The fatty acid desaturase (FADS) gene family at 11q12-13.1 includes FADS1 and FADS2, both known to mediate biosynthesis of omega-3 and omega-6 long-chain polyunsaturated fatty acids (LCPUFA). FADS3 is a putative desaturase due to its sequence similarity with FADS1 and FADS2, but its function is unknown. We have previously described 7 FADS3 alternative transcripts (AT) and 1 FADS2 AT conserved across multiple species. This study examined the effect of dietary LCPUFA levels on liver FADS gene expression in vivo and in vitro, evaluated by qRT-PCR. Fourteen baboon neonates were randomized to three diet groups for their first 12 weeks of life: C: Control, no LCPUFA; L: 0.33% docosahexaenoic acid (DHA)/ 0.67% arachidonic acid (ARA) (w/w); and L3: 1.00% DHA/ 0.67% ARA (w/w). Liver FADS1 and both FADS2 transcripts were downregulated by at least 50% in the L3 group compared to controls. In contrast, FADS3 AT were upregulated (L3>C), with four transcripts significantly upregulated by 40% or more. However, there was no evidence for a shift in liver fatty acids to coincide with increased FADS3 expression. Significant upregulation of FADS3 AT was also observed in human liver-derived HepG2 cells after DHA or ARA treatment. The PPARγ antagonist GW9662 prevented FADS3 upregulation, while downregulation of FADS1 and FADS2 was unaffected. Thus, FADS3 AT were directly upregulated by LCPUFA by a PPARγ-dependent mechanism unrelated to regulation of other desaturases. This opposing pattern and mechanism of regulation suggests a dissimilar function for FADS3 AT compared to other FADS gene products.
docosahexaenoic acid; arachidonic acid; polyunsaturated fatty acids; fatty acid desaturase; FADS3; alternative splicing
Stem cells present an important tool in livestock assisted reproduction and veterinary therapeutic field such as tissue engineering. We report for the first time isolation of pluripotent stem cell-like cells expressing pluripotency markers (alkaline phospahatase, OCT-4, NANOG and SOX-2) from the amnion of water buffalo (Bubalus bubalis). The cells showed no apparent abnormalities in their chromosomal profiles before and after cryopreservation. The cytochemical staining revealed that pluripotent cells were capable of undergoing directed differentiation in vitro into osteocytes. It could be inferred that amnion-derived pluripotent stem cell-like cells can be isolated, cultured for many passages and differentiated into mesoderm lineage, and may be an alternative source to mesenchymal stem cells. These cells can have applications in assisted reproduction, developmental biological and regenerative medicine.
Amnion cells; Buffalo; Differentiation; Pluripotency; Stem cells
The growth conditions for chitinase production by Trichoderma asperellum UTP-16 in solid state fermentation was optimized using response surface methodology based on central composite design. The chitinase production was optimized, using one-factor at a time approach, with six independent variables (temperature, pH, NaCl, incubation period, nitrogen and carbon sources) and 3.31 Units per gram dry substrate (U gds−1) exo-chitinase yield was obtained. A 21.15% increase was recorded in chitinase activity (4.01 U gds−1) through surface response methodology, indicates that it is a powerful and rapid tool for optimization of physical and nutritional variables. Further, efficiency of crude enzyme was evaluated against phytopathogenic Fusarium spp. and a mycelial growth inhibition up to 3.5–6.5 mm was achieved in well diffusion assay. These results could be supplemented as basic information for the development of enzyme based formulation of T. asperellum UTP-16 and its use as a biocontrol agent.
Chitinase; Central composite design; Solid state fermentation; Antifungal activity
Introduction: The dyslipidaemia in Rheumatoid Arthritis (RA) is associated with accelerated atherosclerosis. A prospective clinical evaluation study was undertaken to find out the proportion of the rheumatoid arthritis patients who were suffering from dyslipidaemia, the change in the lipid levels and the disease activity after an intervention with antirheumatic therapy.
