Acupuncture Treatment in the Reduction of Knee Pain Due to Osteoarthritis

Objective: As knee osteoarthritis (KOA) is a very common, progressive, irreversible, and painful disease that affects many as they age, treatment modalities to alleviate symptoms are important to explore. This work aims to gather evidence as to the efficacy of a minimum of 4 weeks of acupuncture for the treatment of patients with knee pain secondary to osteoarthritis. Methods: Google Scholar and Valpo Library Databases were perused with a variety of search phrases to yield potential sources. The date of publication was set to be within the last 5 years. Studies needed to include trials of traditional/ conventional acupuncture. Treatment outcome measures varied between studies but most relied on pain scores and joint functioning protocols. No age, gender or racial exclusions were made. Results: Most studies indicate consensus that acupuncture therapy is beneficial for KOA when done for periods of longer than 4 weeks, however many are concerned with limitations in existing studies regarding variability between execution of trials and quality of reporting. Conclusion: Further research is needed to confirm acupuncture benefits and to determine the optimal frequency, duration, and mode of acupuncture treatment for KOA so that providers can recommend it as a truly effective treatment option for patients. Keywords: knee osteoarthritis, joint, traditional Chinese medicine, acupuncture, pain, needl

Effect of Media and Estrogen on Morphological Change in Candida albicans

Introduction: Candida albicans (C. albicans), an opportunistic pathogen, lives symbiotically within the intestine of its human host. Temperature and chemical factors have been shown to induce a morphological change in C. albicans from yeast to filamentous form turning C. albicans pathogenic. In this study, we investigated the intestinal cues that might be responsible for the change. We found that different solid media impact the morphological phenotype so we focused on characterizing these before further testing. We tested Estradiol (E2) because of its known linkage to sepsis and higher levels during infections. Experiments were conducted to compare solid agar plates of YEPD, Minimal Media (MM), and Spider Media (SP) for C. albicans growth to choose the best one for further testing with E2 and other factors that could be prone to causing morphological changes. Methods: C. albicans was inoculated through streak method on different solid media (YEPD, MM, SP) and incubated at 30℃. The effect of 0.1nM E2 on C. albicans morphology was also tested. Morphological changes were assayed through bright-field microscopy. Results: Using the three different medias, we found three distinctive phenotypes: A, B, and C. Out of 6 experiments of 14 MM plates, the expressed phenotype was 86% A and 14% inconclusive of the time. 8 experiments of 17 SP plates showed 100% of phenotype B. 6 experiments of 14 YEPD plates presented phenotype C 92% of the time and 8% inconclusive. For E2 trials, 2 experiments, 6 MM plates showed 50% phenotype A and 50% inconclusive. 4 experiments, 10 SP plates had phenotype B 100%. YEPD 2 experiments, 2 plates had phenotype C at 100%. Conclusion: We have established experimental conditions of media controls for further testing whether E2 and other cues, such as inflammatory cytokines, have inhibitory or positive effects on the growth of C. albicans

Testing Environmental Cues on Candida albicans Morphology

Introduction C. albicans is a commensal fungus which under certain environmental cues (e.g., pH, oxidative stress) shifts morphology from spores to filamentous and becomes invasive within the human body. This work aims to identify the environmental gut cues responsible for this morphological shift. Estrogen (E2) becomes elevated during sepsis, thus the guiding hypothesis states that E2 may represent a factor responsible for the morphological change in C. albicans. Methods A calibration curve of growth of C. albicans in liquid minimal media (MM) was established using a spectrophotometer and correlating optical density with cell counts measured with a hematocyter. Liquid MM was inoculated in quadruplets of three different amounts of C. albicans. To test the effect of estrogen at 1nM concentration, E2 was added at the time of inoculation to one of each tube set, and fetal bovine serum was the positive control in another tube. All tubes were anaerobically grown over 3 nights in a shaking incubator at 30℃. Morphological changes were assayed using bright-field microscopy. Results C. albicans was inoculated in amounts of 1, 2, and 4 million cells into sets of 4 tubes each based on the established growth curve. The MM relationship between OD and number of cells is described by the following equation: 1.06×106 + 1.83×10 7x + 1.68×10 7x 2 , R 2= 0.867. Adding E2 at 1 nM to the liquid media appeared to induce filamentous growth and budding, as with positive control 10% FBS. Conclusion Our preliminary experiments indicate that regardless of initial cell amount, tubes containing E2 seem to induce more filamentous growth in MM, as observed with FBS (positive control). Further experiments to determine effects of E2 at other concentrations would bring more insight, as well as trials combining E2 and FBS to explore if there is an additive or inhibitory effect on filamentation

Candida albicans Infection Decreases The Expression Of The Na+-K+-2Cl– Cotransporter 1 In T84 and Madin Darby Canine Kidney Cells

