Dark Brown Pigments Research Articles

Streptomyces profundus sp. nov., a novel marine actinobacterium isolated from deep-sea sediment of Madeira Archipelago, Portugal.

A novel strain, MA3_2.13T, was isolated from deep-sea sediment of Madeira Archipelago, Portugal, and characterized using a polyphasic approach. This strain produced dark brown soluble pigments, bronwish black substrate mycelia and an aerial mycelium with yellowish white spores, when grown on GYM 50SW agar. The main respiratory quinones were MK-10(H4), MK-10(H6) and MK-10(H8). Diphosphatidylglycerol, phosphatidylethanolamine, three unidentified phospholipids and two glycophospholipids were identified as the main phospholipids. The major cellular fatty acids were iso-C16 : 1, iso-C16 : 0, anteiso-C17 : 1 and anteiso-C17 : 0. Phylogenetic analyses based on 16S rRNA gene showed that strain MA3_2.13T is a member of the genus Streptomyces and was most closely related to Streptomyces triticirhizae NEAU-YY642T (NR_180032.1; 16S rRNA gene similarity 97.9 %), Streptomyces sedi YIM 65188T (NR_044582.1; 16S rRNA gene similarity 97.4 %), Streptomyces mimosae 3MP-10T (NR_170412.1; 16S rRNA gene similarity 97.3 %) and Streptomyces zhaozhouensis NEAU-LZS-5T (NR_133874.1; 16S rRNA gene similarity 97.0 %). Genome pairwise comparisons with closest related type strains retrieved values below the threshold for species delineation suggesting that strain MA3_2.13T represents a new branch within the genus Streptomyces. Based on these results, strain MA3_2.13T (=DSM 115980T=LMG 33094T) is proposed as the type strain of a novel species of the genus Streptomyces, for which the name Streptomyces profundus sp. nov. is proposed.

Decolorization of Melanoidin Using Sono–Fenton and Photo–Fenton Processes

Ethanol production wastewater contains high quantities of dark–brown pigments (melanoidin) that result in low color removal using conventional biological treatments. Advanced oxidation processes (AOPs) are the most documented methods for reducing the color associated with melanoidin. This study examines the degradation of melanoidin using AOPs based on photo–Fenton, sono–Fenton, and sono–photo–Fenton processes. Their effects on decolorization were investigated based on light intensity, ultrasonic frequency, and the iron concentration (Fe2+)–to–H2O2 ratio. This study showed that ultrasonic waves and UV light result in a higher melanoidin decolorization efficiency than Fenton reactions alone. The initial color values were reduced from 5000–5500 ADMI to below 500 ADMI for both processes because the ultrasonic waves and ultraviolet light induced H2O2 breakdown into the 🞄OH radical. Reducing the color of the melanoidin using the photo–Fenton process resulted in a decolorization rate of 0.1126 min−1, which was higher than the rates of both the sono–Fenton and sono–photo–Fenton processes. These results provide proof that the photo–assisted Fenton process is more applicable to treating dye–contaminated water than are other enhancing approaches.

Esophageal Pseudomelanosis Causing Pseudoachalasia.

