Evaluation of the activity of a novel metabotropic glutamate receptor antagonist (±)-2-amino-2-(3-cis and trans-carboxycyclobutyl-3-(9-thioxanthyl)propionic acid) in the in vitro neonatal spinal cord and in an in vivo pain model

The cyclobutylglycine (±)-2-amino-2-(3-cis and trans-carboxycyclobutyl-3-(9-thioxanthyl)propionic acid) (LY393053) has been identified as a functionally potent metabotropic glutamate receptor antagonist. It is most potent on the two group I metabotropic glutamate receptors, 1α and 5α, with IC₅₀ values of 1.0±0.4 μM and 1.6±1.4 μM, respectively. In this study, LY393053 has also been evaluated electrophysiologically on native group I metabotropic glutamate receptors in an in vitro spinal cord preparation as well as behaviourally, in a mouse model of visceral pain. LY393053 dose-dependently antagonised group I agonist, (RS)-3, 5-dihydroxyphenylglycine, or a broad-spectrum agonist (1S,3R)-amino-1,3-cyclopentanedicarboxylic acid-induced depolarisation of spinal motoneurons. The apparent K_d values were estimated to be 0.3 μM against (RS)-3, 5-dihydroxyphenylglycine-induced depolarisation and 0.5 μM against (1S,3R)-amino-1,3-cyclopentanedicarboxylic acid-induced depolarisation, respectively. On the other hand, the dorsal root–ventral root potential elicited at 8× threshold was depressed by LY393053 with IC₅₀ values of 9.0±0.7 μM and 12.7±1.7 μM on monosynaptic and polysynaptic responses, respectively. When investigated using the mouse acetic acid writhing test, LY393053 showed significant analgesic effects at doses of 1–10 mg/kg intraperitoneally. An ED₅₀ value of 6.0 mg/kg was obtained in this test.By revealing a potent effect of LY393053 in antagonising the native group I metabotropic receptor-mediated responses in the spinal cord in rodents, and an antinociceptive efficacy in a mouse visceral pain model, these results, therefore, provide additional evidence in support of the analgesic potential of metabotropic glutamate receptor antagonists.     

Evaluation of the in vitro and in vivo proinflammatory activities of gold (+) and gold (−) nanoparticles

Objective and design

The aim of this study was to determine potential effects of gold (+) and gold (?) nanoparticles, AuNP(+) and AuNP(?), on neutrophil biology.

Material or subjects

Freshly isolated human neutrophils were used for the in vitro aspects and CD-1 mice were used in the in vivo murine air pouch model of acute neutrophilic inflammation.

Treatment

Human neutrophils were treated with the indicated concentrations of AuNP(+) or AuNP(?) in vitro and mice received 100 or 500 µg/ml AuNP(+) or AuNP(?) into air pouches.

Methods

Cellular uptake of AuNP by neutrophils was confirmed by transmission electron microscopy and the ability of the NP to modulate apoptosis, gelatinase activity, and chemokine production and chemotaxis was determined by cytology, zymography, ELISArray, antibody array, and ELISA and by a micro-chemotaxis chamber, respectively. In vivo, exudates were harvested after 6 h to determine the leukocyte infiltration to detect the production of several cytokines by an antibody array approach and ELISA. One-way analysis of variance was used for statistical analysis.

Results

AuNP possess proinflammatory activities in vitro and induce mainly a neutrophil influx in vivo, albeit at different degrees.

Conclusions

AuNP(+) and AuNP(?) should be added as new candidates into a growing list of NP having proinflammatory activities by themselves.

     

In vitro and in vivo antibacterial activity of tigecycline against Vibrio vulnificus

Background/purpose

The aim of this study is to investigate the role of tigecycline in Vibrio vulnificus infection.