Aims and Objectives: To study the disease activity in Rheumatoid arthritis patients by measuring the serum levels of interleukin-1β (IL-1 β), to find out the proportion of rheumatoid arthritis patients who were suffering from dyslipidaemia, to correlate the disease activity with the lipid profile and to look for the change in the lipid levels and the disease activity after an intervention with antirheumatic therapy.
Material and Methods: This study was done on 30 RA patients (fulfilling the American College of Rheumatology criteria). The lipid profile estimation was done by an enzymatic, colourimetric method and IL-1β was estimated by a chemiluminescence method. Dyslipidaemia was defined by taking the cut-off values of the NCEP-ATPIII guidelines. The patients with other comorbid illnesses and those who were on statins were excluded. The patients were followed up after 12 weeks of starting with the anti-rheumatic therapy.
Results: 36.7% of the patients had high total cholesterols, 53.3% of the patients had high triacylglycerol levels, 73.3% of the patients had decreased HDL-cholesterol and 33.3% of the patients had high LDL-cholesterols. 86.7% of the patients had IL-1β levels which were above the reference range. After the treatment, the number of patients with dyslipidaemia came down, with 23.3% patients having high total cholesterol levels, 43.3% of the patients having elevated triacylglycerol levels, 46.7% patients having low HDL-cholesterol levels and 20% patients having elevated LDL-cholesterol levels. 66.7% of the patients had IL-1β which was above the reference range.
Conclusion: The proportion of dyslipidaemic patients had decreased in the follow up visit, along with a decrease in the disease activity, as were indicated by the decreased levels of IL-1β. The management of dyslipidaemia in RA should be a part of the general cardiovascular risk management. Therefore, a good control of the disease activity should be given priority, so that both the quality of life and the long-term outcomes can be improved.
Cardiovascular risk; Dylipidaemia; Interleukin-1β; Rheumatoid arthritis
We theoretically study the spin-wave spectra in magnonic waveguides periodically patterned with nanoscale square antidots. We show that structural changes breaking the mirror symmetry of the waveguide can close the magnonic bandgap. The effect of these intrinsic symmetry breaking can be compensated by adjusted asymmetric external bias magnetic field, i.e., by an extrinsic factor. This allows for the recovery of the magnonic bandgaps. The described methods can be used for developing parallel models for recovering bandgaps closed due to a fabrication defect. The model developed here is particular to magnonics, an emerging field combining spin dynamics and spintronics. However, the underlying principle of this development is squarely based upon the translational and mirror symmetries, thus, we believe that this idea of correcting an intrinsic defect by extrinsic means, should be applicable to spin-waves in both exchange and dipolar interaction regimes, as well as to other waves in general.
The photothermal effect of single-walled carbon nanotubes (SWCNTs) in combination with the anticancer drug doxorubicin (DOX) for targeting and accelerated destruction of breast cancer cells is demonstrated in this paper. A targeted drug-delivery system was developed for selective killing of breast cancer cells with polyethylene glycol biofunctionalized and DOX-loaded SWCNTs conjugated with folic acid. In our work, in vitro drug-release studies showed that the drug (DOX) binds at physiological pH (pH 7.4) and is released only at a lower pH, ie, lysosomal pH (pH 4.0), which is the characteristic pH of the tumor environment. A sustained release of DOX from the SWCNTs was observed for a period of 3 days. SWCNTs have strong optical absorbance in the near-infrared (NIR) region. In this special spectral window, biological systems are highly transparent. Our study reports that under laser irradiation at 800 nm, SWCNTs exhibited strong light–heat transfer characteristics. These optical properties of SWCNTs open the way for selective photothermal ablation in cancer therapy. It was also observed that internalization and uptake of folate-conjugated NTs into cancer cells was achieved by a receptor-mediated endocytosis mechanism. Results of the in vitro experiments show that laser was effective in destroying the cancer cells, while sparing the normal cells. When the above laser effect was combined with DOX-conjugated SWCNTs, we found enhanced and accelerated killing of breast cancer cells. Thus, this nanodrug-delivery system, consisting of laser, drug, and SWCNTs, looks to be a promising selective modality with high treatment efficacy and low side effects for cancer therapy.