CASE REPORT An 86-year-old White man with a medical history of hypertension managed with hydralazine and dementia presented to our center for further evaluation of dysphagia complicated by weight loss. The patient was diagnosed 6 years ago with achalasia through a barium esophagram and received Botox injections every 6 months. His last esophagogastroduodenoscopy, 6 months before presenting to our center, reported achalasia, and he received balloon dilation and a Botox injection at the lower esophageal sphincter (LES). Esophageal manometry placement was not tolerated. Subsequent evaluation through a barium esophagram showed dysmotility (Figure 1), and endoscopy showed hyperpigmentation (Figure 2), diverticula (Figure 3), and a severely tight LES with a focal stricture but no mass. Biopsies of the distal esophagus demonstrated that the hyperpigmentation was due to dark granules without melanocytes (Figure 4). The patient was diagnosed with pseudoachalasia given endoscopic and esophagram findings of a tight LES with pseudomelanosis on histology. There are reported cases of hydralazine causing pseudomelanosis duodeni, although the patient had minimal response to stopping hydralazine.1 He was managed with serial dilations with an initial endoscopy balloon dilation from 8 to 12 mm, repeat dilation 1 month later from 8 mm to 15, and 4 months later dilation to 16.5 mm.Figure 1.: Barium esophagram demonstrating narrowing at the gastroesophageal junction.Figure 2.: Hyperpigmentation of the distal esophageal.Figure 3.: Two small diverticula in the distal esophagus.Figure 4.: Extensive pseudomelanosis with dark brown granular pigments within histiocytes without melanocytic infiltration with hematoxylin and eosin stain.Esophageal melanocytosis describes hyperpigmentation due to melanocytic proliferation, although pseudomelanosis involves pigmented macrophages without melanocytes.2 Pseudomelanosis has been described in the stomach and small intestine due to chronic medical conditions, such as renal disease, diabetes, and antihypertensive drugs, such as hydralazine and furosemide.3 Treatment is focused on the underlying disorder, cessation of potential causative medications, and symptom management. Pseudomelanosis in the duodenum is the most common location, and the hyperpigmentation is often attributed to iron deposition. This leads to proposed mechanisms involving iron and sulfur deposition in the mucosa secondary to either medication use or impeded clearance.4 Esophageal pseudomelanosis is rare with only case reports in the literature and no clear etiology; it has never been described to cause strictures or pseudoachalasia. Further studies are needed to evaluate esophageal pseudomelanosis and its clinical significance. DISCLOSURES Author contributions: All authors reviewed and approved the final manuscript. A. Jain: study concept and design, revision of the manuscript, and is the article guarantor. AG Issac: acquisition of data and drafting of the manuscript. W. Zheng: pathology image acquisition. Financial disclosure: None to report. Informed consent was obtained for this case report.

First report of Lasiodiplodia theobromae associated with panicle blight of grapes (Vitis vinifera L.) in India.

Grape (Vitis vinefera L.) is a popular horticulture crop in Karnataka, India. A fungal pathogen caused panicle blight on panicles with immature fruit and severity increased subsequently in the grape growing regions of Devanahalli and Doddaballapur, Karnataka, between August and September 2019. The disease incidence varied from 15 to 18 percent in around 45 hectares of grape vineyards surveyed. The associated fungal pathogen was isolated on Potato Dextrose Agar (PDA) medium (HiMedia Laboratory, Mumbai, India) amended with Chloramphenicol. A total of 12 fungal isolates were obtained and identified based on morphology. Fungal cultures obtained from all the panicle blight affected samples were fluffy grayish to black with profuse, dense mycelium. Microscopic examinations revealed that the conidia ellipsoidal, two celled and hyaline when young, and developed dark brown pigments at maturity. Mature conidia measured 18.24±2.35 to 26.62±3.11 μm long and 10.32±1.08 to 12.57±1.82 μm width (n=30). The fungal pathogen was identified as a Lasiodiplodia sp. based on colony morphology and microscopic features. A total of three representative isolates L. theobromae (Vv12, Vv15, and Vv19) were selected for molecular identification based on ITS-rDNA, tub2 and EF-1α gene sequences and phylogenetic analysis. Genomic DNA was isolated from 12 day old cultures and ITS-rDNA, tub2 and EF-1α genes were amplified using ITS1/ITS4; Bt2a/Bt2b and EF1-728F/986R primer pairs, respectively (White et al., 1990; Glass and Donaldson, 1995, Carbone and Kohn, 1999). PCR amplicons were sequenced and the sequences were deposited in GenBank with the accession number ITS: MZ855866.1; MZ855867.1; MZ855868.1; tub2: MZ868708.1; MZ868709.1; MZ868710.1 and EF-1α: OM604750; OM604751; OM604752 respectively. The phylogeny was constructed based on combined ITS, EF-1α and the tub2 regions. Maximum Likelihood (ML) analysis was conducted and an ML tree was constructed with the substitution models (branch support was evaluated by 1,000 bootstrap replications). Combined phylogeny confirmed that the sequences shared a common clade with L. theobromae. Based on micro-morphological features and multi-locus sequence phylogeny, the associated fungal pathogen was identified as L. theobromae. There are no reports on the occurrence of L. theobromae causing panicle blight on grapes from India. Further, the pathogens association was confirmed through pathogenicity assay conducted on field harvested healthy bunches of grapes maintained under humid chamber. A total of 10 grape bunches were inoculated with a mycelial disc on the rachis of the panicle and incubated in a moist chamber for 5 days and control sets were inoculated with only agar plugs. The experiments were conducted in three replicates and repeated twice. A total of 21 panicle bunches developed typical rot symptoms 12-days post inoculation. The identity of the pathogen was confirmed based on micromorphology and cultural features after re-isolation (n=5), thus proving the Koch postulates and confirming the association of L. theobromae with panicle blight of grapes. Lasiodiplodia species are known to cause dieback, stem blight, leaf blights and spots on various crop plants. Mathur (1979) mentioned the occurrence of L. theobromae on grapes, however, no further details are available on the part associated, as well as morphological and molecular confirmation of L. theobromae. This is the first report of the L. theobromae causing panicle blight disease of grapes in India. Further, understanding the host range for L. theobromae and its variation will help to draw suitable disease management strategies.