Effects of a neem seed extract (MiteStop®) on mallophages (featherlings) of chicken: in vivo and in vitro studies

Mallophages of birds (featherlings) are mostly very tiny and can even as adults better be recognized by their movements than by their elongate body shape when using just the naked eye. Since some species (e.g., the “shaft louse” Menopon gallinae, the elongate feather louse Lipeurus caponis, or Columbicola sp.) may pierce the pulp of feathers or the skin by their biting or scratching mandibles and thus lick the excreted blood, they may be extremely dangerous especially to young birds, even if they only feed by nibbling along the feather surface and/or eat epidermal debris. The present paper reports on the successful treatment of different races of fowls being severely infested with both above cited species. This in vivo treatment was done either by a short dipping of the whole fowl into the 1:33 dilution (with tap water) of a neem seed extract (MiteStop®) or by spraying them with the freshly diluted product. It was seen that the dead mallophages dropped down from the feathers as soon as they were dry again. As a precaution, a second treatment was done by some owners 1 week after the first one in order to eliminate all stages, which eventually might have hatched from untouched nits during the time interval between the two treatments. When controlling the treated fowls 4 weeks after the treatment, in no case (treated once or twice), living motile stages were diagnosed indicating the high efficacy of this nontoxic neem seed extract. When treating in vitro cutoff feathers contaminated with L. caponis, it was seen under the stereomicroscope, that the mallophages tried to run away from the 1:33 water-diluted active compound indicating that there is also a repellent effect. Treated L. caponis stopped leg movements within 3 min and died on their feathers within 1–20 min. Then, the last slight trembling movements of their legs and convulsions of their intestine stopped finally.     

In vitro corrosion behavior and in vivo biodegradation of biomedical β-Ca3(PO4)2/Mg-Zn composites

In this study 5, 10 and 15% β-Ca(3)(PO(4))(2)/Mg-Zn composites were prepared through powder metallurgy methods, and their corrosion behavior and mechanical properties were studied in simulated body fluid (SBF) at 37°C. The 10% β-Ca(3)(PO(4))(2)/Mg-Zn composite was selected for cytocompatibility assessment and in vivo biodegradation testing. The results identified the α-Mg, MgZn and β-Ca(3)(PO(4))(2) phases in these sintered composites. The density and elastic modulus of the β-Ca(3)(PO(4))(2)/Mg-6% Zn composite match those of natural bone, and the strength is approximately double that of natural bone. The 10% β-Ca(3)(PO(4))(2)/Mg-6% Zn composites exhibit good corrosion resistance, as determined by a 30 day immersion test and electrochemical measurements in SBF at 37°C. The 10% β-Ca(3)(PO(4))(2)/Mg-6% Zn composite is safe for cellular applications, with a cytotoxicity grade of ～0-1 against L929 cells in in vitro testing. The β-Ca(3)(PO(4))(2)/Mg-6% Zn composite also exhibits good biocompatibility with the tissue and the important visceral organs the heart, kidney and liver of experimental rabbits. The composite has a suitable degradation rate and improves the concrescence of a pre-broken bone. The corrosion products, such as Mg(OH)(2) and Ca(5)(PO(4))(6)(OH)(2), can improve the biocompatibility of the β-Ca(3)(PO(4))(2)/Mg-Zn composite.     

Expression and regulation of αB-crystallin in the kidney in vivo and in vitro

αB-crystallin, a major component of the mammalian eye lens, is a small heat shock protein and molecular chaperone that is also abundant in the mammalian kidney. The present study aimed to characterize more closely the intrarenal expression and regulation of αB-crystallin in vivo and in vitro. In normal rat kidney, the expression of αB-crystallin mRNA and protein were both close to the detection limit in cortex, but increased steeply from the outer to the inner medulla where αB-crystallin constitutes approximately 2% of total tissue protein. Immunohistochemistry disclosed papillary collecting duct cells and thin limbs as the major sites for intrapapillary αB-crystallin immunoreactivity. In rats subjected to sucrose diuresis for 3 days, αB-crystallin mRNA expression was reduced by 27 and 46% in outer and inner medulla, respectively. In agreement with the results obtained in vivo, in Madine–Darby canine kidney cells, αB-crystallin mRNA and protein were induced significantly by elevating the medium osmolality to 500 mosm/kg H₂O by the addition of NaCl and raffinose, and also by urea. The NaCl-induced increase in αB-crystallin expression was concentration-dependently blunted by SP600125, a specific JNK inhibitor. Overexpression of αB-crystallin in 293 cells resulted in increased tolerance to acute osmotic stress. These results indicate that αB-crystallin may be regulated by papillary interstitial tonicity in a JNK-dependent process. Moreover, the high abundance of αB-crystallin in the renal medulla may be important for cell survival in an environment characterized by extreme interstitial solute concentrations as present during antidiuresis.     