cancer; nanotherapy; SWCNTs; targeted drug delivery; photothermal therapy
The fatty acid desaturase genes (FADS1 and FADS2) code for enzymes required for synthesis of omega-3 and omega-6 long-chain polyunsaturated fatty acids (LCPUFA) important in the central nervous system, inflammatory response, and cardiovascular health. SNPs in these genes are associated with numerous health outcomes, but it is unclear how genetic variation affects enzyme function. Here, lymphoblasts obtained from Japanese participants in the International HapMap Project were evaluated for association of expression microarray results with SNPs in the FADS gene cluster. Six SNPs in the first intron of the FADS2 gene were associated with FADS1 expression. A 10-SNP haplotype in FADS2 (rs2727270 to rs2851682) present in 24% of the population was associated with lower expression of FADS1. A highly conserved region coinciding with the most significant SNPs contained predicted binding sites for SREBP and PPARγ. Lymphoblasts homozygous for either the major or minor haplotype were treated with agonists for these transcription factors and expression of FADS1 and FADS2 determined. Simvastatin and the LXR agonist GW3965 both upregulated expression of FADS1 and FADS2; no response was found for PPARγ agonist rosiglitazone. The minor haplotype homozygotes had 20–40% higher induction of FADS1 and FADS2 after simvastatin or GW3965 treatment. A 22 bp polymorphic insertion-deletion (INDEL) was found 137 bp downstream from the putative sterol response element, as well as a 3 or 1 bp INDEL 81–83 bp downstream. All carriers of the minor haplotype had deletions while all carriers of the major haplotype had insertions. Individuals carrying the minor haplotype may be vulnerable to alterations in diet that reduce LCPUFA intake, and especially responsive to statin or marine oil therapy.
FADS2; FADS1; Insertion-deletion; Intron; LCPUFA; LXR; statin; SREBP-1c
The objective of the present review was to determine the effectiveness of maxillary sinus floor augmentation without bone grafts using lateral window technique.
Materials and Methods:
PubMed and Cochrane databases were searched for relevant articles. We also included articles by hand search until June 2012. The analysis included both human and animal studies which satisfied the following criteria: Minimum of 6 months follow-up, no use of bone grafts, and lateral window approach to the sinus.
We included 22 articles in the review. A descriptive analysis of the constructed evidence tables indicated that there is evidence of predictable a mount of bone formation in the maxillary sinus augmentation without the use of bone grafts.
Within the limits of the articles and data available, maxillary sinus augmentation without bone graft might be considered effective inpredictable bone formation.
Augmentation; dental implant; lateral window; maxillary sinus
To analyze whether an association exists between keratometric and pachymetric changes in the cornea, and whether it can be used to create pachymetric cutoff criteria secondary to keratometric criteria.
In this cross-sectional study, 1000 candidates presenting to the refractive surgery services of a tertiary care hospital underwent bilateral Orbscan IIz (Bausch and Lomb) assessment along with other ophthalmic evaluation.
Stepwise regression analysis-based models showed that simulated keratometry (simK) astigmatism was significantly predicted by the minimum corneal thickness (MCT) and difference between central and MCT (δCT), mean SimK by the MCT and δCT, and maximum keratometry in the central 10-mm zone by the MCT and δCT (P<0.001). The mean MCT values were 542.5±39.6, 539.9±39.2, 524.2±49.5, and 449.3±73.7 μm for flatter normal (<44 D), steeper normal (≥44 D), keratoconus suspect and keratoconic eyes, respectively (P<0.001). The mean differences between central corneal thickness and MCT (δCT) were 12.2±7.1 μm, 12.4±7.4 μm, 14.4±8.9 μm and 23.2±10.1 μm for the flatter normal, steeper normal, keratoconus suspect, and keratoconic eyes, respectively (P<0.001). Mean and 2SD cutoff were used to suggest that a cornea having MCT<461 μm or δCT>27 μm has only a 2.5% chance of being normal and not a keratoconus suspect or worse.
Pachymetric diagnostic cutoffs can be used as adjuncts to the existing topographic criteria to screen keratoconus suspect and keratoconic eyes.
keratoconus; keratoconus suspect; cutoffs; pachymetry; keratometry
Objective: To assess the attitude to, the knowledge and practice of contraception and medical abortion in women attending the family planning clinic at the mvj medical college , hosakote , Bangalore, India.
