Split-bolus computed tomography urography (CTU) achieves more than half of radiation dose reduction in females and overweight patients than conventional single-bolus computed tomography urography

ObjectiveTo compare radiation dose between single-bolus and split-bolus computed tomography urography (CTU).Materials and methodsWe prospectively enrolled patients undergoing single-bolus and split-bolus CTU from 2019 June to 2020 June. The age, sex and body mass index (BMI) of each patient was recorded and categorized into BMI classes. The radiation dose indices including volumetric computed dose index, size-specific dose estimate, dose length product and effective dose of each patient were compared between 2 CTU groups with calculation of dose reduction proportions (DRPs).ResultsSeventy-six patients underwent single-bolus (n = 39) and split-bolus (n = 37) CTU. Single-bolus CTU had higher radiation doses than split-bolus CTU and there were statistically significant differences of all radiation dose indices between two CTU groups without and with stratification by sex and BMI classes. The DRPs of volumetric computed dose index, size-specific dose estimate, dose length product and effective dose using split-bolus CTU were 49%, 49%. 50%, and 45%, respectively. Multiple linear regression with an effect size (f²) as 2.24 showed females (p = 0.027) and higher BMI classes (p = 2.38 *10⁻⁹) were associated with higher effective doses; and split-bolus CTU, lower effective doses (p = 5.40 *10⁻¹⁵). Using split-bolus CTU, females had consistently higher DRP of all radiation dose indices than males (54–55% versus 40–42%). Overweight patients had the largest DRP as 55% of effective dose.ConclusionsSplit-bolus CTU could be preferred by its significant radiation dose reduction effect in regard to single-bolus CTU, which was most profound in females and overweight patients.     

Spatial phenotyping of the endocardial endothelium as a function of intracardiac hemodynamic shear stress

Despite substantial evidence for the central role of hemodynamic shear stress in the functional integrity of vascular endothelial cells, hemodynamic and molecular regulation of the endocardial endothelium lining the heart chambers remains understudied. We propose that regional differences in intracardiac hemodynamics influence differential endocardial gene expression leading to phenotypic heterogeneity of this cell layer. Measurement of intracardiac hemodynamics was performed using 4-dimensional flow MRI in healthy humans (n=8) and pigs (n=5). Local wall shear stress (WSS) and oscillatory shear indices (OSI) were calculated in three distinct regions of the LV – base, mid-ventricle (midV), and apex. In both the humans and pigs, WSS values were significantly lower in the apex and midV relative to the base. Additionally, both the apex and midV had greater oscillatory shear indices (OSI) than the base. To investigate regional phenotype, endocardial endothelial cells (EEC) were isolated from an additional 8 pigs and RNA sequencing was performed. A false discovery rate of 0.10 identified 1051 differentially expressed genes between the base and apex, and 321 between base and midV. Pathway analyses revealed apical upregulation of genes associated with translation initiation. Furthermore, tissue factor pathway inhibitor (TFPI; mean 50-fold) and prostacyclin synthase (PTGIS; 5-fold), genes prominently associated with antithrombotic protection, were consistently upregulated in LV apex. These spatio-temporal WSS values in defined regions of the left ventricle link local hemodynamics to regional heterogeneity in endocardial gene expression.     

Double Transgenesis of Humanized fat1 and fat2 Genes Promotes Omega-3 Polyunsaturated Fatty Acids Synthesis in a Zebrafish Model

