Optimization of Formaldehyde Fixative Concentration for Individual Blood Cells to Develop a Stabilized Blood Control for Automated Hematology Analyzers.

In a modern haematology laboratory, the complete and differential counts of blood are performed using complex haematology auto analyzers. In order to ensure the accuracy and reliability of test results, various regulatory authorities have prescribed the use of stabilized blood controls. The major pitfalls of these blood controls are their short shelf life. This could be due to the fact that they are prepared by a common cocktail of fixatives which acts on the discrete cells in various ways and would result in either under-fixation or over-fixation of various cells. Thus, in the present study, we have explored and optimized fixative and buffering for individual cells to achieve stable blood control. Blood cells were isolated using the centrifugation technique and were fixed individually with different concentrations of formaldehyde. After fixation, cells were pooled. Analysis of cell count was done till six months. Cells were also analysed morphologically to see the effect of fixation and storage on cell morphology. In the present study we compared the effect of the concentration of formaldehyde fixative for individual cells in the blood and their role in enhancing the shelf life and maintaining the morphology of the cells when suspended in plasma or suitable buffers post-fixation. It was observed that WBCs can be better fixed with 3 and 3.5% formaldehyde in a buffered solution, whereas RBCs and Platelets can be optimally fixed with 2.5% formaldehyde in a buffered solution.

In vitro and in vivo anti-inflammatory and anti-arthritic effect of Tinospora cordifolia via modulation of JAK/STAT pathway.

BACKGROUND: Rheumatoid arthritis (RA) is a chronic inflammatory disorder causing cartilage and joint degeneration. In spite of the availability of several robust drugs like biologics, most of the patients are unresponsive, and reports of severe adverse effects following long-term use are also there. Subsequently the use of natural plant-based products in RA therapy is broadening over the years. Tinospora cordifolia is a widely used medicinal plant in Ayurveda against various inflammatory disorders including RA. However, there is very limited knowledge regarding the actual molecular events responsible for its therapeutic effect, and this has limited its acceptance among the professionals. PURPOSE: To explore the anti-inflammatory and anti-arthritic effect of hydro-alcoholic extract from Tinospora cordifolia. METHODS: The rich polyphenol nature of the extract was elucidated using HPLC. LPS-stimulated murine macrophage cell line RAW 264.7 was used for in vitro studies, and collagen-induced arthritis (CIA) model was used for in vivo studies. RESULTS: The polyphenols in TCE were identified using HPLC. TCE effectively downregulated the level of pro-inflammatory mediators (IL-6, TNF-α, PGE2, and NO) in LPS-stimulated RAW 264.7 cells. Subsequently the upregulated expression of COX-2 and iNOS following LPS stimulation were also downregulated by TCE. Furthermore, TCE targeted the upstream kinases of the JAK/STAT pathway, a crucial inflammatory pathway. The expression of VEGF, a key angiogenic factor as well as an inflammatory mediator was also decreased following pre-treatment with TCE. The anti-arthritic effect of TCE (150 mg/kg) was evaluated in the CIA model as well. From the results of histopathology, oral administration of TCE was found to be effective in reducing the clinical symptoms of arthritis including paw edema, erythema, and hyperplasia. In vivo results validated the in vitro results and there was a significant reduction in serum level of pro-inflammatory cytokines and mediators (IL-6, TNF-α, IL-17, NO, and PGE2). The phosphorylation of STAT3 and the expression of VEGF were also downregulated following TCE treatment. CONCLUSION: Our study provided a detailed insight into the molecular events associated with anti-inflammatory and anti-arthritic effect of Tinospora cordifolia.

Design and development of a self-propelled indoor wheelchair with direct toilet accessibility.

