First report of Alternaria alternata causing leaf spot on Oat (Avena sativa) in India.

Oat (Avena sativa L.) is an important cereal crop grown worldwide primarily for food and animal feed. In November 2019, a leaf spot disease was observed on the oat plants at Mandya (12.5218° N, 76.8951° E), Karnataka, India. The disease incidence on plants was ranged between 43 to 57 percent. Initially, the symptoms appeared on leaves as small dark-brown spots surrounded by a yellow halo later turned to irregular necrotic spots with a yellow halo. A total of thirty leaves showing typical symptoms were collected from ten plants (three leaves per plant), cut into an area of 4-5 mm pieces at the junction of infected and healthy tissues. Cut tissues were soaked in 75% ethanol for 45 seconds, followed by 1% sodium hypochlorite solution for 1 min, rinsed five times in sterile distilled water, air dried, and placed on PDA and incubated at 25 ± 1 ℃. After 7 days of incubation, greyish fungal colonies appeared on PDA. Single-spore isolation method was employed to recover the pure cultures for five isolates. The colonies initially produced light-greyish aerial mycelia, then turned to dark-greyish upon maturity. Conidia were obclavate to pyriform and measured 17.34 to 46.97 µm long, 5.38 to 14.31 µm wide with 0 to 3 longitudinal, and 1 to 6 transverse septa with short beak (2.73 to 10.17µm) (n = 50.) Based on the morpho-cultural characteristics, the isolates were identified as Alternaria spp., and PCR assay using species-specific primers (AAF2/AAR3; Konstantinova, et al. 2002) confirmed the taxonomic identity of all five isolates as A. alternata. To further confirm the identity, the internal transcribed spacer (ITS), small subunit (SSU), glyceraldehyde-3-phosphate dehydrogenase (gapdh), RNA polymerase second largest subunit (rpb2), Alternaria major allergen (Alt a1), endopolygalacturonase (endoPG), an anonymous gene region OPA10-2, KOG1058 and translation elongation factor 1-alpha (tef1) of two isolates MAAS-1 and MAAS-2 were PCR amplified using the primers described previously (Woudenberg et al. 2015; Praveen et al. 2020) and the resultant PCR products were sequenced and deposited in NCBI GenBank (ITS: MW487388, MW741962, SSU: MW506220, MW752854, gapdh: MW506221, MW752855, rpb2: MW506222, MW752856, Alt a1: MW506223, MW752857, endoPG: MW506224, MW752858, OPA10-2: MW506225, MW752859, KOG1058: MW506226, MW752860, and tef1: MW506227, MW752861) which showed (99.62%, 99.81%), (100%, 100%), (100%, 99.66%), (100%, 100%), (99.58%, 99.15%), (99.55%, 99.32%), (99.53%, 99.68%), (99.23%, 99.56%) and (99.17%, 99.58%) identity with ITS (AF347031), SSU (KC584507), gapdh (AY278808), rpb2 (KC584375), Alt a1 (AY563301), endoPG (JQ811978), OPA10-2 (KP124632), KOG1058 (KP125233) and tef1 (KC584634) genes/genomic regions of type strain CBS916.96 of A. alternata respectively, confirming the identity of MAAS-1 as A. alternata. For pathogenicity assay, the conidial suspension (2 × 106 conidia/ml) of MAAS-1 isolate was artificially sprayed until runoff on ten healthy oat plants (cv. Kent, 35 days old) and ten plants sprayed with sterile water served as control. All plants were covered with polyethylene covers and kept under the greenhouse at 28 ± 1 ℃. The pathogenicity assay was repeated three times. After six days post-inoculation, small dark-brown spots with light-yellow halo appeared on leaves of MAAS-1inoculated plants. In comparison, no symptoms were observed on control plants. The fungal pathogen was re-isolated from the artificially infected plants and confirmed as A. alternata based on morpho-cultural characteristics and PCR assays. The leaf spot disease of Oat caused by A. alternata has already been reported from China (Chen et al. 2020); to our knowledge, it is the first report of A. alternata causing leaf spot on Oat in India. The leaf spot disease is an emerging threat to oat cultivation, and it reduces the grain yield and leaf quality; therefore, its management is essential for increasing productivity.