Omega-3 long-chain polyunsaturated fatty acid (n-3 LC-PUFA), especially eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), are essential nutrients for human health. However, vertebrates, including humans, have lost the abilities to synthesize EPA and DHA de novo, majorly due to the genetic absence of delta-12 desaturase and omega-3 desaturase genes. Fishes, especially those naturally growing marine fish, are major dietary source of EPA and DHA. Because of the severe decline of marine fishery and the decrease in n-3 LC-PUFA content of farmed fishes, it is highly necessary to develop alternative sources of n-3 LC-PUFA. In the present study, we utilized transgenic technology to generate n-3 LC-PUFA-rich fish by using zebrafish as an animal model. Firstly, fat1 was proved to function efficiently in fish culture cells, which showed an effective conversion of n-6 PUFA to n-3 PUFA with the n-6/n-3 ratio that decreased from 7.7 to 1.1. Secondly, expression of fat1 in transgenic zebrafish increased the 20:5n-3 and 22:6n-3 contents to 1.8- and 2.4-fold, respectively. Third, co-expression of fat2, a fish codon-optimized delta-12 desaturase gene, and fat1 in fish culture cell significantly promoted n-3 PUFA synthesis with the decreased n-6/n-3 ratio from 7.7 to 0.7. Finally, co-expression of fat1 and fat2 in double transgenic zebrafish increased the 20:5n-3 and 22:6n-3 contents to 1.7- and 2.8-fold, respectively. Overall, we generated two types of transgenic zebrafish rich in endogenous n-3 LC-PUFA, fat1 transgenic zebrafish and fat1/fat2 double transgenic zebrafish. Our results demonstrate that application of transgenic technology of humanized fat1 and fat2 in farmed fishes can largely improve the n-3 LC-PUFA production.     

Ellenberg's indicator in water resources control: The Jizera Mountains,Czech Republic

Ellenberg's indicator values (simple ordinal classes of plants with a similar realized ecological niche along a gradient) were tested to describe the changing environment and water resources recharge in the upper plateau of the Jizera Mountains. (Czech Republic). In the 1980s, the acid atmospheric deposition led to the decline and clear-cut of spruce plantations (Picea abies): Junco effusi-Calamagrostietum villosae became a new dominant community there. Ellenberg's indices for light (L), moisture (F), nitrogen (N) and acidity (R) were compared with hydrological parameters of a small catchment (soil water content, evapotranspiration, retention, stream-flow pH and nitrate content) in 1982–2008. Ellenberg's indicators F and L describe well the plant succession related to microclimate and hydrology at clear-cut sites. Also, the indices N and R follow trends in the atmospheric deposition and water quality; however, with lower sensitivity. Ellenberg's indices reflect the observed changes in water phenomena with a lag period of 3–10 years.     

Synthesis, in vitro antimalarial activities and cytotoxicities of amino-artemisinin-ferrocene derivatives

Novel derivatives bearing a ferrocene attached via a piperazine linker to C-10 of the artemisinin nucleus were prepared from dihydroartemisinin and screened against chloroquine (CQ) sensitive NF54 and CQ resistant K1 and W2 strains of Plasmodium falciparum (Pf) parasites. The overall aim is to imprint oxidant (from the artemisinin) and redox (from the ferrocene) activities. In a preliminary assessment, these compounds were shown to possess activities in the low nM range with the most active being compound 6 with IC₅₀ values of 2.79?nM against Pf K1 and 3.2?nM against Pf W2. Overall the resistance indices indicate that the compounds have a low potential for cross resistance. Cytotoxicities were determined with Hek293 human embryonic kidney cells and activities against proliferating cells were assessed against A375 human malignant melanoma cells. The selectivity indices of the amino-artemisinin ferrocene derivatives indicate there is overall an appreciably higher selectivity towards the malaria parasite than mammalian cells.     

Mutual entrainment of two pacemaker cells. A study with an electronic parallel conductance model

Repetitive firing of pacemaker cells was simulated with the use of an electronic Hodgkin-Huxley-like membrane model having a capacitance, a sodium-, a potassium- and a leakage conductance connected in parallel. The frequency of firing of the model cells was controlled by the leakage conductance. Two such model cells of sometimes widely different intrinsic frequencies were coupled through a coupling resistance R_c. Mutual synchronization was allowed to occur by decreasing R_c in small steps from high to low values.The lower R_c, the more the ratio of the mean frequencies of both cells approaches unity. Around certain R_c values stable entrainment occurs during which the mean frequencies are related as simple integral values m: n (e.g. 2:1, 3:2, 4:3, 1:1 for decreasing R_c). Within the range of a fixed m: n entrainment the phase difference between the oscillations of the two cells declines with R_c. When R_c is close to the range ofm: n entrainment, “almost entrainment” occurs in which m: n entrainment is periodic. But also further away from the range of m: n entrainment periodic variations in action potential interval occur.The model cells were adjusted in such a way that a depolarizing current pulse can only advance the first occurring and all subsequent action potentials. This property is used to explain effects of R_c on the mean frequency of the two cells both within and without the R_c ranges of entrainment. Furthermore, it is illustrated how in certain cases the synchronized frequency at R_c = 0 ω follows from simple electrical considerations of the properties of the uncoupled cells.Several of the entrainment phenomena observed are well known in the fields of electronic relaxation oscillators, clinical electrocardiography and circadian rhythms, though with different terminologies. The present model results exemplify the general nature of these phenomena and clarify entrainment phenomena, recently observed in coupling experiments with pacemaker cells from embryonic heart tissue.     

