subtilis expression system was investigated. Alpha amylase (from Bacillus amyloliquefaciens) and penicillin acylase (from Bacillus megaterium) were selected as the indicating enzymes. Both the promoterless genes and the promoter-bearing genes were isolated through PCR amplification with properly designed primers, and were inserted into plasmid pSG703 that contains the lacZ-cat expression cartridge. The lysogenic B. subtilis (phi105 MU331) was transformed with the resultant recombinant plasmids, and the heterologous genes were thereby integrated into the chromosommal DNA of B. subtilis via homologous recombination. The transformants were designated as B. subtilis AMY1, B. subtilis AMY2, B. subtilis PA1, and B. subtilis PA2, respectively. In the recombinant B. subtilis strains, the inserted sequences were located down stream of a strong phage promoter that could be activated by thermal induction. In B. subtilis AMY1 and B. subtilis PA1, transcription of the heterologous genes was only initiated by the phage promoter after heat shock, whereas in B. subtilis AMY2 and B. subtilis PA2, transcription of the heterologous genes was initiated by dual promoters, the phage promoter and the native promoter. The application of dual promoters increased the productivity of both enzymes, with 133% enhancement for alpha-amylase production and 113% enhancement for penicillin acylase production.
Bacillus Phages ; genetics ; metabolism ; Bacillus subtilis ; genetics ; metabolism ; Cloning, Molecular ; Penicillin Amidase ; biosynthesis ; genetics ; Promoter Regions, Genetic ; Recombinant Proteins ; biosynthesis ; genetics ; Transformation, Bacterial ; alpha-Amylases ; biosynthesis ; genetics