Comparative evaluation of the effect of two natural collagen cross-linkers on microtensile bond strength of self-etch adhesive system to dentin after contamination with blood and hemostatic agent: An in vitro study.

Background: Cavity preparation often causes gingival bleeding which can be controlled by hemostatic agents (HAs). These along with blood act as contaminants and hamper the bonding mechanism. Collagen cross-linkers (CCLs) are agents known to increase the bond strength (BS) to dentin. Hence, the purpose of this in vitro study was to determine the effect of two different CCLs, proanthocyanidin (grape seed extract [GSE]) and hesperidin on the microtensile BS (µTBS) of a self-etch adhesive (SEA) system to dentin which was contaminated with blood and a HA. Materials and Methods: Thirty-six extracted human molars were collected, and their occlusal surfaces were sectioned to expose the dentin. The teeth were randomly divided into four groups: Group I - Control, Group II - Contamination with blood and HA, Group III - Application of GSE after contamination, and Group IV - Application of hesperidin extract after contamination. The SEA was applied, followed by the use of a nanocomposite. Dentin-composite rods were obtained from each group, and µTBS testing was done. The fracture pattern was visually classified as an adhesive failure at the interface, cohesive failure in composite, or dentin. The scanning electron microscope (SEM) analysis was done for two samples from each group. Statistical analysis was done using the Student's unpaired "t" and ANOVA test. Results: Group II showed a statistically significant reduction in µTBS in comparison to Group I. This was overcome in Groups III and IV. Hesperidin showed marginally better results than GSE. Conclusions: The use of GSE and hesperidin increases the µTBS of composite resin to dentin postcontamination with blood and ViscoStat Clear with Single Bond Universal Adhesive.

Disentangling potential genotypes for macro and micro nutrients and polymorphic markers in Chickpea.

The present investigation was conducted to assess the nutritional diverseness and identify novel genetic resources to be utilized in chickpea breeding for macro and micro nutrients. The plants were grown in randomized block design. Nutritional and phytochemical properties of nine chickpea genotypes were estimated. The EST sequences from NCBI database were downloaded in FASTA format, clustered into contigs using CAP3, mined for novel SSRs using TROLL analysis and primer pairs were designed using Primer 3 software. Jaccard's similarity coefficients were used to compare the nutritional and molecular indexes followed by dendrograms construction employing UPGMA approach. The genotypes PUSA-1103, K-850, PUSA-1108, PUSA-1053 and the EST-SSR markers including the 5 newly designed namely ICCeM0012, ICCeM0049, ICCeM0067, ICCeM0070, ICCeM0078, SVP55, SVP95, SVP96, SVP146, and SVP217 were found as potential donor/marker resources for the macro-micro nutrients. The genotypes differed (p < 0.05) for nutritional properties. Amongst newly designed primers, 6 were found polymorphic with median PIC (0.46). The alleles per primer ranged 1 to 8. Cluster analysis based on nutritional and molecular diversities partially matched to each other in principle. The identified novel genetic resources may be used to widen the germplasm base, prepare maintainable catalogue and identify systematic blueprints for future chickpea breeding strategies targeting macro-micro nutrients.

Life-saving transfusion in autoimmune hemolytic anemia: a case report and procedure review of the dilution method.

A woman with autoimmune hemolytic anemia (AIHA) presented in the emergency department with life-threatening anemia (hemoglobin 3 g/dL). Exaggeration of preexisting chronic anemia to severe anemia after a recent red blood cell (RBC) transfusion led to suspicion of delayed hemolytic transfusion reaction. Given the urgency for transfusion along with a stronger suspicion for coexistence of an alloantibody, the dilution method proposed by Lawrence Petz and George Garratty was used to find an RBC unit for transfusion. An alloantibody with Fyb specificity was identified, which was masked by the coexistent autoantibody. This method is based on the assumption that the titers of an alloantibody are higher than that of autoantibody. Diluting the autoantibody would reveal the alloantibody and, for this purpose, a serial doubling dilution of serum is performed. This method has an important limitation of missing any alloantibodies with titers less than that of the autoantibody. In spite of this, this method may be of use at a resource-poor setting, where trained personnel and other reagents intended for advanced immunohematology methods are unavailable.