Prevalence, intensity, and aneuploidy patterns of disseminated neoplasia in cockles (Cerastoderma edule) from Arcachon Bay: Seasonal variation and position in sediment

The present report presents the first evidence of disseminated neoplasia (DN) in cockles Cerastoderma edule from Arcachon Bay (France). Aneuploidy of neoplastic cells allowed the use of flow cytometry (FCM) to diagnose and stage DN. A 1 year survey (2007) of the prevalence and intensity (% of aneuploid circulating cells in neoplastic cockles) was conducted. Prevalences ranged from 2.2% (June) to 13.6% (May), and disease intensity ranged from 18.7% (June) to 95.5% (September). These percentages were not correlated with seawater temperature, but rather showed unexplained oscillations over the year. Prevalence and intensity of DN were higher in cockles found at the surface of sediment compared to those buried normally (11.8% vs. 6.7% and 53.0% vs. 40.6%, respectively, p < 0.05). DN could thus be one mechanism leading to unexplained presence of cockles at the surface of the sediment in Arcachon Bay. Ploidy characteristics of neoplastic cells were also investigated using FCM, revealing an unusual, broad continuum of ploidy distribution from 1.6 to 9.6n. Ploidy values were not in whole numbers in contrast to the rounded values reported in other studies. Ploidy varied according to DN intensity, with the ploidy distribution of neoplastic cells from lightly-diseased cockles being unimodal (3.7n median). In contrast, highly-diseased cockles showed a bimodal ploidy distribution (3.0n and 4.7n medians). This suggests that, in cockles from Arcachon Bay, mechanisms leading to aneuploidy are complex, developing during disease progression.     

Recommendation for the minimum number of steps to analyze when performing the uncontrolled manifold analysis on walking data

The uncontrolled manifold (UCM) analysis quantifies the extent to which co-variation among a set of variables facilitates consistent performance by partitioning variance in those variables into two components then calculating their normalized difference (i.e., the synergy index). Although UCM-derived measures are thought to depend on the number of data points analyzed, the minimum number needed to reasonably approximate true values of these measures is unknown. For each of two performance variables related to mechanical stability of gait, we evaluated changes in UCM-derived measures when increasing the number of analyzed points, here steps. Fourteen older adults walked on a treadmill while motion capture tracked movement. For each subject, n steps (where n = 2–99) were randomly sampled from the first 100, then used to calculate UCM-derived variables. For each subject, variables were expressed as a percent of the subject-specific value with n = 100 and averaged across 50 simulations. For each n, 95% confidence intervals (CIs) were calculated from group data. The minimum number of steps to “reasonably approximate” a variables was defined as the value of n for which the lower CI was >90% of the value with n = 100. Regardless of performance variable, reasonable approximations of the synergy index were attained with n = 16 steps, whereas n = 50 steps were needed for each of the variance components However, the differences between using 16 steps and 50 steps were small. Collecting 15–20 steps is recommended for a reasonable approximation of the synergy indices considered herein, particularly when data collection is constrained to a limited number of steps.     