CaSiO₃ microstructure modulating the in vitro and in vivo bioactivity of poly(lactide-co-glycolide) microspheres

Poly(lactide-co-glycolide) (PLGA) microspheres have been used for regenerative medicine due to their ability for drug delivery and generally good biocompatibility, but they lack adequate bioactivity for bone repair application. CaSiO? (CS) has been proposed as a new class of material suitable for bone tissue repair due to its excellent bioactivity. In this study, we set out to incorporate CS into PLGA microspheres to investigate how the phase structure (amorphous and crystal) of CS influences the in vitro and in vivo bioactivity of the composite microspheres, with a view to the application for bone regeneration. X-ray diffraction (XRD), N? adsorption-desorption analysis, and scanning electron microscopy (SEM) were used to analyze the phase structure, surface area/pore volume, and microstructure of amorphous CS (aCS) and crystal CS (cCS), as well as their composite microspheres. The in vitro bioactivity of aCS and cCS-PLGA microspheres was evaluated by investigating their apatite-mineralization ability in simulated body fluids (SBF) and the viability of human bone mesenchymal stem cells (BMSCs). The in vivo bioactivity was investigated by measuring their de novo bone-formation ability. The results showed that the incorporation of both aCS and cCS enhanced the in vitro and in vivo bioactivity of PLGA microspheres. cCS/PLGA microspheres improved better in vitro BMSC viability and de novo bone-formation ability in vivo, compared to aCS/PLGA microspheres. Our study indicates that controlling the phase structure of CS is a promising method to modulate the bioactivity of polymer microsphere system for potential bone tissue regeneration.     

Porous alginate/poly(ε-caprolactone) scaffolds: preparation, characterization and in vitro biological activity

In bone tissue engineering, scaffolds with controlled porosity are required to allow cell ingrowth, nutrient diffusion and sufficient formation of vascular networks. The physical properties of synthetic scaffolds are known to be dependent on the biomaterial type and its processing technique. In this study, we demonstrate that the separation phase technique is a useful method to process poly(ε-caprolactone) (PCL) into a desired shape and size. Moreover, using poly(ethylene glycol), sucrose, fructose and Ca2+ alginate as porogen agents, we obtained PCL scaffolds with three-dimensional porous structures characterized by different pore size and geometry. Scanning electron microscopy and porosity analysis indicated that PCL scaffolds prepared with Ca2+ alginate threads resemble the porosity and the homogeneous pore size distribution of native bone. In parallel, MicroCT analysis confirmed the presence of interconnected void spaces suitable to guarantee a biological environment for cellular growth, as demonstrated by a biocompatibility test with MC3T3-E1 murine preosteoblastic cells. In particular, scaffolds prepared with Ca2+ alginate threads increased adhesion and proliferation of MC3T3-E1 cells under basal culture conditions, and upon stimulation with a specific differentiation culture medium they enhanced the early and later differentiated cell functions, including alkaline phosphatase activity and mineralized extracellular matrix production. These results suggest that PCL scaffolds, obtained by separation phase technique and prepared with alginate threads, could be considered as candidates for bone tissue engineering applications, possessing the required physical and biological properties.     

