Biomedical subjects

K Kim

Publications and source records attributed to K Kim.

At least 253 records · Page 14Linked to original sources

PubMed1999

Enhanced splicing of the first intron from the gonadotropin-releasing hormone (GnRH) primary transcript is a prerequisite for mature GnRH messenger RNA: presence of GnRH neuron-specific splicing factors.

The rat GnRH gene consists of four short exons (denoted 1, 2, 3, and 4) and three introns (A, B, and C). All three introns are spliced from the primary transcript, resulting in a mature mRNA. Northern blot and RT-PCR analyses showed that the GnRH primary transcript and its splicing intermediates are more prevalent than the mature GnRH mRNA in a variety of non-GnRH-producing tissues. To delineate the possible splicing mechanism of introns, an in vitro HeLa splicing system was used. Introns B and C were efficiently spliced, while intron A spanning between exon 1 and exon 2 was not. The retention of intron A was relieved when the 5'- and/or 3'-splice sites of intron A were point mutated based on the consensus sequence. The splicing activity was even more strengthened when a putative branchpoint site was moved to the upstream region of the pyrimidine tract of intron A. Intron A could be partially spliced when whole exons (2, 3, and 4) were linked up with intron A. There are two putative exonic splicing enhancers (ESEs) in exon 3 and exon 4. The ESE on exon 4 (ESE4) is much stronger than that on exon 3. The closer the ESE4 to the 3'-splice site of intron A, the better the splicing activity became. However, in the presence of the nuclear extract from GnRH neurons, there was an enhancement in the splicing activity notwithstanding the distance between ESE4 and 3'-splice site of intron A. These results suggest that the ESE4 functions as both the constitutive and regulated enhancer. Collectively, our study provides evidence that enhanced splicing of intron A by putative GnRH neuron-specific splicing factor(s) interacting with the ESEs is a prerequisite for mature GnRH synthesis.

Animals↗

PubMed1999

Homologous upregulation of GnRH receptor mRNA by continuous GnRH in cultured rat pituitary cells.

The present study examined the effects of continuous treatment with gonadotropin-releasing hormone (GnRH) on GnRH receptor (GnRH-R) mRNA levels in dispersed cultures of rat pituitary cells. Pituitary GnRH-R mRNA levels were determined by competitive reverse transcriptase polymerase chain reaction. When pituitary cells were continuously exposed to a low dose of GnRH (0.2 nM), GnRH-R mRNA levels were transiently increased. The levels of GnRH-R mRNA were significantly increased up to 6 h and diminished to untreated levels by 24 h. Luteinizing hormone (LH) release was also increased significantly up to 12 h, maintaining similar levels in LH release thereafter. When GnRH antagonist ([D-pGlu1, D-Phe2, D-Trp3,6]-LH-RH) was added to the cultures together with GnRH (0.2 nM) for 6 h, the stimulatory effect of GnRH on GnRH-R mRNA levels and LH release was significantly diminished in a dose-related manner. In another experiment, pituitary cells were treated with various doses of GnRH (0.02-200 nM) for a relatively short (6 h) or a longer (24 h) period. When pituitary cells were exposed for 6 h, all doses of GnRH (0.02-200 nM) significantly increased GnRH-R mRNA levels in a dose-dependent manner. By contrast, continuous exposure to GnRH for 24 h was ineffective in changing pituitary GnRH-R mRNA levels at any given doses. These results indicate that the duration of GnRH treatment is critical for upregulation of GnRH-R mRNA by continuous GnRH. When pituitary cells were treated for 6 h with either a continuous mode of GnRH (0.2 nM) or an hourly pulsatile mode of GnRH (0.2 nM, 6 min/h), both treatments significantly augmented GnRH-R mRNA levels. Thus, the modes of GnRH application, if treated for a relatively short period, do not appear to make a significant difference in upregulation of GnRH-R mRNA levels. Collectively, our data provide strong evidence that continuous GnRH application is able to upregulate pituitary GnRH-R mRNA levels, if treated for a relatively short period (6 h).

Animals↗

PubMed1999

Anti-proliferating effects of ginsenoside Rh2 on MCF-7 human breast cancer cells.