Materials and Methods: Between 1st of August, 2011 and 31st of July, 2012 200 women attending family planning clinic of the mvj medical college, hosakote, Bangalore India of which 105 requested for medical termination of pregnancy (mtp), 95 for family planning advice, were interrogated on a structured questionnaire. The age of women ranged in between 20-45 years, 71 (35.5%) were illiterate, 30 (15%) had primary school education and 99 (49.5%) had diplomas from high school and above. Patients were grouped into low and high socio-economic status according to modified kuppuswamy socio-economic status scale: (i). upper class, (ii). Upper middle class, (iii). Middle class, (iv). Lower middle class, (v). lower class.consent of both husband and wife was taken. They were counseled about the various contraceptives available and allowed to choose whichever suited them best.
Results: Among the 200 women 85 (42%) did not use contraception; 51 (25.5 %) were on the barrier method; 49 (18.31%) used intrauterine devices (iud); 12 (6%) used oral pills and and 3 (1.5%) used other methods. the request for mtp was on grounds of unplanned pregnancy in 55.25% cases or failure of contraception in 44.7%. there was no eugenic indication of the women, 3 (1.5%) had heard about emergency contraceptives, however none had used them; 20 (10%) had heard of medical abortion and 12 (6%) had previously undergone mtp with satisfaction. the various methods of contraception accepted by the women post abortion were ocps by 11 (10.47%), iuds by 54 (51.5%) and female sterilization by 26 (24.71%). in the other group, 23 (24.2%) had iuds removed and reinserted; 37.8% had iuds inserted; 26 (27.36%) women underwent sterilization operation; and 6 (6.31%) had iuds removed opting for pregnancy. statistical analysis was done using spss software (Chicago) with χ2 test taking p value of 0.05 as significant.
Conclusion: There is lack of awareness of emergency contraception and medical abortion in the women community under study.
Contraception, Methods for family planning services; Medical abortion; Emergency contraception; Medical termination of pregnancy (MTP); Intrauterine devices (idus); Oral contraceptive pills (ocps); Sterilization
This study determined the sensitivity of heart and brain arachidonic acid (ARA) and docosahexaenoic acid (DHA) to the dietary ARA level in a dose-response design with constant, high DHA in neonatal piglets. On day 3 of age, pigs were assigned to 1 of 6 dietary formulas varying in ARA/DHA as follows (% fatty acid, FA/FA): (A1) 0.1/1.0; (A2) 0.53/1.0; (A3-D3) 0.69/1.0; (A4) 1.1/1.0; (D2) 0.67/0.62; (D1) 0.66/0.33. At necropsy (day 28) higher levels of dietary ARA were associated with increased heart and liver ARA, while brain ARA remained unaffected. Dietary ARA had no effect on tissue DHA accretion. Heart was particularly sensitive, with pigs in the intermediate groups having different ARA (A2, 18.6 ± 0.7%; A3, 19.4 ± 1.0%) and a 0.17% increase in dietary ARA resulted in a 0.84% increase in heart ARA. Further investigations are warranted to determine the clinical significance of heart ARA status in developing neonates.
Piglet; Arachidonic acid; Infant nutrition; Docosahexaenoic acid
Arachidonic acid (ARA) and docosahexaenoic acid (DHA) are routinely added to infant formula to support growth and development. We evaluated the bioequivalence and safety of three ARA-rich oils for potential use in infant formula using the neonatal pig model. The primary outcome for bioequivalence was brain accretion of ARA and DHA. Days 3 to 22 of age, domestic pigs fed one of three formulas, each containing ARA at ~0.64% and DHA at ~0.34% total fatty acids (FA). Control diet ARA was provided by ARASCO® and all diets had DHA from DHASCO® (Martek Biosciences Corp., Columbia, MD). The experimental diets a1 and a2 provided ARA from Refined Arachidonic acid-rich Oil (RAO; Cargill, Inc., Wuhan, China) and SUNTGA40S (Nissui, Nippon Suisan Kaisha, Ltd., Tokyo, Japan), respectively. Formula intake and growth were similar across all diets, and ARA was bioequivalent across treatments in the brain, retina, heart, liver and day 21 RBC. DHA levels in the brain, retina and heart were unaffected by diet. Liver sections, clinical chemistry, and hematological parameters were normal. We conclude that RAO and SUNTGA40S, when added to formula to supply ~0.64% ARA are safe and nutritionally bioequivalent to ARASCO in domestic piglets.