PURPOSE: Existing wheelchair (WC) designs lack independent toilet accessibility, transferability and indoor manoeuvrability combined in a WC. In this regard, a new WC model is proposed to simultaneously accommodate these distinct and essential features. In this open-source hardware project, the design, analysis and fabrication of a new active commode WC for indoor applications are performed. MATERIALS AND METHOD: A field survey is conducted to identify the requirements of users so as to choose specific features and design constraints for the WC. In this multipurpose self-propelled model, a temporarily detachable circular section is fitted in the seat position to access the water closet bowl. This hinged portion can be released by gravity before reaching above the closet basin. After the toilet usage, the circular lid can be brought back to the initial seating using a lever mechanism and a spring-loaded lock restrains its downward movement. Thereafter, it can be used as an ordinary WC for indoor applications. Moreover, a small wheel with 22 in diameter and removable hand-rests ensured the transferability between utilities. A CAD model is prepared and numerical simulation is performed to verify the structural stability of the design. RESULTS: The optimized model is fabricated using standard manufacturing practices and is delivered to different persons with disabilities to collect user feedback. A systematic skill test is conducted to validate the compliance of the prototype with the user requirements including transferability and independent restroom access. CONCLUSION: The opinions from most of the users are positive which categorically indicated that the proposed design addresses the multipurpose mobility requirements at indoors.Implications for rehabilitationThe expected outcomes and implications of the current commode WC project with regard to rehabilitation purpose are enlisted below:In the present scenario, a WC user is heavily dependent on external assistance to access private spaces in general and toilets in particular. The introduced model is capable to address this shortcoming by enabling the user to access the toilet utility directly by self propelling.Indeed, for a person with locomotor disability, a WC is the natural extension of his physical body. Thereby, this multipurpose commode WC design offers much Independence and greater flexibility to the indoor movements.The proposed design instils confidence to a WC user to access and transfer between facilities by means of self effort. Consequently it is expected to improve the quality of one's personal life significantly.Importantly, this is an open-source project, those who are interested in rehabilitation motives can modify the present design to suit their local requirements especially in resource-limited settings.

Ferulic acid attenuates high glucose-induced MAM alterations via PACS2/IP3R2/FUNDC1/VDAC1 pathway activating proapoptotic proteins and ameliorates cardiomyopathy in diabetic rats.

BACKGROUND: Diabetic cardiomyopathy (DCM) is one of the severe complications of diabetes with no known biomarkers for early detection. Mitochondria-associated endoplasmic reticulum membranes (MAM) are less studied subcellular targets but an emerging area for exploration in metabolic disorders including DCM. We herein studied the role of MAMs and downstream mitochondrial functions in DCM. We also explored the efficacy of ferulic acid (FeA) against DCM via modulation of MAM and its associated signaling pathway. METHODS: The H9c2 cardiomyoblast cells were incubated with high concentration (33 mM) of d-glucose for 48 h to create a high glucose ambience in vitro. The expression of various critical proteins of MAM, mitochondrial function, oxidative phosphorylation (OxPhos) and the genesis of apoptosis were examined. The rats fed with high fat/high fructose/streptozotocin (single dose, i.p.) were used as a diabetic model and analyzed the insulin resistance and markers of cardiac hypertrophy and apoptosis. RESULTS: High glucose conditions caused the upregulation of MAM formation via PACS2, IP3R2, FUNDC1, and VDAC1 and decreased mitochondrial biogenesis, fusion and OxPhos. The upregulation of mitochondria-driven SMAC-HTRA2-ARTS-XIAP apoptosis and other cell death pathways indicate their critical roles in the genesis of DCM at the molecular level. The diabetic rats also showed cardiomyopathy with increased heart mass index, TNNI3K, troponin, etc. FeA effectively prevented the high glucose-induced MAM alterations and associated cellular anomalies both in vitro and in vivo. CONCLUSION: High glucose-induced MAM distortion and subsequent mitochondrial dysfunctions act as the stem of cardiomyopathy. MAM could be explored as a potential target to treat diabetic cardiomyopathy. Also, the FeA could be an attractive nutraceutical agent for diabetic cardiomyopathy.

Influence of Dy3+ ion concentration on photoluminescence and energy transfer mechanism of promising KBaScSi3O9 phosphors for warm white LEDs.