Ginsenoside Rh2 (G-Rh2) isolated from the root of Panax ginseng has been shown to have anti-cancer proliferation, differentiation and chemopreventive effects in certain cancer cell types. We investigated the mechanism of G-Rh2-induced growth inhibition in MCF-7 human breast carcinoma cells. G-Rh2 significantly inhibited the cell growth in a concentration-dependent manner, which effect was reversible, and induced a G1 arrest in cell cycle progression. G-Rh2 treatment down-regulated the protein level of cyclin D3 but upregulated the expression of cyclin-dependent kinase (Cdk) inhibitor p21WAF1/CIP1. The increased levels of p21 were associated with increased binding of p21 and Cdk2 concomitant with marked decrease in Cdk2 and cyclin E-dependent kinase activities with no changes in Cdk2 and cyclin E expression. G-Rh2 markedly reduced the phosphorylated retinoblastoma protein (pRb) and enchanced association of unphosphorylated pRb and the transcription factor E2F-1. These data suggest that G-Rh2 inhibited the growth of MCF-7 cells, by inducing protein expression of p21 and reducing the protein levels of cyclin D which resulted in the down-regulation of cyclin/Cdk complex kinase activity, decreasing phosphorylation of pRb, and inhibiting E2F release.

Antineoplastic Agents↗

PubMed1999

[Occipital neuralgia caused by the compression of the fenestrated vertebral artery: a case report].

A case is presented of fenestration of the vertebral artery in a 36-year-old woman who had various neurological symptoms with Basedow's disease. We also review 7 cases from the literature. Our patient had had occipital neuralgia and numbness of the left side of the neck and left upper limb several weeks before admission. On admission she had hyperthyroidism. MRI, left vertebral angiography, and 3D-CT scans demonstrated a fenestrated vertebral artery compressing the upper cervical cord. The patient's symptoms gradually improved as her thyroid function was controlled. A possible explanation is that the fenestrated vertebral artery might have compressed the neural structures, resulting in her various symptoms. Occipital neuralgia was apparently caused by the fenestrated artery compressing the C1 and C2 sensory roots. A fenestrated vertebral artery is usually of no clinical significance. The reason for the late onset of symptoms in the present case is unknown, but it may have been due to hemodynamic stress caused by her hyperthyroidism. After her thyroid function was controlled, the hemodynamic stress presumably decreased and the symptoms resolved spontaneously.

Adult↗

PubMed1999

A Rational Approach to Selective Recognition of NH(4)(+) over K(+).

An ion-selective electrode (ISE) based on receptor 1 is highly selective for binding NH(4)(+) over K(+) (lg K(NH(4)(+)/K(+))=-2.6); the three imine nitrogen atoms in 1 are ideally positioned for hydrogen bonding with the tetrahedral NH(4)(+) ion. This selectivity is considerably greater than that found for commercial ISEs based on nonactin (lg K(NH(4)(+)/K(+))=-1.0).

Journal Article↗

PubMed1999

Successful low-dose cytosine arabinoside treatment in a patient with acute leukemia from idiopathic myelofibrosis.

We report a 63-year-old male with acute leukemia developed from idiopathic myelofibrosis. Cytogenetic analysis revealed chromosomal abnormalities; 46,XY,t(3;21)(q21;q22) del(20q). Intensive chemotherapies [combination of adriamycine/vincristine/prednisolone, or idarubicine/cytosine arabinoside (Ara-C)] were unsuccessful. The patient was then treated with continuous intravenous low-dose Ara-C because of his poor physical condition and showed marked hematological improvement. Leukemic cells disappeared from the peripheral blood and the segmented neutrophil count recovered. The dysplastic morphology observed in the segmented neutrophils suggests that the recovery of the neutrophils has resulted mainly from the differentiation activity of Ara-C.

Acute Disease↗

PubMed1998

Dodecanethiol-Derivatized Au/Ag Bimetallic Nanoparticles: TEM, UV/VIS, XPS, and FTIR Analysis.

Dodecanethiol-derivatized Au/Ag bimetallic nanoparticles that can be dissolved in nonpolar solvents were prepared via a two-phase synthetic route in water/toluene mixtures. The position of the surface plasmon band was found to vary linearly between 419 and 522 nm as a function of mole fraction of Au (or Ag) content, implying ready formation of Au-Ag alloy clusters. Transmission electron microscopy images revealed that the bimetallic clusters were monodisperse with an average size of approximately 4 nm. Infrared spectral features indicate that the alkyl chains of the dodecanethiolate species take fully extended trans zigzag conformations on all alloy clusters. The most interesting finding deduced from X-ray photoelectron spectroscopy and FTIR spectroscopic measurements was that Ag atoms are enriched at the outer part of the alloy clusters when compared with the Au atoms. Copyright 1998 Academic Press.

