Viscoelastic and failure properties of spine ligament collagen fascicles

The microstructural volume fractions, orientations, and interactions among components vary widely for different ligament types. If these variations are understood, however, it is conceivable to develop a general ligament model that is based on microstructural properties. This paper presents a part of a much larger effort needed to develop such a model. Viscoelastic and failure properties of porcine posterior longitudinal ligament (PLL) collagen fascicles were determined. A series of subfailure and failure tests were performed at fast and slow strain rates on isolated collagen fascicles from porcine lumbar spine PLLs. A finite strain quasi-linear viscoelastic model was used to fit the fascicle experimental data. There was a significant strain rate effect in fascicle failure strain (P < 0.05), but not in failure force or failure stress. The corresponding average fast-rate and slow-rate failure strains were 0.098 ± 0.062 and 0.209 ± 0.081. The average failure force for combined fast and slow rates was 2.25 ± 1.17 N. The viscoelastic and failure properties in this paper were used to develop a microstructural ligament failure model that will be published in a subsequent paper.     

Apparent elastic modulus and hysteresis of skeletal muscle cells throughout differentiation

The effect of differentiation on thetransverse mechanical properties of mammalian myocytes was determinedby using atomic force microscopy. The apparent elastic modulusincreased from 11.5 ± 1.3 kPa for undifferentiated myoblasts to45.3 ± 4.0 kPa after 8 days of differentiation (P < 0.05). The relative contribution of viscosity, as determined fromthe normalized hysteresis area, ranged from 0.13 ± 0.02 to0.21 ± 0.03 and did not change throughout differentiation. Myosinexpression correlated with the apparent elastic modulus, but neithermyosin nor -tubulin were associated with hysteresis. Microtubulesdid not affect mechanical properties because treatment with colchicinedid not alter the apparent elastic modulus or hysteresis. Treatmentwith cytochalasin D or 2,3-butanedione 2-monoxime led to a significantreduction in the apparent elastic modulus but no change in hysteresis.In summary, skeletal muscle cells exhibited viscoelastic behavior thatchanged during differentiation, yielding an increase in the transverseelastic modulus. Major contributors to changes in the transverseelastic modulus during differentiation were actin and myosin.

     

Survival and population growth of a long-lived threatened snake species, Drymarchon couperi (Eastern Indigo Snake)

Demographic data provide a basis for understanding the life history and ecology of species, factors which are vital for informing conservation efforts; however, little is known regarding the population ecology of most snake species, including the threatened Eastern Indigo Snake (Drymarchon couperi). We used 11 years (1999–2009) of capture-mark-recapture (CMR) and 2.5 years (2003–2005) of radiotelemetry data from southeastern Georgia, USA, in a CMR modeling framework to estimate apparent survival, capture and transition probabilities, and evaluate factors influencing these parameters. The model-averaged estimate of overall apparent annual survival probability was 0.700 (±0.030 SE) and is comparable to that obtained from known fate analysis (radiotelemetry) at the same site. Body size positively influenced survival, regardless of sex. Capture probability differed seasonally by sex, suggesting lower capture probability for females in fall and males in winter. There was no evidence for effect of precipitation or site-specific differences in survival. Model averaged estimate of annual adult survival estimated using multistate CMR models was 0.738 ± 0.030 and 0.515 ± 0.189 for subadults. We estimated population growth rate (λ) and elasticity (proportional sensitivity) of λ to vital rates using a stage-structured matrix population model. Population growth rate ranged from 0.96 to 1.03 depending on the value of the probability of transitioning from subadult to adult stage. The λ was proportionally most sensitive to changes in adult survival rate, followed by subadult survival. Our results suggest that protecting adult snakes and their habitats would result in the highest likelihood of long-term population stability and growth.     

