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DC0-260 - Dell Certified Storage Networking(R) Professional - Dump Information

Vendor	:	DELL
Exam Code	:	DC0-260
Exam Name	:	Dell Certified Storage Networking(R) Professional
Questions and Answers	:	63 Q & A
Updated On	:	December 20, 2017
PDF Download Mirror	:	DC0-260 Brain Dump
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DC0-260 Questions and Answers

Answer: B, C, E

QUESTION: 57

Which statements is correct about switch ports?

If a switch port is connected to an FC-AL port, it acts as an F_Port.
If the switch port is connected to another switch, it acts as an E_Port.
If a switch port is connected to an N_Port, it is configured as an arbitrated loop.
If a universal port on a switch is connected to an N_Port, the universal port works as an FL_Port.

Answer: B

QUESTION: 58

Which two statements about Storage Area Networks (SANs) are true? (Choose two)

SANs provide file level I/O access to storage resources.
SANs provide block level I/O access to storage resources.
SANs enable the sharing of storage resources via IP / Ethernet networking
SANs enable the sharing of storage resources across a high speed, low latency, fiber Channel network

Answer: B, D

QUESTION: 59

Which two statements about implementations for bridges in a Storage AREA Network (SAN) are true? (Choose two)

A bridge can be used to segment Fiber Channel SANs.
A bridge can used to connect Fiber Channel to IP network.
A bridge can connect SCSI devices to Fiber Channel networks
A bridge may reside within the storage device or in an external chassis.

Answer: B, D

QUESTION: 60

Compared to other storage architectures, which two characteristics describe Network Attached Storage (NAS)?

least efficient use of storage resources
simple implementation of centralized storage
removes data management traffic from the network
well suited for heterogeneous computing environments

Answer: B, D

QUESTION: 61

In which three common topologies can Fiber Channel be implemented? (Choose three)

daisy chain
point-to-point
arbitrated loop
switched fabric
hierarchical tree

Answer: B, C, D

QUESTION: 62

Which three protocols can be used in a Storage Area Network (SAN)? (choose three)

NFS
FCIP
IFCP
CIFS
FC-SW

Answer: B, C, E

QUESTION: 63

DRAG DROP

Place the steps for powering down a Dell / EMC storage array in the correct order.

Answer:

Reference:

DELL SAN Functional checklist delivered with my SAN

DELL DC0-260 Exam (Dell Certified Storage Networking(R) Professional) Detailed Information

DELL DC0-260

DC0-260 exam :: Article by ArticleForgeIdaho State Police checking out applicants for Troopers MERIDIAN — The Idaho State Police all started checking out applicants for Troopers Oct. 2 and will continue via Nov. eight. ISP anticipates hiring approximately 20 new Troopers with positions purchasable in all of ISP’s six district workplaces.
ISP is peculiarly hunting for applicants to fill resident positions in faraway and rural communities in Idaho and encourages candidates wishing to reside in these communities to observe. The hiring process takes about 6 months to complete, and successful candidates will start training in may additionally 2016.
candidates might also begin the online utility process and exam starting Oct. 2 however ought to conclude the online examination no later than Nov. 8. applicants from this manner may be selected to take part in an oral board examination and physical health check in order to take place in early December. The right candidates from that system will then undergo an in depth background verify, which comprises reference exams, work heritage verification, crook heritage and credit exams. applicants have to also circulate a collection of psychological, scientific and physical fitness examinations, together with a polygraph exam and substance abuse screening. old legislations enforcement journey isn't required.

Story continues under video

On usual, the working towards manner takes about nine months to comprehensive, which contains ISP’s 20-week advanced working towards classification. Upon successful completion of the academy, the new Troopers will begin on-the-job box training with experienced Troopers. The academy and on-the-job working towards are extreme and require physically and mentally potent individuals.
Trooper applicants should be inclined to work any place within the state of Idaho. Troopers are required to work all shifts including nights, weekends, and vacation trips. beginning pay for a Trooper is $19.48 per hour and possibly better if the candidate is a current peace officer with as a minimum five years adventure. additional pay raises can be found after a probationary length for capabilities and training similar to Spanish fluency, drug focus experts, submit certificates and college degrees.
Have questions concerning the hiring process or what it takes to be a Trooper? ISP has many activities planned to reply questions about the hiring technique and the various profession alternatives purchasable as a Trooper.
Recruiters might be at profession fairs and hobbies to answer questions about these dates at these places:
October eight - college of Western Idaho profession fair - Nampa Campus academic building - 9 am to three pm
October 14 - Boise State college career reasonable - scholar Union constructing - 9-three p.m.
October 27 - profession Enhancement Expo - Mountain domestic Air force Base - militia Personnel handiest
ISP will also hold digital Q&A classes in cooperation with the Idaho branch of Labor. The classes can be broadcast are living from the Meridian branch of Labor workplace and will be linked virtually to branch of Labor offices in Coeur d’Alene, Lewiston, Twin Falls, Pocatello, and Idaho Falls on Oct. 13, Oct. 28 and Nov. 4 from 4-7 p.m.
for people that are unable to make it to an experience in person, ISP will additionally dangle a Q&A session on Twitter and Periscope on Oct. 23 from three-four p.m. ISP’s Twitter deal with is @IdahoStPolice.
ISP invites these attracted to fitting a Trooper, as smartly as the public at tremendous, to comply with ISP and police companies around the globe throughout the 5th world Police Tweet-a-thon on Friday, Oct. 2. beginning at 7 p.m. ISP’s Twitter followers will take a digital journey-along with a Trooper on patrol in the Boise area and be able to enhanced take note what a Trooper encounters throughout a “regular” Friday nighttime shift. All groups collaborating within the Tweet-a-thon will use the hashtag #PolTwt throughout the adventure.

