Search

Background: Many inflammatory responses including chemotaxis, production of nitric oxide, and modulation of pro-inflammatory cytokines in immunological cells are mediated by p38MAPK. Due to its pivotal role, p38MAPK has been extensively explored as a molecular target for inhibition of chronic inflammation; however, it has not been successful so far due to serious toxicity issues. Among several downstream substrates of p38, mitogen-activated protein kinase-activated protein kinase 2 (MK2) has been reported to be a direct and essential downstream component in regulation of innate immune and inflammatory responses. Thus, in this study, we aimed to understand relative molecular differences between p38 and MK2 kinase inhibition in terms of a comparative anti-inflammatory potential along with molecular regulation of toxicity biomarkers such as Phospho c-Jun N-Terminal Kinase (pJNK), caspase-3, and hepatic enzyme levels in relevant human cells in vitro. Results: Both p38 and MK2 inhibitors attenuated lipopolysaccharide-induced pro-inflammatory biomarkers expression. In addition, both these kinase inhibitors inhibited release of Th1 and Th17 cytokines in phytohemagglutinin-induced cells with MK2 inhibitor showing a better potency for inhibition of Th1 cytokine release, interferon-γ. In the mechanistic differentiation studies, p38 inhibitors displayed an increase in pJNK and caspase-3 activity in U937 cells and elevation in aspartate transaminase enzyme in HepG2 cells, whereas MK2 inhibitor did not show such adverse toxic effects. Conclusion: Taken together, inhibition of MK2 kinase can be a relatively preferred strategy as an anti-inflammatory therapy over direct inhibition of p38 kinase in p38MAPK pathway. [ABSTRACT FROM AUTHOR]

Assessment of medical disorders is often aided by objective diagnostic tests which can lead to early intervention and appropriate treatment. In the case of brain dysfunction caused by head injury, there is an urgent need for quantitative evaluation methods to aid in acute triage of those subjects who have sustained traumatic brain injury (TBI). Current clinical tools to detect mild TBI (mTBI/concussion) are limited to subjective reports of symptoms and short neurocognitive batteries, offering little objective evidence for clinical decisions; or computed tomography (CT) scans, with radiation-risk, that are most often negative in mTBI. This paper describes a novel methodology for the development of algorithms to provide multi-class classification in a substantial population of brain injured subjects, across a broad age range and representative subpopulations. The method is based on age-regressed quantitative features (linear and nonlinear) extracted from brain electrical activity recorded from a limited montage of scalp electrodes. These features are used as input to a unique “informed data reduction” method, maximizing confidence of prospective validation and minimizing over-fitting. A training set for supervised learning was used, including: “normal control,” “concussed,” and “structural injury/CT positive (CT+).” The classifier function separating CT+ from the other groups demonstrated a sensitivity of 96% and specificity of 78%; the classifier separating “normal controls” from the other groups demonstrated a sensitivity of 81% and specificity of 74%, suggesting high utility of such classifiers in acute clinical settings. The use of a sequence of classifiers where the desired risk can be stratified further supports clinical utility. [ABSTRACT FROM AUTHOR]

The flavonoid fraction isolated from the ethyl acetate fraction (BF-1) of Butea frondosa (L.) stem bark exhibited distinct antimicrobial activity when tested against 129 bacterial strains belonging to 9 different genera of both gram-positive and gram-negative types. Minimum inhibitory concentration (MIC) of the fraction BF-1 was determined following NCCLS guidelines using the agar dilution method. Twenty-four out of 36 strains of Staphylococcus aureus were inhibited by 50-200 mg/l of the fraction. This fraction also inhibited strains of Bacillus spp., Shigella spp., Salmonella spp. and even a few Pseudomonas at concentrations between 50-200 mg/l. Other bacteria including Escherichia coli, Vibrio cholerae and V. parahaemolyticus were moderately sensitive to BF-1. In the in vivo studies, this fraction offered significant protection to Swiss albino mice at a concentration of 80 microg/mouse (p<0.001) when they were challenged with 50 median lethal dose of Salmonella enteritidis NCTC 74. A fraction named BF-1 that was isolated from an ethyl acetate fraction of Butea frondosa provided protection against an infection from a Salmonella enteritidis NCTC strain.