The influence of topographic microstructures on the initial adhesion of L929 fibroblasts studied by single-cell force spectroscopy

Single-cell force spectroscopy was used to investigate the initial adhesion of L929 fibroblasts onto periodically grooved titanium microstructures (height ~6 μm, groove width 20 μm). The position-dependent local adhesion strength of the cells was correlated with their rheological behavior. Spherical cells exhibited a significantly lower Young’s modulus (<1 kPa) than that reported for spread cells, and their elastic properties can roughly be explained by the Hertz model for an elastic sphere. While in contact with the planar regions of the substrate, the cells started to adapt their shape through slight ventral flattening. The process was found to be independent of the applied contact force for values between 100 and 1,000 pN. The degree of flattening correlated with the adhesion strength during the first 60 s. Adhesion strength can be described by fast exponential kinetics as C₁ [ 1 - exp( - C₂ ·t ) ] C_{1} \left[ {1 - \exp \left( { - C_{2} \cdot t} \right)} \right] with C ₁ = 2.34 ± 0.19 nN and C ₂ = 0.09 ± 0.02 s⁻¹. A significant drop in the adhesion strength of up to 50% was found near the groove edges. The effect can be interpreted by the geometric decrease of the contact area, which indicates the inability of the fibroblasts to adapt to the shape of the substrate. Our results explain the role of the substrate’s topography in contact guidance and suggest that rheological cell properties must be considered in cell adhesion modeling.     

Drug-Cyclodextrin-Vesicles Dual Carrier Approach for Skin Targeting of Anti-acne Agent

In the present study attempt was made for preparation of isotretinoin-hydroxypropyl β cyclodextrin (HP-β-CD) inclusion complex and encapsulate this complex in elastic liposomes to study the effect of dual carrier approach on skin targeting of isotretinoin. The isotretinoin HP-β-CD complex was prepared by freeze-drying method and characterized by IR spectroscopy. The drug and drug-CD complex loaded elastic liposomal formulation were prepared and characterized in vitro, ex-vivo and in vivo for shape, size, entrapment efficiency, no. of vesicles per cubic mm, in vitro skin permeation and deposition study, photodegradation and skin toxicity assay. The transdermal flux for different vesicular formulations was observed between 10.5 ± 0.5 to 13.9 ± 1.6 μg/cm²/h. This is about 15-21 folds higher than that obtained from drug solution (0.7 ± 0.1 μg/cm²/h) and 4-5 folds higher than obtained with drug-CD complex solution (2.7 ± 0.1 μg/cm²/h). The amount of drug deposit was found to increase significantly (p < 0.05) by cyclodextrin complexation (30.1 ± 0.1 μg). The encapsulation of this complex in elastic liposomal formulation further increases its skin deposition (262.2 ± 21 μg). The results of skin irritation study using Draize test also showed the significant reduction in skin irritation potential of isotretinoin elastic liposomal formulation in comparison to free drug. The results of the present study demonstrated that isotretinoin elastic liposomal formulation possesses great potential for skin targeting, prolonging drug release, reduction of photodegradation, reducing skin irritation and improving topical delivery of isotretinoin.     

Aortic Elasticity is Impaired in Patients with Endemic Fluorosis

Sixty-three patients with endemic fluorosis (36 males/27 females; mean age 33.9 ± 8.6 years) and 45 age-, sex-, and body mass index-matched healthy controls (30 males/15 females; mean age 32.7 ± 8.8 years) were included in this study. Aortic stiffness indices, aortic strain (AS), aortic distensibility (AD), and aortic strain index (ASI) were calculated from the aortic diameters measured by echocardiography and blood pressure obtained by sphygmomanometry. The urine fluoride levels of fluorosis patients were significantly higher than control subjects as expected (1.9 ± 0.1 mg/l vs. 0.4 ± 0.1 mg/l, respectively; P < 0.001). AS and AD were significantly lower in fluorosis patients than in the controls (for AS 5.3 ± 3.6 vs. 8.0 ± 3.4%; P < 0.001 and for AD 0.2 ± 0.1 vs. 0.3 ± 0.1 cm² dyn⁻¹ 10⁻³; P < 0.001, respectively). In contrast, signicantly higher ASI was observed in fluorosis patients than in the controls (3.4 ± 0.6 vs. 3.0 ± 0.4; P < 0.001, respectively). The results of our study demonstrate that elastic properties of ascending aorta are impaired in patients with endemic fluorosis.     