Sensitivity changes of photoreceptor cells of Hirudo medicinalis caused by changes in extracellular calcium concentration

Extracellular recordings from the vacoule of photoreceptor cells of Hirudo medicinalis L. were performed using microelectrodes. The cells were adapted by white light flashes given at constant intervals (20 s). Response height versus relative intensity curves obtained from the same cell in physiological saline (PS) and in bathing solutions of either a) lowered calcium contents (2 ΜM/1 or less) or b) raised calcium contents (15 mM/1) were compared. The cells' adaptation state in PS was operationally defined by the ratio Q=h _A /h _S where h _A is the response height evoked by the adapting flashes, and h _S is the corresponding saturation response height. Sensitivity changes were measured by the half saturation intensity shift. Lowering extracellular calcium resulted in:

1. The response height increased and the shape of the response became more rounded and prolonged.
2. The total resistance between the vacuole and outside decreased from 8.2±1.4 MΩ (n=6) in PS to 4.6±0.4 MΩ (n=5). The resistance was independent of the cells' adaptation state.
3. A change of the cells' sensitivity occured either in direction to light adaptation or in direction to dark adaptation. It depended functionally on the ratio Q:

a) if Q was less or equal to about 0.6 the cells' sensitivity increased. b) if Q was greater than 0.6 the cells' sensitivity diminished. Raising extracellular calcium decreased the sensitivity of all cells tested independent of their adaptation states in PS. The results can be interpreted under the assumptions that 1. the sensitivity of leech photoreceptor cells is inversely proportional to the intracellular free calcium concentration and Z. intracellular calcium can interact with extracellular calcium in relatively dark adapted cells whereas in relatively light adapted cells the raise of intracellular free calcium is mainly effected by a release from intracellular stores. It is assumed that a Q value of about 0.6 separates relatively light adapted cells from relatively dark adapted cells.     

Fish gut microbiota analysis differentiates physiology and behavior of invasive Asian carp and indigenous American fish

Gut microbiota of invasive Asian silver carp (SVCP) and indigenous planktivorous gizzard shad (GZSD) in Mississippi river basin were compared using 16S rRNA gene pyrosequencing. Analysis of more than 440 000 quality-filtered sequences obtained from the foregut and hindgut of GZSD and SVCP revealed high microbial diversity in these samples. GZSD hindgut (GZSD_H) samples (n=23) with >7000 operational taxonomy units (OTUs) exhibited the highest alpha-diversity indices followed by SVCP foregut (n=15), GZSD foregut (n=9) and SVCP hindgut (SVCP_H) (n=24). UniFrac distance-based non-metric multidimensional scaling (NMDS) analysis showed that the microbiota of GZSD_H and SVCP_H were clearly separated into two clusters: samples in the GZSD cluster were observed to vary by sampling location and samples in the SVCP cluster by sampling date. NMDS further revealed distinct microbial community between foregut to hindgut for individual GZSD and SVCP. Cyanobacteria, Proteobacteria, Actinobacteria and Bacteroidetes were detected as the predominant phyla regardless of fish or gut type. The high abundance of Cyanobacteria observed was possibly supported by their role as the fish''s major food source. Furthermore, unique and shared OTUs and OTUs in each gut type were identified, three OTUs from the order Bacteroidales, the genus Bacillariophyta and the genus Clostridium were found significantly more abundant in GZSD_H (14.9–22.8%) than in SVCP_H (0.13–4.1%) samples. These differences were presumably caused by the differences in the type of food sources including bacteria ingested, the gut morphology and digestion, and the physiological behavior between GZSD and SVCP.     

Effect of polyamines on palmitoyl-coenzyme A-caused inhibition of glucose-6-phosphate dehydrogenase from baker's yeast

Aliphatic polyamines reversed the inhibition of baker's yeast glucose-6-phosphate dehydrogenase by palmitoyl-CoA. The reversal effects of the polyamines on the inhibition were related not only to the number of amino (imino) groups but also to the carbon chain length of polyamine. The relieving constant (K_r) was calculated to determine the absolute concentration of each examined polyamine required to relieve the enzyme from the inhibition. A smaller K_r value indicates a stronger relieving activity. An ethylamine derivative with a large number of amino groups was more effective in reversing the palmitoyl-CoA inhibition. The K_r values of long-chain aliphatic diamines, triamines, and tetramines were smaller than those of short-chain ones. However, n-butylamine and ornithine, composed of four carbons like putrescine, had markedly larger K_r values than putrescine. Substitution of one or two amino groups by carboxyl or hydroxyl groups also appeared to relieve the enzyme from the inhibition. These results suggest that the reversal effect on palmitoyl-CoA inhibition is one of the important roles of polyamines. Furthermore, it is possible that the interaction of palmitoyl-CoA and polyamines physiologically regulates the pentose monophosphate cycle in baker's yeast.     