Arachidonic acid; ARASCO; DHASCO; infant nutrition; pig
To evaluate the visual acuity and quality-related satisfaction of patients implanted with a refractive design multifocal intraocular lens (IOL), and evaluate the factors predicting it including angle kappa.
Dr Agarwal's Eye Hospital and Eye Research Centre, Chennai.
In this prospective trial, 50 eyes of 44 consecutive patients were included. All patients underwent phacoemulsification with multifocal IOL implantation (Rezoom IOL, Abbott Medical Optics). The preoperative and postoperative assessment included slit lamp biomicroscopy, uncorrected visual acuity (UCVA), best-corrected visual acuity (BCVA) and kappa angle assessment. At 1 year, 37 patients (43 eyes), who finished follow-up, were asked to rate their symptoms on a graded questionnaire (0–5 for five queries).
The decimal scores for UCVA and BCVA were 0.38±0.21 and 0.47±0.17 (preoperative), and 0.75±0.22 and 0.99±0.11 (postoperative), respectively. Symptom scores were haloes 0.98±1.7, glare 0.69±1.48, blurred distance 1.0±1.7, intermediate 1.34±1.6, near 1.06±1.8. On regression analysis haloes depended on angle kappa and distance UCVA (R
2=0.26, P=0.029), and glare on angle kappa (R
2=0.26, P=0.033). Poor satisfactions with distance, intermediate, and near vision were linked with distance UCVA (R
2=0.17, P=2.3 × 10−4), distance UCVA (R
2=0.1, P=0.04), and near UCVA (R
2=0.12, P=0.03), respectively. The strongest predictor, however, for overall visual discomfort was distance UCVA (R
Our study suggests that there may be a role of misalignment between the visual and pupillary axis (angle kappa) in the occurrence of photic phenomenon after refractive multifocal IOL implantation.
multifocal intraocular lens; visual satisfaction; kappa angle
This paper presents the synthesis of aqueous cadmium sulfide (CdS) quantum dots (QDs) and silica-encapsulated CdS QDs by reverse microemulsion method and utilized as targeted bio-optical probes. We report the role of CdS as an efficient cell tag with fluorescence on par with previously documented cadmium telluride and cadmium selenide QDs, which have been considered to impart high levels of toxicity. In this study, the toxicity of bare QDs was efficiently quenched by encapsulating them in a biocompatible coat of silica. The toxicity profile and uptake of bare CdS QDs and silica-coated QDs, along with the CD31-labeled, silica-coated CdS QDs on human umbilical vein endothelial cells and glioma cells, were investigated. The effect of size, along with the time-dependent cellular uptake of the nanomaterials, has also been emphasized. Enhanced, high-specificity imaging toward endothelial cell lines in comparison with glioma cells was achieved with CD31 antibody-conjugated nanoparticles. The silica-coated nanomaterials exhibited excellent biocompatibility and greater photostability inside live cells, in addition to possessing an extended shelf life. In vivo biocompatibility and localization study of silica-coated CdS QDs in medaka fish embryos, following direct nanoparticle exposure for 24 hours, authenticated the nanomaterials’ high potential for in vivo imaging, augmented with superior biocompatibility. As expected, CdS QD-treated embryos showed 100% mortality, whereas the silica-coated QD-treated embryos stayed viable and healthy throughout and after the experiments, devoid of any deformities. We provide highly cogent and convincing evidence for such silica-coated QDs as a model nanoparticle in practice, to achieve in vitro and in vivo precision targeted imaging.
endothelial imaging; CD31; silica nanoparticle; CdS QDs; medaka embryos; biocompatibility
Prasarani is one of the drugs used in Vata Rogas such as Amavata, Avabahuka, etc. Among the different source plants of Prasarani, the plant Merremia tridentata Haller.f. is mostly used in South India and the plant Paderia foetida Linn. in North India, hence taken in the present work for comparative analytical study. It was observed that there is a common constituent (having 350 mm absorbance maxima) present in both the drug samples indicating that both works on a similar disease. It was also found that the whole plant powder sample of P. foetida has more number of constituents than that of M. tridentata which indicates P. foetida may have a better efficacy than M. tridentata.