Acid digestion of carbonates using break seal method for clumped isotope analysis.

RATIONALE: Acid digestion of carbonates to release CO2 is a crucial and sensitive step in sample preparation for clumped isotope analysis. In addition to data reduction and instrumental artefacts, many other uncertainties in the clumped isotope analysis of carbonates arise from the method used for the preparation of CO2 . We describe here an in-house-designed reaction vessel that circumvents degassing and contamination problems commonly associated with the McCrea-type digestion protocols. METHODS: We designed a leak-free break seal reaction vessel (made of Pyrex™) suitable for prolonged acid digestion at 25°C. Using this new vessel, several carbonate reference materials widely used in the clumped isotope community and other in-house laboratory standards were acid-digested and analysed for their δ13 C, δ18 O and Δ47 values with a dual inlet MAT 253 isotope ratio mass spectrometer following standard gas chromatography purification and data evaluation protocols. RESULTS: Long-term reproducibility in Δ47 determination was established using international references and in-house working standards as follows (mean and SE): Carrara-1 (0.395 ± 0.002‰, n = 43), Carrara-2 (0.441 ± 0.003‰, n = 22), OMC (0.587 ± 0.004‰, n = 16), NBS 19 (0.393 ± 0.005‰, n = 10), NBS 18 (0.473 ± 0.003‰, n = 5), ETH 1 (0.271 ± 0.005‰, n = 7), ETH 3 (0.698 ± 0.005‰, n = 3), MZ (0.715 ± 0.002‰, n = 3) and several others. CONCLUSIONS: A new method using a break seal tube was found to be efficient for the clumped isotope analysis of carbonates that require longer reaction time at 25°C. This method yields good precision in Δ47 analysis and was found to be suitable for acid digestions at any desired temperature.

Foxp3 promoter polymorphism (rs3761548) in breast cancer progression: a study from India.

Breast cancer is the most common female neoplasm that drives the transformation of normal mammary epithelial cells into highly malignant derivatives. Forkhead Box Protein3 (Foxp3), a tumor suppressor/immunomodulatory gene, which controls the function of Treg cells and oncogenes is down regulated in breast cancer. The main aim of the present study is to evaluate the potential influence of Foxp3-3279 C>A polymorphism (rs3761548) and -2383 C>T polymorphism (rs3761549) in 202 breast cancer patients and 130 normal healthy women of Indian origin. The genotypes were determined using ARMS-PCR for rs3761548 and PCR-RFLP method for rs3761549 using specific primers. The results revealed lack of association of these two polymorphisms with breast cancer susceptibility. However, with respect to AA genotype of rs3761548, we found highly significant association with the advanced stage (T3-4) of the tumor (OR = 3.90; 95% confidence interval (CI) = 1.56-9.70; p = 0.03). Stratified data also revealed an association of homozygous mutant genotype with advanced stage of tumor in premenopausal women (OR = 4.56; 95% CI = 1.07-19.38; p = 0.04) with disease duration of <6 months (OR = .10; 95% CI = 1.80-20.50; p = 0.002) suggestive of modulating effect of rs3761548 in tumor progression. We conclude that Foxp3 rs37161548 has a potential to be a polymorphic marker for tumor progression in premenopausal breast cancer patients in Indian women.

CYP11A1 microsatellite (tttta)n polymorphism in PCOS women from South India.

BACKGROUND: Polycystic ovary syndrome (PCOS) is a condition with central feature of hyperandrogensism that affects 5-12 % of women worldwide. P450sec the cholesterol side chain cleavage enzyme encoded by CYP11A1 gene is instrumental in the synthesis of sex hormones. A promoter pentanucleotide repeat (tttta)(n) polymorphism of this gene is reported to be associated with several hormone related diseases including PCOS. Here we aimed to examine the involvement of CYP11A1 polymorphism with PCOS susceptibility in a case-control study conducted among South Indian women. METHODS: A total of 542 subjects comprised of 267 PCOS patients and 275 controls were recruited. DNA was extracted from blood and CYP11A1 (tttta)(n) polymorphism was genotyped by PCR-PAGE. RESULTS: Fifteen different alleles ranging between 2-16 repeats were identified in the studied group and the most frequent allele observed in controls was of 8 repeats. The presence of >8 repeat allele was common in patients (64 % vs. 38 %) and showed a three-fold risk for PCOS susceptibility than controls (OR = 2.93; p < 0.05). PCOS women with higher BMI were markedly elevated in early quartile (p < 0.05). CONCLUSION: CYP11A1 (tttta)(n) repeat polymorphism appeared to be a potential molecular marker for PCOS risk in our population. Gene-gene and gene-environmental interactions with respect to obesity may play a role in the early onset of this multifactorial condition. This is the first report from South India; however, replicative studies considering other probable causative factors for PCOS risk are warranted.