Journal Article↗

PubMed1998

An inhibitor of inducible nitric oxide synthase ameliorates experimental autoimmune myocarditis in Lewis rats.

We studied the effect of nitric oxide (NO) on experimental autoimmune myocarditis (EAC) in rats. We examined the role of inducible nitric oxide synthase (iNOS), an enzyme that produces NO, on hearts affected with EAC, by testing the effects of aminoguanidine (AG), a selective iNOS inhibitor, on the course of EAC. Western blotting detected iNOS in the affected cardiac tissues, but not in CFA immunized cases. Immunohistochemically, the majority of ED1+ macrophages in the EAC lesions were positive for iNOS and nitrotyrosine. A high dose of AG (200 mg/kg/day) significantly reduced the incidence of EAC (p < 0.05) and ameliorated the histological score for the cardiac inflammation (p < 0.01) compared with the low dose AG (100 mg/kg/day) and vehicle treated groups. The immunoblot analysis showed that a high dose of AG effectively suppressed iNOS in hearts affected with EAC. An iNOS band was barely detected in the high dose AG (200 mg/kg) treated group, while it was distinctively visualized in the vehicle and low dose AG (100 mg/kg) treated groups. These results suggest that iNOS is upregulated in EAC lesions and increased NO production plays an important role in the development of EAC. In addition, selective iNOS inhibitors may have a therapeutic role in treating certain autoimmune diseases including EAC.

Animals↗

PubMed1998

Insulin-like growth factor binding proteins localize to discrete cell culture compartments in periosteal and osteoblast cultures from fetal rat bone.

Insulin-like growth factor (IGF)-I and IGF-II are expressed at biologically effective levels by bone cells. Their stability and activity are modulated by coexpression of IGF binding proteins (IGFBPs). Secreted IGFBPs may partition to soluble, cell-associated, and matrix-bound compartments. Extracellular localization may sequester, store, or present IGFs to appropriate receptors. Of the six IGFBPs known, rat osteoblasts synthesize all but IGFBP-1. Of these, IGFBP-3, -4, and -5 mRNAs are induced by an increase in cAMP. Little is known about extracellular IGFBP localization in bone and nothing about IGFBP expression by nonosteoblastic periosteal bone cells. We compared basal IGFBP expression in periosteal and osteoblast bone cell cultures and assessed the effects of changes in cAMP-dependent protein kinase A or protein kinase C. Basal IGFBP gene expression differed principally in that more IGFBP-2 and -5 occurred in osteoblast cultures, and more IGFBP-3 and -6 occurred in periosteal cultures. An increase in cAMP enhanced IGFBP-3, -4, and -5 mRNAand accordingly increased soluble IGFBP-3, -4, and -5 and matrix-bound IGFBP-3 and -5 in both bone cell populations. In contrast, protein kinase C activators suppressed IGFBP-5 mRNA, and its basal protein levels remained very low. We also detected low Mr bands reactive with antisera to IGFBP-2, -3, and -5, suggesting proteolytic processing or degradation. Our studies reveal that various bone cell populations secrete and bind IGFBPs in selective ways. Importantly, inhibitory IGFBP-4 does not significantly accumulate in cell-associated compartments, even though its secretion is enhanced by cAMP. Because IGFBPs bind IGFs less tightly in cell-bound compartments, they may prolong anabolic effects by agents that increase bone cell cAMP.

Animals↗

PubMed1998

Tissue-specific expression of beta-catenin in normal mesenchyme and uveal melanomas and its effect on invasiveness.

