Natural and induced bactericidal activities in the hemolymph of the lobster, Homarus americanus: products of hemocyte–plasma interaction

1972 ◽  
Vol 18 (9) ◽  
pp. 1499-1509 ◽  
Author(s):  
James E. Stewart ◽  
B. M. Zwicker

The salient features of this study of the enhancement of bactericidal activity in the hemolymph of the American lobster were as follows: (a) increase in response to a number of non-pathogens isolated from the lobster's intestinal tract (several pseudomonads, an Achromobacter and Sarcina lutea), (b) one isolate identified as Pseudomonas perolens was used for the bulk of the studies, (c) apparent bactericidal activity of the hemolymph increased severalfold with reduction of the pH of the assay system from the physiological value of 7.6 to a value of 6.0, (d) the extent of the enhancement in vivo was roughly proportional to the concentration of the vaccine, (e) the bactericidal activity's enhancement in vivo was temperature dependent, (f) heat-stability trials indicated the probable presence of more than one bactericidin, (g) the bactericidal principle(s) exists in vivo in an inactive form until activated by material contained within the hemocytes, (h) no protection against Gaffkya homari was conferred on the lobster by prior treatment with vaccines prepared from P. perolens, G. homari, or S. lutea.

1997 ◽  
Vol 41 (11) ◽  
pp. 2527-2532 ◽  
Author(s):  
M Manduru ◽  
L B Mihm ◽  
R L White ◽  
L V Friedrich ◽  
P A Flume ◽  
...  

Bactericidal activity, historically assessed by in vitro tests which employ fixed drug concentrations, may also be evaluated in in vitro pharmacodynamic models in which in vivo pharmacokinetics and bacterial growth conditions can be simulated. However, systematic comparisons between the two methods are lacking. We evaluated the bactericidal activities of ceftazidime, at two different concentration/MIC ratios (C/MICs), against 10 clinical isolates of Pseudomonas aeruginosa in a two-compartment model with continuous-infusion conditions and a 2-h half-life. These values were compared to those determined by traditional 24-h time-kill (TTK) methods at the same C/MICs. Bactericidal activities were compared by using area under the colony count-time curves. Antibiotic exposure (area under the drug concentration-time curve) was also evaluated. Although bactericidal activity appeared greater by the TTK method (P = 0.05), when it was normalized for drug exposure, these differences disappeared (P = 0.2). This disparity was likely due to differences in drug exposure in the TTK method and in the peripheral compartment of the model (site of bacteria) over the first 8 h of the experiment, during which the antibiotic accumulated to target concentrations. This suggests that the bactericidal effects with constant antibiotic concentrations are similar in the two methods; however, this may not hold true with fluctuating drug concentrations. Further, results from the pharmacodynamic model may theoretically be more relevant, as in vivo pharmacokinetics and bacterial growth conditions call be more faithfully simulated.


1997 ◽  
Vol 41 (4) ◽  
pp. 781-784 ◽  
Author(s):  
K Fuursted ◽  
J D Knudsen ◽  
M B Petersen ◽  
R L Poulsen ◽  
D Rehm

In this report, we present MIC, bactericidal activity, postantibiotic effect (PAE), and in vivo infectivity data for postantibiotic-phase pneumococci. We compared and evaluated penicillin G and six macrolides, erythromycin, azithromycin, clarithromycin, dirithromycin, roxithromycin, and spiramycin, against 10 strains of pneumococci with various levels of susceptibility to penicillin. All of the agents, except azithromycin, exhibited a bactericidal effect (a > or = 3 log10 decrease in the number of CFU per milliliter) after 4 h of exposure to a concentration equal to 10 times the MIC, displaying the following hierarchy: spiramycin = penicillin G = erythromycin = dirithromycin = clarithromycin = roxithromycin > azithromycin. The bactericidal rate of penicillin G was significantly lower for resistant strains (MIC, > or = 2 microg/ml), while bactericidal rates of macrolides were unaffected by penicillin susceptibility. A PAE was induced in all of the strains by all of the antibiotics after exposure for 1 h to a concentration equivalent to 10 times the MIC. The mean duration of PAEs varied between 2.3 and 3.9 h, showing the following hierarchy: spiramycin = dirithromycin = clarithromycin = erythromycin = roxithromycin > azithromycin > penicillin G. Virulence studies were performed with immunocompetent mice by intraperitoneal inoculation of virulent, penicillin-susceptible serotype 3 pneumococci which had been pre-exposed to penicillin G or a macrolide for 1 h. A significant decrease in the virulence of postantibiotic-phase pneumococci was induced only by erythromycin, azithromycin, dirithromycin, and spiramycin, displaying 5.9-, 7.1-, 4.2-, and 3.6-fold increases in the 50% lethal dose (LD50) compared to a control suspension, respectively. No significant correlation could be demonstrated between the LD50 and the MIC, bactericidal activity, or PAE duration. These results suggest that antimicrobial interaction with host defenses in terms of virulence might be a significant parameter that could influence the drug or drug regimen of choice.