Delayed haemolytic transfusion reaction due to Kidd antibodies.

A delayed haemolytic transfusion reaction (DHTR) encompasses a positive direct antiglobulin test (DAT) developed anytime between 24 hours to 28days after cessation of transfusion, a positive eluate or a newly identified alloantibody in the plasma or serum along with features of haemolysis in the patient. Routinely, it is expected that with the transfusion of one unit of packed red cells in a patient of average height and weight, the haemoglobin level and hematocrit increase by 1 g/dL and 3% respectively. However, in a patient with DHTR, an inadequate rise of post-transfusion haemoglobin (<1 g/dL) or rapid fall in haemoglobin back to pre-transfusion levels is observed. Kidd antibodies are particularly known to cause DHTR, maybe alone or in unison with other antibodies. Detection of these alloantibodies is consequential in providing good transfusion support to these patients. These events may be difficult to detect as they may present as varied clinical features or immunological nuisances. In this case series, we have presented three cases of DHTR with special emphasis on their clinical presentations, immunohaematological evaluations, laboratory parameters and the role of proper transfusion support in these patients to avoid further complications.

Effect of total-etch and self-etch adhesives on the bond strength of composite to glass-ionomer cement/resin-modified glass-ionomer cement in the sandwich technique - A systematic review.

The success of the sandwich technique depends on the bond strength of composite to glass-ionomer cement (GIC)/resin-modified glass-ionomer cement (RMGIC). Currently used adhesives employ the total-etch and the self-etch techniques. The total-etch system is technique sensitive, whereas the self-etch system is popular for its ease of use. The aim of this systematic review is to compare the effect of total-etch and self-etch adhesives (SEAs) on the bond strength of composite to GIC/(RMGIC) in the sandwich technique. A literature search was conducted using electronic databases (PubMed, Web of Science, Ebscohost, and Scopus) limiting the year of publications from January 1, 2000, to September 30, 2018, to identify the relevant studies. All the cross-references of the selected studies were also screened. In vitro studies on extracted human teeth were selected. A total of 10 articles were included in this review. A conclusion was drawn that SEAs when used in the sandwich technique resulted in greater bond strength in comparison to total-etch adhesives. Moreover, increased bond strength was achieved when the primer was employed on unset GIC as compared to set GIC. Furthermore, the application of SEAs over uncured RMGIC (co-curing technique) resulted in better bond strengths as compared to their application over cured RMGIC.

Should treatment of low-level rifampicin mono-resistant tuberculosis be different?

BACKGROUND: Rifampicin resistant tuberculosis (RR-TB) was frequently detected in Suriname after the introduction of Xpert MTB/RIF in 2012. Subsequent phenotypic drug-susceptibility testing (DST) was not conclusive at that moment, while RR-TB patients treated with first-line tuberculostatics had good treatment outcome. In our study, we analysed this interesting observation. METHODS: We collected demographic and clinical characteristics and treatment outcome of TB patients from May 2012-December 2018 and performed a univariate and multivariate analysis to assess possible associations with resistance to rifampicin. Secondly, we conducted whole genome sequencing on all available Mycobacterium tuberculosis isolates that had a rifampicin resistance in the Xpert MTB/RIF test and performed phenotypic DST on selected isolates. FINDINGS: RR-TB was detected in 59 (9.6%) patients confirmed by Xpert. These patients were treated with rifampicin-containing regimens in most (88%) of the cases. In all 32 samples examined, a D435Y mutation in the rpoB gene was identified; only one isolate revealed an additional isoniazid mutation. Phenotypic DST indicated low-level rifampicin resistance. In multivariate analysis, the Creole ethnicity was a factor associated with rifampicin resistance (aOR 3.5; 95%CI 1.9-6.4). The treatment success rate for patients with RR-TB (78.0%) was comparable to the treatment outcome in non-RR-TB patients 77.8%. INTERPRETATION: This study confirms a low-level rifampicin mono-resistance in TB patients of Suriname. These patients could benefit from a first-line regimen with high dose rifampicin (or rifabutin), rather than from the lengthy treatment regimens for rifampicin-resistant and multi-drug resistant TB, a concept of stratified medicine also advocated for the treatment of TB. FUNDING: None.

