The region-dependent biphasic viscoelastic properties of human temporomandibular joint discs under confined compression

The objective of this study was to determine the biphasic viscoelastic properties of human temporomandibular joint (TMJ) discs, correlate these properties with disc biochemical composition, and examine the relationship between these properties and disc dynamic behavior in confined compression. The equilibrium aggregate modulus (H_A), hydraulic permeability (k), and dynamic modulus were examined between five disc regions. Biochemical assays were conducted to quantify the amount of water, collagen, and glycosaminoglycan (GAG) content in each region. The creep tests showed that the average equilibrium moduli of the intermediate, lateral, and medial regions were significantly higher than for the anterior and posterior regions (69.75±11.47 kPa compared to 22.0±5.15 kPa). Permeability showed the inverse trend with the largest values in the anterior and posterior regions (8.51±1.36×10^?15 m⁴/Ns compared with 3.75±0.72×10^?15 m⁴/Ns). Discs were 74.5% water by wet weight, 62% collagen, and 3.2% GAG by dry weight. Regional variations were only observed for water content which likely results in the regional variation in biphasic mechanical properties. The dynamic modulus of samples during confined compression is related to the aggregate modulus and hydraulic permeability of the tissue. The anterior and posterior regions displayed lower complex moduli over all frequencies (0.01–3 Hz) with average moduli of 171.8–609.3 kPa compared with 454.6–1613.0 kPa for the 3 central regions. The region of the TMJ disc with higher aggregate modulus and lower permeability had higher dynamic modulus. Our results suggested that fluid pressurization plays a significant role in the load support of the TMJ disc under dynamic loading conditions.     

Evidence against proteoglycan mediated collagen fibril load transmission and dynamic viscoelasticity in tendon

The glycosaminoglycan (GAG) dermatan sulfate and chondroitin sulfate side-chains of small leucine-rich proteoglycans have been increasingly posited to act as molecular cross links between adjacent collagen fibrils and to directly contribute to tendon elasticity. GAGs have also been implicated in tendon viscoelasticity, supposedly affecting frictional loss during elongation or fluid flow through the extra cellular matrix. The current study sought to systematically test these theories of tendon structure–function by investigating the mechanical repercussions of enzymatic depletion of GAG complexes by chondroitinase ABC in a reproducible tendon structure–function model (rat tail tendon fascicles). The extent of GAG removal (at least 93%) was verified by relevant spectrophotometric assays and transmission electron microscopy. Dynamic viscoelastic tensile tests on GAG depleted rat tail tendon fascicle were not mechanically different from controls in storage modulus (elastic behavior) over a wide range of strain-rates (0.05, 0.5, and 5% change in length per second) in either the linear or nonlinear regions of the material curve. Loss modulus (viscoelastic behavior) was only affected in the nonlinear region at the highest strain-rate, and even this effect was marginal (19% increased loss modulus, p = 0.035). Thus glycosaminoglycan chains of small leucine-rich proteoglycans do not appear to mediate dynamic elastic behavior nor do they appear to regulate the dynamic viscoelastic properties in rat tail tendon fascicles.     

