Assessment of bioflocculant production by Bacillus sp. Gilbert, a marine bacterium isolated from the bottom sediment of Algoa Bay

The bioflocculant-producing potentials of a marine bacteria isolated from the bottom sediment of Algoa Bay was investigated using standard methods. The 16S rDNA sequence analysis revealed 98% similarity to that of Bacillus sp. HXG-C1 and the nucleotide sequence was deposited in GenBank as Bacillus sp. Gilbert with accession number {"type":"entrez-nucleotide","attrs":{"text":"HQ537128","term_id":"312064208","term_text":"HQ537128"}}HQ537128. Bioflocculant was optimally produced when sucrose (72% flocculating activity) and ammonium chloride (91% flocculating activity) were used as sole sources of carbon and nitrogen, respectively; an initial pH 6.2 of the production medium; and Mg²⁺ as cation. Chemical analysis of the purified bioflocculant revealed the compound to be a polysaccharide.     

Genome Analysis of a Novel Polysaccharide-Degrading Bacterium Paenibacillus algicola and Determination of Alginate Lyases

Carbohydrate-active enzymes (CAZymes) are an important characteristic of bacteria in marine systems. We herein describe the CAZymes of Paenibacillus algicola {"type":"entrez-nucleotide","attrs":{"text":"HB172198","term_id":"239334117"}}HB172198^T, a novel type species isolated from brown algae in Qishui Bay, Hainan, China. The genome of strain {"type":"entrez-nucleotide","attrs":{"text":"HB172198","term_id":"239334117"}}HB172198^T is a 4,475,055 bp circular chromosome with an average GC content of 51.2%. Analysis of the nucleotide sequences of the predicted genes shows that strain {"type":"entrez-nucleotide","attrs":{"text":"HB172198","term_id":"239334117"}}HB172198^T encodes 191 CAZymes. Abundant putative enzymes involved in the degradation of polysaccharides were identified, such as alginate lyase, agarase, carrageenase, xanthanase, xylanase, amylases, cellulase, chitinase, fucosidase and glucanase. Four of the putative polysaccharide lyases from families 7, 15 and 38 were involved in alginate degradation. The alginate lyases of strain {"type":"entrez-nucleotide","attrs":{"text":"HB172198","term_id":"239334117"}}HB172198^T exhibited the maximum activity 152 U/mL at 50 °C and pH 8.0, and were relatively stable at pH 7.0 and temperatures lower than 40 °C. The average degree of polymerization (DP) of the sodium alginate oligosaccharide (AOS) degraded by the partially purified alginate lyases remained around 14.2, and the thin layer chromatography (TCL) analysis indicated that it contained DP2-DP8 oligosaccharides. The complete genome sequence of P. algicola {"type":"entrez-nucleotide","attrs":{"text":"HB172198","term_id":"239334117"}}HB172198^T will enrich our knowledge of the mechanism of polysaccharide lyase production and provide insights into its potential applications in the degradation of polysaccharides such as alginate.     

Different Morphologies of Leishmania major Amastigotes with No Molecular Diversity in a Neglected Endemic Area of Zoonotic Cutaneous Leishmaniasis in Iran

Background:

Molecular diversity of Leishmania major and its morphological changes have become a controversial issue among researchers. Some aspects of polymorphic shapes of amastigotes in clinical manifestations along with molecular variation were evaluated among suspected patients of some exceptional zoonotic cutaneous leishmaniasis locations in Northern Khuzestan, Southwestern Iran.

Methods:

Suspected patients (n = 165) were sampled in zoonotic cutaneous leishmaniasis foci over two consecutive years during 2012-2014. Prepared smears were stained, scaled and measured by ocular micrometer. DNA was extracted from smears; ITS-rDNA and Cytochrome b (Cyt b) markers were amplified, and PCR products were digested by BsuR1 restriction enzyme. Then the RFLP and sequencing were employed.

