Effect of salt concentration on microbial communities, physicochemical properties and metabolite profile during spontaneous fermentation of Chinese northeast sauerkraut.

AIM: The aim of this study was to study the effects of salt concentrations on the microbial communities, physicochemical properties, metabolome profiles and sensory characteristics during the fermentation of traditional northeast sauerkraut. METHODS AND RESULTS: Northeast sauerkraut was spontaneously fermented under four salt concentrations (0·5, 1·5, 2·5 and 3·5%, w/w). The result of microbiological analysis showed that the population of lactic acid bacteria in 2·5%-salted sauerkraut was significantly higher than that in the other samples. Correspondingly, the speed of decrease in pH and accumulation of acids were the highest in 2·5%-salted sauerkraut. The glucose (analysed by HPLC) in 2·5%-salted sauerkraut was consumed more completely to produce higher levels of organic acids compared to those in the other samples. Principle component analysis showed clear differences in the metabolites of sauerkraut according to different salt concentrations. A higher level of volatiles (detected by HS-SPME/GC-MS) was identified in 2·5%-salted sauerkraut, and sensory evaluation demonstrated that 2·5%-salted sauerkraut had the best sensory characteristics. CONCLUSION: The best quality of sauerkraut was obtained from fermented under 2·5% salt concentration. SIGNIFICANCE AND IMPACT OF THE STUDY: This study facilitated the understanding of the effects of salt on the sauerkraut fermentation and may be useful for developing the quality of sauerkraut.

Autophagy inhibitor sensitizes MCF-7 breast cancer cells to novel cyclic tetrapeptide CTS203-induced caspase-9-dependent apoptotic cell death.

UNLABELLED: Histone deacetylase (HDAC) inhibitors have been demonstrated to be effective anti-cancer candidates against aggressive malignancies. In previous study, a novel hydroxamic acid derivate, CTS203 (cyclo(-l-Asu(NHOH)-l-A3mc6c-l-Phe-d-Pro-)), demonstrated promising HDAC inhibitory activity. Herein, more biological evaluations including cell viability, cell cycle distribution, cellular morphology, expression quantification as well as protein-protein interactions were measured to investigate its cytotoxic mechanism. Corresponding with its significant HDAC inhibitory activity, CTS203 led to increased acetylation of H3K14, cell cycle arrest as well as consequent apoptotic cell death, with bearable influence on the viability of normal cells. However, schedule-dependent cytotoxicity against MCF-7 breast cancer cells revealed a delayed cellular response to chemo-stimuli. Within this corresponding period, autophagy was rapidly triggered once exposure started, whereas autophagy inhibitor sensitized MCF-7 cells to CTS203, exhibiting synergistically anti-proliferative effects. The expression variation in MCF-7 cells revealed that the cleavage of Beclin 1 mediated by caspase-8 resulted in disabled autophagy, thus ultimately facilitated and fastened caspase-9-dependent apoptotic cell death. Taken together, these findings elucidated the mechanism of CTS203-induced cytotoxicity as well as suggested that appropriate manipulation of autophagy would be an adjunctive strategy to enhance HDAC inhibitor-induced cell death. KEYWORDS: cyclic tetrapeptide, apoptosis, autophagy, HDAC inhibitor, Beclin 1 cleavage.

Spray-as-you-go airway topical anesthesia in patients with a difficult airway: a randomized, double-blind comparison of 2% and 4% lidocaine.

BACKGROUND: We designed this randomized, double-blind clinical study to compare the safety and efficacy of 2% and 4% lidocaine during airway topical anesthesia with a spray-as-you-go technique via the fiberoptic bronchoscope. METHODS: Fifty-two adult patients with a difficult airway were randomly assigned to 1 of 2 study groups to receive 2% (Group 1) or 4% lidocaine (Group 2) by a spray-as-you-go technique with the fiberoptic bronchoscope, in a double-blind manner. After airway topical anesthesia, awake fiberoptic orotracheal intubation (FOI) was performed. Level of sedation, time for each lidocaine spray in different targeted areas, total times for airway sprays, total dosages of lidocaine used for airway sprays, intubation times, and number of intubation attempts were noted. An independent investigator scored patients' comfort during airway topical anesthesia, patients' reaction, coughing severity, and intubating condition during awake FOI, and observed changes of arterial blood pressure and heart rate during each stage in the airway manipulation process. Serial blood samples were obtained for analysis of plasma lidocaine concentrations. RESULTS: Except for the total dosages and plasma concentrations of lidocaine, there were no significant differences in any of the observed variables between groups. All patients exhibited excellent or acceptable intubating conditions. The total dosages of lidocaine were significantly smaller in Group 1 (3.4 +/- 0.6 mg/kg) than in Group 2 (7.1 +/- 2.1 mg/kg). The plasma lidocaine concentrations in all observed points after the supraglottic sprays were larger in Group 2 than in Group 1. CONCLUSIONS: Both 2% and 4% lidocaine administered topically by a spray-as-you-go technique can provide clinically acceptable intubating conditions for awake FOI in sedated patients with a difficult airway. As compared with 4% lidocaine, however, 2% lidocaine requires a smaller dosage and results in lower plasma concentrations.