Synthesis and biology of oligoethylene glycol linked naphthoxylosides

Proteoglycans (PGs) are important macromolecules in mammalian cells, consisting of a core protein substituted with carbohydrate chains, known as glycosaminoglycans (GAGs). Simple xylosides carrying hydrophobic aglycons can enter cells and act as primers for GAG chain synthesis, independent of the core protein. Previously it has been shown that aromatic aglycons can be separated from the sugar residue by short linkers without affecting the GAG priming ability. To further investigate the effects of the xylose–aglycon distance on the GAG priming ability, we have synthesized xyloside derivatives with 2-naphthyl and 2-(6-hydroxynaphthyl) moieties connected to xylose, directly, via a methylene bridge, or with oligoethylene glycol linkers of three different lengths. The GAG priming ability and the antiproliferative activity of the xylosides, as well as the composition of the xyloside-primed GAG chains were investigated in a matched pair of human breast fibroblasts and human breast carcinoma cells. An increase of the xylose–aglycon distance from 0.24 to 0.37 nm resulted in an increased GAG priming ability in both cell lines. Further increase of the xylose–aglycon distance did not result in any pronounced effects. We speculate that by increasing the xylose–aglycon distance, and thereby the surface area of the xyloside, to a certain level would make it more accessible for enzymes involved in the GAG synthesis. The compositions of the primed GAG chains varied with different xylosides, independent of the xylose–aglycon distance, probably due to various affinities for enzymes and/or different cellular uptake. Furthermore, no correlations between the antiproliferative activities, the xylose–aglycon distances, and the amounts or compositions of the GAG chains were detected suggesting involvement of other factors such as fine structure of the GAG chains, effects on endogenous PG synthesis, or other unknown factors for the antiproliferative activity.     

Effects of a viscosupplementation therapy on rabbit menisci in an anterior cruciate ligament transection model of osteoarthritis

The aim of this study was to evaluate the morphological, microstructural, and mechanical effects of a viscosupplementation therapy on rabbit menisci at an early stage of osteoarthritis (OA). Anterior cruciate ligament transection (ACLT) was performed in twelve male New-Zealand White rabbits on the right knee joint. Six of these twelve rabbits received a mono intra-articular injection of high molecular weight hyaluronic acid (HA) two weeks after ACLT. Six additional healthy rabbits served as controls. Medial menisci were removed from all right knees (n = 18) six weeks after ACLT and were graded macroscopically. Indentation-relaxation tests were performed in the anterior and posterior regions of the menisci. Collagen fiber organization and glycosaminoglycan (GAG) content were assessed by biphotonic confocal microscopy and histology, respectively. Viscosupplementation significantly (p = 0.002) improved the surface integrity of the medial menisci compared to the operated non-treated group. Moreover, the injection seems to have an effect on the GAG distribution in the anterior region of the menisci. However, the viscoelastic properties of both operated groups were similar and significantly lower than those of the healthy group, which was explained by their modified collagen fiber organization. They displayed disruption of the tie fibers due to structural alterations of the superficial layers from which they emanate, leading to modifications in the deep zone. To conclude, the viscosupplementation therapy prevents macroscopic lesions of the menisci, but it fails to restore their collagen fiber organization and their viscoelastic properties. This finding supports the role of this treatment in improving the lubrication over the knee.     

Resistance exercise modulates lipid plasma profile and cytokine content in the adipose tissue of tumour-bearing rats