n-Butylidenephthalide Protects against Dopaminergic Neuron Degeneration and α-Synuclein Accumulation in Caenorhabditis elegans Models of Parkinson's Disease

Background

Parkinson''s disease (PD) is the second most common degenerative disorder of the central nervous system that impairs motor skills and cognitive function. To date, the disease has no effective therapies. The identification of new drugs that provide benefit in arresting the decline seen in PD patients is the focus of much recent study. However, the lengthy time frame for the progression of neurodegeneration in PD increases both the time and cost of examining potential therapeutic compounds in mammalian models. An alternative is to first evaluate the efficacy of compounds in Caenorhabditis elegans models, which reduces examination time from months to days. n-Butylidenephthalide is the naturally-occurring component derived from the chloroform extract of Angelica sinensis. It has been shown to have anti-tumor and anti-inflammatory properties, but no reports have yet described the effects of n-butylidenephthalide on PD. The aim of this study was to assess the potential for n-butylidenephthalide to improve PD in C. elegans models.

Methodology/Principal Findings

In the current study, we employed a pharmacological strain that expresses green fluorescent protein specifically in dopaminergic neurons (BZ555) and a transgenic strain that expresses human α-synuclein in muscle cells (OW13) to investigate the antiparkinsonian activities of n-butylidenephthalide. Our results demonstrate that in PD animal models, n-butylidenephthalide significantly attenuates dopaminergic neuron degeneration induced by 6-hydroxydopamine; reduces α-synuclein accumulation; recovers lipid content, food-sensing behavior, and dopamine levels; and prolongs life-span of 6-hydroxydopamine treatment, thus revealing its potential as a possible antiparkinsonian drug. n-Butylidenephthalide may exert its effects by blocking egl-1 expression to inhibit apoptosis pathways and by raising rpn-6 expression to enhance the activity of proteasomes.

Conclusions/Significance

n-Butylidenephthalide may be one of the effective neuroprotective agents for PD.     

Ultrasound-Microbubble Transplantation of Bone Marrow Stromal Cells Improves Neurological Function after Forebrain Ischemia in Adult Mice

In this study, bone marrow stromal cells (MSCs) were transplanted into the brain of adult rats after forebrain ischemia induced by 4VO. SD rats (n = 60) were randomly divided into three groups: (I) rats (n = 20) were subjected to 4VO and transplanted with MSCs into the ischemic region using ultrasound-microbubble method, (2) rats (n = 20) were subjected to 4VO and transplanted with MSCs into the ischemic region (n = 20), and (3) 4VO alone (n = 20). Rats were sacrificed 28 days after treatment. Neurological functions of rats were evaluated by Morris Water Maze. The current findings suggest that the ultrasound microbubble transplanted MSCs survived in the ischemic brain and significantly improved functional recovery of adult rats compared to regular transplantation.     

DHA status of vegetarians

The aim of the present study was to evaluate the influence of pregnancy in adolescents on the fatty acid composition of the erythrocyte membrane, which was used as a proxy for status of n-3 and n-6 polyunsaturated fatty acids (PUFA), and also on the composition of plasma non-esterified fatty acids (NEFA) mobilized from the adipose tissue. Two matched groups of healthy adolescents (14–19 years) from Rio de Janeiro, Brazil, were compared: pregnant (n=26; 32.7±3.9 weeks of gestation, mean±SD) and non-pregnant (n=20). Blood samples were collected after an overnight fast. Mean dietary intakes of total fat and n-3 and n-6 PUFA (energy %) were not different between pregnant and non-pregnant adolescents, and the consumption of food sources of docosahexaenoic acid (DHA) was low. Fasting total NEFA and NEFA 18:2n-6, 18:3n-6 and 20:4n-6 (g/100 g fatty acids) were higher in pregnant than in non-pregnant adolescents. Although erythrocyte 20:4n-6 was lower in pregnant adolescents, there were no differences in DHA (g/100 g fatty acids), in DHA status indices (22:5n-6/22:4n-6 and 22:6n-3/22:5n-6 ratios) and in the index of n-3+n-6 PUFA status ([Σn-3+Σn-6]/[Σn-7+Σn-9]) in erythrocytes as compared with those of non-pregnant adolescents. In conclusion, pregnancy did not have an adverse effect on erythrocyte DHA content or on DHA and n-3+n-6 PUFA status indices in the adolescents studied.     