Purpose: To determine the in vitro and in vivo antibacterial activity of a cardiovascular drug dobutamine hydrochloride., Methods: The minimum inhibitory concentration (MIC) of dobutamine was determined both by agar and broth dilution methods against 331 strains of bacteria from three gram positive and 13 gram negative genera. The antibacterial action of dobutamine was further tested in animal models., Results: Dobutamine was seen to possess powerful inhibitory action (5-200mg/mL) against most test bacteria in in vitro studies. It was bacteriostatic in nature. In vivo studies showed that the drug offered significant protection (p< 0.001) to mice challenged with a virulent bacterium., Conclusion: Dobutamine showed remarkable antibacterial property against several pathogenic bacteria. Its potential as an antibacterial agent may be confirmed after further pharmacological studies.

The antiinflammatory agent diclofenac sodium (Dc) exhibited remarkable antibacterial effects both in vitro and in vivo. Fifteen different bacteria sensitive to Dc as well as to a number of common antibiotics were tested for synergistic effects in vitro. Disc diffusion test with Dc and aminoglycosides assessed by stringent computation showed clear-cut synergism. Synergism between Dc and streptomycin (Sm) was found to be statistically significant (p < or = 0.01) when compared with their individual effects. By the checkerboard assessment procedure, the fractional inhibitory concentration (FIC) index of this combination was found to be 0.49, confirming synergism. The mouse protective capacity of this combination was then evaluated in vivo against S. typhimurium as the virulent infecting bacterium, and the size of bacterial load determined from infected autopsied animals. Statistical analysis by Student's 't' test suggested this drug combination is highly synergistic; synergism was also noted between Dc and other aminoglycosides.

Among two flavanones [YS01, YS02] and eight prenylflavanones [YS03-YS10], preliminary screening with fifteen test bacterial strains showed that YS06 was the most active agent. YS06 exhibited highly significant antimicrobial action when tested against 228 bacterial strains comprising two Gram-positive and six Gram-negative genera. The in vitro susceptibility test was carried out by determining the minimum inhibitory concentration (MIC) of YS06 by agar dilution technique. Twenty-two out of fifty strains of Staphylococcus aureus were inhibited at 25 to 50 micrograms/mL of the agent. YS06 also inhibited strains of Salmonella, Shigella and a few strains of Escherichia coli strains were also highly sensitive to YS06, while Kleibsiella spp. and Pseudomonas aeruginosa were much less sensitive. In in vivo study, YS06 offered significant protection (p < 0.001 according to chi-square test) to Swiss albino mice (challenged with 50 minimum lethal dose (MLD, virulent bacterium) at concentrations of 160 and 80 micrograms/mouse.

Leprosy bacillus (LB) and leprosy derived in vitro culture forms, the chemoautotrophic nocardioform (CAN) bacteria, showed an extremely close homology and identity with each other as regards a chemoautotrophic nutritional pattern, a nocardioform morphology, a weak acid-fastness coupled with Gram and Gomori's stain positivity, an exclusive mycolate and lipid profile, a phenolic glycolipid (PGL-I) and a highly sequestrated DNA characteristic, namely, a unique small size, a low G+C % mole, an exceptionally high gamma and UV radiation resistance, and a high thermal resistance. LB/CAN bacteria (CANb) gave positive signals for 36 kDa protein PCR, as well as, for 65 kDa epitope, and hybridisation with two or more probes and also by RFLP-analysis. Both LB/and CAN bacteria exhibited bacillary multiplication in the mouse footpads (MFP), nerve infiltration and evidences for local pathogenicity associated with pronounced systemic invasion. A highly reproducible mutilation model could be established which enabled a successful application of the postulates of Koch. The proof of their total identity was their anergic reactions in LL cases counterpoised against Mitsuda type strong nodular responses, mirroring the reactions of leprosy bacilli in TT cases, in accordance with the dictum of XIth International Leprosy Congress (1978). Thus, the chemoautotrophic nutritional requirements of LB, entirely unsuspected for a medically important pathogenic bacterium, having dimorphic (both bacillary and mycelial) characters with spores, mycelia and granules and unique pathogenicity of multilation manifested through the virulence factor, the enzyme collagenase, made LB or M leprae the highly enigmatic bacterium for so long.