Effect of Melanization on Thallus Microstructure in the Lichen Lobaria pulmonaria.

Lichens often grow in microhabitats where they experience severe abiotic stresses. Some species respond to high UV radiation by synthesizing dark brown melanic pigments in the upper cortex. However, unlike the melanized structures of non-lichenized fungi, the morphology of the melanic layer in lichens remains unstudied. Here, we analyzed the morphology, ultrastructure, and elemental composition of the melanized layer in UV-exposed thalli of the lichen Lobaria pulmonaria (L.) Hoffm. Using light microscopy, we detected a pigmented layer sensitive to staining with 3,4-L-dihydroxyphenylalanine, a precursor of eumelanin, in the upper cortex of melanized thalli. Analysis of cross-sections of melanized thalli using scanning electron microscopy revealed that melanin-like granules are deposited into the hyphal lumens. Melanized thalli also possessed thicker hyphal cell walls compared to pale thalli. Energy-dispersive X-ray spectroscopy analysis of the elemental composition of the hyphal walls and extracted melanin indicated that the type of melanin synthesized by L. pulmonaria is eumelanin. Transmission electron microscopy was used to show that during melanization melanosome-like dark vesicles are transported to the cell surface and secreted into the cell walls of the fungal hyphae. Results from this study provide new insights into the effects of melanin synthesis on the microstructure of lichen thalli.

Histological analysis of tattoo removal by water cavitation bubbles and jet formation using Nd: YAG and nanosecond laser pulses

Background: For efficient laser tattoo removal, photodisruption of tissue can ensure a very powerful means to shutter ink granules. At very high laser intensity level, photodisruption dominates and selective photothermolysis will have secondary effect in clearing the tattoo ink. Lower laser fluence is all what to trigger non-linear photons absorption and the generation of exploding cavitation bubbles that can tremendously hammer the ink granules.Subjects, materials, and methods: Three domestic white rabbits; each received simultaneous injections of a color pigment tattoo under general anesthesia, followed by a single session of (1064) Q.S Nd: YAG nanosecond laser pulses for tattoo removal.Results and Discussion: Spectroscopic properties of black, dark brown and red tattoo inks were studied. Near threshold laser fluence was selected to select the optimum conditions for obtaining scar-free treatment. Histological images of the biopsies, taken after thirty days of laser treatment of black, dark brown and red tattoos showed a marked reduction in pigment granules size with no appearance of hyperplasia or inflammatory cells. Coexistence of macrophages was suggested to be responsible for actively phagocytizing the laser-dispersed tattoo fragments.Conclusion: skin biopsies have demonstrated ink granules local redistribution. Photodisruption at 1064nm laser effectively targeted black and dark brown tattoo pigments by the generation of cavitation bubbles. The weaker laser light absorption of red pigments at 1064nm only showed tattoo clearance when using 532nm wavelength.