Subtyping of a Large Collection of Historical Listeria monocytogenes Strains from Ontario,Canada, by an Improved Multilocus Variable-Number Tandem-Repeat Analysis (MLVA)

Listeria monocytogenes is responsible for severe and often fatal food-borne infections in humans. A collection of 2,421 L. monocytogenes isolates originating from Ontario''s food chain between 1993 and 2010, along with Ontario clinical isolates collected from 2004 to 2010, was characterized using an improved multilocus variable-number tandem-repeat analysis (MLVA). The MLVA method was established based on eight primer pairs targeting seven variable-number tandem-repeat (VNTR) loci in two 4-plex fluorescent PCRs. Diversity indices and amplification rates of the individual VNTR loci ranged from 0.38 to 0.92 and from 0.64 to 0.99, respectively. MLVA types and pulsed-field gel electrophoresis (PFGE) patterns were compared using Comparative Partitions analysis involving 336 clinical and 99 food and environmental isolates. The analysis yielded Simpson''s diversity index values of 0.998 and 0.992 for MLVA and PFGE, respectively, and adjusted Wallace coefficients of 0.318 when MLVA was used as a primary subtyping method and 0.088 when PFGE was a primary typing method. Statistical data analysis using BioNumerics allowed for identification of at least 8 predominant and persistent L. monocytogenes MLVA types in Ontario''s food chain. The MLVA method correctly clustered epidemiologically related outbreak strains and separated unrelated strains in a subset analysis. An MLVA database was established for the 2,421 L. monocytogenes isolates, which allows for comparison of data among historical and new isolates of different sources. The subtyping method coupled with the MLVA database will help in effective monitoring/prevention approaches to identify environmental contamination by pathogenic strains of L. monocytogenes and investigation of outbreaks.     

Prolamin storage proteins and alloploidy in wild populations of the small grass Brachypodium distachyon (L.) P. Beauv.

This paper reports the glutenin diversity of a collection of 23 wild populations of the grass Brachypodium distachyon collected in the Mediterranean and southern areas of the Iberian Peninsula. The plant material studied included the three different cytotypes of this species: 2n?=?10, 2n?=?20 and 2n?=?30. A specific method of extraction was used to isolate the glutenin subunits from the caryopsis. Separation by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) showed them to correspond to wheat low-molecular-weight glutenin subunits (LMW-GS). Twenty-two LMW-GS?Clike were identified that showed great diversity within and between populations. All the populations investigated were polymorphic for the endosperm proteins studied. The 2n?=?30 forms had the largest number of subunits; these were also more diverse than those of the 2n?=?10 or 2n?=?20 forms. The 2n?=?10 forms, the most common in the higher, interior areas of the Iberian Peninsula, showed the smallest subunit variation. In fact, negative correlations were found between subunit diversity and altitude and longitude. In contrast, a positive correlation was detected with the annual average temperature and the indices of thermicity and Mediterraneity. The similarity between the populations was estimated using the Sorensen?CDice coefficient, calculated on the basis of the presence/absence of the 22 LMW-GS proteins. The similarity indices were used to produce a dendrogram using the unweighted pair-group method with arithmetic means (UPGMA). This method produced three main groups corresponding to the three cytotypes. An analysis was made of the possible correlation between eco-geographic and climatic factors and the gene diversity and polymorphism shown in the populations. The diversity correlated positively with the number of chromosomes (2n), annual mean temperature, index of thermicity and index of Mediterraneity. In contrast, diversity correlated negatively with altitude, longitude and index of precipitation during the summer. The number of chromosomes correlated negatively with altitude, longitude and precipitation during summer and positively with all the other climatic indices. These results are coherent with the fact that diploid forms were more common in areas of the interior and at higher altitude, where the climate is more extreme.     