HPLC; Merremia tridentata; Paederia foetida; phytochemistry; Prasarani
Le but de cette étude était de déterminer les caractéristiques biomécaniques des membranes fœtales humaines (FM) durant la gestation. Ces propriétés notamment la force et le travail produisant la rupture ont été déterminées pour 115 d’entre elles de 23–41 semaines de gestation. Les domaines de FM à proximité immédiate des zones les plus et les moins résistantes testées ont été échantillonnés pour l’analyse histomorphométriques. Les données cliniques sur les patientes dont les FM ont été examinées ont également été recueillies. Les FM de ≤ 28 semaines de gestation sont associées à une incidence plus élevée de décollement placentaire et de chorioamniotite. Sur le plan topographique, les FM à tous les stades de gestations ont des caractéristiques biomécaniques hétérogènes distinctes des zones fragiles quand à leur résistance. La plupart des membranes de préterme ont une plus forte résistance. La force et le travail de rupture des FM diminuent avec l’âge gestationnel. Cette diminution est spectaculaire à ≥ 38 semaines de gestation. La couche amnio-choriale est plus mince dans les zones fragiles par rapport aux zones résistantes. Les FM de préterme sont plus résistantes mais elles sont aussi très hétérogènes. L’augmentation progressive de la fragilité des FM avec la gestation atteint un maximum à 38 semaines. Les zones de faiblesses sont plus minces que les parties les plus résistantes. On ne peut affirmer que la différence d’épaisseur soit suffisante pour expliquer des différences de résistance des MF.
Membranes fœtales; grossesse; rupture; prématurité; terme; biomécanique; Fetal membranes; pregnancy; rupture; prematurity; term; biomechanics
A phyllodes tumour of the breast converting to fibrosarcoma of the breast is a rare entity. Prognosis of fibrosarcoma of the breast is poor and the role of various treatment modalities is not clearly defined due to the rarity of the disease. One such case, which was treated successfully with a combination of surgery, radiotherapy and chemotherapy, is presented here.
Alzheimer's disease is a growing concern in the modern world. As the currently available medications are not very promising, there is an increased need for the fabrication of newer drugs. Curcumin is a plant derived compound which has potential activities beneficial for the treatment of Alzheimer's disease. Anti-amyloid activity and anti-oxidant activity of curcumin is highly beneficial for the treatment of Alzheimer's disease. The insolubility of curcumin in water restricts its use to a great extend, which can be overcome by the synthesis of curcumin nanoparticles. In our work, we have successfully synthesized water-soluble PLGA coated- curcumin nanoparticles and characterized it using different techniques. As drug targeting to diseases of cerebral origin are difficult due to the stringency of blood-brain barrier, we have coupled the nanoparticle with Tet-1 peptide, which has the affinity to neurons and possess retrograde transportation properties. Our results suggest that curcumin encapsulated-PLGA nanoparticles are able to destroy amyloid aggregates, exhibit anti-oxidative property and are non-cytotoxic. The encapsulation of the curcumin in PLGA does not destroy its inherent properties and so, the PLGA-curcumin nanoparticles can be used as a drug with multiple functions in treating Alzheimer's disease proving it to be a potential therapeutic tool against this dreaded disease.