A preliminary exploration of coumarin analogs as novel antimicrobial agents was carried out to determine the basic features of the structure responsible for the observed biological activity. The substituents ester or carboxylic acid on the coumarin ring were needed to have potent inhibitory activity against both Gram-positive and Gram-negative bacteria. The presence of phenolic hydroxyl group and/or carboxylic acid was necessary to possess higher activity against Helicobacter pylori.

Leprosy is believed to be transmitted only through human contacts. However, many anomalous observations had gradually accumulated which had weakened such beliefs. These are: only 1/3 rd cases of leprosy give a definite history of being transmitted from other known cases; life-long spouses, in whom only one has leprosy, seldom lead to leprosy to others; while MDT applied intensively in most leprosy endemic countries, could successfully reduce incidence of leprosy, however, simultaneously new cases arise unabated. Besides, a close look at animal leprosies also suggested a mode of transmission other than human-type contact. Thus, a search for alternative hypothesis led to the findings that leprosy bacillus (LB) could be a soil chemoautotroph and could facultatively live both in the human body and the soil which could serve as an alternative source of infection. Evaluation of accumulated evidences points to this possibility.

The biological activity of ten prenylflavanones purified from Sophora tomentosa L., and Sophora moorcroftiana Benth. ex Baker (Leguminosae) was investigated. The flavanones with prenyl-, lavandulyl- or geranyl groups on A ring, and two bioactive flavonostilbenes on ring B and stilbene (resveratrol) showed tumor-specific cytotoxic activity, antimicrobial activity, and anti-HIV activity, radical generation, and O2- scavenging activity. There was a positive relationship between radical generation and O2- scavenging activity in these prenylflavanones. These data suggest the medicinal significance of prenylflavanones.

We have attempted a new evaluation of the process of conjugation in bacteria, because of some basic dissimilarities observed between this and that of eukaryotes, or plants and animals. Reference donor and recipient strains, widely used to prove conjugation in bacteria, were chosen; addition of DNase during the conjugation process, led to an unexpected but highly reproducible increase in the transconjugant colony counts (TCC; ca. > or = 1 log), when compared with that of the controls without DNase. Transconjugants were also obtained when the same live donors were substituted with the UV-killed ones although the TCC was very low initially. Contrarily, donors treated with DNA-intercalating agents, e.g. acridine orange or ethidium bromide, resulted in a complete failure to produce transconjugants. There was a quantitative relationship between the DNase used on donors and levels of DNA sugars/nucleotides/DNA, which possibly resulted from interaction between the DNase and DNA being present/produced on the donor surface. This may be indicative of what may actually happen in the donor-recipient mixtures in the conjugation test proper, where the recipient DNase may activate a donor DNA production cycle. The evidences presented did not suggest that the donor DNA in the conjugation process is actually vestibuled through any intercellular conjugation passages, and is susceptible to the action of DNase or the intercalating dyes.

The antihistaminic phenothiazine trimeprazine (Tz) was found to exhibit significant antibacterial activity on the basis of in vitro and in vivo tests. For the study of synergism due to a combination between Tz and trimethoprim (Tm), drug soaked filter paper discs were placed on young culture lawns of sensitive bacteria on nutrient agar plates. Calculation of the area of inhibition zones for determining the degree of synergism between Tz and Tm showed the increase to be statistically significant (p<0.01) when compared with their individual effects. By the checkerboard assessment procedure, the fractional inhibitory concentration (FIC) index was found to be 0.18, confirming synergism. The protective capacity of this combination was then assessed in Swiss white mice using S. typhimurium as the challenge bacterium, and the level of bacterial load was determined from infected autopsied animals. Statistical analysis of the data by students 't' test finally proved that a combination of Tz+Tm was highly synergistic.