Comparative regulatory network of transcripts behind radicle emergence and seedling stage of maize (Zea mays L.).

The transition from radicle emergence to seedling growth in maize is a crucial phase in the plant's life cycle, where rapid physiological and biochemical changes occur to facilitate successful development. In this study, we conducted a comparative transcriptomic analysis to gain a deeper understanding of the molecular processes driving this critical transition. The early divergence in gene expression patterns highlighted the upregulation of a substantial number of genes during radicle emergence. During radicle emergence, gene ontology (GO) term enrichment analysis unveiled active participation in biological processes such as chromatin assembly, cellular response to abiotic stress, and hormone signaling. This indicates that the initial stages of growth are marked by cellular expansion and adaptation to environmental stimuli. Conversely, in the seedling growth stage, GO analysis demonstrated a shift toward processes such as photosynthesis, nitrogen metabolism, and secondary metabolite biosynthesis, reflecting a transition to energy production and enhanced growth. In contrast, seedling growth was characterized by pathways related to photosynthesis and the production of gibberellins, crucial for robust seedling development. Hormonal regulation and starch metabolism were also prominent during radicle emergence, with various hormones, including auxins, diterpenoids, and brassinosteroids, driving processes like cell enlargement and stem growth. Moreover, starch and sucrose metabolism genes were expressed to mobilize stored reserves for energy during this stage. These findings offer valuable insights into the dynamic regulation of genes and pathways during this critical phase of maize development.

Do ACE I/D gene polymorphism serve as a predictive marker for age at onset in PCOS?

BACKGROUND: Polycystic ovary syndrome (PCOS) is an endocrine disorder exhibiting variable age at onset of clinical features allied with complex diseases in the later life. ACE is a pleiotropic molecule associated with various pathophysiological functions. The present study was aimed to establish the frequency of ACE I/D gene polymorphism in patients and controls and to assess the influence of this polymorphism on anthropometric and various clinical features of the condition. METHODS: ACE I/D genotyping was carried out in 259 PCOS patients and 315 healthy ultrasound scanned women of South Indian origin. RESULTS: The distribution of DD, ID and II genotypes in patients was 39, 37 and 24 %, whereas in the controls it was 31, 51 and 18 % respectively. Significant difference was observed in the genotypic frequency distributions between the patients and controls, however the allelic frequencies did not vary between the groups (p>0.05). Quartile analysis revealed preponderance of DD genotype in the first two quartiles and a linear increase of II genotype from first to the last quartiles. Further, Multiple Logistic regression analysis revealed significant association of ACE I/D gene polymorphism with acanthosis and age at onset (AAO) of the syndrome (p<0.05). CONCLUSION: The present study is the first report to highlight the predisposing role of DD and protective role of ID genotype towards PCOS. Patients with single or double dose of D allele may develop PCOS symptoms at an early age and also significantly associated with acanthosis, a marker of insulin resistance.

TNF-α haplotype association with polycystic ovary syndrome - a South Indian study.