Histamine release “in vivo” and “in vitro” induced by hypnotics in normal and atopic patients

The “in vitro” test of histamine release induced in the leukocytes of atopic subjects selected according to specific criteria would seem to be much more accurate to study the histamine releasing characteristics of intravenous agents than the “in vivo” study in patients who have to be anaesthetised. Moreover, different concentrations of the test drug may be used, and thus the threshold for histamine release may be compared. It is the test which we recommend for investigating new drugs. However, it is lengthy and expensive; it can therefore not be recommended as a routine preoperative investigation.     

In vivo and in vitro characteristic of HIF-1α and relative genes in ischemic femoral head necrosis

Background: Legg-Calvé-Perthes Disease (Perthes’ disease) is a childhood hip disorder initiated by ischemic necrosis of the growing femoral head. So far, the etiology and pathogenesis of Perthes’ disease is poorly understood. Materials and methods: Avascular osteonecrosis rat model was established to mimic the pathophysiological changes of femoral head necrosis. The chondrocytes of newborn Sprague-Dawley rats were isolated and cultured in hypoxic and normoxic condition. The expression characteristic of the hypoxia-inducible factor-1 alpha (HIF-1α) was evaluated both in vivo and in vitro models. Vascular endothelial growth factor (VEGF) and apoptotic genes in chondrocytes treated with normoxia and hypoxia were also studied. Results: HIF-1α expression increased greatly after ischemic operation and kept at relative high level in the arthromeningitis stage and declined in the stages of osteonecrosis and reconstruction. The HIF-1α mRNA levels of chondrocytes incubated at hypoxia were significantly higher than the cells treated with normoxia at 24 and 72 hours. Hypoxia inhibited VEGF expression; chondrocytes could oppose this inhibition manifested by the increasing of VEGF mRNA level after 72 hours hypoxia. The expression of apoptotic genes, Casp3, Casp8 and Casp9, elevated in chondrocytes after hypoxia with time differences. Conclusion: Hypoxia might be an etiological factor for femoral head necrosis, HIF-1α, VEGF as well as apoptotic genes participated the pathophysiological process of ischemic osteonecrosis.     

Inducible expression of TGF��, Snail and Zeb1 recapitulates EMT in vitro and in vivo in a NSCLC model

The progression of cancer from non-metastatic to metastatic is the critical transition in the course of the disease. The epithelial to mesenchymal transition (EMT) is a mechanism by which tumor cells acquire characteristics that improve metastatic efficiency. Targeting EMT processes in patients is therefore a potential strategy to block the transition to metastatic cancer and improve patient outcome. To develop models of EMT applicable to in vitro and in vivo settings, we engineered NCI-H358 non-small cell lung carcinoma cells to inducibly express three well-established drivers of EMT: activated transforming growth factor β (aTGFβ), Snail or Zeb1. We characterized the morphological, molecular and phenotypic changes induced by each of the drivers and compared the different end-states of EMT between the models. Both in vitro and in vivo, induction of the transgenes Snail and Zeb1 resulted in downregulation of epithelial markers and upregulation of mesenchymal markers, and reduced the ability of the cells to proliferate. Induced autocrine expression of aTGFβ caused marker and phenotypic changes consistent with EMT, a modest effect on growth rate, and a shift to a more invasive phenotype. In vivo, this manifested as tumor cell infiltration of the surrounding mouse stromal tissue. Overall, Snail and Zeb1 were sufficient to induce EMT in the cells, but aTGFβ induced a more complex EMT, in which changes in extracellular matrix remodeling components were pronounced.     

In vivo anti-influenza virus activity of Kampo (Japanese herbal) medicine “Sho-seiryu-to” and its mode of action