Chromosomes ofLauxaniidae andChamaemyiidae (Diptera)

Chromosome numbers of 28 species ofLauxaniidae range from 2n=8 to 2n=12 with one having 2n=8, three having 2n=10 and 24 having 2n=12. Total complement length averaged 58.1 μ for 71 complements measured. The species studied in detail are classified according to chromosome number and morphology. The two Chamaemyiid species studied have 2n=6 and 2n=15, the latter including several microchromosomes. The karyotypical reorganizations have clearly involved inversions and translocations and a progressive reduction of chromosome numbers.     

Terpenoids inhibit Candida albicans growth by affecting membrane integrity and arrest of cell cycle

Anti-Candida potential of six terpenoids were evaluated in this study against various isolates of Candida albicans (n = 39) and non-C. albicans (n = 9) that are differentially susceptible to fluconazole. All the six terpenoids tested, showed excellent activity and were equally effective against isolates of Candida sps., tested in this study. Linalool and citral were the most effective ones, inhibiting all the isolates at ≤0.064% (v/v). Five among the six terpenoids tested were fungicidal. Time dependent kill curve assay showed that MFCs of linalool and eugenol were highly toxic to C. albicans, killing 99.9% inoculum within seven min of exposure, while that of citronellal, linalyl acetate and citral required 15 min, 1 h and 2 h, respectively. FIC index values (Linalool - 0.140, benzyl benzoate - 0.156, eugenol - 0.265, citral - 0.281 and 0.312 for linalyl acetate and citronellal) and isobologram obtained by checker board assay showed that all the six terpenoids tested exhibit excellent synergistic activity with fluconazole against a fluconazole resistant strain of C. albicans. Terpenoids tested arrested C. albicans cells at different phases of the cell cycle i.e. linalool and LA at G1, citral and citronellal at S phase and benzyl benzoate at G2-M phase and induced apoptosis. Linalool, citral, citronellal and benzyl benzoate caused more than 50% inhibition of germ tube induction at 0.008%, while eugenol and LA required 0.032 and 0.016% (v/v) concentrations, respectively. MICs of all the terpenoids for the C. albicans growth were non toxic to HeLa cells. Terpenoids tested exhibited excellent activity against C. albicans yeast and hyphal form growth at the concentrations that are non toxic to HeLa cells. Terpenoids tested in this study may find use in antifungal chemotherapy, not only as antifungal agents but also as synergistic agents along with conventional drugs like fluconazole.     

Evaluation of reflectance spectroscopy indices for estimation of chlorophyll content in leaves of a tropical tree species

The effectiveness of eight spectral reflectance indices for estimating chlorophyll (Chl) content in leaves of Eugenia uniflora L., a tropical tree species widely distributed throughout the world and a key species for ecosystem restoration projects, was evaluated. Spectral reflectance indices were tested using sun and shade leaves with a broad variation in leaf mass per area (LMA). Shortly after plants were exposed to chilling temperatures, there was a dramatic visible change in some sun leaves from green to red. Prior to testing Chl-related reflectance indices, the green and red leaves were separated according to the anthocyanin reflectance index (ARI). Slightly green to dark green leaves corresponded to an ARI value less than 0.11 (n = 107), whereas slightly red to red leaves corresponded to an ARI value greater than 0.11 (n = 35). To estimate leaf Chl, two simple reflectance indices (SR₆₈₀ and SR₇₀₅), two normalized difference indices (ND₆₈₀ and ND₇₀₅), two modified reflectance indices (mSR₇₀₅ and mND705), a modified Chl absorption ratio index (mCARI₇₀₅) and an index insensitive to the presence of anthocyanins (CI_re) were evaluated. Good estimates of leaf Chl content were obtained using the reflectance indices tested regardless of the presence of anthocyanins and changes in LMA. Based on the coefficients of determination (r ²) and the root mean square errors (RMS?_c) the best results were obtained with reflectance indices measured at wavelengths of 750 and 705 nm. Considering the performance of the models the best reflectance indices to estimate Chl contents in E. uniflora leaves with a broad variation in LMA and anthocyanin contents was SR₇₀₅ and mCARI₇₀₅.