BACKGROUND: Tumor Necrosis Factor Alpha (TNF-α), is a proinflammatory cytokine in the pathogenesis of Polycystic Ovary Syndrome (PCOS). In order to investigate the role of rs1800629 and rs1799964 polymorphisms in relation to anthropometric measures, family history of complex diseases, diet and clinical features, we performed a case control study in PCOS women from South India. METHODS: A total of 589 samples comprising of 283 patients and 306 controls were enrolled in the present study. Patients were selected based on Rotterdam criteria and ultrasound scanned normal women were selected as controls. Following extraction of DNA, genotyping for rs1800629 and rs1799964 was performed by polymerase chain reaction using tetra primers and PCR-RFLP respectively. RESULTS: The distribution of genotypes for rs1799964 was significantly different between the groups (p = 0.001), however it was not for rs1800629. Haplotype analysis revealed a significant difference between patients and controls. The predisposing and protective role of haplotype with mutant allele at both loci (combination 3) and haplotype with mutant allele at either loci was reflected by the over representation of combination 3 in patients and combination 2 in controls respectively. In addition, rs1799964 showed an association with dietary habit, clinical hyperandrogenism and AAO. The modifying role of TT genotype on age at onset was noted in quartile analysis. CONCLUSION: Replicative studies on the influence of TNF-α polymorphism in different ethnic groups may identify the potentiality of these polymorphisms as markers of inflammation and in turn may help the clinicians for the better management of the condition.

Seed nutritional quality in lentil (Lens culinaris) under different moisture regimes.

The world's most challenging environmental issue is climate change. Agricultural productivity and nutritional quality are both substantially threatened by extreme and unpredicted climate events. To develop climate resilient cultivars, stress tolerance along with the grain quality needs to be prioritized. Present study was planned to assess the effect of water limitation on seed quality in lentil, a cool season legume crop. A pot experiment was carried out with 20 diverse lentil genotypes grown under normal (80% field capacity) and limited (25% field capacity) soil moisture. Seed protein, Fe, Zn, phytate, protein and yield were recorded in both the conditions. Seed yield and weight were reduced by 38.9 and 12.1%, respectively, in response to stress. Seed protein, Fe, Zn, its availability as well as antioxidant properties also reduced considerably, while genotype dependent variation was noted with respect to seed size traits. Positive correlation was observed between seed yield and antioxidant activity, seed weight and Zn content and availability in stress. Based on principal component analysis and clustering, IG129185, IC559845, IC599829, IC282863, IC361417, IG334, IC560037, P8114 and L5126 were promising genotypes for seed size, Fe and protein content, while, FLIP-96-51, P3211 and IC398019 were promising for yield, Zn and antioxidant capacity. Identified lentil genotypes can be utilized as trait donors for quality improvement in lentil breeding.

The interferon stimulated gene 54 promotes apoptosis.

The ability of interferons (IFNs) to inhibit viral replication and cellular proliferation is well established, but the specific contribution of each IFN-stimulated gene (ISG) to these biological responses remains to be completely understood. In this report we demonstrate that ISG54, also known as IFN-induced protein with tetratricopeptide repeats 2 (IFIT2), is a mediator of apoptosis. Expression of ISG54, independent of IFN stimulation, elicits apoptotic cell death. Cell death and apoptosis were quantified by propidium iodide uptake and annexin-V staining, respectively. The activation of caspase-3, a key mediator of the execution phase of apoptosis, was clearly apparent in cells expressing ISG54. The anti-apoptotic B cell lymphoma-xl (Bcl-xl) protein inhibited the apoptotic effects of ISG54 as did the anti-apoptotic adenoviral E1B-19K protein. In addition, ISG54 was not able to promote cell death in the absence of pro-apoptotic Bcl family members, Bax and Bak. Analyses of binding partners of ISG54 revealed association with two homologous proteins, ISG56/IFIT1 and ISG60/IFIT3. In addition, ISG60 binding negatively regulates the apoptotic effects of ISG54. The results reveal a previously unidentified role of ISG54 in the induction of apoptosis via a mitochondrial pathway and shed new light on the mechanism by which IFN elicits anti-viral and anti-cancer effects.

Fractured T Tube Fragment in Common Bile Duct during Retrieval: An Unforeseen Mishap.

Common bile duct (CBD) exploration by surgical method-open or laparoscopic, traditionally involved using a T tube to take care of postoperative intraluminal pressure and edema. The complications of T tube include bile leak after removal, formation of biliary fistula, excoriation of the skin, dehydration, saline depletion, retained T tube fragment, CBD obstruction, cholangitis, pancreatitis, and duodenal erosion. Here, we report a case of retained T tube fragment after an attempted removal in an operated case of choledocholithiasis, which was managed by endoscopic retrograde cholangiopancreatography and balloon catheter removal of the remnant.