This paper is the first in a series aimed at understanding the role of beta-catenin in epithelial-mesenchymal transformation (EMT) and acquisition of mesenchymal invasive motility. Here, we compare the expression of this and related molecules in the two major tissue phenotypes, epithelial and mesenchymal, the latter including normal avian and mammalian fibroblasts and malignant human uveal melanoma cells. Previously, it was proposed that src initiates EMT by tyrosine phosphorylation of the cadherin/catenin complex resulting in a negative effect on epithelial gene expression. On the contrary, we found that although beta-catenin becomes diffuse in the cytoplasm during embryonic EMT, the cytoplasmic beta-catenin of the embryonic and adult mesenchymal cells we examined is not tyrosine phosphorylated. Pervanadate experiments indicate that cytoplasmic PTPases maintain this dephosphorylation. GSK-3beta is present, but little or no APC occurs in normal and neoplastic mesenchymal cells. The function of the nonphosphorylated cytoplasmic beta-catenin in mesenchyme may be related to invasive motility. Indeed, in order to invade extracellular matrix, transitional (Mel 252) melanoma cells transform from an epithelial to a mesenchymal phenotype with increased cytoplasmic beta-catenin. Moreover, antisense beta-catenin and plakoglobin ODNs inhibit Mel 252 and corneal fibroblast invasion of collagen. All fibroblastic, transitional, and spindle melanoma cells contain nuclear as well as cytoplasmic beta-catenin, but they are not significantly more invasive than normal fibroblasts that contain only cytoplasmic beta-catenin.

3T3 Cells↗

PubMed1998

Harvey ras results in a higher frequency of mammary carcinomas than Kirsten ras after direct retroviral transfer into the rat mammary gland.

Exclusive activation of either the Harvey-, Kirsten-, or N-ras gene is often found in human and rodent cancers, although the mechanisms responsible for tissue-specific ras gene activation are poorly understood. In this study, the contribution of ras gene expression and Ras protein activity to the tissue-specificity of ras gene activation was investigated using the rat mammary carcinogenesis model where ras activation, when it occurs, is exclusively in the Harvey ras gene. Differential ras gene expression was examined in mammary tissue from virgin, pregnant, and lactating rats. Harvey ras expression was 1.5-2-fold higher than Kirsten ras or N-ras at each adult stage of development, with the highest ras levels expressed during pregnancy. The modest difference in total mRNA expression found between the independent members of the ras gene family is unlikely to fully account for the exclusive tissue-specificity of Harvey ras activation observed in rat mammary carcinogenesis. Thus, the role of Ras protein specificity was studied by infecting the mammary gland of virgin rats in situ with replication-defective retroviral vectors expressing either the activated or wild-type forms of Harvey- or Kirsten-ras. A 7-14-fold higher number of mammary carcinomas was observed after infection with vectors expressing the G35 to A activated Harvey ras gene product compared with those expressing G35 to A activated Kirsten ras. Mammary carcinomas also developed from infusion of vectors expressing wild-type Harvey ras, but not wild-type Kirsten ras. These data suggest the importance of the Ras protein itself in determining the specificity of the highly homologous Ras family members in organ-specific carcinogenesis.

Animals↗

PubMed1998

Gonadotropin-releasing hormone (GnRH) gene regulation by N-methyl-D-aspartic acid in GT1-1 neuronal cells: differential involvement of c-fos and c-jun protooncogenes.

The present study examined the regulatory mechanisms of GnRH gene expression by N-methyl-d-aspartic acid (NMDA) in immortalized hypothalamic GnRH neurons (GT1-1 cells). NMDA (100 microM) stimulated GnRH mRNA levels transiently at 2 h after treatment. Dose-response experiment showed that there was a biphasic action of NMDA on GnRH mRNA levels: GnRH mRNA levels were increased by NMDA at lower concentrations (10 and 100 microM), but not at higher concentrations (1 and 10 mM). NMDA (100 microM)-induced GnRH mRNA levels were efficiently blocked by pre-treatment with NMDA receptor antagonists, MK-801 and AP-5. We next examined the signal transduction pathways involved in NMDA-induced GnRH gene expression based on previous findings that NMDA signal propagates into the cell through Ca2+ and nitric oxide (NO) pathways in many neurons. While ionomycin, a Ca2+ ionopore, application failed to alter GnRH gene expression, treatment of GT1-1 cells with sodium nitroprusside (SNP), an NO donor, increased GnRH gene expression with a similar time course to NMDA treatment. Moreover, application of GT1-1 cells with nitric oxide synthase (NOS) inhibitors (l-NAME, d-NAME, and NA) prior to NMDA treatment, inhibited NMDA-induced GnRH gene expression. These results indicate that the effect of NMDA is mediated by the NO signalling cascade. The mouse GnRH promoter activity was also increased by NMDA at low concentration (100 microM), but not at high concentration (1 microM), confirming the biphasic action of NMDA on GnRH mRNA levels. Since NMDA (100 microM) and SNP (1 microM) markedly induced c-jun expression, but not c-fos expression, we hypothesized that Jun activation is responsible for the transcriptional activation of GnRH gene expression. To examine this, we performed two different experiments. Treatment of NMDA greatly increased the activity of heterologous promoter of Fos/Jun responsive sequence (-187/-69) from the mouse GnRH promoter fused to hsv-tk minimal promoter. Moreover, overexpression of c-jun induced GnRH promoter activity, while c-fos overexpression decreased GnRH promoter activity. Taken together, this study indicates that NMDA regulates GnRH gene expression in GT1-1 cells through the NO-Jun signal transduction pathway.