Susceptibility of <Emphasis Type="Italic">Candida</Emphasis> biofilms to histatin 5 and fluconazole

Candida-associated denture stomatitis has a high rate of recurrence. Candida biofilms formed on denture acrylic are more resistant to antifungals than planktonic yeasts. Histatins, a family of basic peptides secreted by the major salivary glands in humans, especially histatin 5, possess significant antifungal properties. We examined antifungal activities of histatin 5 against planktonic or biofilm Candida albicans and Candida glabrata. Candida biofilms were developed on poly(methyl methacrylate) discs and treated with histatin 5 (0.01–100 μM) or fluconazole (1–200 μM). The metabolic activity of the biofilms was measured by the XTT reduction assay. The fungicidal activity of histatin 5 against planktonic Candida was tested by microdilution plate assay. Biofilm and planktonic C. albicans GDH18, UTR-14 and 6122/06 were highly susceptible to histatin 5, with 50% RMA (concentration of the agent causing 50% reduction in the metabolic activity; biofilm) of 4.6 ± 2.2, 6.9 ± 3.7 and 1.7 ± 1.5 μM, and IC₅₀ (planktonic cells) of 3.0 ± 0.5, 2.6 ± 0.1 and 4.8 ± 0.5, respectively. Biofilms of C. glabrata GDH1407 and 6115/06 were less susceptible to histatin 5, with 50% RMA of 31.2 ± 4.8 and 62.5 ± 0.7 μM, respectively. Planktonic C. glabrata was insensitive to histatin 5 (IC₅₀ > 100 μM). Biofilm-associated Candida was highly resistant to fluconazole in the range 1–200 μM; e.g. at 100 μM only ~20% inhibition was observed for C. albicans, and ~30% inhibition for C. glabrata. These results indicate that histatin 5 exhibits antifungal activity against biofilms of C. albicans and C. glabrata developed on denture acrylic. C. glabrata is significantly less sensitive to histatin 5 than C. albicans.     

Ciprofloxacin-Induced Antibacterial Activity is Reversed by Vitamin E and Vitamin C

In the present study, we investigated the possible involvement of oxidative stress in ciprofloxacin-induced cytotoxicity against several reference bacteria including Pseudomonas aeruginosa ATCC 27853, Escherichia coli ATCC 25922, Staphylococcus aureus ATCC 29213, and clinical isolate of methicillin-resistant Staphylococcus aureus (MRSA). Oxidative stress was assessed by measurement of hydrogen peroxide generation using a FACScan flow cytometer. The antibacterial activity of ciprofloxacin was assessed using the disk diffusion method and by measuring the minimum inhibitory concentration (MIC). Ciprofloxacin induced a dose-dependent antibacterial activity against all bacteria where the highest tested concentration was 100 ug/ml. Results revealed that E. coli cells were highly sensitive to ciprofloxacin (MIC = 0.21 μg/mL ± 0.087), P. aeruginosa and S. aureus cells were intermediately sensitive (MIC = 5.40 μg/mL ± 0.14; MIC = 3.42 μg/mL ± 0.377, respectively), and MRSA cells were highly resistant (MIC = 16.76 μg/mL ± 2.1). Pretreatment of E. coli cells with either vitamin E or vitamin C has significantly protected cells against ciprofloxacin-induced cytotoxicity. These results indicate the possible antagonistic properties for vitamins C or E when they are used concurrently with ciprofloxacin.     