1947 ◽  
Vol 85 (2) ◽  
pp. 175-186 ◽  
Author(s):  
Harry Eagle ◽  

1. The relative bactericidal activities of penicillins F, G, K, and X against Type I pneumococcus in vitro were 60, 100, 180, and 135. The corresponding activities against Streptococcus pyogenes, strain C-203, were 75, 100, 115, and 145, respectively. 2. The total curative doses (CD50) of penicillins F, G, K, and X in pneumococcal infections of white mice (ten injections at 3 hour intervals) were 4.6, 3.8, 20, and 2.4 mg. per kg., respectively, or relative activities of 83, 100, 19, and 160, referred to G as 100. 3. The corresponding curative doses in streptococcal infections of white mice were 2.6, 1.3, 14.0, and 0.5 mg. per kg., or relative activities of 50, 100, 9, and 260. 4. Penicillin K was therefore one-tenth as active in vivo as would be implied by its bactericidal activity in vitro. This probably reflects its rapid inactivation in vivo, evidenced by the low and evanescent blood levels observed in both rabbits and man, and the low urinary recovery of this species of penicillin. 5. Penicillin X was significantly more active therapeutically than its bactericidal activity in vitro would imply. This probably reflects its slower inactivation in vivo, evidenced by the somewhat higher and more prolonged blood levels afforded by this penicillin in comparison with penicillin G. Judged by the mouse infections with the strains here used, penicillin X is the penicillin of choice in the treatment of infections with pneumococcus Type I and hemolytic streptococci. 6. The curative dose of penicillin in streptococcal and pneumococcal infections paralleled the varying susceptibility of these organisms to penicillin in vitro.


2011 ◽  
Vol 18 (5) ◽  
pp. 825-834 ◽  
Author(s):  
Shimin Zhang ◽  
Shaw-Huey Feng ◽  
Bingjie Li ◽  
Hyung-Yong Kim ◽  
Joe Rodriguez ◽  
...  

ABSTRACTOur laboratory has developed more than a hundred mouse monoclonal antibodies (MAbs) againstBurkholderia pseudomalleiandBurkholderia mallei. These antibodies have been categorized into different groups based on their specificities and the biochemical natures of their target antigens. The current study first examined the bactericidal activities of a number of these MAbs by anin vitroopsonic assay. Then, thein vivoprotective efficacy of selected MAbs was evaluated using BALB/c mice challenged intranasally with a lethal dose of the bacteria. The opsonic assay using dimethyl sulfoxide-treated human HL-60 cells as phagocytes revealed that 19 out of 47 tested MAbs (40%) have prominent bactericidal activities againstB. pseudomalleiand/orB. mallei. Interestingly, all MAbs with strong opsonic activities are those with specificity against either the capsular polysaccharides (PS) or the lipopolysaccharides (LPS) of the bacteria. On the other hand, none of the MAbs reacting to bacterial proteins or glycoproteins showed prominent bactericidal activity. Further study revealed that the antigenic epitopes on either the capsular PS or LPS molecules were readily available for binding in intact bacteria, while the epitopes on proteins/glycoproteins were less accessible to the MAbs. Ourin vivostudy showed that four MAbs reactive to either the capsular PS or LPS were highly effective in protecting mice against lethal bacterial challenge. The result is compatible with that of ourin vitrostudy. The MAbs with the highest protective efficacy are those reactive to either the capsular PS or LPS of theBurkholderiabacteria.