Animals↗

PubMed1998

On-line sample preparation of paraquat in human serum samples using high-performance liquid chromatography with column switching.

A new ion-pair high-performance liquid chromatographic method with column-switching has been developed for the determination of paraquat in human serum samples. The diluted serum sample was injected onto a precolumn packed with LiChroprep RP-8 (25-40 microm) and polar serum components were washed out by 3% acetonitrile in 0.05 M phosphate buffer (pH 2.0) containing 5 mM sodium octanesulfonate. After valve switching to inject position, concentrated compounds were eluted in the back-flush mode and separated on an Inertsil ODS-2 column with 17% acetonitrile in 0.05 M phosphate buffer (pH 2.0) containing 10 mM sodium octanesulfonate. The total analysis time per sample was about 30 min and mean recovery was 98.5+/-2.8% with a linear range of 0.1-100 microg/ml. This method has been successfully applied to serum samples from incidents by paraquat poisoning.

Acetonitriles↗

PubMed1998

Presence of activated ras correlates with increased cysteine proteinase activities in human colorectal carcinomas.

The metastatic potential of ras-transfected cells has been attributed in part to significant ras induction of proteinase expression. To determine whether primary cancers also demonstrate higher cysteine proteinase activities in the presence of activated ras genes or altered ras protein expression, we have analyzed 60 primary human colorectal carcinomas for ras gene or protein changes together with the expression of cathepsins B and L. Cancers containing K-ras mutations (47% of 60 carcinomas) demonstrated greater increases in cathepsin L activity than cancers without K-ras mutations (p = 0.029), with particularly significant correlations for earlier stage cancers (Dukes' A and B carcinomas, p = 0.006). Western blots used to characterize ras protein patterns in the same cancer/normal pairs have demonstrated that N-ras protein is more highly expressed in colon tissues than H- or K-ras proteins and that N-ras overexpression occurs in almost 70% of colorectal cancers, with or without a concurrent change in electrophoretic mobility of N-ras protein. Our current study has now shown that N-ras protein overexpression alone does not significantly induce cathepsin B or L activity levels in colon cancers. However, carcinomas demonstrating altered N-ras protein forms, in the absence of any K- or N-ras mutations, expressed significantly higher levels of cathepsin B and L activities compared with carcinomas with normal N-ras protein banding patterns. Our data suggest that colorectal carcinomas with either K-ras mutations or altered forms of N-ras protein may increase their tumorigenic potential via the induction of cathepsin L or B expression levels. Our results also confirm that ras oncogene up-regulation of cathepsin B and L activities, previously reported in cultured cells, is a frequent event in primary human colorectal carcinomas.

Blotting, Western↗

PubMed1998

FMRFamide-related gene family in the nematode, Caenorhabditis elegans.

Many organisms, including mammals, use short peptides as neurotransmitters. The family of FMRFamide (Phe-Met-Arg-Phe-NH2)-like neuropeptides, which all share an -RFamide sequence at their C-termini, has been shown to have diverse functions, including neuromodulation and stimulation or inhibition of muscle contraction. In the nematode, Caenorhabditis elegans, FMRFamide-like peptides (FaRPs) are expressed in approximately 10% of the neurons, including motor, sensory, and interneurons that are involved in movement, feeding, defecation, and reproduction. At least 14 genes, designated flp-1 through flp-14, encode FaRPs in C. elegans. Here, we present data that all 14 flp genes are transcribed in C. elegans, and several of these genes are alternatively spliced. Each flp gene encodes a different set of FaRPs, yielding a predicted total of 44 distinct FaRPs. Using staged RNA for reverse-transcription/polymerase chain reactions (RT/PCR), we determined that most flp genes are expressed throughout development. These results suggest that a complex family of FaRPs have varied roles through all stages of development and in adulthood in C. elegans.