Results:

Only L. major was identified in patients containing regular amastigotes'' shapes (oval or round) with a size of 2-4 µm in each of classical wet, dry, mixed lesions. Meanwhile, irregular shapes (spindle, pear, or cigarette) were observed separately in non-classical wet lesions with more than 4 µm. Interestingly, a few amastigotes with an external flagellum were observed in some lesions. All sequenced ITS-rDNA and Cyt b genes of L. major did not show any molecular variation (χ ² P > 0.05), including only one common haplotype (GenBank access no. {"type":"entrez-nucleotide","attrs":{"text":"EF413075","term_id":"151936076","term_text":"EF413075"}}EF413075).

Conclusion:

Findings proved that unlike other endemic foci, there is not a meaningful correlation between phenotypic and genotypic features of L. major isolates. This study is considered as the first comprehensive report to incriminate morphometric shapes of L. major amastigotes, which enhances our knowledge concerning their relevance with various clinical appearances and genotypic traits. Key Words: Leishmania major, Nuclear gene, Mitochondrial gene, Amastigote shapes, Iran     

Three New Asperentin Derivatives from the Algicolous Fungus Aspergillus sp. F00785

Three new asperentin-type compounds, 6-O-α-d-ribosylasperentin (1) and 6-O-α-d-ribosyl-8-O-methylasperentin (2) and 5-hydroxyl-6-O-methylasperentin (3), along with asperentin (4) and its known analogues (5–9), were isolated from a halotolerant Aspergillus sp. strain {"type":"entrez-nucleotide","attrs":{"text":"F00785","term_id":"707638","term_text":"F00785"}}F00785, an endotrophic fungus from marine alga. Their structures were determined using extensive NMR and HRESIMS spectroscopic analysis, including the X-ray crystallographic data for the assignment of the absolute configurations of compound 9. Compound 4 exhibited highly potent inhibitory activity against crop pathogens, Colletotrichum gleosporioides Penz. and Colletotrichum gleosporioides (Penz.) Sacc.     

Fucoidan and Derived Oligo-Fucoses: Structural Features with Relevance in Competitive Inhibition of Gastrointestinal Norovirus Binding

Norovirus infections belong to the most common causes of human gastroenteritis worldwide and epidemic outbreaks are responsible for hundreds of thousands of deaths annually. In humans, noroviruses are known to bind to gastrointestinal epithelia via recognition of blood-group active mucin-type O-glycans. Considering the involvement of l-α-fucose residues in these glycans, their high valency on epithelial surfaces far surpasses the low affinity, though specific interactions of monovalent milk oligosaccharides. Based on these findings, we attempted to identify polyfucoses (fucans) with the capacity to block binding of the currently most prevalent norovirus strain GII.4 (Sydney, 2012, {"type":"entrez-nucleotide","attrs":{"text":"JX459908","term_id":"409032931"}}JX459908) to human and animal gastrointestinal mucins. We provide evidence that inhibitory effects on capsid binding are exerted in a competitive manner by α-fucosyl residues on Fucus vesiculosus fucoidan, but also on the galacto-fucan from Undaria pinnatifida and their oligo-fucose processing products. Insight into novel structural aspects of fucoidan and derived oligosaccharides from low-mass Undaria pinnatifida were revealed by GCMS and MALDI mass spectrometry. In targeting noroviral spread attenuation, this study provides first steps towards a prophylactic food additive that is produced from algal species.     

Astaxanthin Alleviates Early Brain Injury Following Subarachnoid Hemorrhage in Rats: Possible Involvement of Akt/Bad Signaling