When BALB/c mice were treated with a Kampo (Japasese herbal) medicine “Sho-seiryu-to” (SST) (2 g/kg, 10 times) orally from 7 days before to 4 days after the infection and infected with mouse-adapted influenza virus A/PR/8/34 by nasal site-restricted infection, replication of the virus in the nasal cavity and spread of the virus to the lung were efficiently inhibited at 5 days after infection in comparison with water-treated mice. However, another Kampo medicine “Kakkon-to” showed no anti-influenza virus activity in the same condition. The antiviral IgA antibody in the nasal and broncho-alveolar washes of the SST treated mice increased significantly in comparison with that of water-treated control. Oral administration of SST (2 g/kg, 18 times) from 7 days before to 13 days after vaccination also significantly augmented serum hemagglutination-inhibiting antibody by nasal inoculation of influenza HA vaccine (5 μg/mouse) that was insufficient to induce antiviral antibody. SST did not inhibit the replication of mouse-adapted influenza virus A/PR/8/34 in Madin-Darby canine kidney cells. SST also did not inhibit the influenza virus sialidase activity against sodium p-nitrophenyl-N-acetyl-α-D-neuraminate and hemagglutination by mouse-adapted influenza virus A/PR/8/34. SST showed no influence on interferon production in nasal wash of mice at 5 days after the virus infection. These results suggest that SST confers better protection against influenza virus infection through augmentation of production of antiviral IgA antibody but not direct action to the virus, and can be used as an adjuvant to nasally inoculated influenza HA vaccine.     

Diverse roles of TGF-β receptor II in renal fibrosis and inflammation in vivo and in vitro

TGF-β1 binds receptor II (TβRII) to exert its biological activities but its functional importance in kidney diseases remains largely unclear. In the present study, we hypothesized that TβRII may function to initiate the downstream TGF-β signalling and determine the diverse role of TGF-β1 in kidney injury. The hypothesis was examined in a model of unilateral ureteral obstructive (UUO) nephropathy and in kidney fibroblasts and tubular epithelial cells in which the TβRII was deleted conditionally. We found that disruption of TβRII inhibited severe tubulointerstitial fibrosis in the UUO kidney, which was associated with the impairment of TGF-β/Smad3 signalling, but not with the ERK/p38 MAP kinase pathway. In contrast, deletion of TβRII enhanced NF-κB signalling and renal inflammation including up-regulation of Il-1β and Tnfα in the UUO kidney. Similarly, in vitro disruption of TβRII from kidney fibroblasts or tubular epithelial cells inhibited TGF-β1-induced Smad signalling and fibrosis but impaired the anti-inflammatory effect of TGF-β1 on IL-1β-stimulated NF-κB activation and pro-inflammatory cytokine expression. In conclusion, TβRII plays an important but diverse role in regulating renal fibrosis and inflammation. Impaired TGF-β/Smad3, but not the non-canonical TGF-β signalling pathway, may be a key mechanism by which disruption of TβRII protects against renal fibrosis. In addition, deletion of TβRII also enhances NF-κB signalling along with up-regulation of renal pro-inflammatory cytokines, which may be associated with the impairment of anti-inflammatory properties of TGF-β1.     

Comparison of in vitro activity of metronidazole and garlic-based product (Tomex®) on Trichomonas vaginalis

Trichomonas vaginalis is a common sexually transmitted parasite in humans. Metronidazole has been the gold standard for treatment of trichomoniasis. The prevalence of metronidazole resistance and its unpleasant adverse effects drew the attention to the investigation of other lines of treatment, as that of herbal medicine. Garlic has been proven to have antibacterial, antiprotozoal, and antihelminthic activity. The current study was carried out to evaluate the efficacy of commercially available garlic (Tomex®) on T. vaginalis in vitro. The effect of different concentrations of garlic (12.5, 25, 50, and 100 μg/ml) was determined on multiplication and motility of trophozoites at different time points (after 24, 48, 72, and 96 h) in comparison to the same concentrations of metronidazole at the same different time points. The results showed that parasite multiplication inhibition was noticed in proportion of concentration of Tomex and incubation time. The minimal lethal concentration of Tomex was 100 μg/ml after 24 h, 50 μg/ml after 48 h, 25 μg/ml after 72 h, and 12.5 μg/ml after 96 h. These results were similar to that of metronidazole as its minimal lethal concentration was 50 μg/ml after 24 and 48 h and 12.5 μg/ml after 72 and 96 h. Garlic had completely inhibited the motility of trophozoites with concentration of 100 μg/ml after 24 h, 50 μg/ml after 48 h, 25 μg/ml after 72 h, and 12.5 μg/ml after 96 h while metronidazole had completely inhibited the motility of trophozoites with concentration of 50 μg/ml after 24 h, 25 μg/ml after 48 h, and 12.5 μg/ml after 72 and 96 h. This suggests that commercially available garlic (Tomex®) may be a promising phytotherapeutic agent for trichomoniasis.     