Amino Acid Sequence↗

PubMed1998

The trophic action of IL-7 on pro-T cells: inhibition of apoptosis of pro-T1, -T2, and -T3 cells correlates with Bcl-2 and Bax levels and is independent of Fas and p53 pathways.

Signals from the IL-7R are essential for normal thymocyte development. We isolated thymocytes from early developmental stages and observed that suspensions of pro-T1, -T2, and -T3 cells rapidly died in culture. Addition of IL-7 promoted their survival, but did not induce cell division. Pro-T4 cells did not undergo rapid cell death, and their survival was therefore independent of IL-7. Death in the absence of IL-7 showed the hallmarks of apoptosis, including DNA fragmentation and annexin V binding; however, caspase inhibitors blocked DNA fragmentation, but did not block cell death. The trophic effect of IL-7 was partially inhibited by blocking protein synthesis. The p53 pathway was not involved in this death pathway, since pro-T cells from p53-/- mice also underwent cell death in the absence of IL-7. The Fas/Fas ligand pathway was not involved in cell death, since Fas-deficient pro-T cells died normally in the absence of IL-7, anti-Fas Abs did not protect cells from death in the absence of IL-7, and Fas expression was undetectable on cells at these stages. The IL-7 trophic affect correlated with increased intracellular levels of Bcl-2 and decreased levels of Bax, whereas no Bcl-X(L), Bcl-w, or Bad was detectable. Thus, maintaining a favorable Bcl-2/Bax ratio may account for the trophic action of IL-7.

Animals↗

PubMed1998

Cloning and in vitro expression of TPK3, a Toxoplasma gondii homologue of shaggy/glycogen synthase kinase-3 kinases.

As an initial effort to dissect the signaling pathways responsible for pathogenesis of Toxoplasma gondii infection, we report the cloning and in vitro functional studies of TPK3 (Toxoplasma protein kinase-3), a homologue of shaggy/glycogen synthase kinase-3 (GSK-3) kinases. The shaggy/GSK-3 family of kinases are highly conserved protein kinases that play important roles in cell fate determination, nuclear signaling and hormonal regulation. The TPK3 gene was isolated by RT-PCR with degenerate primers corresponding to highly conserved regions of serine/threonine protein kinases. The complete sequences of genomic and cDNA clones indicated the open reading frame, 1185 bp in size, is interrupted by five introns. The predicted protein sequence of TPK3 shows 54% identity to shaggy/GSK-3 over the catalytic domains. Southern analysis revealed TPK3 is a single copy locus in the Toxoplasma genome. Antisera to other GSK-3 proteins from other species recognized GST-TPK3 and a protein of the predicted size in parasites lysates. In vitro kinase assays with GST-TPK3 indicated that TPK3 autophosphorylates and phosphorylates protein phosphatase inhibitor-2 (I-2), a specific substrate of GSK-3 kinase.

Amino Acid Sequence↗

PubMed1998

Anhydrolide macrolides. 1. Synthesis and antibacterial activity of 2,3-anhydro-6-O-methyl 11,12-carbamate erythromycin A analogues.

A series of 3-descladinosyl-2,3-anhydro-6-O-methylerythromycin A 11, 12-carbamate analogues have been synthesized and evaluated for antibacterial activity. These compounds were found to be potent antibacterial agents against Gram-positive organisms in vitro, many having MIC values below 1 microg/mL for the macrolide-susceptible Staphylococcus aureus, Streptococcus pyogenes, and Streptococcus pneumoniae, as well as improved activity compared to erythromycin A against the inducibly MLS (macrolide, lincosamide, and streptogramin B)-resistant organisms. Structure-activity studies revealed that arylalkyl carbamates with two and four carbon atoms between the aromatic moiety and carbamate nitrogen have the best in vitro activity. All of the C-10 epi analogues evaluated were found to have substantially less activity than the corresponding natural C-10 isomer. Several analogues demonstrated moderate antibacterial activity against the constitutively resistant S.aureus A-5278, S. pneumoniae5979, and S.pyogenes 930. However, despite potent in vitro activity, these analogues showed only moderate in vivo activity in mouse protection studies.

Animals↗