The commensal human fungal pathogen, Candida albicans, prior to infect the human body, must penetrate the intestinal mucosal barrier. To do so, it needs to bypass the different protective mechanisms such as fluid secretion. The basolateral Na+-K+-2Cl– cotransporter 1 (NKCC1) is a key protein regulating fluid secretion in the intestine. We hypothesize that C. albicans decreases fluid secretion prior to invasion by inducing NKCC1 internalization. In our experiments, we used Madin Darby canine kidney (MDCK) cells expressing a GFP-NKCC1 fluorescent tag and T84 cells, a human colonic cell line. Cells were infected with 100,000 C. albicans for varying lengths of time, fixed, stained and mounted for fluorescence microscopy. The number of internalized vesicles was evaluated using FIJI. Our results show that in MDCK cells, C. albicans only increased NKCC1 internalization at the 30-minute time point (P\u3c0.05), all subsequent time points were not significant. Similarly, infecting T84 cells with C. albicans significantly induced NKCC1 internalization at the 30-minute (P\u3c0.05), 1 hour (P\u3c0.05), and 90-minute (P\u3c0.05) time points. Past 90 minutes, we observed a sharp decline in the number of internalized vesicles that continued to decrease through 6 hours of exposure to C. albicans. Finally, in C. albicans-T84 infected cells, using an immunoblot approach, we found that total NKCC1 protein expression was decreased by ~20% (P\u3c0.05) compared to uninfected cells. Our results suggest that C. albicans induces internalization of NKCC1, and subsequent degradation of NKCC1, which would decrease fluid secretion and allow adhesion, and invasion of the epithelium

Effect of Media and Estrogen on Morphological Change in Candida albicans

Introduction: Candida albicans (C. albicans), an opportunistic pathogen, lives symbiotically within the intestine of its human host. Temperature and chemical factors have been shown to induce a morphological change in C. albicans from yeast to filamentous form turning C. albicans pathogenic. In this study, we investigated the intestinal cues that might be responsible for the change. We found that different solid media impact the morphological phenotype so we focused on characterizing these before further testing. We tested Estradiol (E2) because of its known linkage to sepsis and higher levels during infections. Experiments were conducted to compare solid agar plates of YEPD, Minimal Media (MM), and Spider Media (SP) for C. albicans growth to choose the best one for further testing with E2 and other factors that could be prone to causing morphological changes. Methods: C. albicans was inoculated through streak method on different solid media (YEPD, MM, SP) and incubated at 30℃. The effect of 0.1nM E2 on C. albicans morphology was also tested. Morphological changes were assayed through bright-field microscopy. Results: Using the three different medias, we found three distinctive phenotypes: A, B, and C. Out of 6 experiments of 14 MM plates, the expressed phenotype was 86% A and 14% inconclusive of the time. 8 experiments of 17 SP plates showed 100% of phenotype B. 6 experiments of 14 YEPD plates presented phenotype C 92% of the time and 8% inconclusive. For E2 trials, 2 experiments, 6 MM plates showed 50% phenotype A and 50% inconclusive. 4 experiments, 10 SP plates had phenotype B 100%. YEPD 2 experiments, 2 plates had phenotype C at 100%. Conclusion: We have established experimental conditions of media controls for further testing whether E2 and other cues, such as inflammatory cytokines, have inhibitory or positive effects on the growth of C. albicans

Effects of Environmental Factors on Candida albicans Morphology: A Focus On Estrogen and Microgravity

C. albicans is one of the commensal fungi living in the human intestinal tract in a harmless spore form. In its filamentous form, C. albicans becomes invasive and penetrates the human body, which can cause serious health issues. In vitro factors such as change in temperature or pH are known to induce morphology shift in C. albicans. Interestingly, microgravity has been reported to decrease the human immunity and increase gene virulence expression in C. albicans. During sepsis, high levels of estrogen are reported and the risk of candidiasis also increases. Within present work, we tested the effect of microgravity and estrogen on the shift of morphology (spore to filamentous). C. albicans were grown in minimum media for 3 days in presence or absence of 0.1 nM estrogen. In addition, two other groups of C. albicans were subjected to microgravity for 3 days, using a clinostat, in presence or in absence of estrogen. For each condition, 5 random pictures were taken and scored 1 for the presence and 0 for absence of filament. Experiments were conducted in duplicate. Our results show that subjecting C. albicans to microgravity significantly increase the number of filaments compared to control (9.59±2.77 versus 1.68±1.93, P\u3c0.001, unpaired t-test), whereas estrogen did not significantly affect the number of filaments compared to control (2.66±1.61 versus 1.68±1.93, p=0.6, unpaired t-test). Finally, there was no significant effect of estrogen found on the number of filament when C. albicans was exposed to microgravity plus estrogen versus microgravity alone (8.0±2.76 versus 9.59±2.77). In conclusion, we have found that simulated microgravity dramatically increases the number of filaments, and estrogen at 0.1 nM has no effect on the number of filaments in our experimental conditions

Effects of Environmental Factors on Candida albicans Morphology: A Focus on Estrogen and Microgravity