Cancer cachexia is a multifactorial syndrome characterised by progressive weight loss, frequently accompanied by anorexia, sarcopenia, and chronic systemic inflammation. The white adipose tissue is markedly affected by cachexia and contributes to this syndrome throught the secretion of pro-inflammatory factors which reach the adjacent tissues and the circulation. A nonpharmacologic intervention that may attenuate cancer cachexia is chronic physical activity, but the effect of resistance training upon adipose tissue inflammation in cachexia has never been examined. For that purpose we designed a protocol in which animals were randomly assigned to a control group (CT, n = 7), a Tumour bearing group (TB, n = 7), a Resistance Trained group (RT, n = 7) and a Resistance Trained tumour bearing group (RTTB, n = 7). Trained rats climbed a vertical ladder with an extra load attached to the tail, representing 75–90% of total body mass, 3 times per week, for 8 weeks. In the 6th week of resistance training, tumour cells (3 × 10⁷ Walker 256 carcinosarcoma) were inoculated in the tumour groups. Body, adipose tissue, muscle and tumour mass was determined, as well a blood biochemical parameters, and the hormone and cytokine profile assessed. The glycogen content of the liver and muscle was measured. IL-10, IL-6 and TNF-α protein expression was evaluated in the mesenteric adipose tissue (MEAT) examined. Resistance training increased by 9% body weight gain in RTTB (final weight 310.8 ± 9.8 g), when compared with TB (final weight 288.3 ± 4.9 g). LDL-c levels were decreased in RTTB (0.28 ± 0.9 mmol/L) by 43% when compared with TB (0.57 ± 0.1 mmol/L). HDL-c levels were increased in RTTB (1.31 ± 0.12 mmol/L) by 15% in regard to CT (1.13 ± 0.7 mmol/L) and 22% as compared with TB (1.07 ± 0.07 mmol/L). RTTB testosterone levels (577 ± 131 ng/mL) were 55% higher when compared with CT (254 ± 41.3 ng/mL) and 63% higher when compared with TB (221 ± 23.1 ng/mL). Adiponectin levels were augmented in RT (23 μg/mL) by 43% when compared with TB (11 μg/mL). Protein expression of IL-6 was increased 38% in TB MEAT (5.95 pg/μg), as compared with CT (3.64 pg/μg) and 50% compared with RTTB (2.91 pg/μg). Similar results with respect to TNF-α TB (7.18 pg/μg) were observed: 39% and 46%, higher protein expression in comparison with CT (4.63 pg/μg) and RTTB (3.8 pg/μg), respectively. IL-10 protein expression was found to be increased in TB (4.4 pg/μg) and RTTB (3.2 pg/μg) 50% and 47%, respectively, in comparison with CT (1.2 pu/μg). The IL-10/TNF-α ratio was higher in RTTB in relation to all others experimental groups. The results show a robust effect of resistance exercise training in preventing important symptoms of cancer cachexia, thus strongly suggesting it may appear as an alternative to endurance exercise as a non-pharmacological therapy in the management of this syndrome.     

Comparison of the lipid properties of captive,healthy wild,and pansteatitis-affected wild Nile crocodiles (Crocodylus niloticus)

The results presented describe and compare the fatty acid composition and melting properties of captive, healthy wild, and pansteatitis-affected wild crocodiles (Crocodylus niloticus). Differences in fatty acid composition between intramuscular and adipose fat is noted in captive crocodiles, and the latter differs from wild crocodiles as a result of different diets. Adipose fat of healthy wild crocodiles differs minimally from diseased ones, respectively with 37.3 ± 2.6% vs. 43.2 ± 2.3% monounsaturated fatty acids, and 43.2 ± 2.9% in dead crocodiles, while polyunsaturated fatty acids decrease from 27.3 ± 1.9% to as low as 21.9 ± 3.6% respectively. Of the unsaturated fatty acids 18:2n? 6 decreased from 6.5 ± 2.6% in unaffected crocodiles to 3.5 ± 0.6% in highly affected and 3.2 ± 0.4% in dead crocodiles, and 22:5n?3 from 2.8 ± 0.6% to 1.8 ± 0.3% and 2.2 ± 0.3% respectively. The melting properties as determined by differential scanning calorimetry show that extracted adipose fat is a small degree softer in pansteatitis-affected tissue, specifically in the temperature range 7–36 °C, and does not contribute to the hard texture noted for adipose fat tissue of pansteatitis-affected animals. A high moisture content of 51.0 ± 19.7% of the fat tissue of pansteatitis-affected animals vs.17.1 ± 8.0% of healthy ones, suggests that physiological changes due to interstitial inflammation may contribute to the hard texture.     