A novel white light emitting silicate-based phosphor of Dy3+ activated KBaScSi3O9 (xDyKBS) was prepared by a conventional solid state method. Powder X-ray diffraction patterns represent the pure phase of synthesized materials that was formed with monoclinic structure. The metal-ligand bonding nature and electronic band structure have been examined optical absorption spectra. The luminescence emission curves of the silicate phosphors display an intense yellow emission peak at 579 nm and a blue emission peak at 491 nm. Among the emission bands, the band observed in the yellow region due to 4F9/2→6H13/2 transition was found to be higher intensity. The radiative parameters like transition probabilities (AR), branching ratios (ßR) and stimulated emission cross-section (σPE) values were calculated using Judd-Ofelt parameters and refractive index values for the observed transitions in emission spectra. The life time measurements were made for 4F9/2 → 6H13/2 transition of all the studied samples by keeping an excitation at 350 nm and emission at 579 nm and decay curves were fitted to bi-exponential fitting method. The CCT values obtained from the color coordinates suggested that present xDyKBS phosphors can emit warm and neutral white light depending upon the dopant concentration under near-UV excitation. Our results demonstrated that the optimum concentration 0.05DyKBS phosphor can be successfully utilized as a promising and potential candidate for various innovative photonic applications like warm white LEDs, solar cells, optical sensors and lasers.

In vitro and in vivo studies reveal the beneficial effects of chlorogenic acid against ER stress mediated ER-phagy and associated apoptosis in the heart of diabetic rat.

Endoplasmic reticulum (ER) and associated signaling pathways are involved in diabetic cardiomyopathy (DCM) however, detailed studies are not available. The present study investigated the role of ER stress and related pathways such as ER-phagy, apoptosis and their underlying mechanisms using appropriate models. Beneficial effect of chlorogenic acid was also evaluated against ER stress mediated DCM. H9c2 cells with high glucose (33 mM, in vitro model of hyperglycemia) showed significant activation of ER stress response (GRP78, PERK, IRE1α, ATF6α) and altered its regulatory proteins (PDI, ERO1α). Also, it enhanced ER-phagy through upregulation of Sec62, RTN3 and downregulation of FAM134B. High glucose caused apoptosis via increased levels of CHOP, caspase 12 and calnexin. All these proteins (PERK, IRE1α, ATF6α, RTN3, Sec62 and FAM134B) have been found to have a significant role in the functioning of heart such as excitation contraction coupling and we expect these alterations to induce cardiomyopathy during diabetes. This was confirmed in in vivo study too. High fat, high fructose diet with mild streptozotocin induced diabetic rats showed an increased expression of BNP confirming cardiac injury. We also noticed severe ER stress in the heart of diabetic animals. All these have contributed significantly into alterations in histopathology and increase of weight of the hearts. These findings clearly show that ER stress plays a vital protagonist in the progression of DCM. We also found chlorogenic acid is effective against hyperglycemia induced pathological alteration both in vitro as well as in vivo.

Mixing characterization of binary-coalesced droplets in microchannels using deep neural network.

Real-time object identification and classification are essential in many microfluidic applications especially in the droplet microfluidics. This paper discusses the application of convolutional neural networks to detect the merged microdroplet in the flow field and classify them in an on-the-go manner based on the extent of mixing. The droplets are generated in PMMA microfluidic devices employing flow-focusing and cross-flow configurations. The visualization of binary coalescence of droplets is performed by a CCD camera attached to a microscope, and the sequence of images is recorded. Different real-time object localization and classification networks such as You Only Look Once and Singleshot Multibox Detector are deployed for droplet detection and characterization. A custom dataset to train these deep neural networks to detect and classify is created from the captured images and labeled manually. The merged droplets are segregated based on the degree of mixing into three categories: low mixing, intermediate mixing, and high mixing. The trained model is tested against images taken at different ambient conditions, droplet shapes, droplet sizes, and binary-fluid combinations, which indeed exhibited high accuracy and precision in predictions. In addition, it is demonstrated that these schemes are efficient in localization of coalesced binary droplets from the recorded video or image and classify them based on grade of mixing irrespective of experimental conditions in real time.

Anorectal melanoma: experience from a tertiary cancer care centre in South India.

Introduction Mucosal malignant melanoma of the anorectum is a rare and aggressive disease, in which early diagnosis is difficult. The prognosis remains extremely poor, irrespective of the treatment. We share our experience in treating this malignancy at our centre in South India. Methods This study describes a retrospective analysis of 31 cases of anorectal melanoma presented to our centre between January 2001 and December 2013. Results Twenty-two patients (71%) presented with metastasis and had a median overall survival of nine months. None of the 22 patients survived for two years. Nine patients (29%) had curative surgery, in the form of abdominoperineal resection (six patients), abdominoperineal resection with bilateral inguinal node dissection (one patient), abdominoperineal resection with liver resection (one patient) and posterior exenteration (one patient). In patients who underwent curative surgery, the median overall survival was 15 months and disease-free survival was nine months, with a two-year overall survival of 22%. Conclusions Anorectal melanoma is an aggressive disease with a poor prognosis. The majority of patients present with distant metastases. Prognosis depends on stage at presentation. Early diagnosis and surgical resection may improve the overall outcome. Newer modalities such as immunotherapy and targeted therapies such as anti-CTLA4 monoclonal antibody and anti-programmed cell death protein 1 monoclonal antibodies have radically changed the management of mucosal melanoma and may, in the future, improve the overall prognosis of anorectal melanoma.