During body invasion, C.albicans change their morphology from yeast to filamentous, but the environmental factors responsible for the change in morphology are not well characterized. During Sepsis, high levels of estrogen (E2) are recorded (~0.1 nM), in addition during spaceflight it has been shown that C.albicans become virulent. In this study, we aimed at characterizing the effect of estrogen and microgravity as environmental factors inducing filamentous growth. In our experiments, we grow C.albicans in minimum liquid media and use brightfield microscopy to observe the morphology. Microgravity is simulated using a clinostat. In an experimental series, we tested the effect of FBS (positive control) and estrogen on filamentous growth. We found that in the control only 1 out of 10 slides showed filaments. In the presence of FBS, we found filamentous growth in 10 out of 10 slides. In the presence of estrogen, filaments were observed in 8 out of 10 slides. In addition to FBS, the combination of FBS + E2 we found filament growth in 10 out 10 of the slides. However, in the presence of microgravity, we observed filaments in 9 out of 10 slides, meanwhile, we only observed 4 out of 10 slides without microgravity. In addition to microgravity, the combination of E2 + Microgravity, filaments were observed in 3 out of 10 slides. Meanwhile, we only observed 4 out of 10 slides with just E2. In conclusion, estrogen does not inhibit filament growth stimulated by FBS, but it prevents filamentous growth in microgravity

Candida albicans induces internalization of the Na + -K + -2Cl – cotransporter 1 expressed in Madin-Darby Canine Cells

Candida albicans (C. albicans) is one of the most common human fungal pathogens, causing life threatening systemic infections in critically ill and immunocompromised patients. In the gut, fluid secretion participates in the body defense by flushing microorganisms and by maintaining the surface mucus hydrated. It is still unknown whether C. albicans decreases the host defense mechanism prior to invading intestinal cells. Previous work in the lab has demonstrated that the basolateral Na+ -K+ -2Cl - cotransporter 1 (NKCC1) plays a key role in the regulation of fluid secretion, and internalization of NKCC1 represents a potent pathway to blunt fluid secretion. We hypothesize that C. albicans, before invading intestinal cells, decreases fluid secretion by causing NKCC1 internalization. To test our hypothesis, we utilized Madin-Darby Canine Kidney (MDCK) cells stably expressing eGFP-NKCC1. MDCK were cultured on coverslips in 6 well plate. On the day of the experiment, MDCK were exposed to 50,000 C. albicans, for 15, 30 min,1 and 2 h hours. MDCK were also exposed to 100 nM phorbol 12-myristate 13-acetate (PMA) for 15 min as a positive control for NKCC1 internalization. Coverslips were fixed in 1% paraformaldehyde for 30 min and mounted for fluorescence microscopy. Fluorescent images were acquired using an inverted IX83 Olympus microscope equipped with a CCD camera. Exposing MDCK cells to C. albicans for 15 and 30 min did not result in NKCC1 internalization as compared to 100 nM PMA. Conversely, after 1- and 2-hours exposure to C. albicans, NKCC1 internalization was observed in some cells. Our preliminary experiments suggest that C. albicans induces NKCC1 internalization and may represent one mechanism by which C. albicans lower the intestinal defense mechanism. Gaining a better understanding of how C. albicans invades the mucosal barrier will greatly help developing new strategies for fighting candidiasis

Effect of Candida albicans infection on the plasma membrane expression of the Na+-K+-2Cl– cotransporter 1 (NKCC1) in T84 and Madin Darby Canine Kidney cells (MDCK)

C. albicans is a commensal human fungal pathogens. To infect the human body, it must penetrate the intestinal mucosal barrier. Fluid secretion is one of the intestinal defense mechanisms and NKCC1 is a key protein regulating fluid secretion in the colon. We hypothesize that, C. albicans, before invasion decreases fluid secretion by causing NKCC1 internalization. In our experiments, we used MDCK cells expressing a GFP-NKCC1, and T84 a human colonic. Cells were infected with 100,000 C. albicans for different times, fixed, stained and mounted for fluorescence microscopy. Images were acquired using an Olympus IX83 microscope equipped with a DP80 CCD camera. The number of vesicles was evaluated using FIJI. Our preliminary results show that in MDCK cells, phorbol 12 myristate 13 acetate (PMA), a positive control, induced a significant increase of vesicles containing NKCC1 (P\u3c0.001), whereas C. albicans did not significantly increase NKCC1 internalization at all time points tested (ANOVA, Dunnett\u27s Multiple Comparison). Similarly, PMA induced a significant increase of NKCC1 internalization (P=0.007) in T84 Cells. Infecting T84 cells with C. albicans, significantly induced NKCC1 internalization only at 90 min (P=0.01), but not at other time points. Our results suggest that C. albicans causes internalization of NKCC1 which would decrease fluid secretion. The non-significance at some time points may be due to the low number of replicates at our early time points, whereas at later time points we suspect that NKCC1 is already degraded and cannot be detected

El porvenir segoviano : periódico literario y de intereses materiales: Año I Número 97 - 02 Septiembre 1864

Copia digital. Madrid : Ministerio de Cultura. Subdirección General de Coordinación Bibliotecaria, 200