Tolerance of hairy roots of carrots to U chronic exposure in a standardized in vitro device

In soil, high variability of U bioavailability results in large range of apparent U toxic levels for plants. U toxicity on hairy roots of carrot was studied in nutrient gel with a standardized in vitro device. After exposure to 2.5 and 20 mg U L^?1 for 34 days, U concentration ranged between 4 and 563 mg U kg^?1 fresh weight which was in good accordance with U accumulation by roots of plant from contaminated soils. Threshold of U toxicity for root length decreased with time and a transient hormesis occurred for exposure to 2.5 and 5 mg U L^?1. After 34 days and with root length as endpoint, significant toxicity appeared at a gel contamination level above 7.5 mg U L^?1 corresponding to a maximum U concentration in the liquid phase of 0.8 mg L^?1. The calculated EC50 for root length as a function of gel contamination was 9.4 mg U L^?1. Lower threshold and EC50 were observed for biomass as endpoint (resp. 5 and 7.3 mg U L^?1). The low values observed in this study could result from high sensitivity of carrot to U, high bioavailability of U in gel or absence of interferences with microorganisms. This in vitro device appeared adapted to study toxicity of U to plant roots in optimal conditions of both exposure and observations and is recommended to examine further physiological processes and the influence of microorganism interactions.     

2-Heteroarylidene-1-indanone derivatives as inhibitors of monoamine oxidase

In the present study a series of fifteen 2-heteroarylidene-1-indanone derivatives were synthesised and evaluated as inhibitors of recombinant human monoamine oxidase (MAO) A and B. These compounds are structurally related to series of heterocyclic chalcone derivatives which have previously been shown to act as MAO-B specific inhibitors. The results document that the 2-heteroarylidene-1-indanones are in vitro inhibitors of MAO-B, displaying IC₅₀ values of 0.0044–1.53 μM. Although with lower potencies, the derivatives also inhibit the MAO-A isoform with IC₅₀ values as low as 0.061 μM. An analysis of the structure-activity relationships for MAO-B inhibition indicates that substitution with the methoxy group on the A-ring leads to a significant enhancement in MAO-B inhibition compared to the unsubstituted homologues while the effect of the heteroaromatic substituent on activity, in decreasing order is: 5-bromo-2-furan > 5-methyl-2-furan > 2-pyridine ≈ 2-thiophene > cyclohexyl > 3-pyridine ≈ 2-furan. It may therefore be concluded that 2-heteroarylidene-1-indanone derivatives are promising leads for the design of MAO inhibitors for the treatment of Parkinson’s disease and possibly other neurodegenerative disorders.     

SAR development of a series of 8-azabicyclo[3.2.1]octan-3-yloxy-benzamides as kappa opioid receptor antagonists. Part 2

Further structure activity relationship studies on a previously reported 8-azabicyclo[3.2.1]octan-3-yloxy-benzamide series of potent and selective kappa opioid receptor antagonists is discussed. Modification of the pendant N-substitution to include a cyclohexylurea moiety produced analogs with greater in vitro opioid and hERG selectivity such as 12 (κ IC₅₀ = 172 nM, μ:κ ratio = 93, δ:κ ratio = >174, hERG IC₅₀ = >33 μM). Changes to the linker conformation and identity as well as to the benzamide ring moiety were also investigated.     

Metabolomic markers of fatigue: Association between circulating metabolome and fatigue in women with chronic widespread pain

Background

Fatigue is a sensation of unbearable tiredness that frequently accompanies chronic widespread musculoskeletal pain (CWP) and inflammatory joint disease. Its mechanisms are poorly understood and there is a lack of effective biomarkers for diagnosis and onset prediction. We studied the circulating metabolome in a population sample characterised for CWP to identify biomarkers showing specificity for fatigue.