A whole genome scan to map QTL for milk production traits and somatic cell score in Canadian Holstein bulls.

The detection and mapping of genetic markers linked to quantitative trait loci (QTL) can be utilized to enhance genetic improvement of livestock populations. With the completion of the bovine genome sequence assembly, single nucleotide polymorphisms (SNP) assays spanning the whole bovine genome and research work on large scale identification, validation and analysis of genotypic variation in cattle has become possible. The objective of the present study was to perform a whole genome scan to identify and map QTL affecting milk production traits and somatic cell scores using linkage disequilibrium (LD) regression and 1536 SNP markers. Three and 18 SNP were found to be associated with only milk yield (MY) at a genome and chromosome wise significance (p < 0.05) level respectively. Among the 21 significant SNP, 16 were in a region reported to have QTL for MY in other dairy cattle populations and while the rest five were new QTL finding. Four SNP out of 21 are significant for the milk production traits (MY, fat yield, protein yield (PY), and milk contents) in the present study. Six and nine SNP were associated with PY at a genome and chromosome wise significant (p < 0.05) level respectively. Three and 17 SNP were found to be associated with FY at a genome and chromosome wise significant (p < 0.05) level. Five and seven SNP were mapped with somatic cell score at a genome and chromosome wise significant (p < 0.05) level respectively. The results of this study have revealed QTL for MY, PY, protein percentage, FY, fat percentage, somatic cell score and persistency of milk in the Canadian dairy cattle population. The chromosome regions identified in this study should be further investigated to potentially identify the causative mutations underlying the QTL.

A whole-genome scan to map quantitative trait loci for conformation and functional traits in Canadian Holstein bulls.

Genetic improvement of livestock populations can be achieved through detection and mapping of genetic markers linked to quantitative trait loci (QTL). With the completion of the bovine genome sequence assembly, single nucleotide polymorphism (SNP) assays spanning the whole bovine genome and research work on large-scale identification, validation, and analysis of genotypic variation in cattle has become possible. A total of 462 Canadian Holstein Bulls were used to test the association between SNP and QTL. Single locus linkage disequilibrium regression model was implemented to perform a whole genome scan to identify and map QTL affecting conformation and functional traits. One thousand five hundred thirty-six SNP markers from introns and exons of potential QTL regions for economically important traits across the bovine genome were selected for association analysis. A total of 45 and 151 SNP were found to be associated with 17 conformation and functional traits at a genome- and chromosome-wise significance level, respectively. Among the 196 significant SNP, 169 of them are newly detected in this study, whereas 27 of them have been reported in previous literature and 161 of these were located in genes and are worth further investigating to potentially identify the causative mutations underlying the QTL. The single locus linkage disequilibrium regression method using SNP marker genotypes has proven to be a successful methodology for detecting and mapping QTL in dairy cattle populations.

Impaired response of perforating arteries to hypercapnia in chronic hyperglycemia.

Diabetes mellitus increases the risk of cerebrovascular disease, the effects of hypercapnia on CBF (cerebral blood flow) and cerebrovascular reactivity during diabetes are still inconsistent. Here, we have established a new microangiographic technique using synchrotron radiation (SPring-8, Japan), which enabled us to visualize rat cerebral vessels with high spatial resolution in real time. The goal of the study presented here was to identify the effects of chronic hyperglycemia on hypercapnia-induced vascular responses (endothelium-dependent vasodilatation) and nitric oxide (NO) donor- induced vascular responses (endothelium-independent) of perforating arteries and of the deeply located large cerebral arteries. We found a significant vasodilatation of rat perforating arteries after hypercapnia with a maximum diameter of approximately 140% of baseline in normal Wistar rats. Chronic hyperglycemia impaired vasodilatation of perforating arteries in genetically diabetic GK rats. SNP (sodium nitroprusside) caused a similar vasodilatation of perforating vessels in normal and chronic hyperglycemia, indicating that endothelium-dependent vasodilatation of perforating arteries may be specifically impaired in chronic hyperglycemia. Possible impairment of endothelium-dependent vasodilatation in perforating vessels during chronic hyperglycemia may cause decreased vascular reserve capacity of perforating artery, resulting in the increased ischemic insults and cerebrovascular diseases in diabetes.

[Molecular evolution of AdoMet synthetase by DNA recombination with a novel separate-mixing method].