Apoptosis has been proven to play a crucial role in early brain injury pathogenesis and to represent a target for the treatment of subarachnoid hemorrhage (SAH). Previously, we demonstrated that astaxanthin (ATX) administration markedly reduced neuronal apoptosis in the early period after SAH. However, the underlying molecular mechanisms remain obscure. In the present study, we tried to investigate whether ATX administration is associated with the phosphatidylinositol 3-kinase-Akt (PI3K/Akt) pathway, which can play an important role in the signaling of apoptosis. Our results showed that post-SAH treatment with ATX could cause a significant increase of phosphorylated Akt and Bad levels, along with a significant decrease of cleaved caspase-3 levels in the cortex after SAH. In addition to the reduced neuronal apoptosis, treatment with ATX could also significantly reduce secondary brain injury characterized by neurological dysfunction, cerebral edema and blood-brain barrier disruption. In contrast, the PI3K/Akt inhibitor, {"type":"entrez-nucleotide","attrs":{"text":"LY294002","term_id":"1257998346","term_text":"LY294002"}}LY294002, could partially reverse the neuroprotection of ATX in the early period after SAH by downregulating ATX-induced activation of Akt/Bad and upregulating cleaved caspase-3 levels. These results provided the evidence that ATX could attenuate apoptosis in a rat SAH model, potentially, in part, through modulating the Akt/Bad pathway.     

Properties of thermoplastic starch from cassava bagasse and cassava starch and their blends with poly (lactic acid)

Cassava bagasse is an inexpensive and broadly available waste byproduct from cassava starch production. It contains roughly 50% cassava starch along with mostly fiber and could be a valuable feedstock for various bioproducts. Cassava bagasse and cassava starch were used in this study to make fiber-reinforced thermoplastic starch (TPS_B and TPS_I, respectively). In addition, blends of poly (lactic acid) and TPS_I (20%) and TPS_B (5, 10, 15, 20%) were prepared as a means of producing low cost composite materials with good performance. The TPS and PLA blends were prepared by extrusion and their morphological, mechanical, spectral, and thermal properties were evaluated. The results showed the feasibility of obtaining thermoplastic starches from cassava bagasse. The presence of fiber in the bagasse acted as reinforcement in the TPS matrix and increased the maximum tensile strength (0.60 MPa) and the tensile modulus (41.6 MPa) compared to cassava starch TPS (0.40 and 2.04 MPa, respectively). As expected, blending TPS with PLA reduced the tensile strength (55.4 MPa) and modulus (2.4 GPa) of neat PLA. At higher TPS_B content (20%) the maximum strength (19.9 MPa) and tensile modulus (1.7 GPa) were reduced about 64% and 32%, respectively, compared to the PLA matrix. In comparison, the tensile strength (16.7) and modulus (1.2 GPa) of PLA blends made with TPS_I were reduced 70% and 51% respectively. The fiber from the cassava bagasse was considered a filler since no increase in tensile strength of PLA/TPS blends was observed. The TPS_I (33.1%) had higher elongation to break compared to both TPS_B (4.9%) and PLA (2.6%). The elongation to break increased from 2.6% to 14.5% by blending TPS_I with PLA. In contrast, elongation to break decreased slightly by blending TPS_B with PLA. Thermal analysis indicated there was some low level of interaction between PLA and TPS. In PLA/TPS_B blends, the TPS_B increased the crystallinity of the PLA component compared to neat PLA. The fiber component of TPS_B appeared to have a nucleating effect favoring PLA crystallization.     

Astaxanthin Attenuates Early Acute Kidney Injury Following Severe Burns in Rats by Ameliorating Oxidative Stress and Mitochondrial-Related Apoptosis

Early acute kidney injury (AKI) is a devastating complication in critical burn patients, and it is associated with severe morbidity and mortality. The mechanism of AKI is multifactorial. Astaxanthin (ATX) is a natural compound that is widely distributed in marine organisms; it is a strong antioxidant and exhibits other biological effects that have been well studied in various traumatic injuries and diseases. Hence, we attempted to explore the potential protection of ATX against early post burn AKI and its possible mechanisms of action. The classic severe burn rat model was utilized for the histological and biochemical assessments of the therapeutic value and mechanisms of action of ATX. Upon ATX treatment, renal tubular injury and the levels of serum creatinine and neutrophil gelatinase-associated lipocalin were improved. Furthermore, relief of oxidative stress and tubular apoptosis in rat kidneys post burn was also observed. Additionally, ATX administration increased Akt and Bad phosphorylation and further down-regulated the expression of other downstream pro-apoptotic proteins (cytochrome c and caspase-3/9); these effects were reversed by the PI3K inhibitor {"type":"entrez-nucleotide","attrs":{"text":"LY294002","term_id":"1257998346","term_text":"LY294002"}}LY294002. Moreover, the protective effect of ATX presents a dose-dependent enhancement. The data above suggested that ATX protects against early AKI following severe burns in rats, which was attributed to its ability to ameliorate oxidative stress and inhibit apoptosis by modulating the mitochondrial-apoptotic pathway, regarded as the Akt/Bad/Caspases signalling cascade.     