1983 ◽  
Vol 50 (1) ◽  
pp. 3-8
Author(s):  
A. John Bramley ◽  
Elizabeth M. Hogben

SummaryThe bactericidal activities of various concentrations of 3 disinfectants commonly used for teat disinfection were compared in vitro using small discs of pig skin previously contaminated with Staphylococcus aureus. Solutions containing 40 g/1 Na hypochlorite, 25 or 50 g/1 chlorhexidine digluconate or iodophor containing 5 g/1 iodine were found to have equivalent or superior bactericidal activity to a solution containing 10 g/1 Na hypochlorite. This in vitro technique offers a convenient alternative to in vivo tests for teat disinfectants and with minor modifications could be applicable to the testing of skin antiseptics in general.


2021 ◽  
Vol 22 (5) ◽  
pp. 2530
Author(s):  
Bijean D. Ford ◽  
Diego Moncada Giraldo ◽  
Camilla Margaroli ◽  
Vincent D. Giacalone ◽  
Milton R. Brown ◽  
...  

Cystic fibrosis (CF) lung disease is dominated by the recruitment of myeloid cells (neutrophils and monocytes) from the blood which fail to clear the lung of colonizing microbes. In prior in vitro studies, we showed that blood neutrophils migrated through the well-differentiated lung epithelium into the CF airway fluid supernatant (ASN) mimic the dysfunction of CF airway neutrophils in vivo, including decreased bactericidal activity despite an increased metabolism. Here, we hypothesized that, in a similar manner to neutrophils, blood monocytes undergo significant adaptations upon recruitment to CFASN. To test this hypothesis, primary human blood monocytes were transmigrated in our in vitro model into the ASN from healthy control (HC) or CF subjects to mimic in vivo recruitment to normal or CF airways, respectively. Surface phenotype, metabolic and bacterial killing activities, and transcriptomic profile by RNA sequencing were quantified post-transmigration. Unlike neutrophils, monocytes were not metabolically activated, nor did they show broad differences in activation and scavenger receptor expression upon recruitment to the CFASN compared to HCASN. However, monocytes recruited to CFASN showed decreased bactericidal activity. RNASeq analysis showed strong effects of transmigration on monocyte RNA profile, with differences between CFASN and HCASN conditions, notably in immune signaling, including lower expression in the former of the antimicrobial factor ISG15, defensin-like chemokine CXCL11, and nitric oxide-producing enzyme NOS3. While monocytes undergo qualitatively different adaptations from those seen in neutrophils upon recruitment to the CF airway microenvironment, their bactericidal activity is also dysregulated, which could explain why they also fail to protect CF airways from infection.


2020 ◽  
Vol 15 (1) ◽  
Author(s):  
Mingming Yang ◽  
Longlong Wang ◽  
Xiaofen Qiao ◽  
Yi Liu ◽  
Yufan Liu ◽  
...  

Abstract The defects into the hexagonal network of a sp2-hybridized carbon atom have been demonstrated to have a significant influence on intrinsic properties of graphene systems. In this paper, we presented a study of temperature-dependent Raman spectra of G peak and D’ band at low temperatures from 78 to 318 K in defective monolayer to few-layer graphene induced by ion C+ bombardment under the determination of vacancy uniformity. Defects lead to the increase of the negative temperature coefficient of G peak, with a value almost identical to that of D’ band. However, the variation of frequency and linewidth of G peak with layer number is contrary to D’ band. It derives from the related electron-phonon interaction in G and D’ phonon in the disorder-induced Raman scattering process. Our results are helpful to understand the mechanism of temperature-dependent phonons in graphene-based materials and provide valuable information on thermal properties of defects for the application of graphene-based devices.


2021 ◽  
Vol 22 (1) ◽  
Author(s):  
Olanrewaju Ayodeji Durojaye ◽  
Nkwachukwu Oziamara Okoro ◽  
Arome Solomon Odiba

