High Mitochondrial DNA Diversity with Little Structure Within and Among Leaf Monkey Populations (Trachypithecus cristatus and Trachypithecus auratus)

We used analyses of mitochondrial DNA restriction site polymorphisms to estimate population genetic structure and phylogenetic relationships among 42 individuals from two Asian leaf monkey species (Trachypithecus auratus and T. cristatus) and to compare them to the geographically proximate species, Presbytis comata. We amplified a 2300-base pair fragment spanning the mitochondrial NADH 3 and NADH 4 genes, including their tRNA flanking subunits, glycine and leucine, and digested it with a battery of 22 restriction endonucleases, yielding 21 unique multienzyme haplotypes and 60 variable restriction sites. Presbytis comata is clearly divergent from both Trachypithecus species. Within the Javan T. auratus, our analysis does not support the distinction of two subspecies currently recognized on the basis of morphological features (Weitzel and Groves, 1985). T. auratus and T. cristatus are not internally monophyletic with respect to each other in our phylogenetic analysis. These results indicate either a recent speciation event with the retention of ancestral polymorphisms or that the two taxa are not separate species. Therefore with respect to conserving genetic diversity within the leaf monkey, we would have to consider T. auratus and T. cristatus as essentially one large polymorphic, conservation unit. However, within that conservation unit, T. auratus of Java represent a separate management unit from T. auratus/T. cristatus of Sumatra and Peninsular Malaysia.     

Microbial production of d‐lactic acid from dried distiller's grains with solubles

d ‐Lactic acid production is gaining increasing attention due to the thermostable properties of its polymer, poly‐d ‐lactic acid . In this study, Lactobacillus coryniformis subsp. torquens, was evaluated for its ability to produce d ‐lactic acid using Dried Distiller's Grains with Solubles (DDGS) hydrolysate as the substrate. DDGS was first subjected to alkaline pretreatment with sodium hydroxide to remove the hemicellulose component and the generated carbohydrate‐rich solids were then subjected to enzymatic hydrolysis using cellulase mixture Accellerase® 1500. When comparing separate hydrolysis and fermentation and simultaneous saccharification and fermentation (SSF) of L. coryniformis on DDGS hydrolysate, the latter method demonstrated higher d ‐lactic acid production (27.9 g/L, 99.9% optical purity of d ‐lactic acid), with a higher glucose to d ‐lactic acid conversion yield (84.5%) compared to the former one (24.1 g/L, 99.9% optical purity of d ‐lactic acid). In addition, the effect of increasing the DDGS concentration in the fermentation system was investigated via a fed‐batch SSF approach, where it was shown that the d ‐lactic acid production increased to 38.1 g/L and the conversion yield decreased to 70%. In conclusion, the SSF approach proved to be an efficient strategy for the production of d ‐lactic acid from DDGS as it reduced the overall processing time and yielded high d ‐lactic acid concentrations.     

Responses of the antioxidative enzymes in Malaysian rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.) cultivars under submergence condition

The potential involvement of activated oxygen species by submergence stress was studied in two Malaysian rice cultivars, MR219-4 and MR219-9, and cultivar FR13A that is known to be tolerant to submergence. Seedlings of these three rice cultivars were subjected to different submergence periods (4, 8, and 12 days). Under 8 days of complete submergence, FR13A cultivar showed higher lipid peroxidation in terms of malondialdehyde level and activities of antioxidative enzymes, superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), and glutathione reductase (GR) when compared to the MR219-4 and MR219-9 cultivars. MR219-9 showed higher SOD, APX, and GR activities after 12 days of submergence. The levels of SOD activity indicated that detoxification of O₂^·− to H₂O₂ was maintained at a stable level throughout the submergence stress until up to 8 days and increased rapidly at 12 days of submergence. The results indicated that tolerance to submergence in rice is associated until 8 days submergence for MR219-4 and FR13A cultivars. These findings suggested that tolerance to submergence stress in rice might be proven by increased the capacity of antioxidative system. In addition, CAT activity has much higher affinity for scavenges H₂O₂ than APX. Therefore, ascorbate glutathione cycle might be more efficient to scavenge H₂O₂.     

Mixed Herbs Drugs: Inhibitory Effect on Growth of the Endogenous Mycoflora and Aflatoxin Production

Twenty commercial mixed herbal drugs were examined for mycological profile. Aspergillus species were the predominant fungi found in the drugs. Other fungi harboured in the drugs with less frequency were Paecilomyces species, Eurotium species, Monascus species, Acremonium species, Penicillium species, Cladosporium species, Scopulariopsis species, Phialophora species and Fonseceae species. Fungal count was between 1.0 log₁₀ CFU and 2.4 log₁₀ CFU per gram of sample. When the drugs were incubated in 85% humidity at 25°C, fungal colonies grew on only two of the drugs. The mixed herbal drugs were extracted with water and the extracts were used to grow Aspergillus parasiticus. All extracts reduced aflatoxin B₁ and aflatoxin G₁ production by 62–97%. All but two of the extracts reduced aflatoxin B₂ and aflatoxin G₂ production by 39–95%. It can be concluded that the commercial powdered mixed herbal drugs contained low number of endogenous fungi, and these drugs are inhibitory to the growth of its endogenous fungi and aflatoxins production by aflatoxigenic fungi.     