We describe a new approach to in vitro DNA recombination termed Separate-Mixing method in this study. The reaction process of this method consists of two stages: at the first stage the reaction was implemented in two parallel teams, which generated random recombination by template-switching of growing polynucleotides from primers in the presence of unidirectional single-stranded DNA fragments used as templates, and then both teams were mixed together for further extension and recombination of DNA sequences at the second stage. Because of the particular strategy, the reaction process was also accompanied by the other two processes of DNA shuffling and StEP simultaneously. Two AdoMet synthetase genes sam2 from Saccharomyces cerevisiae and metK from Escherichia coli, which have only 56% homology on the DNA level were used for recombination with Separate-Mixing method. DNA recombination was available after a single round of reaction. With sequencing of 10 randomly selected recombinants, no unshuffled parental clone was found, and also no unexpected insertion, deletion or rearrangement was detected. An evolved gene sam' was obtained after screen and selection, which could obviously increase the accumulation of AdoMet in S. cerevisiae.

Preparation and characterization of the monoclonal antibodies against Japanese encephalitis virus.

Six mouse monoclonal antibodies (MoAbs) against Japanese encephalitis virus (JEV) were prepared and analyzed with indirect immunofluorescence assay (IFA), enzyme linked immunosorbent assay (ELISA), haemagglutination inhibition test (HI), neutralization test (NT), antibody dependent cell mediated cytotoxicity (ADCC) assay, antigenic site specific analysis and relative affinity measurement. These MoAbs could be divided into three classes by indirect immunofluorescence cross reactivity among four flaviviruses, 2H4, 2F2, and nG2 were type specific; 2D2 and mC3 were subgroup specific; and mG9 was family specific. 2H4 and 2F2 had higher neutralization activity, 2D2 and mC3 had the function of inducing ADCC effect, mG9 had higher titer in HI. The six MoAbs recognized five antigenic sites on JEV envelope glycoprotein, 2H4 and 2F2 recognized the same or very similar antigenic site and their relative affinity was ranked as: nG2 > 2H4 > 2D2 > mG9 > 2F2 > mC3.

[A preliminary cytogenetic study of uterine leiomyoma].

Thirty fresh samples of uterine leiomyoma were disaggregated in collagenase (200kU/L) and cultured for 3-10 days with incubator. Of 30 cases, 27 cases of uterine leiomyoma obtained karyotypes and the rate of successful cultured was 90%, 9 cases showed abnormal karyotypes. Structural changes were frequently seen. Breakpoint of chromosome were 12q13-15 (7 cases), 14q24 (6 cases), 7q31-32 (3 cases). Specific chromosomal rearrangements were t(12;14) (q13-15; q24) and del (7) (q31-32). Our data suggested that chromosome 12q13-15, 14q24, 7q31-32 may be related to the proliferation of uterine leiomyoma.

[Cardiac metal foreign body: analysis of 21 cases].

OBJECTIVE: To improve early diagnosis and clinical treatment of metal foreign body in the heart. METHODS: In 21 patients, case history, clinical manifestation, chest X-ray film, and echocardiograph were reviewed. 20 patients received operation: emergency exploration (13) and elective operation (7). Four patients were subjected to cardiopulmonary bypass. The other one was not operated on because of absence of symptoms and small foreign body. Foreign bodies mostly bullets, were usually located in the right ventricle. Ventricular fibrillation occurred in 3 patients during operation, two of them were resuscitated. RESULTS: 20 patients but one recovered, a mortality rate of 5%. CONCLUSIONS: Early diagnosis, rapid management, and localization of foreign body are essential to the selection of treatment.

Effect of TiN coating on microstructure of Tif/Al composite.

In the present work, Ti fibre reinforced Al matrix composites (Ti(f)/Al) were fabricated by pressure infiltration method. In order to suppress the severe Ti-Al reaction and reduce the formation of brittle TiAl(3) phase, a TiN layer was coated on Ti fibres by an arc ion plating method before composite preparation. A thin TiN layer was coated on the Ti fibre surface, and the maximum and minimum thickness values of layer were about 3.5 and 1µm, respectively. Prefer orientation of TiN on (111) and (200) was found by XRD analysis. A thin and uniform TiAl(3) layer was observed in Ti(f)/Al composite. However, after coated with TiN layer, no significant reaction layer was found in (Ti(f)+TiN)/Al composite. Segregation of Mg element was found in Ti(f)/Al composite, and the presence of TiN layer showed little effect on this behaviour. Due to the large CTE difference between Ti fibre and Al matrix, high density dislocations were observed in the Al matrix. Meanwhile, fine dispersed Mg(2)Al(3) phases were also found in Al matrix. Ti fibre is mainly composed of α- and ß-Ti. Small discontinuous needle-like TiAl(3) phases were detected at TiN/Al interface, which implies that the presence of TiN layer between the Ti fibre and Al matrix could effectively hinder the formation of TiAl(3) phases.