Effects of Pulsed Electric Field on the Viscoelastic Properties of Potato Tissue

We have investigated whether transient permeabilization caused by the application of pulsed electric field would give rise to transient changes in the potato tissue viscoelastic properties. Potato tissue was subjected to nominal field strengths (E) ranging from 30 to 500 V/cm, with a single rectangular pulse of 10⁻⁵, 10⁻⁴, or 10⁻³ s. The changes on the viscoelastic properties of potato tissue during pulsed electric fields (PEF) were monitored through small amplitude oscillatory dynamic rheological measurements. The elastic (G′) and viscous moduli (G″) were measured every 30 s after the delivery of the pulse and the loss tangent change (tan-δ) was calculated. The results were correlated with measurements of changes on electrical resistance during the delivery of the pulse. Results show a drastic increase of tan-δ in the first 30 s after the application of the pulse, followed by a decrease 1 min after pulsation. This response is strongly influenced by pulsing conditions and is independent of the total permeabilization achieved by the pulse. Our results, supported by similar measurements on osmotically dehydrated control samples, clearly show that PEF causes a rapid change of the viscoelastic properties of the tissue that could be attributed to a partial loss in turgor pressure. This would be an expected consequence of electroporation. The recovery of tan-δ to values similar to those before pulsation strongly suggests recovery of cell membrane properties and turgor, pointing at reversible permeabilization of the cells. A slight increase of stiffness traduced by a negative change of tan-δ after application of certain PEF conditions may also give an indication of events occurring on cell wall structure due to stress responses. This study set the basis for further investigations on the complex cell stress physiology involving both cell membrane functional properties and cell wall structure that would influence tissue physical properties upon PEF application.     

Electrophysiologic Characteristics of the Crista Terminalis and Implications on Atrial Tachycardia in Rabbits

The objective of this study was to evaluate the electrophysiologic characteristics of Crista terminalis (CT) and their implication in the pathogenesis of atrial tachycardia in rabbits. For this purpose, 27 New Zealand rabbits were used. Using standard glass microelectrode technique, cellular action potentials (APs) of CT and pectinate muscle (PM) were recorded in normal Tyrode’s perfusion and Tyrode’s perfusion with 4 μM isoproterenol. Longitudinal conduction velocity (V _L) and transverse conduction velocity (V _T) of CT were measured. As our data show, CT tissue had a trend of spontaneous phase IV depolarization. Conduction anisotropy (V _L/V _T) of CT was 4.53 ± 0.91. The duration of the AP of CT was longer than that of PM cells. APD₂₀ and APD₉₀ for CT were 28.1 ± 3.5 and 145.3 ± 7.1 ms; and for PM cells were 21.8 ± 4.1 and 125.3 ± 6.3 ms, respectively (all P values < 0.01). The early and delayed action depolarizations were recorded after isoproterenol perfusion. A fast paroxysmal irregular rhythm was recorded which could be arrested by 0.1 mmol/l Isoptin. It was, therefore, concluded that the latent autorhythmicity, trigger activity, and conduction properties of CT might provide the electrophysiologic basis for the occurrence and sustenance of atrial arrhythmia.     

Plasma Leptin,Plasma Zinc,and Plasma Copper Are Associated in Elite Female and Male Judo Athletes

The purpose of this study was to compare plasma leptin, plasma zinc, and plasma copper levels and their relationship in trained female and male judo athletes (n = 10 women; n = 8 men). Blood samples were obtained 24 h after training to measure plasma zinc, copper, and leptin levels. Subjects presented similar values to age (22 ± 2 years old), body mass index (24 ± 1 kg/m²), plasma zinc (17.2 ± 2 μmol/L), copper (12.5 ± 2 μmol/L), and leptin (5.6 ± 1.3 μg/L). However, height, total body mass, lean mass, fat mass, and sum of ten-skinfold thickness were higher in male than female. Plasma leptin was associated with sum of ten skinfolds in male (r = 0.91; p < 0.001) and female athletes (r = 0.84; p < 0.003). Plasma zinc was associated with leptin in males (r = 0.82; p < 0.05) while copper was associated with plasma leptin in females (r = 0.66; p < 0.05). Our results suggest that young judo athletes lost sex-related differences in leptin levels. Plasma zinc, plasma copper, and energy homeostasis may be involved in regulation of plasma leptin.     