Skeletal muscle pericyte subtypes differ in their differentiation potential

Neural progenitor cells have been proposed as a therapy for central nervous system disorders, including neurodegenerative diseases and trauma injuries, however their accessibility is a major limitation. We recently isolated Tuj1 + cells from skeletal muscle culture of Nestin–GFP transgenic mice however whether they form functional neurons in the brain is not yet known. Additionally, their isolation from nontransgenic species and identification of their ancestors is unknown. This gap of knowledge precludes us from studying their role as a valuable alternative to neural progenitors. Here, we identified two pericyte subtypes, type-1 and type-2, using a double transgenic Nestin–GFP/NG2–DsRed mouse and demonstrated that Nestin–GFP +/Tuj1 + cells derive from type-2 Nestin–GFP +/NG2–DsRed +/CD146 + pericytes located in the skeletal muscle interstitium. These cells are bipotential as they generate either Tuj1 + cells when cultured with muscle cells or become “classical” α-SMA + pericytes when cultured alone. In contrast, type-1 Nestin–GFP ?/NG2–DsRed +/CD146 + pericytes generate α-SMA + pericytes but not Tuj1 + cells. Interestingly, type-2 pericyte derived Tuj1 + cells retain some pericytic markers (CD146 +/PDGFRβ +/NG2 +). Given the potential application of Nestin–GFP +/NG2–DsRed +/Tuj1 + cells for cell therapy, we found a surface marker, the nerve growth factor receptor, which is expressed exclusively in these cells and can be used to identify and isolate them from mixed cell populations in nontransgenic species for clinical purposes.     

Plasma renin and aldosterone levels in obese and non-obese women with polycystic ovary syndrome

ObjetiveTo assess plasma renin and aldosterone levels in obese and non-obese women with polycystic ovary syndrome (PCOS).MethodsObese women (body mass index [BMI] > 30 kg/m2; group A, n = 34) and non-obese women (BMI < 25 kg/m2; group B, n = 13) with PCOS were selected. The control group (group C, n =47) consisted of age-matched women with regular menses and normal ultrasonographic ovaries. Luteinizing hormone, follicle-stimulating hormone, androstenedione, testosterone, sex hormone-binding globulin, serum glucose, insulin, renin, plasma renin activity, and aldosterone levels were measured.ResultsObese and non-obese women with PCOS had higher luteinizing hormone, follicle-stimulating hormone, androstenedione, testosterone, and insulin levels as compared to women in the control group (p < 0.05). Women with PCOS had significantly higher renin levels (group A: 50.2 ± 4.9 picoU/mL, group B: 39.9 ± 2.7 picoU/mL, and group C: 24.6 ± 2.6 picoU/mL), plasma renin activity (group A: 3.7 ± 0.3 ng/mL/h, group B: 3.6 ± 0.3 ng/mL/h, and group C: 2.2 ± 0.4 ng/mL/h), and aldosterone levels (group A: 31.2 ± 3.3 ng/dL, group B: 29.3 ± 2.9 ng/dL, and group C: 22.2 ± 3.9 ng/dL) as compared with controls.ConclusionSignificant differences exist in plasma renin and aldosterone levels between obese and non-obese women as compared with polycystic ovary syndrome and normal controls.     

Stress as adaptation? A test of the adaptive boost hypothesis among Batswana men

Many studies have found elevated cortisol linked to negative events (“stress”) and subsequent negative outcomes, such as reduced immunity and stunted growth, leading to the conclusion that high cortisol is “bad.” However, a growing number of studies have found more advantaged groups showing relatively elevated cortisol. For example, higher morning cortisol followed by a steeper diurnal decline among Caucasians compared to ethnic minorities has been interpreted as a context-specific “adaptive boost” to meet daily demands. We tested the adaptive boost hypothesis using data on socioeconomic status, depressive affect and salivary cortisol among adult men (n = 32) in Botswana. Three findings emerged: (i) depressive affect was associated with lower morning cortisol (r = ? 0.43, p = 0.014); (ii) depressive affect was associated with a diurnal increase in cortisol when comparing morning and evening samples (r = 0.49, p = 0.004); and (iii) depressive affect was associated with lower income (r = ? 0.55, p = 0.001). Findings are consistent with the adaptive boost hypothesis and add to a growing body of evidence that elevated cortisol is not universally bad. Hypothalamic–pituitary adrenal axis (HPAA) activity, such as cortisol, may be adaptive depending on a person's contextual circumstances. Based on our findings and those of previous studies, we develop a “person-in-context” model of the threat appraisal process. Integrated with life history theory, our model facilitates testable hypotheses about intra- and inter-individual variability in HPAA and adaptive consequences.     