Root Trait Variation in Lentil (Lens culinaris Medikus) Germplasm under Drought Stress.

Drought is the most critical environmental factor across the continents affecting food security. Roots are the prime organs for water and nutrient uptake. Fine tuning between water uptake, efficient use and loss determines the genotypic response to water limitations. Targeted breeding for root system architecture needs to be explored to improve water use efficiency in legumes. Hence, the present study was designed to explore root system architecture in lentil germplasm in response to drought. A set of 119 lentil (Lens culinaris Medik.) genotypes was screened in controlled conditions to assess the variability in root traits in relation to drought tolerance at seedling stage. We reported significant variation for different root traits in lentil germplasm. Total root length, surface area, root volume and root diameter were correlated to the survival and growth under drought. Among the studied genotypes, the stress tolerance index varied 0.19-1.0 for survival and 0.09-0.90 for biomass. Based on seedling survival and biomass under control and drought conditions, 11 drought tolerant genotypes were identified, which may be investigated further at a physiological and molecular level for the identification of the genes involved in drought tolerance. Identified lines may also be utilised in a lentil breeding program.

Cross-Talk Signaling in Rice During Combined Drought and Bacterial Blight Stress.

Due to climatic changes, rice crop is affected by moisture deficit stress and pathogens. Tissue water limitation besides reducing growth rates, also renders the crop susceptible to the infection by Xanthomonas oryzae pv. oryzae (Xoo) that causes bacterial leaf blight. Independently, both drought adaptation and Xoo resistance have been extensively studied. Though the cross-talk between drought and Xoo stress responses have been explored from individual stress studies, examining the combinatorial stress response is limited in rice. Recently published combined stress studies showed that under the combined stress, maintenance of carbon assimilation is hindered and such response is regulated by overlapping cellular mechanisms that are different from either of the individual stresses. Several receptors, MAP kinases, transcription factors, and ribosomal proteins, are predicted for playing a role in cellular homeostasis and protects cells from combined stress effects. Here we provide a critical analysis of these aspects using information from the recently published combined stress literature. This review is useful for researchers to comprehend combinatorial stress response of rice plants to drought and Xoo.

Purification and characterization of a novel protease from the latex of Pedilanthus tithymaloids.

A novel protease was purified to homogeneity from the latex of Pedilanthus tithymaloids by a simple purification procedure involving ammonium sulfate precipitation and cation-exchange chromatography. The molecular weight of the protease was estimated to be approximately 63.1 kDa and the extinction coefficient (epsilon(1%)(280nm)) was 28.4. The enzyme hydrolyzes denatured natural substrates like casein, azoalbumin and azocasein with a high specific activity but little activity towards synthetic substrates. The pH and temperature optima were pH 8.0-9.5 and 65-70 degrees C, respectively. The proteolytic activity of the enzyme was inhibited by different protease-specific inhibitors (e.g., thiol, serine, metallo, etc.) up to a certain extent but not completely by any class of inhibitors. The enzyme was relatively stable towards pH change, temperature, denaturants and organic solvents. The enzyme consists of five disulfide bridges compared to three observed in most plant cysteine proteases. Overall, the striking features of this protease are its high molecular weight, high cysteine content and only partial inhibition of activity by different classes of protease inhibitors contrary to known proteases from other plant sources. The enzyme is named as pedilanthin as per the protease nomenclature.

Analysis of oxidative stress status through MN test and serum MDA levels in PCOS women.