Synthesis and antitumor activity of goniofufurone analogues 15, 16, 17, 33, and 46 is reported. Key step in the synthesis is Pd (II) mediated oxidative cyclisation of vinyl-(hydroxy) furans 18, 19 to the corresponding lactols 32, 43. Cytotoxicities of 15, 16, 17, 33, and 46 tested against six human cancer cell lines are reported. Change of stereochemistry at C-5, C-6 and C-7 position of goniofufurone (1) did not enhance the cytotoxicities significantly.

Thioridazine (Th), which is therapeutically used in psychiatric patients, was found to possess conspicuous antimicrobial activity when tested against 316 strains belonging to a number of Gram positive and Gram negative bacteria. Although Staphylococcus aureus, Vibrio chloerae and V. parahaemolyticus were found to be most sensitive, Th was highly bactericidal against S. aureus and bacteriostatic for vibrios and other Gram negative organisms. In the study of antiplasmid/curing effect of Th on twelve multiply antibiotic and Th resistant bacteria, it was observed that elimination of R plasmids was facilitated by choice of optimal concentration of Th. Significant elimination of single and combined antibiotic resistance occurred in E. coli and Shigella flexneri and not in S. aureus.

Anergy, or contrarily, Mitsuda-type responses towards 4 chemoautotrophic nocardioform antigens (CAN-Ags) and a control standard lepromin were tested in 73 LL, TT and borderline cases of leprosy. The antigens injected per patient varied from a maximum of 5 to a minimum of 2. Complete anergy to CAN-Ags was seen in 92/92 instances tested on 24 LL cases. The anergy was weakly modified or unmodified in 3 other LL cases which had been vaccinated before. Concurrent studies with the same antigens tested on 33 TT cases showed clear-cut, dose-dependent, Mitsuda-type late responses in 80/81 instances. The CAN bacteria, therefore, despite their origin from different unrelated leprous human, mouse footpad (MFP) and armadillo tissues, appeared to be identical with each other and also probably related to the leprosy bacillus, on the basis of these parameters.

Antimicrobial property of ten antiinflammatory drugs was tested with eleven sensitive bacteria belonging to both Gram positive and Gram negative types. Since most of the bacteria were moderate to highly sensitive to diclofenac (Dc), this compound was tested in vitro against 397 bacteria, most of which were inhibited by Dc at 50-100 micrograms/ml level. When tested in vivo, Dc at 1.5 and 3.0 micrograms/g body weight of a Swiss strain of white mice, could significantly protect the animals challenged with 50 MLD of Salmonella typhimurium NCTC 74. According to chi 2 test the in vivo data were highly significant (P < 0.001).

Antimicrobial action of penicillin and some of its derivatives including fosfomycin was studied with respect to 225 strains of Gram-positive and Gram negative bacteria. Fosfomycin was found to possess somewhat less activity against Staphylococcus aureus compared with other penicillins; however, it showed powerful activity towards Escherichia coli, Klebsiella spp. and Proteus mirabilis.

Purpose: Our purpose was to verify our earlier observation of an altered endometrial echo character during the periimplantation period, signifying the earliest detection of implantation by ultrasound., Methods: The transvaginal ultrasound study used a 7.5-MHz probe on day 6 following preembryo transfer., Results: Of 31 embryo transfers studied. 8 (25.8%) were positive for this sign on day 6, whereas 9 patients later showed a gestational sac, giving an accuracy of this sign of 88.8% (8 of 9)., Conclusions: The normal secretory-phase endometrium shows a typical, evenly distributed hyperechogenecity described earlier by Hackeloer (1). In some cases of proliferative phases, too, this echo pattern has been reported by others (2, 3), and observed by us in stimulated and unstimulated cycles, and has been named type A endometrium by Gonen et al. (4) Both these varieties are seen to have an even, unbroken outline at the anterior and posterior myometrial-endometrial interphase. We report what we believe to be the earliest ultrasound picture of implantation, resembling this variety of endometrial echo but having an irregular outline (Fig. 2).