Rapid determination of 25-hydroxy vitamin D3 in swine tissue using an isotope dilution HPLC-MS assay

A rapid method for quantification of 25-hydroxy vitamin D3 in different swine tissues based on isotope dilution HPLC-MS has been developed and validated. Six times deuterated analyte is used as internal standard. The method is fast and can be performed with only 1 g sample. Sample preparation for kidney, liver, muscle and spleen requires only homogenisation and extraction with methanol. An additional enzymatic digest is required for skin, and clean-up of the extract by solid-phase extraction (SPE) is used for adipose tissue and skin. The lower limit of detection varies from 1 ng/g (muscle) to 5 ng/g (adipose and skin). The method has been successfully applied to various tissue samples of pigs fed for 119 days either 2000 IU of vitamin D3 or 50 μg of 25-hydroxy vitamin D3 per kg feed. For animals ingesting 25-OH-D3 supplements the highest tissue contents were observed in the skin (24.8 ± 3.5 ng/g), followed by kidney (14.2 ± 1.5 ng/g), liver and muscle (5.7 ± 0.6 ng/g). The 25-OH-D3 content in the skin was significantly higher in animals ingesting 2000 IU/kg of vitamin D3 (39.5 ± 13.4 ng/g). Levels in selected tissues of some animals were below the lower limit of quantification. No measurable amounts of 25-OH-D3 were found in spleen, abdominal fat and subcutaneous fat of the animals of both groups as well as in the liver, kidney and muscle of the animals ingesting 2000 IU/kg of vitamin D3.     

Patients with chronic,but not episodic,migraine display altered activity of their neck extensor muscles

The current study aimed to investigate differences in activity of neck flexor and extensor muscles in women with migraine considering the chronicity of their condition. Thirty-one subjects with episodic migraine, 21 with chronic migraine and 31 healthy controls participated. Surface electromyography signals were recorded bilaterally from the sternocleidomastoid, anterior scalene, splenius capitis and upper trapezius muscles as subjects performed 5 stages of cranio-cervical flexion (CCF), representing a progressive increase in range of CCF motion. Comparison of normalized root-mean-square among groups was conducted with 3 × 5 ANCOVA with task level as the within-subject variable, group as the between-subject variable, and the presence of neck pain and disability as co-variates. The group with chronic migraine exhibited increased activity of their extensor muscles compared to the control and episodic migraine groups (splenius capitis: F = 3.149, P = 0.045; upper trapezius: F = 3.369, P = 0.041). No significant between-group differences were found for the superficial neck flexors (sternocleidomastoid: F = 1.161, P = 0.320; anterior scalene: F = 0.135, P = 0.874). In conclusion, women with chronic migraine exhibit increased activity of their superficial neck extensor muscles when acting as antagonists during low-load isometric CCF contractions in comparison to non-headache subjects.     

Estimation of the phosphorus requirement of weanling pigs fed supplemental phytase