Evaluation of nuisance removal for functional MRI of rodent brain.

Functional MRI (fMRI) has become an important translational tool for studying brain activity and connectivity in animal models and humans. For accurate and reliable measurement of functional connectivity, nuisance removal strategies developed for human brain, such as regressing motion parameters, cerebrospinal fluid (CSF)/white matter-derived signals and the global signal, have been applied to rodent. However, due to the very different anatomy, with the majority of the rodent brain being gray matter, and experimental conditions, in which animals are anesthetized and head-fixed, these methods may not be suitable for rodent fMRI. In this study, we assessed various nuisance regression methods and the effects of motion correction on a large dataset of both task and resting fMRI of anesthetized rat brain. Sensitivity and specificity were assessed in the somatosensory pathway under forepaw stimulation and resting state. Reproducibility at various sample sizes was simulated by randomly subsampling the dataset. To overcome the difficulty in extracting nuisance from the brain, a method using principal components estimated from tissues outside the brain was evaluated. Our results showed that neither detrend, motion correction, motion regression nor CSF signal regression could improve specificity despite increasing temporal signal-to-noise ratios. Although global signal regression increased the specificity of task activation and functional connectivity, the sensitivity and connectivity strength was drastically reduced, likely due to its strong correlation with the cortical signal. Motion parameters also correlated with task activation and the global signal, indicating that motion correction detected intensity variations in the brain. The nuisance estimated from tissues outside the brain produced a moderate improvement in specificity. In conclusion, nuisance removal suitable for human fMRI may not be optimal for rodents. While further development is needed, estimating nuisance from tissues outside the brain may be an alternative.

Stochastic resonance in a proton pumping Complex I of mitochondria membranes.

We make use of the physical mechanism of proton pumping in the so-called Complex I within mitochondria membranes. Our model is based on sequential charge transfer assisted by conformational changes which facilitate the indirect electron-proton coupling. The equations of motion for the proton operators are derived and solved numerically in combination with the phenomenological Langevin equation describing the periodic conformational changes. We show that with an appropriate set of parameters, protons can be transferred against an applied voltage. In addition, we demonstrate that only the joint action of the periodic energy modulation and thermal noise leads to efficient uphill proton transfer, being a manifestation of stochastic resonance.

GABAergic effect on resting-state functional connectivity: Dynamics under pharmacological antagonism.

Resting state functional connectivity MRI measures synchronous activity among brain regions although the mechanisms governing the temporally coherent BOLD signals remain unclear. Recent studies suggest that γ-amino butyric acid (GABA) levels are correlated with functional connectivity. To understand whether changes in GABA transmission alter functional connectivity, we modulated the GABAergic activity by a GABAA receptor antagonist, bicuculline. Resting and evoked electrophysiology and BOLD signals were measured in isoflurane-anesthetized rats under infusion of low-dose bicuculline or vehicle individually. Both somatosensory BOLD activations and evoked potentials induced by forepaw stimulation were increased significantly under bicuculline compared to vehicle, indicating increased excitability. Gradually elevated resting BOLD correlation within and between the somatosensory and visual cortices, as well as between somatosensory and caudate putamen but not within subcortical areas were found with the infusion of bicuculline. Increased cerebral blood flow was observed throughout the cortical and subcortical areas where the receptor density is high, but it didn't correlate with BOLD connectivity except in the primary somatosensory cortex. Furthermore, resting EEG coherence in the alpha and beta bands exhibited consistent change with the BOLD correlation. The increased cortico-cortical and cortico-striatal connectivity without dependence on the receptor distribution indicate that the functional connectivity may be mediated by long-range projection via the cortical and striatal GABAergic inter-neurons. Our results indicate an important role of the GABAergic system on neural and hemodynamic oscillations, which further supports the neuronal basis of functional connectivity MRI and its correlation with neurotransmission.

Caesarian Scar Pregnancy- a Diagnostic Dilemma.