Biotransformation of piceid in Polygonum cuspidatum to resveratrol by Aspergillus oryzae.

Biotransformation of piceid in Polygonum cuspidatum to resveratrol by Aspergillus oryzae was investigated in this study. Resveratrol is widely used in medicine, food, and cosmetic because of its pharmacological properties. However, it has a much lower content in plants compared with its glucoside piceid, which has a much lower bioavailability. Traditionally, the aglycone is acquired by acid or enzymatic hydrolysis of its glucoside, but the violent condition and the acid pollution in hydrolytic reaction and the high cost of the enzyme limit their industrial development. In this paper, fermentation of P. cuspidatum by A. oryzae was successfully performed, during which, piceid was converted to resveratrol with the highest yield of trans-resveratrol 1.35%, 3.6 times higher than that obtained from raw herb by microwave-assisted extraction. Scale-up production was also performed and the yield of trans-resveratrol was 3.1 times higher after 24 h incubation. Therefore, biotransformation is a better method to increase the yield of resveratrol because of its high yield and mild conditions.

[Experimental study of mechanism of survival of venous flaps].

The mechanism of survival of venous flaps was studied by observations employed in the study of microcirculation and blood vessel perfusion in rabbits. Initially, the flap was nourished by venous blood (within 72 hours): venous blood flowed slowly from the principal vein to its tributary veins, and then through the venous anastomoses (including small venous anastomoses, venular anastomoses and terminal venule anastomoses), reaching other tributary veins. No backflow from venules into capillaries was seen. The second stage was arterial nourishing and vessel reconstructive period (72 hours to 6 weeks): new vessels began to grow into the flaps at 72 hours postoperatively. 4 days postoperation, arterial flow with low velocity appeared in the primary flap arteries and the backflow of venous blood disappeared. 7 days postoperation, the abnormal, flow in 'unphysiologic' channels was replaced by the normal flow in 'physiologic' channels completely. 6 weeks postoperation, the vessel structure of venous flaps was the same as that of normal skin. These results suggested that rich venous network in venous flaps and early invasion of new blood vessels are the mainstay of venous flaps survival.

[Transplantation of a minute artery to bridge over the defect of an artery of significantly larger calibre. An experimental study].

Based on our clinical experience for years of "unequal bite" suturing technique to perform end-to-end anastomosis of vessels with great discrepancy in diameter, the animal experimental study was carried out. An artery defect, 1.0-1.4 mm in diameter, was reunited with, an autogenous arterial graft of 0.3-0.4 mm in diameter. A high patency rate (96% in total) was obtained postoperatively. Histological examination by light microscope and scanning electron microscope showed that the unequal bite suturing technique can produce very smooth anastomotic junction and the minute arterial grafts healed well on a large artery. The unequal bite suturing technique is a very valuable development of the basic end-to-end anastomotic technique. A small autogenous arterial graft may be used to repair defect of an artery with large calibre clinically.

[Experimental study on revascularization of subdermal vascular network thin flap].

OBJECTIVE: To investigate the survival mechanism of subdermal vascular network thin flap (SVNF). METHODS: 24 rabbits were divided into three groups, in which the SVNF, subdermal vascular network skin graft and the traditional random flap were created respectively. Tissue hyalinization technique was used to find out the differences of vascular architecture between the three groups at the 1st-7th day, and the 14th day postoperatively. RESULTS: The results showed that the blood supply reestablished earlier in SVNF group than the other two groups. CONCLUSION: There is a zone of rapid and intensive angiogenesis in the mid-part of SVNF known as "intermediary pedicle". This is the anatomic basis of the early pedicle division and superior survival ratio of SVNF.

[A long-term study of a small-caliber arterial autograft used for repair of a large-caliber arterial defect].

OBJECTIVE: The long-term fate of a small arterial graft used to repair a large diameter artery was studied in this study. METHODS: An end segment of the tail artery (0.3-0.4 mm in diameter) was transplanted to the defect of the carotid artery (1.0-1.4 mm in diameter) in 50 rats. The vessel grafts were examined histologically at 3 weeks and 5, 10 and 12 months after implantation. RESULTS: At 3 weeks, intimal thickening of the grafts was mild and present only in regions adjacent to the anastomosis. At 5 months, severe intimal thickening was seen along the entire graft wall. At 10 and 12 months, the intimal thickening reduced. Regression of the thickening was uneven within the graft wall. The adventitia thickened at the early postoperative period and reduced at the end of this study. The patency rate of the grafts at 12 months postoperation was 95% (19/20). Angiography indicated that the grafts stopped expanding after 5 months and were only slightly smaller than the recipient arteries at 12 months postoperation. CONCLUSION: These results suggest that a small, autogenous arterial graft may effectively repair a large-calibre arterial defect.