Polycystic Ovary Syndrome (PCOS) is a multifactorial reproductive healthcare problem affecting 4-12% of women and a leading cause of female infertility worldwide. The potential genetic contributors of PCOS are unclear. However, over the past decade emerging evidence has shown that increased Oxidative Stress (OS) and decreased antioxidant status were often linked with PCOS. The present case-control study was aimed to assess the reactive oxygen species induced OS in women from South India. A total of 164 individuals comprising of 89 patients and 75 controls were enrolled in the present study. For all the subjects, the frequency of micronucleated cells (MNC) in epithelial samples and serum Malondialdehyde (MDA) levels were estimated to assess genomic instability and cytotoxicity respectively. A statistically significant difference between the groups were identified with respect to Body Mass Index, Waist to Hip Ratio, luteinizing hormone and prolactin levels (< 0.05), however the mean follicle stimulating hormone was not different between the groups (p = 0.055). The frequency of MN cells (5.89 ± 4.86 vs. 2.24 ± 2.01) and mean serum MDA (360.84 ± 87.08 vs. 301.70 ± 82.82) levels were considerably higher in patients than controls (p = < 0.0001), furthermore, a positive correlation was observed between MNC and MDA levels in patients (r = 0.349, p = 0.0008) and not in controls (r = 0.104, p = 0.37), suggest high OS in PCOS women. Therefore, MN assay and serum MDA levels may serve together or individually as biomarkers of OS in PCOS women.

Investigation of the ex vivo and in vivo iontophoretic delivery of aceclofenac from topical gels in Albino rats.

INTRODUCTION: Iontophoresis was used to enhance the delivery of aceclofenac (ACF) from topical gels formulated with various polymers for the purpose of relieving pain and inflammation. MATERIALS AND METHODS: Gels were formulated from hydroxypropyl methyl cellulose (HPMC), carbopol 934P, and sodium carboxymethyl cellulose (NaCMC). The formulations were evaluated for cathodal iontophoretic delivery of ACF through excised rat abdominal skin at three levels of current density of 0.5, 0.6 and 0.7 mA/cm(2). The in vivo effectiveness of the drug delivered passively as well as under the influence of iontophoresis at pH 7.4 at a current density of 0.5 mA/cm(2) was also investigated using male Albino rats with carrageenan induced paw edema. RESULTS AND DISCUSSION: In the ex vivo studies, though it was clear that iontophoresis significantly increased drug permeation through the excised skin from all formulations; the percentage drug permeated from HPMC gels was superior to that from carbopol 934P or NaCMC gels but increased with an increase in the current density only for the former. The steady state flux, permeability coefficient, enhancement factor were significantly greater from HPMC gels than from the gels of the ionic polymers due to the interference of competitive ions. With iontophoresis, the carrageenan induced paw edema was significantly reduced by 61.53% (P < 0.01) for HPMC gels as compared to the control although passive permeation without iontophoresis showed a 54.6% reduction (P < 0.05) at the end of 4 h. CONCLUSION: The results of the study indicate that ACF could be administered topically by using iontophoresis from a suitably formulated gel for effective control of pain and inflammation.

Histoid leprosy - unusual presentation.

A 39-year-old man presented with a chronic nonhealing toe ulcer and multiple skin-colored papules on the back, arms, and knees. From an initial small erosion, the toe lesion ulcerated over a 1.5-year period, while the papules progressed over a 6-month period, first appearing on the back and then spreading to the arms and knees. The past medical and family history were non-contributory. Pertinent findings included the aforementioned well-defined, asymmetric, shiny papules (Fig. 1). There were multiple, discrete, erythematous, pedunculated, tumor-like masses of various sizes over the right thigh (Fig. 2) and a well-defined 4 cm x 3 cm ulcer on the second toe of the left foot, the floor of which was covered by necrotic slough. There was distal loss of sensation to temperature, touch, and pain. The greater auricular, ulnar, radial cutaneous and common peroneal nerves were thickened bilaterally, but non-tender. A clinical diagnosis of histoid leprosy was made. The differential diagnosis for the tumor-like thigh masses included dermatofibroma, neurofibroma, and Kaposi's sarcoma. The hemogram, liver/renal function tests, chest X-ray, and abdominal ultrasound were normal. Human immunodeficiency virus enzyme-linked immunosorbent assay (HIV ELISA) was negative. The ear lobe smear (ELS) for acid-fast bacilli showed a bacterial index (BI) of 6+[> 1000 organisms/oil immersion field (oif)] and a morphological index (MI) of 50%. The skin-colored papules on the back and the pedunculated masses showed a BI of 5+ (100-1,000 organisms/oif) and an MI of 50%. The normal skin showed a BI of 4+ (10-100 organisms/oif) and an MI of 5%. Biopsy of the papules and pedunculated tumors showed a well-circumscribed area of the dermis packed with many acid-fast organisms and foamy macrophages, consistent with histoid leprosy (Fig. 3). Fite-Faraco stain demonstrated cells packed with lepra bacilli. A final diagnosis of lepromatous leprosy, histoid variant, was made. Dapsone, clofazimine, rifampicin, and, later, ofloxacin were started.