Abruption-induced thrombin generation and inflammation/infection induced cytokine production have both been associated with fetal membrane (FM) weakening and preterm premature rupture of the fetal membranes (PPROM). Using our in vitro model system we have demonstrated that thrombin, and separately the cytokines, tumor necrosis factor-alpha (TNFα) and interleukin-1-beta (IL-1β), remodel and weaken full thickness FM. Additionally, we have reported that the anti-oxidant and NFκB inhibitor, alpha-lipoic acid (LA), blocks these thrombin and cytokine induced effects. The purpose of these studies was to determine whether thrombin and cytokines directly weaken the amnion membrane (AM), the major load-bearing component of FM. Isolated AM or full thickness FM fragments from unlabored Cesarean deliveries were incubated with thrombin, TNFα, or IL-1β, for 48h. Rupture strength (breaking force) of each fragment was thereafter determined using our published methodology. Biochemical evidence of remodeling and apoptosis; immunoreactive Matrix Metalloproteinase 9 (MMP9), Tissue Inhibitor of Matrix Metalloproteinase 3 (TIMP3) and cleaved poly (ADP-ribose) polymerase (C-PARP) levels in tissue extracts, were determined by western blot and densitometry. Thrombin induced a dose-dependent weakening of isolated AM (P<0.001) coupled with dose-dependent increases in PARP cleavage, and reciprocal increases and decreases, respectively, in MMP9 and TIMP3 protein (all P<0.01). Thrombin receptor activating peptide-6 (TRAP) also weakened isolated AM. Neither TNFα nor IL-1β weakened isolated AM. However, both cytokines weakened AM when it was incubated together with the choriodecidua as part of full thickness FM (P<0.001). Cytokine conditioned choriodecidua medium also weakened isolated AM (P<0.001). Under conditions in which cytokines weakened the AM, the changes in MMP9, TIMP3 and PARP cleavage were consistent with those seen after thrombin incubation. LA blocked the FM weakening and remodeling effects. In summary, thrombin weakens AM directly whereas cytokines weaken AM indirectly by causing the release of soluble intermediates from the choriodecidua.
The male germ cell-specific fatty acid binding protein 9 (FABP9/PERF15) is the major component of the murine sperm perforatorium and perinuclear theca. Based on its cytoskeletal association and sequence homology to myelin P2 (FABP8), it has been suggested that FABP9 tethers sperm membranes to the underlying cytoskeleton. Furthermore, its upregulation in apoptotic testicular germ cells and its increased phosphorylation status during capacitation suggested multiple important functions for FABP9. Therefore, we investigated specific functions for FABP9 by means of targeted gene disruption in mice. FABP9−/− mice were viable and fertile. Phenotypic analysis showed that FABP9−/− mice had significant increases in sperm head abnormalities (~8% greater than their WT cohorts); in particular, we observed the reduction or absence of the characteristic structural element known as the “ventral spur” in ~10% of FABP9−/− sperm. However, deficiency of FABP9 neither affected membrane tethering to the perinuclear theca nor the fatty acid composition of sperm. Moreover, epididymal sperm numbers were not affected in FABP9−/− mice. Therefore, we conclude that FABP9 plays only a minor role in providing the murine sperm head its characteristic shape and is not absolutely required for spermatogenesis or sperm function.
Fatty acid binding protein; Germ cell; Sperm; Testis; Lipid; Perforatorium
Genes coding for the fatty acid desaturases (FADS1, 2, 3) localized at the cancer genomic hotspot 11q13 locus are required for the biosynthesis of 20 carbon polyunsaturated fatty acids (PUFA) that are direct eicosanoid precursors. In several cancer cell lines, FADS2 encoded Δ6 and Δ8 desaturation is not functional.
Analyzing MCF7 cell fatty acids with detailed structural mass spectrometry, we show that in the absence of FADS2 activity, the FADS1 product Δ5-desaturase operates to produce 5,11,14–20∶3 and 5,11,14,17–20∶4. These PUFA are missing the 8–9 double bond of the eicosanoid signaling precursors arachidonic acid (5,8,11,14–20∶4) and eicosapentaenoic acid (5,8,11,14,17–20∶5). Heterologous expression of FADS2 restores Δ6 and Δ8-desaturase activity and normal eicosanoid precursor synthesis.