The antibacterial and bactericidal activities of the antihistamine trimeprazine were studied against 243 strains of bacteria which included both Gram positive and Gram negative types. The susceptibility of these bacterial strains to trimeprazine was assessed by determining their minimum inhibitory concentration (MIC) which was found to be between 10 and 100 micrograms/ml. Nineteen strains of Staphylococcus spp. and Salmonella spp. were trimeprazine. Most of the strains belonging to Bacillus spp. and Salmonella spp. were inhibited by less than 100 micrograms/ml. Trimeprazine could also inhibit strains of Shigella spp. Vibrio cholerae and V. parahaemolyticus at 10-100 micrograms/ml. Strains of klebsiella, proteus, pseudomonas and citrobacter were moderately sensitive to trimeprazine. In in vivo studies it was seen that when trimeprazine was used at a concentration of 0.75 and 0.4 micrograms/gm body weight of the mouse both levels offered significant protection to Swiss mice challenged with 50LD50 of virulent strain of S. typhimurium 74. Statistical analysis of the data was found to be significant, p < 0.001 according to chi 2 test.

The effect of augmentin alone and in combination with various beta-lactam antibiotics was studied against a pathogenic Mycobacterium, M. marinum. The in vitro studies did not reveal any additional advantage over that found with augmentin alone and this antibiotic seemed considerably inhibitory to M. marinum at < 1 microgram/ml concentration. In vivo, the effects of augmentin on experimentally produced lesions in the mouse foot pads (MFPs) showed a significant regression of the lesions, which was compatible with an early disappearance of M. marinum from the MFP, in contrast with those of the untreated, control animals.

The transmission electron microscopic (TEM) studies of the human leprosy derived chemoautotrophic nocardio-form (CAN) bacteria and EUS derived CAN bacteria showed presence of double contoured cell-walls consisting of an electron transparent and a dense layer. The fibrillar structures on the surface of these CAN bacterial cells also suggested their similarity to the human tissue derived Mycobacterium leprae cells. These EM studies further revealed mycelial and coccoid bodies in all these bacteria as was observed originally.

The splenic tissue of a mouse experimentally infected with M. lepraemurium (Hawaiian strain, M-65) and developing 'rat leprosy', yielded a pure culture of an acid - fast bacterium having all the characteristics of M. lepraemurium on mineral salt minimal medium supplemented with simple sources of C and N, e.g., NH4 -salts, liquid paraffin, urea, gelatin etc. This could be maintained, by serial passages in vitro with good growth. Its indefinite propagation with tissue - free washed, small inoculum on complex media including Ogawa medium was difficult, and its serial sub-culture was practically impossible. The in vitro isolate from supplemented minimal medium could produce pathological lesions in mice typical of rat leprosy.

Penicillin resistance plasmid was transferred from Staphylococcus aureus B4 (PcrKms, donor) to S. aureus ML351 (PcsKmr, recipient) by co-cultivation of the donor with the recipient in nutrient broth with or without the modifying effects of CaCl2 or sodium dodecyl sulfate. It was found that the transfer of drug-resistance occurred maximally between 6 and 18 hr postinoculation; however, addition of DNase (200 micrograms/ml) could totally prevent such a transfer up to 6 hr and significantly reduce it thereafter. Cell-free filtrate of the donor culture when mixed with the recipient was ineffective in bringing about the transfer of Pcr.

The antipsychotic drug fluphenazine was obtained in a dry powder form and was screened with respect to 482 strains of bacteria, which included 170 Gram-positive and 326 Gram-negative strains. Nutrient agar plates containing increasing concentrations of fluphenazine (0-200 micrograms/ml) were used for the determination of the minimum inhibitory concentration (MIC) which was demonstrated by inoculating a loopful of an overnight peptone water culture of the organism on nutrient agar plates and determining the MIC against a control. Fluphenazine was detected to possess pronounced action against both Gram-positive and Gram-negative bacteria at 20-100 micrograms/ml. In the in vivo studies it was seen that when fluphenazine was used at a concentration of 1.5 micrograms/g and 3 micrograms/g mouse body weight both the levels offered significant protection to Swiss strain of white mice when challenged with 50 minimum lethal dose (MLD) of a virulent strain of Salmonella typhimurium 74. The in vivo data with fluphenazine were highly significant (p < 0.001) according to the chi-square test.