Induction of cell cycle changes and modulation of apoptogenic/anti-apoptotic and extracellular signaling regulatory protein expression by water extracts of I'm-Yunity™ (PSP)

BACKGROUND: I'm-Yunity™ (PSP) is a mushroom extract derived from deep-layer cultivated mycelia of the patented Cov-1 strain of Coriolus versicolor (CV), which contains as its main bioactive ingredient a family of polysaccharo-peptide with heterogeneous charge properties and molecular sizes. I'm-Yunity™ (PSP) is used as a dietary supplement by cancer patients and by individuals diagnosed with various chronic diseases. Laboratory studies have shown that I'm-Yunity™ (PSP) enhances immune functions and also modulates cellular responses to external challenges. Recently, I'm-Yunity™ (PSP) was also reported to exert potent anti-tumorigenic effects, evident by suppression of cell proliferation and induction of apoptosis in malignant cells. We investigate the mechanisms by which I'm-Yunity™ (PSP) elicits these effects. METHODS: Human leukemia HL-60 and U-937 cells were incubated with increasing doses of aqueous extracts of I'm-Yunity™ (PSP). Control and treated cells were harvested at various times and analyzed for changes in: (1) cell proliferation and viability, (2) cell cycle phase transition, (3) induction of apoptosis, (4) expression of cell cycle, apoptogenic/anti-apoptotic, and extracellular regulatory proteins. RESULTS: Aqueous extracts of I'm-Yunity™ (PSP) inhibited cell proliferation and induced apoptosis in HL-60 and U-937 cells, accompanied by a cell type-dependent disruption of the G(1)/S and G(2)/M phases of cell cycle progression. A more pronounced growth suppression was observed in treated HL-60 cells, which was correlated with time- and dose-dependent down regulation of the retinoblastoma protein Rb, diminution in the expression of anti-apoptotic proteins bcl-2 and survivin, increase in apoptogenic proteins bax and cytochrome c, and cleavage of poly(ADP-ribose) polymerase (PARP) from its native 112-kDa form to the 89-kDa truncated product. Moreover, I'm-Yunity™ (PSP)-treated HL-60 cells also showed a substantial decrease in p65 and to a lesser degree p50 forms of transcription factor NF-κB, which was accompanied by a reduction in the expression of cyclooxygenase 2 (COX2). I'm-Yunity™ (PSP) also elicited an increase in STAT1 (signal transducer and activator of transcription) and correspondingly, decrease in the expression of activated form of ERK (extracellular signal-regulated kinase). CONCLUSION: Aqueous extracts of I'm-Yunity™ (PSP) induces cell cycle arrest and alterations in the expression of apoptogenic/anti-apoptotic and extracellular signaling regulatory proteins in human leukemia cells, the net result being suppression of proliferation and increase in apoptosis. These findings may contribute to the reported clinical and overall health effects of I'm-Yunity™ (PSP)

Survey of Borreliae in ticks, canines, and white-tailed deer from Arkansas, U.S.A.

<p>Abstract</p> <p>Background</p> <p>In the Eastern and Upper Midwestern regions of North America, <it>Ixodes scapularis</it> (L.) is the most abundant tick species encountered by humans and the primary vector of <it>B. burgdorferi,</it> whereas in the southeastern region <it>Amblyomma americanum</it> (Say) is the most abundant tick species encountered by humans but cannot transmit <it>B. burgdorferi.</it> Surveys of Borreliae in ticks have been conducted in the southeastern United States and often these surveys identify <it>B. lonestari</it> as the primary <it>Borrelia</it> species, surveys have not included Arkansas ticks, canines, or white-tailed deer and <it>B. lonestari</it> is not considered pathogenic. The objective of this study was to identify <it>Borrelia</it> species within Arkansas by screening ticks (n = 2123), canines (n = 173), and white-tailed deer (n = 228) to determine the identity and locations of Borreliae endemic to Arkansas using PCR amplification of the flagellin (<it>flaB)</it> gene.</p> <p>Methods</p> <p>Field collected ticks from canines and from hunter-killed white-tailed were identified to species and life stage. After which, ticks and their hosts were screened for the presence of <it>Borrelia</it> using PCR to amplify the <it>flaB</it> gene. A subset of the positive samples was confirmed with bidirectional sequencing.</p> <p>Results</p> <p>In total 53 (21.2%) white-tailed deer, ten (6%) canines, and 583 (27.5%) Ixodid ticks (252 <it>Ixodes scapularis</it>, 161 <it>A. americanum</it>, 88 <it>Rhipicephalus sanguineus</it>, 50 <it>Amblyomma maculatum,</it> 19 <it>Dermacentor variabilis,</it> and 13 unidentified <it>Amblyomma</it> species) produced a <it>Borrelia flaB</it> amplicon. Of the positive ticks, 324 (22.7%) were collected from canines (151 <it>A. americanum,</it> 78 <it>R. sanguineus</it>, 43 <it>I. scapularis,</it> 26 <it>A. maculatum,</it> 18 <it>D. variabilis</it>, and 8 <it>Amblyomma</it> species) and 259 (37.2%) were collected from white-tailed deer (209 <it>I. scapularis,</it> 24 <it>A. maculatum,</it> 10 <it>A. americanum,</it> 10 <it>R. sanguineus</it>, 1 <it>D. variabilis</it>, and 5 <it>Amblyomma</it> species). None of the larvae were PCR positive. A majority of the <it>flaB</it> amplicons were homologous with <it>B. lonestari</it> sequences: 281 of the 296 sequenced ticks, 3 canines, and 27 deer. Only 22 deer, 7 canines, and 15 tick <it>flaB</it> amplicons (12 <it>I. scapularis</it>, 2 <it>A. maculatum</it>, and 1 <it>Amblyomma</it> species) were homologous with <it>B. burgdorferi</it> sequences.</p> <p>Conclusions</p> <p>Data from this study identified multiple Borreliae genotypes in Arkansas ticks, canines and deer including <it>B. burgdorferi</it> and <it>B. lonestari;</it> however, <it>B. lonestari</it> was significantly more prevalent in the tick population than <it>B. burgdorferi</it>. Results from this study suggest that the majority of tick-borne diseases in Arkansas are not <it>B. burgdorferi.</it></p