Species and material considerations in the formation and development of microalgal biofilms

The development of microalgal biofilms has received very limited study despite its relevance in the design of photobioreactors where film growth may be advantageous for biomass separation or disadvantageous in fouling surfaces. Here, the effects of species selection, species control, and substrate properties on biofilms of Scenedesmus obliquus and Chlorella vulgaris were investigated. Experiments were conducted in batch culture and in continuous culture modes in a flow cell. Cell growth was monitored using confocal laser scanning microscopy and gravimetrically. Species selection and species control had significant effects on biofilm development. On non-sterile wastewater, C. vulgaris shifted from primarily planktonic (23.7% attachment) to primarily sessile (79.8% attachment) growth. The biofilms that developed in non-sterile conditions were thicker (52 ± 19 μm) than those grown in sterile conditions (7 ± 6 μm). By contrast, S. obliquus attained similar thicknesses (54 ± 31 and 53 ± 38 μm) in both sterile and non-sterile conditions. Neither species was able to dominate a non-sterile biofilm. The effect of substrate surface properties was minimal. Both species grew films of similar thickness (∼30 μm for S. obliquus, <10 μm for C. vulgaris) on materials ranging from hydrophilic (glass) to hydrophobic (polytetrafluoroethylene). Surface roughness created by micropatterning the surface with 10 μm grooves did not translate into long-term increases in biofilm thickness. The results indicate that species selection and control are more important than surface properties in the development of microalgal biofilms.     

Energy gain and loss during lactation and postweaning in southern elephant seal pups (Mirounga leonina) at King George Island

Deuterium-labeled water was used to measure changes in the proximate body composition during the lactation period and after weaning in southern elephant seal pups at King George Island, South Shetland Islands, Antarctica. During the lactation period (23.0 ± 1.4 days) pups gained a mean of 4.9 ± 0.5 kg/day (n=7). Of the total mass gain (112 ± 8 kg), 38% was water, 48% was fat, and 11% was protein. This represented an increase in total body gross energy of 2437 ± 145 MJ. The proportion of body mass represented by fat was less than 2% at birth, increasing to 35 ± 2% at weaning. We followed the pups during a mean period of 36 ± 3 days after weaning. During this period, pups had a mean loss of 1.21 ± 0.10 kg/day (n=7) comprising 39% water, 48% fat, and 12% protein. The energy cost over this period was 952 ± 168 MJ, which represented, on average, 39% of the total energy gained during the suckling period. Accepted: 3 January 2000     

Hypocalcemia in Pregnant Women

The objective of this study is to estimate the prevalence of hypocalcemia and its correlation with dietary intake of calcium (DICa) and urinary calcium excretion (UCaE) in pregnant women. Healthy pregnant women (n = 543) were enrolled consecutively. DICa was calculated form dietary history. Serum calcium (SCa) and 24-h UCaE was measured. Student t test and Chi-square tests were used to compare the continuous and categorical data in women with and without hypocalcemia (SCa ≤ 8.7 mg/dL). Linear regression was applied for determining the independent variables for hypocalcemia. The age and gestation (mean ± SD) were 21.9 ± 2.5 years and 18.0 ± 3.5 weeks, respectively. The body mass index (BMI; mean ± SD) was 23.3 ± 3.9 kg/m². Seventy-one percent women had an education of less than 10 years. The DICa and SCa were 325 ± 198 mg and 8.1 ± 1.5 mg/dL, respectively. The prevalence of hypocalcemia was 66.4% (362/545); all being asymptomatic. There was no significant difference in women with and without hypocalcemia in terms of weight, BMI, monthly family income, DICa, UCaE, and their obstetric outcome. Daily dietary calcium intake was less than the recommended dietary allowances. There was a high prevalence of asymptomatic hypocalcemia in pregnant women of low socio-economic status which was unrelated to their overall nutritional status and daily calcium intake. It did not have any adverse effect on the immediate pregnancy outcome.     