Caesarean scar pregnancy is one of the rarest forms of ectopic pregnancy. Transvaginal ultrasound and color flow Doppler provides a high diagnostic accuracy. A delay in diagnosis and treatment can lead to uterine rupture, major hemorrhage, hysterectomy and serious maternal morbidity and mortality. Early diagnosis can offer treatment options of avoiding uterine rupture and hemorrhage, thus preserving the uterus and future fertility. Primary health care provider should know about this rare entity, because if diagnosed timely, and referral to specialized centre is done without delay will definitely save maternal morbidity and mortality. Management plan should be individualized. Termination of pregnancy is the treatment of choice in the first trimester. Expectant treatment has a poor prognosis because of risk of rupture. In this case report we aim to discuss the associated diagnostic dilemma, most appropriate methods of diagnosis and management, with their implications in clinical practice.

Incidence of Buxtonella sulcata in bovines in R.S. Pura, Jammu.

Incidence of Buxtonella sulcata was recorded as 20.9 % in bovines (n = 374) of R.S. Pura, Jammu with cattle showing higher infection rate (23.6 %) than the buffaloes (18.5 %). The infection was significantly (p < 0.0001) higher in young animals (33.1 %) than the adults (13.9 %). Animals with diarrhoeic faeces had a significantly higher (p < 0.0001) infection rate (38.5 %) than the animals with normal faeces (9.9 %), suggesting the influence of protozoan on the incidence of diarrhoeal symptoms in bovines.

Occurrence of Setaria digitata in a cow.

The present investigation reports the occurrence of filarial worm, Setaria digitata, recovered during the surgery of a cow suffering from intussusception. The worms were thread like, milky white, long with tapering ends especially towards the hind end with an average size of 62.8 ± 9.89 mm in length. On examination of anterior end, the cuticular rings surrounding the mouth region with dorsal, ventral and lateral prominences were observed. The posterior end had few spines along with a pair of appendages near the tip of the tail. On the basis of morphological characteristics the worms were identified as female S. digitata.

Concordance of Mycobacterium tuberculosis fluoroquinolone resistance testing: implications for treatment.

Fluoroquinolone (FQ) drug susceptibility testing (DST) is an important step in the design of effective treatment regimens for multidrug-resistant tuberculosis. Here we compare ciprofloxacin, ofloxacin and moxifloxacin (MFX) resistance results from 226 multidrug-resistant samples. The low level of concordance observed suggests that DST should be performed for the specific FQ planned for clinical use. The results also support the new World Health Organization recommendation for testing MFX at a critical concentration of 2.0 µg/ml.

IL-33 drives airway hyper-responsiveness through IL-13-mediated mast cell: airway smooth muscle crosstalk.

BACKGROUND: Mast cell localization within the airway smooth muscle (ASM)-bundle plays an important role in the development of airway hyper-responsiveness (AHR). Genomewide association studies implicate the 'alarmin' IL-33 in asthma, but its role in mast cell-ASM interactions is unknown. OBJECTIVES: We examined the expression and functional role of IL-33 in bronchial biopsies of patients with and without asthma, ex vivo ASM, mast cells, cocultured cells and in a mouse model system. METHODS: IL-33 protein expression was assessed in human bronchial tissue from 9 healthy controls, and 18 mild-to-moderate and 12 severe asthmatic patients by immunohistochemistry. IL-33 and ST2 mRNA and protein expression in human-derived ASM, epithelial and mast cells were assessed by qPCR, immunofluorescence and/or flow cytometry and ELISA. Functional assays were used to assess calcium signalling, wound repair, proliferation, apoptosis and contraction. AHR and inflammation were assessed in a mouse model. RESULTS: Bronchial epithelium and ASM expressed IL-33 with the latter in asthma correlating with AHR. ASM and mast cells expressed intracellular IL-33 and ST2. IL-33 stimulated mast cell IL-13 and histamine secretion independent of FcεR1 cross-linking and directly promoted ASM wound repair. Coculture of mast cells with ASM activated by IL-33 increased agonist-induced ASM contraction, and in vivo IL-33 induced AHR in a mouse cytokine installation model; both effects were IL-13 dependent. CONCLUSION: IL-33 directly promotes mast cell activation and ASM wound repair but indirectly promotes ASM contraction via upregulation of mast cell-derived IL-13. This suggests that IL-33 may present an important target to modulate mast cell-ASM crosstalk in asthma.