Materials and techniques used in the manufacture of painted sculptures at Fushan Temple (1595–1882 AD), Heyang County, Shaanxi Province, China

The painted sculptures in ancient Chinese folk temples are important traditional folk painting art forms, and it is of great significance for painted sculptures conservation to study the materials and techniques employed in their production. In this study, firstly, the samples from eight different colored painted sculptures were collected at Fushan Temple (1595–1882 AD) in Heyang County, Shaanxi Province, China. Secondly, the surface morphology and cross-sectional structure of the painted sculptures were investigated using optical microscopy and scanning electron microscopy (SEM). In addition, qualitative and semi-quantitative studies of the elemental and physical phase compositions of the materials used in the manufacture of the painted sculptures were conducted by X-ray diffraction (XRD), Raman spectroscopy, and energy-dispersive X-ray spectroscopy (EDS). The results showed that the painted sculpture samples from Fushan Temple comprised a pigment layer, white powder layer, fine clay layer, and coarse clay layer from the outside to the inside. The white powder layer was made of chalk, with functions in color development and leveling. The fine clay layer comprised quartz and other sand particles with small grain sizes. The coarse clay layer comprised feldspar and cotton silk, with larger grain size. In addition, the pigments used in the painted sculptures were chrome red, lapis lazuli, phthalocyanine green, earth green, amorphous carbon black, iron oxide red, and pyrite. The green pigment was earth green in the early stages but it was later repainted with phthalocyanine green mixed with some lapis lazuli pigments for toning. In addition, the dark green and brown pigments comprised mixtures of amorphous carbon black and phthalocyanine green, and amorphous carbon black and iron oxide red, respectively.

Morphological differentiation of bovine and equine hair for species’ identification in forensic veterinary investigations

In this paper a morphological differentiation tool of bovine and equine hair for their identification in forensic investigations is provided. 30 animals were used, fifteen adult bovines of the “Siboney de Cuba” breed and fifteen adult equines of the “Criolla” breed. These species are the most bred in Cuba and, at the same time, the most exposed to poaching and theft crimes. Hair from the cows was sampled from the dorsal region, the lateral region and the ventral region; on the other hand, the horses’ hair was sampled from the dorsal region of the body, of the mane and of the tail. The sampled areas, different for bovine and equine, have been chosen because they represent the different types of hairs present in the two species under examination. Getty et al. (2004), infact, reported that both in cattle and horses the length and thickness of the hair is reduced in a dorsal-ventral direction towards the abdomen, increasing its density. The samples were observed at an optical microscope with LCD display, at lens 4X, 10X, 40X and 100X, to determine cuticle scales, cortex, medulla and medullary index. As a result, it was noticed that both bovine and equine hair show a morphological and histomorphometric pattern based on the characteristics of the constitutive structures: cuticle scales, cortex and medulla. The bovines show a thin cuticle, coronal scales in mosaic with serrated edges and flattened irregularly, while in the equines it is thin and imbricated with irregular waves and intermediate spaces. The cortex in the bovines is thin crust dark brown and not pigmented; in the equines the cortex is dark brown, with light texture and isolated dark brown and black pigments. In both species, the white hairs do not show pigments. Through this investigation it can be noticed that one of the most important aspects in the species identification in forensic analysis is the medullary index, since it allows differentiation between animal and human species. In the bovine it is shown with an average of 0.60, while the equine shows an average medullary index of 0.49.

Unraveling Melanin Biosynthesis and Signaling Networks in Cryptococcus neoformans.