The loss of FADS2-encoded activities in cancer cells shuts down normal PUFA biosynthesis, deleting the endogenous supply of eicosanoid and downstream docosanoid precursors, and replacing them with unusual butylene-interrupted fatty acids. If recapitulated in vivo, the normal eicosanoid and docosanoid cell signaling milieu would be depleted and altered due to reduction and substitution of normal substrates with unusual substrates, with unpredictable consequences for cellular communication.
Cytokine-mediated inflammation and abruption-induced thrombin generation are separately implicated in matrix metalloproteinase (MMP)-mediated weakening of fetal membranes (FM) leading to preterm premature rupture of the fetal membranes (PPROM). At term, FM of both labored vaginal and unlabored caesarian deliveries exhibit a weak zone overlying the cervix exhibiting ECM remodeling characterized by increased MMP9 protein and activity. We have reproduced these biochemical changes as well as FM weakening in vitro using tumor necrosis factor-alpha (TNF) and interleukin (IL)-1β, inflammatory cytokines implicated in PPROM. Additionally, we have reported that the antioxidant and NFκB inhibitor alpha-lipoic Acid (LA) blocks these TNF-induced effects. We now present the first direct evidence that thrombin also can induce FM weakening in vitro, and LA treatment inhibits this thrombin-induced weakening. Full thickness FM fragments from unlabored caesarian deliveries were incubated with increasing doses of thrombin (0–100 u/ml) for 48h. Fragments were then strength tested (breaking force and work to rupture) using our published methodology. MMP3 and 9 levels in tissue extracts were determined by Western blot and densitometry. To determine the effect of LA, FM fragments were incubated with control medium or 10 u/ml thrombin, with or without 0.25mM LA. Strength testing and MMP induction was determined. Thrombin induced a dose-dependent decrease in FM strength (42% baseline rupture force and 45% work to rupture) coupled with a dose-dependent increase in MMP3 and 9 expression (all p<.001). Treatment of FM with 0.25mM LA completely inhibited thrombin-induced FM weakening and MMP expression (all p<.001). Thrombin treatment of cultured FM induces mechanical weakening and increased MMP3 and 9. Treatment of FM with LA inhibits these thrombin-induced effects. We speculate LA may prove clinically useful in prevention of PPROM associated with abruption.
In the title molecule, C21H14N2O2, the anthracenyl system is approximately planar [maximum deviation = 0.056 (4) Å] and is oriented at a dihedral angle of 73.6 (1)° with respect to the benzene ring. An intramolecular C—H⋯N hydrogen bond generates an S(6) ring motif. The crystal packing is stabilized by C—H⋯π and π–π interactions [centroid–centroid distances of 3.688 (2), 3.656 (1) and 3.716 (2) Å].
Bacillus cereus, Bacillus thuringiensis and Bacillus anthracis are the major concerns for the food safety in terms of frequency and/or seriousness of the disease. Being members of the same group and sharing DNA homology to a larger extent, they do create problems when their specific detection/identification is attempted from different food and environmental sources. Numerous individual polymerase chain reaction (PCR) and few multiplex PCR (mPCR) methods have been employed to detect these organisms by targeting toxin genes but with lack of internal amplification control (IAC). Therefore, we attempted a mPCR with IAC for the detection of enterotoxic B. cereus group strains by selecting hbl A, nhe A and cyt K genes from B. cereus, indicative of the diarrheal potential and cry I A and pag genes, the plasmid borne phenotypic markers specific to B. thuringiensis and B. anthracis strains, respectively. Multiplex PCR assay validation was performed by simultaneous comparison with the results of single-target PCR assays and correlated to the classical conventional and biochemical identification of the organisms. The mPCR was able to detect as low as 101–102 organisms per ml following overnight enrichment of spiked food samples (vegetable biriyani and milk) in buffered peptone water (BPW). The presence of these organisms could also be detected by mPCR in naturally contaminated samples of rice based dishes and milk. The high throughput and cost-effective mPCR method described could provide a powerful tool for simultaneous, rapid and reliable detection of enterotoxic B. cereus group organisms.
Bacillus cereus; Bacillus thuringiensis; Bacillus anthracis; Toxins; mPCR