Decreased trophic position as a function of increasing body size of a benthic omnivorous fish from the largest freshwater lake in China

Potential body size-trophic position relationships of the Darkbarbel catfish Pelteobagrus vachelli (Richardson 1846) were examined using stable isotope analysis. Pelteobagrus vachelli is a benthic feeding fish from Lake Poyang, the largest freshwater lake in China. Two-source mixing model with mussel (Corbicula fluminea) and snail (Bellamya aeruginosa) as baseline primary consumers of planktonic and benthic food webs, respectively, was used to estimate contribution of carbon derived from planktonic vs. benthic food web. Results showed that as an indicator of trophic position, δ¹⁵N was negatively correlated with the body length and weight of the fish; on the other hand, as an indicator of the end-member food sources, δ¹³C was not correlated with fish size. The mixing model results showed that the averaged trophic position of our sampled 3.3–12.7 cm Pelteobagrus vachelli was 3.1 ± 0.2 and derived 68 ± 27% of their food from the benthic food web, confirming that the feeding behavior of the catfish favors benthic food sources.     

Recovery dynamics and viability of the white-tailed eagle (<Emphasis Type="Italic">Haliaeetus albicilla</Emphasis>) in Germany

Understanding the effect of protection measures on recovery of endangered populations is crucial for assessing the efficiency of management plans. Following the ban of DDT, PCB and other detrimental chemicals in the 1970s, the German white-tailed eagle population recovered rapidly. Using nest monitoring data, dead recovery data and population dynamics models, we examined both short and long-term viabilities of the population. Between 1991 and 2005 the proportion of breeding pairs with an active nest among territorial pairs was α = 0.88 ± 0.007. Annual breeding success was γ = 0.62 ± 0.014. Mean fledged brood size was b = 1.50 ± 0.010 fledged/successful breeding pair. The analysis of dead recovery data of 1,273 individuals ringed as nestling between 1991 and 2006 allowed survival rates to be estimated. We needed to distinguish between “young” (5 years old) and “old” (older than 5 years) adults. Furthermore the annual survival rate was higher for juvenile and immature birds (0.921 ± 0.024) than for adults (s _“young” = 0.760 ± 0.097 and s _“old” = 0.875 ± 0.079), a pattern presumably explained by territorial fights. Finally, an age structured model was used to examine the joint effect of population regulation, environmental and demographic stochasticities. All results indicated that only pessimistic scenarios could lead to substantial extinction probabilities within 100 years. We conclude that management measures were successful in removing the main extinction factors of the population and that future conservation efforts need to focus on the remaining ultimate extinction causes.     

Challenges using stable isotopes for estimating trophic levels in marine amphipods

In marine food web studies, stable isotopes of nitrogen (δ¹⁵N) and carbon (δ¹³C) are widely used to estimate organisms’ trophic levels (TL) and carbon sources, respectively. For smaller organisms, whole specimens are commonly analyzed. However, this “bulk method” involves several pitfalls since different tissues may fractionate stable isotopes differently. We compared the δ¹⁵N and δ¹³C values of exoskeleton versus soft tissue, in relation to whole specimens, of three common Arctic amphipods in Svalbard waters: the benthic Anonyx nugax, the sympagic (ice-associated) Gammarus wilkitzkii and the pelagic Themisto libellula. The δ¹⁵N values of the exoskeletons were significantly lower than those of the soft tissues for A. nugax (10.5 ± 0.7‰ vs. 15.7 ± 0.7‰), G. wilkitzkii (3.3 ± 0.3‰ vs. 8.3 ± 0.4‰) and T. libellula (8.6 ± 0.3‰ vs.10.8 ± 0.3‰). The differences in δ¹³C values between exoskeletons and soft tissues were insignificant, except for A. nugax (−21.2 ± 0.2‰ vs. −20.3 ± 0.2‰, respectively). The δ¹⁵N values of whole organisms were between those of the exoskeletons and the soft tissues, being similarly enriched in ¹⁵N as the exoskeletons (except G. wilkitzkii) and depleted in ¹⁵N by 1.2–3.7‰ compared to the soft tissues. The δ¹⁵N-derived TLs of the soft tissues agreed best with the known feeding preferences of the three amphipods, which suggest a potential underestimation of 0.5–1.0 TL when stable isotope analyses are performed on whole crustaceans with thick exoskeletons. The insignificant or small differences in δ¹³C values among exoskeletons, soft tissues and whole specimens, however, suggest low probability for misinterpretations of crustaceans’ primary carbon source in marine ecosystems with distinctly different δ¹³C-carbon sources.     