Melanin is an antioxidant polyphenol pigment required for the pathogenicity of many fungal pathogens, but comprehensive regulatory mechanisms remain unidentified. In this study, we systematically analyzed melanin-regulating signaling pathways in Cryptococcus neoformans and identified four melanin-regulating core transcription factors (TFs), Bzp4, Usv101, Mbs1, and Hob1, required for induction of the laccase gene (LAC1). Bzp4, Usv101, and Mbs1 independently regulate LAC1 induction, whereas Hob1 controls Bzp4 and Usv101 expression. Both Bzp4 and Usv101 are localized in the cytoplasm under nutrient-rich conditions (i.e., in the presence of yeast extract-peptone-dextrose [YPD] medium) but translocate into the nucleus upon nutrient starvation (i.e., in the presence of yeast nitrogen base [YNB] medium without glucose), and Mbs1 is constitutively localized in the nucleus. Notably, the cAMP pathway is not involved in regulation of the four TFs, but the high-osmolarity glycerol response (HOG) pathway negatively regulates induction of BZP4 and LAC1 Next, we searched for potential kinases upstream of the core TFs and identified nine core kinases; their deletion led to defective melanin production and LAC1 induction. Deletion of GSK3 or KIC1 abolished induction of LAC1 and BZP4 and perturbed nuclear translocation of Bzp4. Notably, Gsk3 also regulated expression of HOB1, USV101, and MBS1, indicating that it is a critical melanin-regulating kinase. Finally, an RNA sequencing-based transcriptome analysis of the wild-type strain and of bzp4Δ, usv101Δ, hob1Δ, and mbs1Δ strains under nutrient-rich and nutrient-starved conditions revealed that the melanin-regulating core TFs govern redundant and distinct classes of genes involved in a variety of biological processes.IMPORTANCE Melanins are dark green, brown, or black pigments that serve as antioxidant, reactive oxygen species (ROS) scavengers that protect fungal pathogens from radiation and host immune responses. Cryptococcus neoformans, the major etiological agent of fungal meningoencephalitis, also utilizes melanin as a key virulence factor. In this basidiomycete pathogen, melanin production is regulated by the cAMP and high-osmolarity glycerol response (HOG) pathways, and yet its complex signaling networks remain poorly described. In this study, we uncovered novel melanin synthesis regulatory networks consisting of core transcription factors (TFs), including Bzp4, Usv101, Hob1, and Mbs1, and core kinases Gsk3 and Kic1. These networks were identified through coupling systematic analyses of the expression and epistatic relationships of TF and kinase mutant libraries in the presence of diverse melanin substrates with transcriptome profiling of the core TF mutants. Thus, this report provides comprehensive insight into the melanin-regulating pathways in C. neoformans and other fungal pathogens.

Assessment of the environmental impacts of solous municipal solid waste landfills using soil properties and earthworms (<i>Eudrilus eugeniae</i>)

This study assessed the effects of Solous Municipal Solid Waste Landfills on soil properties, heavy metal contamination and macro-invertebrates (earthworms). Soil and earthworm samples were collected from Solous municipal solid waste landfills with reference samples obtained from the Botanical Garden, University of Lagos and taken to the laboratory for analyses using standard methods. Results showed that pH and organic matter values of soil samples from the landfills ranged from 8.51±0.88 to 9.85±0.02 and 5.79±0.53 % to 7.11±1.19 % respectively, and they were statistically ( P < .05) higher than the values of the reference samples. Most of the assessed heavy metals in soil and earthworms from the landfills were significantly ( P < .05) higher when compared to the reference samples. The antioxidant enzymes and lipoperoxidation product levels were significantly ( P < .05) higher in earthworms from most of the landfills in relation to the reference samples. The mean total protein concentrations in annelids from the landfills were lower than the mean concentration in the reference samples, however, only those from two of the landfills (Solous B and C) were significant ( P < .05). Histological studies showed that the clitella of earthworms from the landfills had abnormalities such as disruption of body wall muscle layers, cellular degeneration, oedematous longitudinal muscle, and dark brown pigments. Solous Landfills had an obvious effect on the assessed abiotic and biotic parameters, and could be a source of heavy metal pollution in nearby vicinities especially their groundwater sources and surface water. Keywords: Biochemical markers, Bioindicator, Eudrilus eugeniae , Heavy metals, Landfill, Soil properties

Effect of Co and Pr doping on the properties of solar-reflective ZnFe2O4 dark pigment

High NIR-reflective Co-doped ZnFe2O4 black pigments were synthesized by a simple solid-state reaction of ZnO and Fe2O3 in the presence of 3 to 30 wt.% Co3O4 at 1000 and 1100 °C. A series of black pigments with low L* values in a range of 25.5-26.5 and NIR reflectance of 39.5-48.7% were obtained at 1000 °C while the pigments with comparable L* values and slightly lower NIR reflectance were obtained at 1100 °C. A change of the pigment property could be attributed to substitution of Co2+ for Zn2+ as revealed by XRD analysis as well as reflectance spectra. Calcination of ZnO and Fe2O3 in the presence of 3 to 30 wt.% Pr6O11 at 1100 °C resulted in dark brown composite pigments consisting of ZnFe2O4, PrFe2O3 and unreacted Fe2O3 with L* value around 40 ±1 and NIR reflectance in a range of 48-50%.