Mesoscopic simulation of single DNA dynamics in rotational flows.

In this numerical study, the transport and dynamics of an isolated DNA in rotational flow generated in a microchannel have been investigated using dissipative particle dynamics. Often, inertial flow through microchannels with a sudden change in surface structure facilitates a re-circulation or vortex region. The conformation and mobility of the bio-polymer under the influence of such rotating fluid inside a square cavity of the microchannel is analyzed. The flexible polymer chain is found to migrate towards the rotating region and follows the vortex streamline. The orientation, size and tumbling period of polymer strands are affected by the strength of the microvortex. At elevated flow rates, the macromolecule prefers to remain inside the vortex and a hydrodynamic trap is formed. Moreover, residence time of the single molecule in the microcavity is significantly influenced by the chain length and flow strength. Further, it has been demonstrated that, such entrapment duration can be strategically altered by modifying the hydrophobicity of the microchannel.

Crystal structure of piperazine-1,4-diium bis-(4-amino-benzene-sulfonate).

The asymmetric unit of the title salt, C4H12N2 (2+)·2C6H6NO3S(-), consists of half a piperazindiium dication, located about an inversion centre, and a 4-amino-benzene-sulfonate anion. The piperazine ring adopts a chair conformation. In the crystal, the cations and anions are linked via N-H⋯O and C-H⋯O hydrogen bonds, forming a three-dimensional framework. Within the framework there are C-H⋯π inter-actions and the N-H⋯O hydrogen bonds result in the formation of R 4 (4)(22) and R 3 (4)(13) ring motifs.

Crystal structure of 1-cyclo-propane-carbon-yl-3-methyl-2,6-di-p-tolyl-piperidin-4-one.

The title compound, C24H27NO2, crystallizes with two independent mol-ecules (A and B) in the asymmetric unit. The two mol-ecules have very similar conformations and each exhibits an intra-molecular C-H⋯π inter-action. The central piperidine rings adopt boat conformations and the p-tolyl rings are inclined to the mean plane of the piperidine ring by 71.21 (11) and 89.86 (12)° in mol-ecule A and by 68.01 (12) and 89.33 (12)° in mol-ecule B. The cyclopropanecarbonyl group is oriented at an angle of 68.5 (2)° with respect to the mean plane of the piperidine ring in mol-ecule A and 66.2 (2)° in mol-ecule B. In the crystal, the A and B mol-ecules are linked by C-H⋯O hydrogen bonds, enclosing R 2 (1)(6) ring motifs, forming ribbons running along the a-axis direction.

N'-[(E)-3-Bromo-5-chloro-2-hy-droxy-benzyl-idene]furan-2-carbohydrazide.

In the title compound, C12H8BrClN2O3, the furan ring makes a dihedral angle of 17.2 (2)° with the six-membered ring. An intra-molecular O-H⋯N hydrogen bond stabilizes the mol-ecular conformation. In the crystal, N-H⋯O hydrogen bonds connect the mol-ecules into chains running along the c-axis direction. The crystal packing is additionally stabilized by C-H⋯O inter-actions into a three-dimensional supramolecular architecture.

Transport of DNA in hydrophobic microchannels: a dissipative particle dynamics simulation.

In this work, we numerically study a new means of manipulating single DNA chains in microchannels. The method is based on the effect of finite slip at hydrophobic walls on the hydrodynamics and, consequently, on the dynamics of the DNA in microchannels. We use dissipative particle dynamics to study DNA transport as a function of chain length and the Reynolds number in two dimensional parallel plate channels. We show how an asymmetric velocity profile in a channel with hydrophobic and hydrophilic walls can be used to manipulate the location of the DNA molecules. Using this effect, we propose a simple arrangement of hydrophobic and hydrophilic strips which can be exploited to separate long and short DNA chains.