Display of α-amylase on the surface of <Emphasis Type="Italic">Corynebacterium glutamicum</Emphasis> cells by using NCgl1221 as the anchoring protein,and production of glutamate from starch

We developed a new cell surface display system in Corynebacterium glutamicum based on the C-terminally truncated NCgl1221 anchor protein to increase l-glutamate production from starch directly. The C-terminally truncated NCgl1221 protein is a mutant NCgl1221 and leads to the constitutive export of l-glutamate. The N terminus of α-amylase (AmyA) was fused to truncated NCgl1221, and the resulting fusion protein was expressed on the cell surface by IPTG induction. Localization of the fusion protein was confirmed by immunofluorescence microscopy and flow cytometric analysis. The results of l-glutamate fermentation showed that the soluble starch was utilized to grow and produce l-glutamate by the recombinant strain displaying AmyA. The amount of soluble starch was reduced from 30.0 ± 2.8 to 4.5 ± 0.7 g/l under non-inducing condition and from 50.0 ± 2.4 to 12.5 ± 1.1 g/l under biotin limitation in 36 h. The glutamate concentration in the medium was transiently increased in 14 h under no induction, while under biotin-limiting condition, glutamate production was continuously elevated during fermentation. The amount of glutamate reached 19.3 ± 2.1 g/l after 26 h of fermentation with biotin limitation, which was greater than that produced by the strain using PgsA, one of the poly-γ-glutamate synthetase complexes, as the anchor protein under the same condition. Therefore, the truncated NCgl1221 anchor protein has more advantages than the PgsA anchor protein in glutamate fermentation because truncated NCgl1221 leads to the constitutive export of l-glutamate without any treatments.     

A new manganese superoxide dismutase identified from Beauveria bassiana enhances virulence and stress tolerance when overexpressed in the fungal pathogen

A superoxide dismutase (SOD) was characterized from Beauveria bassiana, a fungal entomopathogen widely applied to insect control. This 209-aa enzyme (BbSod2) showed no more than 71% sequence identity to other fungal Mn-SODs, sharing all conserved residues with the Mn-SOD family and lacking a mitochondrial signal. The SOD activity of purified BbSod2 was significantly elevated by Mn²⁺, suppressed by Cu²⁺ and Zn²⁺ but inhibited by Fe³⁺. Overexpressing the enzyme in a BbSod2-absent B. bassiana strain enhanced its SOD activity (107.2 ± 6.1 U mg⁻¹ protein) by 4–10-fold in different transformants analyzed. The best BbSod2-transformed strain with the SOD activity of 1,157.9 ± 74.7 U mg⁻¹ was 93% and 61% more tolerant to superoxide-generating menadione in both colony growth (EC₅₀ = 2.41 ± 0.03 versus 1.25 ± 0.01 mM) and conidial germination (EC₅₀ = 0.89 ± 0.06 versus 0.55 ± 0.07 mM), and 23% more tolerant to UV-B irradiation (LD₅₀ = 0.49 ± 0.02 versus 0.39 ± 0.01 J cm⁻²). Its virulence to Spodoptera litura larvae was enhanced by 26% [LT₅₀ = 4.5 (4.2–4.8) versus 5.7 (5.2–6.4) days]. Our study highlights for the first time that the Mn²⁺-cofactored, cytosolic BbSod2 contributes significantly to the virulence and stress tolerance of B. bassiana and reveals possible means to improving field persistence and efficacy of a fungal formulation by manipulating the antioxidant enzymes of a candidate strain.