Rapid Detection of Oculopathogenic Adenovirus in Conjunctivitis

Ocular adenovirus (Ad) infections occur throughout the world in both sporadic and epidemic forms. Accurate laboratory diagnosis of Ad in conjunctival samples is always valuable. The present study was carried out to explore the presence of Ad as a causative agent in clinically suspected viral conjunctivitis and to compare the performance of conventional virus isolation on cell cultures, direct detection of Ad antigens in conjunctival cells by a direct fluorescence assay, Ad DNA detection by polymerase chain reaction (PCR), and specific IgM measurement by ELISA. Samples included scrapes from conjunctiva. Scrapes were subjected to study by direct immunofluorescence stain, culture on the Hep-2 cell line, and PCR for Ad detection. Blood samples were also taken and subjected to study for specific anti-Ad IgM determination. The culture for Ad was positive in 77.8%, direct antigen detection by fluorescent stain was positive in 72.2%, PCR was positive in 83.3%, and serology was positive in 88.9% of patients. Both determination of antibody IgM and PCR correctly identified a larger group of patients compared to cell culture. The most sensitive and specific method for diagnosis of Ad compared to culture was PCR (100%), followed by IgM detection (92.9%) then direct antigen detection by fluorescent stain (85.8%). From this study, we conclude that Ad is a common pathogen in sporadic cases of conjunctivitis. Screening of adenoviral conjunctivitis is possible by using specific IgM due to its high sensitivity. A confirmatory test can be done by PCR for diagnosis of Ad, as it is a rapid, specific, and accurate method.     

Understanding Thermostability Factors of <Emphasis Type="Italic">Aspergillus niger</Emphasis> PhyA Phytase: A Molecular Dynamics Study

Molecular dynamics simulation was used to study the dynamic differences between native Aspergillus niger PhyA phytase and a mutant with 20 % greater thermostability. Atomic root mean square deviation, radius of gyration, and number of hydrogen bonds and salt bridges are examined to determine thermostability factors. The results suggest that, among secondary structure elements, loops have the most impact on the thermal stability of A. niger phytase. In addition, the location rather than the number of hydrogen bonds is found to have an important contribution to thermostability. The results also show that salt bridges may have stabilizing or destabilizing effect on the enzyme and influence its thermostability accordingly.     

Gentianella tumailica (Gentianaceae) – A new species from cold deserts of northwest Himalaya,India

A new species of Gentianella, G. tumailica sp. nov., from Tumail in the Kargil district of Ladakh, northwest Himalaya, India is described and illustrated. This novelity is morphologically similar to the sympatric G. stoliczka in inflorescence pattern, calyx and corolla shapes, sagitate anthers, more or less rounded seeds, but differs in smaller size of vegetative and floral structures, larger basal leaves, few cauline leaves, leaf apex obtuse to rounded, calyx lobes unequal (2 longer, 3 shorter), inflorescence enveloped by 4 foliaceous bracts, capsule elliptic to oblong and seed coat smooth. In addition, a diagnostic key to all Indian species of Gentianella is presented.     

Targeted Delivery of Lovastatin and Tocotrienol to Fracture Site Promotes Fracture Healing in Osteoporosis Model: Micro-Computed Tomography and Biomechanical Evaluation

Osteoporosis is becoming a major health problem that is associated with increased fracture risk. Previous studies have shown that osteoporosis could delay fracture healing. Although there are potential agents available to promote fracture healing of osteoporotic bone such as statins and tocotrienol, studies on direct delivery of these agents to the fracture site are limited. This study was designed to investigate the effects of two potential agents, lovastatin and tocotrienol using targeted drug delivery system on fracture healing of postmenopausal osteoporosis rats. The fracture healing was evaluated using micro CT and biomechanical parameters. Forty-eight Sprague-Dawley female rats were divided into 6 groups. The first group was sham-operated (SO), while the others were ovariectomized (OVx). After two months, the right tibiae of all rats were fractured at metaphysis region using pulsed ultrasound and were fixed with plates and screws. The SO and OVxC groups were given two single injections of lovastatin and tocotrienol carriers. The estrogen group (OVx+EST) was given daily oral gavages of Premarin (64.5 µg/kg). The Lovastatin treatment group (OVx+Lov) was given a single injection of 750 µg/kg lovastatin particles. The tocotrienol group (OVx+TT) was given a single injection of 60 mg/kg tocotrienol particles. The combination treatment group (OVx+Lov+TT) was given two single injections of 750 µg/kg lovastatin particles and 60 mg/kg tocotrienol particles. After 4 weeks of treatment, the fractured tibiae were dissected out for micro-CT and biomechanical assessments. The combined treatment group (OVx+Lov+TT) showed significantly higher callus volume and callus strength than the OVxC group (p<0.05). Both the OVx+Lov and OVx+TT groups showed significantly higher callus strength than the OVxC group (p<0.05), but not for callus volume. In conclusion, combined lovastatin and tocotrienol may promote better fracture healing of osteoporotic bone.     

Heterotrophic cultivation of microalgae for production of biodiesel

High cell density cultivation of microalgae via heterotrophic growth mechanism could effectively address the issues of low productivity and operational constraints presently affecting the solar driven biodiesel production. This paper reviews the progress made so far in the development of commercial-scale heterotrophic microalgae cultivation processes. The review also discusses on patentable concepts and innovations disclosed in the past four years with regards to new approaches to microalgal cultivation technique, improvisation on the process flow designs to economically produced biodiesel and genetic manipulation to confer desirable traits leading to much valued high lipid-bearing microalgae strains.