Forkhead containing transcription factor Albino controls tetrapyrrole-based body pigmentation in planarian.

Pigmentation processes occur from invertebrates to mammals. Owing to the complexity of the pigmentary system, in vivo animal models for pigmentation study are limited. Planarians are capable of regenerating any missing part including the dark-brown pigments, providing a promising model for pigmentation study. However, the molecular mechanism of planarian body pigmentation is poorly understood. We found in an RNA interference screen that a forkhead containing transcription factor, Albino, was required for pigmentation without affecting survival or other regeneration processes. In addition, the body color recovered after termination of Albino double stranded RNA feeding owing to the robust stem cell system. Further expression analysis revealed a spatial and temporal correlation between Albino and pigmentation process. Gene expression arrays revealed that the expression of three tetrapyrrole biosynthesis enzymes, ALAD, ALAS and PBGD, was impaired upon Albino RNA interference. RNA interference of PBGD led to a similar albinism phenotype caused by Albino RNA interference. Moreover, PBGD was specifically expressed in pigment cells and can serve as a pigment cell molecular marker. Our results revealed that Albino controls planarian body color pigmentation dominantly via regulating tetrapyrrole biogenesis. These results identified Albino as the key regulator of the tetrapyrrole-based planarian body pigmentation, suggesting a role of Albino during stem cell-pigment cell fate decision and provided new insights into porphyria pathogenesis.

Invertase production and molasses decolourization by cold-adapted filamentous fungus Cladosporium herbarum ER-25 in non-sterile molasses medium

This study was undertaken to remove the coloring compounds of molasses as well as produce extracellular (exo) invertase in sterile and non-sterile molasses medium by using cold-adapted filamentous fungus Cladosporium herbarum ER-25. It was determined that a combination of low culture pH (5.5), temperature (20°C) and high molasses concentration (6%) could completely prevent undesired bacterial contamination during the cultivation of C. herbarum ER-25. Under the optimized non-sterile culture conditions, the maximum invertase activity (36.1U/mL) was attained after 72h. On the other hand, the fungus could remove toxical dark brown pigments (melanoidins) in non-sterilized molasses medium through biodegradation and bioadsorption mechanisms. A color removal rate of 64.8% in non-sterile medium could be achieved at the end of 144-h cultivation period. It was found that laccase and manganese peroxidase were responsible for biodegradation. No ligninase activity was detected for the fungus during the cultivation. Maximum laccase (4.6U/mL) and manganese peroxidase (3.5U/mL) activities could be reached after 120h. Higher invertase activity and color removal rate were achieved in non-sterilized medium compared to sterilized one. This is the first report on invertase production from cold-adapted microorganisms under non-sterile culture conditions. As an additional contribution, use of cold-adapted fungi for molasses decolourization was investigated for the first time in the present study.

Functional characterization of a veA-dependent polyketide synthase gene in Aspergillus flavus necessary for the synthesis of asparasone, a sclerotium-specific pigment

The filamentous fungus, Aspergillus flavus, produces the toxic and carcinogenic, polyketide synthase (PKS)-derived family of secondary metabolites termed aflatoxins. While analysis of the A. flavus genome has identified many other PKSs capable of producing secondary metabolites, to date, only a few other metabolites have been identified. In the process of studying how the developmental regulator, VeA, affects A. flavus secondary metabolism we discovered that mutation of veA caused a dramatic down-regulation of transcription of a polyketide synthase gene belonging to cluster 27 and the loss of the ability of the fungi to produce sclerotia. Inactivation of the cluster 27 pks (pks27) resulted in formation of greyish-yellow sclerotia rather than the dark brown sclerotia normally produced by A. flavus while conidial pigmentation was unaffected. One metabolite produced by Pks27 was identified by thin layer chromatography and mass spectral analysis as the known anthraquinone, asparasone A. Sclerotia produced by pks27 mutants were significantly less resistant to insect predation than were the sclerotia produced by the wild-type and more susceptible to the deleterious effects of ultraviolet light and heat. Normal sclerotia were previously thought to be resistant to damage because of a process of melanization similar to that known for pigmentation of conidia. Our results show that the dark brown pigments in sclerotia derive from anthraquinones produced by Pks27 rather than from the typical tetrahydronapthalene melanin production pathway. To our knowledge this is the first report on the genes involved in the biosynthesis of pigments important for sclerotial survival.