Studies were conducted with crossbred weanling pigs to determine the level of phosphorus needed to be fed when a maize–soyabean meal–whey diet was supplemented with exogenous phytase (Natuphos™). In Trial 1, phytase was added at 1200 phytase units (PTU) kg⁻¹ as phosphorus decreased. The control diet in Phase I (0–14 days) contained 7.3 g kg⁻¹ phosphorus and in Phase II (14–28 days) contained 6.5 g kg⁻¹ phosphorus. Dietary phosphorus was calculated to decrease by 0.8, 1.6 or 2.4 g kg⁻¹ when phytase was supplemented. Chromic oxide was added for estimation of apparent absorption of phosphorus. Performance was optimum when 5.7 and 4.8 g kg⁻¹ phosphorus (analysed levels) were fed with 1200 PTU kg⁻¹ phytase in Phases I and II, respectively. The lowest dietary phosphorus levels did not reduce performance for the overall 28-day period. Apparent phosphorus digestibility was increased by phytase in Phase I when 5.7 g kg⁻¹ phosphorus was fed compared to the control diet and in Phase II when 6.0 g kg⁻¹ phosphorus was fed with phytase. Faecal phosphorus excretion decreased in both phases as dietary phosphorus decreased. Faecal phosphorus excretion was minimized at the lowest phosphorus level with no decrease in performance. The estimated requirement for dietary phosphorus, as determined by the NLIN procedure, is 5.0 g kg⁻¹ in Phase I and 4.3 g kg⁻¹ in Phase II when 1200 PTU kg⁻¹ is used. In Trial 2, phytase was supplemented at 500 PTU kg⁻¹ when phosphorus was decreased in the diet. The control diet contained 6.6 and 6.0 g kg⁻¹ phosphorus in Phases I and II, respectively, and phosphorus was calculated to decrease by 0.5, 1.0, 1.5, or 2.0 g kg⁻¹ when phytase was added. Daily gain decreased when 5.0 g kg⁻¹ phosphorus was fed in Phase I and when 4.6 or 4.2 g kg⁻¹ (analysed levels) phosphorus was fed in Phase II with 500 PTU kg⁻¹. Faecal phosphorus excretion decreased as dietary phosphorus decreased, but there were no treatment effects on apparent phosphorus digestibility. The dietary phosphorus requirement was estimated to be 5.7 and 5.0 g kg⁻¹ in Phases I and II, respectively, when phytase is fed at 500 PTU kg⁻¹. At the present recommendation of 500 PTU kg⁻¹ in starter feed, phosphorus can be decreased by 0.10 g kg⁻¹. However, higher levels of phytase are needed to actually increase apparent phosphorus digestibility.     

Alleviating the adverse effects of NaCl stress in maize seedlings by pretreating seeds with salicylic acid and 24-epibrassinolide

The effects of bio-regulators salicylic acid (SA) and 24-epibrassinolide (EBL) as seed soaking treatment on the growth traits, content of photosynthetic pigments, proline, relative water content (RWC), electrolyte leakage percent (EC%), antioxidative enzymes and leaf anatomy of Zea mays L. seedlings grown under 60 or 120 mM NaCl saline stress were studied. A greenhouse experiment was performed in a completely randomized design with nine treatments [control (treated with tap water); 60 mM NaCl; 120 mM NaCl; 10^− 4 M SA; 60 mM NaCl + 10^− 4 M SA; 120 mM NaCl + 10^− 4 M SA; 10 μM EBL; 60 mM NaCl + 10 μMEBL or 120 mM NaCl + 10 μM EBL] each with four replicates. The results indicated that NaCl stress significantly reduced plant growth traits, leaf photosynthetic pigment, soluble sugars, RWC%, and activities of catalase (CAT), peroxidase (POX) as well as leaf anatomy. However, the application of SA or EBL mitigated the toxic effects of NaCl stress on maize seedlings and considerably improved growth traits, photosynthetic pigments, proline, RWC%, CAT and POX enzyme activities as well as leaf anatomy. This study highlights the potential ameliorative effects of SA or EBL in mitigating the phytotoxicity of NaCl stress in seeds and growing seedlings.     

Association of TP53 codon 72 and CDH1 genetic polymorphisms with colorectal cancer risk in Bangladeshi population