Echinococcus multilocularis: immunological study on the “Em2-positive” laminated layer during in vitro and in vivo post-oncospheral and larval development

Echinococcus multilocularis oncospheres, primary vesicular cysts, and protoscolices were assessed in vitro and in vivo for their potential to synthesize a PAS-positive laminated layer containing monoclonal antibody (mAb) G11-binding Em2 antigen. The presence of Em2 antigen in developed oncospheres and cysts was subsequently correlated to the potential of in vivo development into a secondary metacestode in recipient host mice, which also responded by anti-Em2 serum antibody formation. In contrast, protoscolices failed to develop the Em2-positive layer in vitro under the selected experimental conditions. The failure to develop subsequently in vivo into a secondary metacestode was underlined by a lack of anti-Em2 serum antibody formation by the hosts. We furthermore developed a technique to obtainE. multilocularis clones by inoculating single oncospheres into recipient mice.This work was supported by grant 31-29651.90 from the Swiss National Science Foundation     

Mycobacterium bovis bacillus Calmette-Guérin (BCG) induces human CC- and CXC-chemokines in vitro and in vivo

Both CC- and CXC-chemokines are known to be potent leucocyte activators and chemoattractants and play important roles in inflammatory responses. However, chemokine response to bacillus Calmette-Guérin (BCG) infection remains incompletely defined. In this study, we investigated human CC- [macrophage-derived chemokine (MDC), monocyte chemoattractant protein (MCP)-1, macrophage inflammatory protein (MIP)-1alpha and eosinophil chemoattractant activity (eotaxin)] and CXC-interferon-inducible protein (IP)-10 chemokine production in response to BCG stimulation. BCG efficiently induced all chemokines tested in the urine of four bladder cancer patients undergoing intravesical BCG immunotherapy. The peak urinary chemokine responses occurred generally between the fourth and sixth weekly treatment, except eotaxin, which was less predictable. To evaluate the effect of BCG on induction of chemokines in vitro, urothelial cell lines and peripheral blood mononuclear cells (PBMCs) were used. Although BCG induced no or marginal chemokines from urothelial SV-HUC-1, RT4 and T24 cells, BCG-derived cytokines [interleukin (IL)-1beta, interferon (IFN)-gamma and tumour necrosis factor (TNF)-alpha] induced all chemokines tested except eotaxin from these cell lines. BCG also efficiently induced all chemokines tested except eotaxin from PBMCs of both BCG-naive and BCG-vaccinated subjects. MCP-1 and MIP-1alpha emerged at 4-5 h post-BCG exposure (early chemokines); IP-10 elevated at day 1 and peaked at day 2 (intermediate chemokine); and MDC elevated at day 1 and peaked at day 7 (late chemokine). This kinetic pattern was paralleled with that of BCG-induced cytokines [early: TNF-alpha; intermediate: IL-6 and IL-10; and late: IFN-gamma and granulocyte-macrophage colony-stimulating factor (GM-CSF)]. Taken together, these results indicate that BCG directly or indirectly induces human CC- and CXC-chemokine production, which may represent one of the mechanisms by which BCG exerts its anti-tumour activity.     