Condensed and hydrolyzable tannins in two soybean (Glycine max L. cv. Madarin) cell lines that differ in their ploidy levels

Comparison of SB&lt;sub&gt;1&lt;/sub&gt; and SB&lt;sub&gt;3&lt;/sub&gt; cell lines in both suspension and callus cultures showed that the latter, which produce much more dark brown pigments, had higher levels of condensed tannins. They also had higher total phenolics and flavonoid contents and L-phenylalanine ammonia lyase activity.

Phenolic compounds and phenylalanine ammonia lyase activity in two soybean (Glycine max L. cv. Mandarin) cell lines that differ in their ploidy levels

Through this preliminary experiment examining different color expressions in two soybean cell lines, SB&lt;sub&gt;1&lt;/sub&gt;, and SB&lt;sub&gt;3&lt;/sub&gt; several important features were observed. SB&lt;sub&gt;3&lt;/sub&gt; cells, which produce much more dark brown pigments in both suspension and callus cultures, had higher total phenolics and flavonoid contents as well as higher L-phenylalanine ammonia lyase activity.

Lignocellulolytic Activities among Trichoderma Isolates from Lahad Datu, Sabah and Deception Island, Antarctic

Trichoderma species have the potential for application in composting as biological control agents in controlling disease and increasing yield of production in the agricultural industry. The prevalent soil fungus of Trichoderma produces lignocellulolytic enzymes that assist the degradation of woody lignocellulose materials. The aim of the experimental work was to check the potential of lignocellulolytic Trichoderma fungi for the use of rapid composting of oil palm empty fruit bunches fibers. Fifty-two of Trichoderma isolates from Sabah and seven isolates from Antarctic were examined for in-vitro lignocellulolytic activity by assaying their ability to develop dark brown pigments, yellow halo zone, and clear white zone on tannic acid media (TAM) for lignin; Jensen Media (JM) for cellulose; and modified Melin–Nokrans media (MMNM) for starch. The best six Sabah Trichoderma isolates (5D, 10L2, 10P, 5E, 10X, and 10E2) were found to be potential lignocellulolytic agents based on their diameter of halo zone formed on amended media for further testing of in vitro bioconversion of oil palm empty fruit bunches. The diameters of halo zones were measured for the analysis of their ability in degrading lignin, cellulose, and starch. In contrast, Antarctic Trichoderma isolates consistently exhibited lower lignocellulolytic activities based on their smaller diameter of halo zone formed on TAM, JM, and MMNM. Most of the Trichoderma isolates are found to synthesize polyphenol oxidase, endoglucanases, and are able to hydrolyze starch to glucose in the three different media. Thus, the finding shows the potential of these isolates for use in large-scale composting of oil palm empty fruit bunches.

Facile Method for Site-specific Gene Integration in Lysobacter enzymogenes for Yield Improvement of the Anti-MRSA Antibiotics WAP-8294A and the Antifungal Antibiotic HSAF

Lysobacter is a genus of Gram-negative gliding bacteria that are emerged as novel biocontrol agents and new sources of bioactive natural products. The bacteria are naturally resistant to many antibiotics commonly used in transformant selection, which has hampered the genetic manipulations. Here, we described a facile method for quick-and-easy identification of the target transformants from a large population of the wild type and nontarget transformants. The method is based on a distinct yellow-to-black color change as a visual selection marker for site-specific integration of the gene of interest. Through transposon random mutagenesis, we identified a black-colored strain from the yellow-colored L. enzymogenes . The black strain was resulted from a disruption of hmgA, a gene required for tyrosine/phenylalanine metabolism. The disruption of hmgA led to accumulation of dark brown pigments. As proof of principle, we constructed a series of expression vectors for a regulator gene found within the WAP-8294A biosynthetic gene cluster. The yield of WAP-8294A in the black strains increased by 2 fold compared to the wild type. Interestingly, the yield of another antibiotic (HSAF) increased up to 7 fold in the black strains. WAP-8294A is a family of potent anti-MRSA antibiotics and is currently in clinical studies, and HSAF is an antifungal compound with distinct structural features and a novel mode of action. This work represents the first successful metabolic engineering in Lysobacter. The development of this facile method opens a way toward manipulating antibiotic production in the largely unexplored sources.