Acaricidal activity of ethanolic extract of Artemisia absinthium against Hyalomma anatolicum ticks.

The aim of the study was to evaluate the in vitro efficacy of different concentrations of ethanolic extract obtained from the aerial parts of Artemisia absinthium in comparison to amitraz on adults, eggs and larvae of Hyalomma anatolicum using the adult immersion test (AIT), egg hatchability test and larval packet test (LPT), respectively. Four concentrations of the extract (2.5, 5, 10 and 20%) with three replications for each concentration were used in all the bioassays. In AIT, the mortality rates at 2.5, 5 and 10% were significantly different (p < 0.05) in comparison to the control group; however, at 20%, it was similar to the positive control group. Maximum mortality of 86.7% was recorded at 20%. The LC50 and LC95 values were calculated as 6.51 and 55.43%, respectively. The oviposition was reduced significantly by 36.8 and 59.1% at concentrations of 10 and 20%, respectively. Egg hatchability was reduced significantly at all concentrations (2.5-20%) in comparison to the control. In LPT, the extract caused 100% mortality of larvae at all the concentrations after 24 h. The results show that ethanolic extract obtained from the aerial parts of A. absinthium has acaricidal properties and could be useful in controlling H. anatolicum.

CCL2 release by airway smooth muscle is increased in asthma and promotes fibrocyte migration.

BACKGROUND: Asthma is characterized by variable airflow obstruction, airway inflammation, airway hyper-responsiveness and airway remodelling. Airway smooth muscle (ASM) hyperplasia is a feature of airway remodelling and contributes to bronchial wall thickening. We sought to investigate the expression levels of chemokines in primary cultures of ASM cells from asthmatics vs healthy controls and to assess whether differentially expressed chemokines (i) promote fibrocyte (FC) migration towards ASM and (ii) are increased in blood from subjects with asthma and in sputum samples from those asthmatics with bronchial wall thickening. METHODS: Chemokine concentrations released by primary ASM were measured by MesoScale Discovery platform. The chemokine most highly expressed by ASM from asthmatics compared with healthy controls was confirmed by ELISA, and expression of its cognate chemokine receptor by FCs was examined by immunofluorescence and flow cytometry. The role of this chemokine in FC migration towards ASM was investigated by chemotaxis assays. RESULTS: Chemokine (C-C motif) ligand 2 (CCL2) levels were increased in primary ASM supernatants from asthmatics compared with healthy controls. CCR2 was expressed on FCs. Fibrocytes migrated towards recombinant CCL2 and ASM supernatants. These effects were inhibited by CCL2 neutralization. CCL2 levels were increased in blood from asthmatics compared with healthy controls, and sputum CCL2 was increased in asthmatics with bronchial wall thickening. CONCLUSIONS: Airway smooth muscle-derived CCL2 mediates FC migration and potentially contributes to the development of ASM hyperplasia in asthma.

Acaricidal activity of extract of Artemisia absinthium against Rhipicephalus sanguineus of dogs.

The objective of the study was to evaluate the in vitro efficacy of different concentrations of chloroform extract obtained from the aerial parts of Artemisia absinthium in comparison to amitraz on adults, eggs and larvae of the dog tick Rhipicephalus sanguineus using the adult immersion test (AIT), egg hatchability test (EHT) and larval packet test (LPT), respectively. Five concentrations of the extract (1.25, 2.5, 5, 10 and 20 %) with three replications for each concentration were used in all the bioassays. A control group was established (water + dimethylsulphoxide) together with a positive control group (amitraz), with three repetitions each. In AIT, the mortality rates were 0.0, 13.3, 16.7, 33.3 and 93.3 % in concentrations of 1.25, 2.5, 5, 10 and 20 %, respectively, and the variation was significant (p=0.0151). The LC50 (CI) and LC95 (CI) values were calculated as 8.793 % (8.217-9.408) and 34.59 % (29.71-40.26), respectively. The egg production was reduced by 6.6, 6.6, 18.3, 42.5 and 85.1 % in the concentrations of 1.25, 2.5, 5, 10 and 20 %, respectively, and it was statistically significant (p=0.0274). In EHT, hatching was completely inhibited at 5, 10 and 20 % displaying 100 % ovicidal action while at the concentrations of 1.25 and 2.5 %, the inhibition rates were 20 and 60 %, respectively. In LPT, the extract caused 100 % mortality of larvae in the concentrations of 5, 10 and 20 % after 24 h while at the concentrations of 1.25 and 2.5 %, the mortality rates were 54.3 and 96.7 %, respectively. The LC50 (CI) and LC95 (CI) values were determined to be 1.11 % (1.099-1.121) and 2.37 % (2.328-2.423), respectively. The results show that the extract of A. absinthium has acaricidal properties and could be useful in controlling R. sanguineus which is an efficient vector of pathogens both in dogs and humans.