甜菜Actin基因片段的克隆及序列分析

根据其它植物Actin基因设计一对简并性引物,以甜菜根总RNA为模板,采用RT-PCR的方法扩增出Actin基因片段并克隆到pUCm-T载体上,阳性克隆经PCR鉴定测序。序列分析表明,克隆得到Actin基因家族的两个成员,其片段长度均为598 bp,编码198个氨基酸,它们间的同源性为87%。将其分别命名为BvACT1和BvACT2,已在GenBank中注册,登录号为KF214784和KF214785。多重序列比较表明,Bv A CT1氨基酸序列与其他植物Actin基因的同源性在92%以上,而Bv A CT2则在93%以上。     

Differential Regulatory Mechanisms of CBF Regulon Between Nipponbare (Japonica) and 93-11 (Indica) During Cold Acclimation

Nine CBF/DREB1 homologous genes in rice were obtained by BLAST search in the NCBI database,which share conserved amino acid sequences with DREB1 protein in Arabidopsis.Three CBF genes organized in tandem,named OsCBF1,OsCBF2 and OsCBF3,showed a transient induction in the process of cold acclimation,much stronger in indica rice 93-11 compared with japonica rice Nipponbare.The candidate downstream genes OsLIP5 and OsLIP9 were induced in 93-11 but not in Nipponbare.The differential expression of CBF regulon might be caused by polymorphisms within promoter sequences between these two rice varieties.These results could be useful for utilization of CBF/DREB1 genes and illustration of differences in chilling tolerance between indica and japonica rice varieties.     

菜豆几丁质酶基因的克隆、序列分析及其表达

根据GenBank中收录的菜豆几丁质酶基因mRNA序列设计一对引物,以菜豆品种五常油豆总DNA为模板,通过PCR扩增获得一个约1.1kb的DNA片段Bchi,将其克隆入pUC18载体.测序和序列分析结果表明,该序列全长1088bp,含有一个981bp的完整开放读码框,无内含子,编码327个氨基酸,编码产物在结构上具有Class Ia几丁质酶的结构特征:N-端具有一个26个氨基酸的信号肽,其后是富含8个半胱氨酸、长41个氨基酸的几丁质结合区和由249个氨基酸组成的高度保守的催化区,C-端为由11个氨基酸组成的液泡定位多肽.序列与结构上的同源性分析表明Bchi基因是菜豆Class Ia几丁质酶基因.此序列已在GenBank中注册,登录号为AY357300.以pQE-30为原核表达载体,构建成pQE-Bchi重组表达载体,转化表达受体菌E.coliM15,经IPTG诱导后表达出一个约35kD的蛋白,与推测的Bchi基因编码产物的大小一致,表明Bchi基因在大肠杆菌中能够表达,是一个具有表达功能的基因.     

龙眼体胚Myb转录因子基因的克隆及序列分析

以龙眼同步化调控的鱼雷形胚为材料,设计特异上下游引物,采用RT-PCR法,获得了797 bp的基因序列。通过NC-BI的BLAST软件进行核苷酸序列和氨基酸序列的同源性搜索及相似度分析,结果表明,所获得的核苷酸序列和翻译的氨基酸序列与其他物种的Myb转录因子有较高的同源性,推断该基因序列属于龙眼Myb转录因子基因。已在GenBank登录,登录号为HQ661039。