Melanotic Schwannoma of the Cervical Spine Progressing With Pulmonary Metastasis: Case Report

Melanotic schwannoma (MS) is an unusual variant of nerve sheath neoplasm. Only 10% of these tumors will undergo malignant degeneration, with exceedingly rare reported metastasis. We present a 32-year-old woman with a 6-month history of cervical pain and left arm progressive weakness. Neurological examination showed a left upper limb radicular pain, with pyramidal syndrome at C5 level. The magnetic resonance imaging (MRI) study highlighted an intradural extramedullary heterogeneous mass along the spinal cord at the C4–C5 level, slightly hyperintense with T1 and hypointense with T2-weighted sequences, invading the left neural foramen. The patient underwent C3–C5 laminectomy with total resection of a black tumor. In the postoperative period, a patent deficit of shoulder abduction ensued related to the nervous section. Microscopically, compactly fascicles of spindle-shaped cells with pleomorphic and hypercromatic nuclei, dark brown intracellular pigments, as well as some mitotic figures were seen. Immunohistochemical stains for S-100, Human Melanoma Black-45 (HMB-45), and vimentin were positive, with Ki-67 labelling index (LI) of 15% compatible with MS. Six months after radiotherapy she presents local recurrence and lung metastatic dissemination of the MS. She underwent left pulmonary segmentectomy, followed by chemotherapy and radiosurgery. The patient developed a febrile neutropenia and worsening of general status, and died after 3 months due to respiratory complications. MS are rare tumors with potential for local recurrence and distal metastasis. Complete surgical resection remains as the treatment of choice, once the uncommon cases with malignant progression shows low response to chemo and radiotherapy.

A bHLH protein partially controls proanthocyanidin and phytomelanin pigmentation in the seed coats of morning glory Ipomoea tricolor

In the morning glory Ipomoea tricolor, the ivory-seed (IVS) protein, which bears a basic helix-loop-helix (bHLH) domain homologous to Arabidopsis transparent testa 8 (TT8) and petunia anthocyanin 1 (AN1) that activate various epidermal traits, was reported to regulate anthocyanin pigmentation in flowers and the dark-brown pigmentation in seeds. In this study, additional roles of the Ipomoea tricolor IVS (ItIVS) gene were elucidated and seed pigments of I. tricolor were also analyzed. Microscopic observations showed that trichomes on seed surface were reduced in seeds of the ivs mutant Blue Star (BS), closely resembling ivs mutant seeds of I. purpurea. Different from petunia AN1, the ItIVS gene appears not to be related to expression of the I. tricolor Na+/H+exchanger (ItNHX1) gene that increases petal pH during flower opening, because the expression profile of ItNHX1 in BS was virtually indistinguishable from that of the wild type Heavenly Blue (HB). The dark-brown pigments of HB were mainly composed of proanthocyanidins and phytomelanins. As phytomelanin content in BS seeds was about 22% of wild type level, and vanillin and ρ-dimethylaminocinnamaldehyde (DMACA) treatments for its immature seeds also showed weak positive reactions, it seems that the ItIVS gene partially activates the accumulation of both pigments. Some biosynthetic steps in anthocyanin and proanthocyanidin biosynthesis are thought to be common in Arabidopsis. However, the chalconesynthase D (CHS-D) and the dihydroflavonol 4-reductase B (DFR-B) genes in Ipomoea, which are essential for anthocyanin biosynthesis, were not transcribed in seed coats of HB, and a dfr-b mutant Pearly Gates (PG) still produces proanthocyanidins suggesting that ItIVS may activate different sets of structural genes for anthocyanin biosynthesis in flowers and proanthocyanidin production in seeds. Our results suggest that proanthocyanidin biosynthesis in morning glory is not accomplished by a unique system like it is in Arabidopsis, and more complicated processes are involved.