Summertime Bacillus cereus colonization of hospital newborns traced to contaminated, laundered linen.

BACKGROUND: Routine screening of premature newborns for haemolytic streptococci, Staphylococcus aureus and enteric Gram-negative bacteria done at birth using umbilical swabs identified clustering of babies colonized with Bacillus cereus in summers of 2009 and 2010 at a 400-bedded UK general hospital. AIM: To determine the source of this organism by focusing on the clinical environment. METHODS: Umbilical swab screening was extended to all newborns and the labour ward environment, including construction-related dust, was sampled for B. cereus. FINDINGS: During the summer of 2009, 65% of newborns had umbilical swabs which were culture positive for B. cereus. Blood agar and B. cereus selective agar impression plates of unused labour ward linen, and freshly received linen from the hospital's external laundry, gave mainly confluent growth of B. cereus in >85% of items sampled. In-use and exposed healthcare products including liquid handwashing agents, paper hand-towels, vaginal lubricants, labour ward dust and air were culture negative. Linen contamination and umbilical swab culture positivity both approached zero in autumn. B. cereus colonization of newborn umbilici recurred in summer 2010 and unused laundered linen was again found to be as contaminated. Washing linen at the laundry in a washer-extractor, with higher dilution than the continuous tunnel washer normally used, coincided with lowering of detectable B. cereus numbers in unused washed linen and no clustering in newborns the following summer (2011). CONCLUSION: Freshly laundered linen can be contaminated with B. cereus with subsequent spread and colonization of newborns. This contamination appears to be associated with low-dilution washing and high ambient temperatures.

Over-expression of gene encoding heat shock protein 70 from Mycobacterium tuberculosis and its evaluation as vaccine adjuvant.

BACKGROUND: Heat shock proteins (Hsps) are evolutionary ancient and highly conserved molecular chaperons found in prokaryotes as well as eukaryotes. Hsp70 is a predominant member of Hsp family. Microbial Hsp70s (mHsp70s) have acquired special significance in immunity since they have been shown to be potent activators of the innate immune system and generate specific immune responses against tumours and infectious agents. OBJECTIVES: The present study was aimed to clone express and purify recombinant Hsp70 from the Mycobacterium tuberculosis and characterise it immunologically. The study also aimed at determining the potential of recombinant M. tuberculosis heat shock protein (rMTB-Hsp70) as adjuvant or antigen carrier. MATERIALS AND METHODS: Cloning of M. tuberculosis heat shock protein (MTB-Hsp70) amplicon was carried out using the pGEMT-Easy vector although for expression, pProExHTb prokaryotic expression vector was used. Purification of recombinant Hsp70 was carried out by nickel-nitrilotriacetic acid (Ni-NTA) affinity chromatography. For immunological characterization and determining the adjuvant effect of MTB-Hsp70, BALB/c mice were used. The data obtained was statistically analysed. RESULTS: Hsp70 gene was cloned, sequenced and the sequence data were submitted to National Center for Biotechnology Information (NCBI). Recombinant MTB-Hsp70 was successfully over-expressed using the prokaryotic expression system and purified to homogeneity. The protein was found to be immunodominant. Significant adjuvant effect was produced by the rMTB-Hsp70 when inoculated with recombinant outer membrane protein 31; however, effect was less than the conventionally used the Freund's adjuvant. CONCLUSION: Protocol standardised can be followed for bulk production of rHsp70 in a cost-effective manner. Significant adjuvant effect was produced by rMTB-Hsp70; however, the effect was than Freund's adjuvant. Further, studies need to be carried out to explore its applicability as carrier of antigen.