Differential activation of IFN regulatory factor (IRF)-3 and IRF-5 transcription factors during viral infection.

Members of the IFN regulatory factor (IRF) family regulate gene expression critical to immune response, hemopoiesis, and proliferation. Although related by homology at their N-terminal DNA-binding domain, they display individual functional properties. The distinct properties result from differences in regulated expression, response to activating signals, and interaction with DNA regulatory elements. IRF-3 is expressed ubiquitously and is activated by serine phosphorylation in response to viral infection or TLR signaling. Evidence indicates that the kinases TANK-binding kinase 1 and inhibitor of NF-kappaB kinase-epsilon specifically phosphorylate and thereby activate IRF-3. We evaluated the contribution of another member of the IRF family, IRF-5, during viral infection since prior studies provided varied results. Analysis of phosphorylation, nuclear translocation, dimerization, binding to CREB-binding protein, recognition of DNA, and induction of gene expression were used comparatively with IRF-3 as a measure of IRF-5 activation. IRF-5 was not activated by viral infection; however, expression of TANK-binding kinase 1 or inhibitor of NF-kappaB kinase-epsilon did provide clear activation of IRF-5. IRF-5 is therefore distinct in its activation profile from IRF-3. However, similar to the biological effects of IRF-3 activation, a constitutively active mutation of IRF-5 promoted apoptosis. The apoptosis was inhibited by expression of Bcl-x(L) but not a dominant-negative mutation of the Fas-associated death domain. These studies support the distinct activation profiles of IRF-3 in comparison to IRF-5, but reveal a potential shared biological effect.

In situ EPR spectroscopy of aromatic diyne cyclopolymerization.

Electron paramagnetic resonance (EPR) spectroscopy was successfully used for the first time to follow the Bergman cyclization of bis-ortho-diynyl arene (BODA) compounds. Five BODA monomers with different spacer (X) and terminal groups (R) were compared. In situ polymerization via EPR spectroscopy yielded first-order rate expressions. Monomers with spacer -O- or -C(CF(3))(2) and terminal group R = Ph exhibited similar kinetic behavior upon thermal polymerization, whereas monomers with pyridine and thiophene terminal groups gave significantly higher rates of polymerization over phenyl-terminated derivatives. A model compound, 1,2-bis(phenylethynyl)benzene, was used to probe the polymerization mechanism, and radical intermediates were found to be stable indefinitely at room temperature.

Three bis-ortho-diynylarenes (BODA).

The molecule of 3,3',4,4'-tetrakis(phenylethynyl)biphenyl, C(44)H(26), is approximately planar and is located on a crystallographic inversion center. Bis[3,4-bis(phenylethynyl)phenyl] ether, C(44)H(26)O, has molecules located on twofold symmetry axes, whereas the molecule of 2,2-bis[3,4-bis(phenylethynyl)phenyl]propane, C(47)H(32), does not exhibit any molecular symmetry.

Amido-modified polylactide for potential tissue engineering applications.

Poly(ester amide) copolymers based on L-lactide (2) and a new depsipeptide (1) were prepared by ring opening polymerization in the presence of Sn(Oct)2 as the catalyst. Variable monomer feed ratios up to 2.3 mol% 1 afforded copolymers containing ester and amido functional groups in the backbone. Lower glass transition temperatures and reduced crystallization kinetics and crystallinity compared to homo-polylactide (PLA) was achieved with low levels of amido incorporation. A reactivity comparison between enchainment of 2 and 1 was determined using in situ infrared spectroscopy. An increase in shear viscosity was observed with the increase of 1 content as determined by rheology studies. Cellular compatibility of the co-polymers was investigated by seeding D1 mouse stem cells onto films and characterizing cell morphology by optical microscopy. Preliminary results indicate that these novel materials exhibit reduced cell attachment compared to PLA and, pending further exploration, may have potential use in biomedical applications.