Mitochondrial respiratory inhibition and oxidative stress elevate β-secretase (BACE1) proteins and activity in vivo in the rat retina

Cerebral hypometabolism, oxidative stress and β-amyloid peptide (Aβ) accumulation are key pathological events in Alzheimer’s disease (AD). Beta-secretase (BACE, i.e., BACE1), a prerequisite for Aβ genesis, is elevated in sporadic AD. Recent studies show BACE upregulation in experimental conditions likely associated with energy insufficiency and/or oxidative stress. We investigated the effect of sublethal doses of mitochondrial respiratory inhibitors and potential endogenous oxidative substances on BACE expression in vivo using the retina as a model. Retinas were analyzed biochemically and anatomically 48 h following intraocular applications of mitochondrial complex I, II and IV inhibitors including rotenone, 3-nitropropionic acid and sodium azide, and plaque-containing oxidants including Fe³⁺ and Aβ42 fibrils (Aβ42f). All agents caused elevations of BACE proteins and β-site amyloid precursor protein (APP) cleavage product, β-CTF, in retinal lysates in a dose-dependant manner. BACE activity and Aβ40 levels were also increased in agent-treated retinas relative to vehicle controls. BACE immunoreactivity in normal adult rat retina was present mostly in the plexiform layers, indicating a localization of the enzyme to synaptic terminals. No apparent change in laminar or cellular distribution of BACE labeling was detected in the experimental retinas. However, signs of neuronal stress including glial activation were observed in agent-treated retinas especially in high dosage groups. Our data suggest that mitochondrial respiratory inhibition and oxidative stress facilitate BACE expression in vivo. In addition, plaque constituents such as Fe³⁺ and Aβ42f may participate in a self-enforcing cycle of amyloidogenesis via BACE upregulation.     

Respiratory rhythm generation: converging concepts from in vitro and in vivo approaches?

The timing and activation pattern of breathing movements are determined by the respiratory network. This network is amenable to a variety of in vivo and in vitro approaches, which offers a unique opportunity to investigate multiple organizational levels. It is only recently, however, that concepts obtained under in vivo and in vitro conditions are being integrated into a coherent model of breathing behavior. For example, the pre-B?tzinger complex as an essential site for rhythm generation was first identified in vitro, but has since been verified in vivo. Conversely, timing signals provided by other central and peripheral neuronal areas have so far been investigated in vivo, but it is now possible to address these issues with more complex in vitro preparations. Several key issues remain unresolved. For example, to what extent is the respiratory pattern controlled independently of the underlying rhythm? Answers to this and other questions require a dissection of mechanisms that is only possible through a complementary combination of experimental approaches.     

New hemostatic technique with combined use of Hydrofit® and Surgicel®: an in vitro and in vivo study

Journal of Artificial Organs - We developed an effective hemostatic technique using Hydrofit® and Surgicel® simultaneously. The aim of this study was to demonstrate the hemostatic...     

Anti-fungal activity of cold and hot water extracts of spices against fungal pathogens of Roselle (Hibiscus sabdariffa) in vitro

Crude extracts of seven spices, viz. cardamom, chilli, coriander, onion, garlic, ginger, and galangale were made using cold water and hot water extraction and they were tested for their anti-fungal effects against the three Roselle pathogens i.e. Phoma exigua, Fusarium nygamai and Rhizoctonia solani using the ‘poisoned food technique’. All seven spices studied showed significant anti-fungal activity at three concentrations (10, 20 and 30% of the crude extract) in-vitro. The cold water extract of garlic exhibited good anti-fungal activity against all three tested fungi. In the case of the hot water extracts, garlic and ginger showed the best anti-fungal activity. Of the two extraction methods, cold water extraction was generally more effective than hot water extraction in controlling the pathogens. Against P. exigua, the 10% cold water extracts of galangale, ginger, coriander and cardamom achieved total (100%) inhibition of pathogen mycelial growth. Total inhibition of F. nygamai mycelial growth was similarly achieved with the 10% cold water extracts garlic. Against R. solani, the 10% cold water extract of galangale was effective in imposing 100% inhibition. Accordingly, the 10% galangale extract effectively controlled both P. exigua and R. solani in vitro. None of the hot water extracts of the spices succeeded in achieving 100% inhibition of the pathogen mycelial growth.