Is it possible to detect dendrite currents using presently available magnetic resonance imaging techniques?

The action currents of a dendrite, peripheral nerve or skeletal muscle create their own magnetic field. Many investigators have attempted to detect neural and dendritic currents directly using magnetic resonance imaging that can cause the phase of the spins to change. Our goal in this paper is to use the calculated magnetic field of a dendrite to estimate the resulting phase shift in the magnetic resonance signal. The field produced by a dense collection of simultaneously active dendrites may be just detectable under the most ideal circumstances, but in almost every realistic case the field cannot be detected using current MRI technology.

1-Benzyl-2,5-diphenyl-3-tosylimidazol-idin-4-one.

In the title compound, C(29)H(26)N(2)O(3)S, the central imidazolidine ring adopts an envelope conformation with the N atom bearing the benzyl ring at the flap. The S atom has distorted tetra-hedral geometry. The benzyl and tosyl rings are oriented at a dihedral angle of 52.1 (1)°. The phenyl rings connected to the imidazolidine ring form a dihedral angle of 28.7 (1)°.

2-(4-Chloro-phen-yl)-5-(cyclo-hex-1-en-1-yl)-3-(4-methyl-phenyl-sulfon-yl)-1-phenyl-imidazolidin-4-one.

In the title compound, C(28)H(27)ClN(2)O(3)S, the central imidazolidine ring adopts an envelope conformation with the C atom bearing the chloro-phenyl ring at the flap. The geometry around the S atom is distorted tetra-hedral. Three methyl-ene groups of the cyclo-hexene ring are disordered over two sets of sites [site occupancies = 0.562 (10) and 0.438 (10)]. The crystal packing is stabilized by C-H⋯π inter-actions.

Diethyl 2-{[3-(2-meth-oxy-benz-yl)thio-phen-2-yl]methyl-idene}malonate.

In the title compound, C(20)H(22)O(5)S, the dihedral angle between the mean planes through the thio-phene and benzene rings is 75.2 (1)°. The meth-oxy group is essentially coplanar with the benzene ring, the largest deviation from the mean plane being 0.019 (2) Šfor the O atom. The malonate group assumes an extended conformation.

3-Ethyl-5-(4-meth-oxy-phen-oxy)-2-(pyridin-4-yl)-3H-imidazo[4,5-b]pyridine.

In the title compound, C(20)H(18)N(4)O(2), the imidazopyridine fused ring system is almost perpendicular to the benzene ring [dihedral angle = 87.6 (5)°]. The pyridine ring makes a dihedral angle of 35.5 (5)° with the mean plane of the imidazopyridine fragment. The crystal structure is stabilized by an aromatic π-π stacking inter-action between the phenyl rings of neighbouring mol-ecules [centroid-centroid distance = 3.772 (2) Å, inter-planar distance = 3.546 (2) Šand slippage = 1.286 (2) Å].

2-(4-Chloro-phen-yl)-1,5-diphenyl-3-tosyl-imidazolidin-4-one.

In the title compound, C(28)H(23)ClN(2)O(3)S, the central imidazolidine ring adopts a twisted conformation and the S atom has distorted tetra-hedral geometry. The crystal packing is stabilized by C-H⋯O, C-H⋯π and π-π inter-actions [centroid-centroid distance = 3.8302 (10) Å].

Phen-yl(1-phenyl-sulfonyl-1H-indol-2-yl)methanone.

The asymmetric unit of the title compound, C(21)H(15)NO(3)S, contains two crystallographically independent mol-ecules. As a result of the electron-withdrawing character of the phenyl-sulfonyl groups, the N-Csp(2) bond lengths are slightly longer than the anti-cipated value of approximately 1.35 Šfor N atoms with planar configurations. Both unique S atoms have a distorted tetra-hedral configuration. In each mol-ecule, the indole ring system is essentially planar (r.m.s. deviations for all non-H atoms of 0.020 and 0.023 Å). In one mol-ecule, the indole ring system makes dihedral angles of 65.7 (8) and 73.4 (8)°, respectively, with the benzene and phenyl rings [62.2 (7) and 72.1 (7)°, respectively, in the other mol-ecule].