Abstract Background The novel coronavirus SARS-CoV-2 is currently a global threat to health and economies. Therapeutics and vaccines are in rapid development; however, none of these therapeutics are considered as absolute cure, and the potential to mutate makes it necessary to find therapeutics that target a highly conserved regions of the viral structure. Results In this study, we characterized an essential but poorly understood coronavirus accessory X4 protein, a core and stable component of the SARS-CoV family. Sequence analysis shows a conserved ~ 90% identity between the SARS-CoV-2 and previously characterized X4 protein in the database. QMEAN Z score of the model protein shows a value of around 0.5, within the acceptable range 0–1. A MolProbity score of 2.96 was obtained for the model protein and indicates a good quality model. The model has Ramachandran values of φ = − 57o and ψ = − 47o for α-helices and values of φ = − 130o and ψ = + 140o for twisted sheets. Conclusions The protein data obtained from this study provides robust information for further in vitro and in vivo experiment, targeted at devising therapeutics against the virus. Phylogenetic analysis further supports previous evidence that the SARS-CoV-2 is positioned with the SL-CoVZC45, BtRs-BetaCoV/YN2018B and the RS4231 Bat SARS-like corona viruses.


2004 ◽  
Vol 378 (1) ◽  
pp. 93-103 ◽  
Author(s):  
Nathan A. LOCKWOOD ◽  
Judith R. HASEMAN ◽  
Matthew V. TIRRELL ◽  
Kevin H. MAYO

We have conjugated dodecyl and octadecyl fatty acids to the N-terminus of SC4, a potently bactericidal, helix-forming peptide 12-mer (KLFKRHLKWKII), and examined the bactericidal activities of the resultant SC4 ‘peptide-amphiphile’ molecules. SC4 peptide-amphiphiles showed up to a 30-fold increase in bactericidal activity against Gram-positive strains (Staphylococcus aureus, Streptococcus pyogenes and Bacillus anthracis), including S. aureus strains resistant to conventional antibiotics, but little or no increase in bactericidal activity against Gram-negative bacteria (Escherichia coli and Pseudomonas aeruginosa). Fatty acid conjugation improved endotoxin (lipopolysaccharide) neutralization by 3- to 6-fold. Although acylation somewhat increased lysis of human erythrocytes, it did not increase lysis of endothelial cells, and the haemolytic effects occurred at concentrations 10- to 100-fold higher than those required for bacterial cell lysis. For insight into the mechanism of action of SC4 peptide-amphiphiles, CD, NMR and fluorescence spectroscopy studies were performed in micelle and liposome models of eukaryotic and bacterial cell membranes. CD indicated that SC4 peptide-amphiphiles had the strongest helical tendencies in liposomes mimicking bacterial membranes, and strong membrane integration of the SC4 peptide-amphiphiles was observed using tryptophan fluorescence spectroscopy under these conditions; results that correlated with the increased bactericidal activities of SC4 peptide-amphiphiles. NMR structural analysis in micelles demonstrated that the two-thirds of the peptide closest to the fatty acid tail exhibited a helical conformation, with the positively-charged side of the amphipathic helix interacting more with the model membrane surface. These results indicate that conjugation of a fatty acid chain to the SC4 peptide enhances membrane interactions, stabilizes helical structure in the membrane-bound state and increases bactericidal potency.


2015 ◽  
Vol 2015 ◽  
pp. 1-9 ◽  
Author(s):  
Saiful Azmi Johari ◽  
Mastura Mohtar ◽  
Sharifah Aminah Syed Mohammad ◽  
Rohana Sahdan ◽  
Zurina Shaameri ◽  
...  

28 new pyrrolidine types of compounds as analogues for natural polyhydroxy alkaloids of codonopsinine were evaluated for their anti-MRSA activity using MIC and MBC value determination assay against a panel ofS. aureusisolates. One pyrrolidine compound, MFM 501, exhibited good inhibitory activity with MIC value of 15.6 to 31.3 μg/mL against 55S. aureusisolates (43 MRSA and 12 MSSA isolates). The active compound also displayed MBC values between 250 and 500 μg/mL against 58S. aureusisolates (45 MRSA and 13 MSSA isolates) implying that MFM 501 has a bacteriostatic rather than bactericidal effect against both MRSA and MSSA isolates. In addition, MFM 501 showed no apparent cytotoxicity activity towards three normal cell lines (WRL-68, Vero, and 3T3) with IC50values of >625 µg/mL. Selectivity index (SI) of MFM 501 gave a value of >10 suggesting that MFM 501 is significant and suitable for furtherin vivoinvestigations. These results suggested that synthetically derived intermediate compounds based on natural products may play an important role in the discovery of new anti-infective agents against MRSA.


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