Till now no pharmacogenetic study of TP53 codon 72 (Arg72Pro) and CDH1 rs16260 (-160C<A) genes has been reported on Bangladeshi population relating those with colorectal cancer. So the aim of the study is to determine whether there is an elevated risk of colorectal cancer development with TP53 codon 72 and CDH1 rs16260 genetic polymorphism in Bangladeshi population for the first time. To investigate the association of these two SNPs, we conducted a case-control study with 288 colorectal cancer patients and 295 healthy volunteers by using polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) method. We found an increased risk of association between Arg/Pro heterozygosity (adjusted OR = 2.58, 95% CI = 1.77–3.77, p < 0.05) and Pro/Pro mutant homozygosity (adjusted OR = 2.92, 95% CI = 1.78–4.78, p < 0.05) along with the combined genotype (Arg/Pro + Pro/Pro) (adjusted OR = 2.70, 95% CI = 1.90–3.82, p < 0.05) and colorectal cancer predisposition. In case of CDH1 rs16260 polymorphism, C/A heterozygous and A/A mutant homozygous are significantly (p < 0.05) found to be associated with colorectal cancer risk with adjusted OR of 1.94 and 2.63, respectively. The combined genotype of C/A and A/A was also found to be strongly associated with colorectal cancer risk compared to C/C genotype (adjusted OR = 2.02, 95% CI = 1.42–2.87, p < 0.05). In conclusion, heterozygosity and mutant homozygosity as well as the combination of both TP53 Arg72Pro and CDH1 rs16260 polymorphisms are responsible to increase the risk of colorectal cancer development in Bangladeshi population.     

Molecular distinction of C × R hybrid (Coffea congensis × Coffea canephora) from morphologically resembling male parent using rbcL and matK gene sequences

Interspecific C × R hybrid (Coffea congensis × Coffea canephora) in India is cultivated as mixed population with male parent C. canephora as this species is an efficient pollen donor for enhanced yield. But distinction of C × R hybrid from C. canephora in old plantation is difficult due to varying plant sizes of C × R hybrid and often resembles with C. canephora. C × R hybrid cultivated under different agroclimatic conditions show distinct vegetative growth pattern with varying yields. Thus development of DNA marker for identification of C × R hybrid is important for clonal propagation and seed preparation from selective individuals. In this study, two DNA bar coding loci of chloroplast genome (rbcL and matK) of parents, F1 hybrid and its back cross progeny were partially sequenced to identify SNPs as DNA marker for distinction of C × R hybrid from C. canephora. Seven SNPs in the matK gene sequence and three nucleotides in the rbcL gene sequence were identified as DNA markers for the genetic identity of C. congensis. These SNPs were found in F1 and advanced progenies of C × R hybrid due to maternal inheritance. Large number of samples of C × R hybrids with varying morphological features revealed no polymorphism among C × R hybrid and C. congensis. Thus, the SNPs in C. congensis can be used as DNA markers for precise identification of C × R hybrid for production of clones besides tagging the chloroplast inheritance in advanced progenies.     

Discovery of (R)-5-(benzo[d][1,3]dioxol-5-yl)-7-((1-(vinylsulfonyl)pyrrolidin-2-yl)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine (B6) as a potent Bmx inhibitor for the treatment of NSCLC

Described as a Btk inhibitor, ibrutinib also potently inhibits Bmx and EGFR, two good targets for lung cancer. Owing to its high CLogP (4.07) and low aqueous solubility (<0.01 mg/ml), resulting in unfavorable bioavailability, ibrutinib requires high dosages to achieve good clinical response in the treatment of non-small cell lung cancer (NSCLC). In our effort to improve the CLogP of ibrutinib by structural optimization led to the discovery of a potent anti-cancer agent B6, with beneficial physicochemical parameters (CLogP = 2.56, solubility in water ≈ 0.1 mg/ml) meeting the principles of oral drugs. B6 exhibited anti-proliferation activities against EGFR-expressing cells, especially the mutant ones, such as H1975 (L858R/T790M, IC₅₀ = 0.92 ± 0.19 μM) and HCC827 (Del119 IC₅₀ = 0.014 ± 0.01 μM). Moreover, B6 significantly slowed down H1975 tumor growth with anti-tumor rate of 73.9% (p < 0.01). Enzyme potencies assay demonstrated B6 moderately selectively inhibited Bmx (IC₅₀ = 35.7 ± 0.1 nM) over other kinases. So, as a potent Bmx inhibitor, B6 has the potential to be an efficacious treatment for NSCLC with acquired drug resistance.