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medical institution profitability in Florida: a revisitation.sanatorium fiscal efficiency and its determinants has been a vital field of empirical research and of incredible interest to traders, board participants, creditors, score agencies, and regulators. Sear (1991) examined sanatorium profitability in a sample of 50 investor-owned or for-earnings (FP) and 60 no longer-for-income (NP) hospitals located in Florida for the duration 1982 to 1988. The criminal big difference between FP and NP hospitals lies in issues regarding tax exempt repute--the potential to get hold of tax-deductible donations, and trouble within the distribution of gains and belongings. His statistical consequences point out that FP hospitals are greater ecocnomic than NP hospitals, and average length of stay and wages per adjusted patient day are the variables with tremendous explanatory vigor in assessing health facility profitability. Walker (1993), the usage of a Logit regression model, found that financial variables, through themselves, don't seem to be enough to discriminate between profitable and non-profitable hospitals. Watt, Renn, Hahn, Derzon & Schramm (1986) said that FP hospitals had better earnings than their NP counterparts. Herzlinger & Krasker (1987) suggested that NP hospitals do not function as neatly financially as FP hospitals nor do they make amends for this by using returning greater social merits. although, Haddock, Arrington & Skelton (1989), Arrington & Haddock (1990), and Fitzgerald & Jacobsen (1987) disputed these findings on conceptual and methodological grounds. in keeping with a pattern of hospitals observed in Florida in 1980, Sloan & Vraciu (1983) discovered that FP and NP hospitals had been very nearly similar in terms of profitability. Forgione (1987) and Forgione, Schiff & Crumbley (1996) found that FP hospitals are typically half the size of NP hospitals, with lower case mix, shorter length of stay, greater charge per day, lower can charge per case, and improved profitability.during this study DC0-260, we reexamine issues regarding hospital profitability via enhancing upon previous analysis in a few instructions. First, not like most outdated studies which use statistics from earlier than or all the way through the section-in of the Medicare operating cost potential price equipment (PPS), the records set employed during this study DC0-260 is obtained from the submit-PPS working cost length and is for this reason much more representative of the price methods presently facing US hospitals. in contrast to the previous charge-based compensation mechanism, price below PPS is in accordance with a predetermined charge expense. PPS operating can charge charge turned into initiated in 1983 and turned into phased-in until 1987 providing hospitals with a transitional period to alter to the new charge gadget. PPS capital can charge price was phased-in over the 10-yr period 1991-2001. 2nd, we comprise in our empirical evaluation economic and financial variables (e.g. diploma of competition and financial indebtedness) and additionally examine the results of conversion to FP repute on hospital profitability. Third, we make use of piecewise linear regression evaluation which allows for a parsimonious modeling of definite styles of nonlinearity within the variables. Our empirical proof suggests that vicinity, dimension, occupancy cost, earnings popularity, competitors, instructing repute, and monetary indebtedness proceed to be statistically enormous in explaining health center profitability within the put up-PPS period. additionally, mattress measurement and occupancy rate are discovered to be non-linearly regarding clinic profitability.The the rest of the paper is constructed as follows. The subsequent part presents the data and gives descriptive summary data. The empirical outcomes are offered in here. We conclude with a summary of our main findings within the remaining part.
information AND ECONOMETRIC METHODOLOGY
The data set includes seventy nine FP hospitals, 70 NP hospitals, and 29 hospitals that converted from NP to FP status, masking the years 1991 and 1995. The information source is the Medicare cost file statistics offered via HCIA, Inc., Baltimore, Maryland.
The measure of profitability in this examine is the Return on belongings (ROA), a continual economic fame variable described as web revenue divided through total property. ROA reflects a type of efficiency ratio because it relates medical institution output to non-labor inputs. right here regression model is estimated to clarify model in health center profitability:
ROA = f(BEDS, OCCUPANCY, ownership, LOS, DEBT, personnel, educating, SOLE, yr, CONVERT)the place,
personnel = variety of full-time equal employees per 100 admissions adjusted for case combine index.
ownership = dummy variable indicating class of possession handle (equals 1 for NP reputation, 0 for FP status).
teaching = dummy variable, taking the price of 1 if the medical institution gives educating and intern training, 0 otherwise.
SOLE = dummy variable shooting the degree of competitors dealing with a hospital (equals 1 if a medical institution is the sole Medicare company, 0 in any other case).
yr = dummy variable, taking the price of 1 if yr is 1991 and 0 if 12 months is 1995.
BEDS = variety of beds in carrier, which enters the regression equation in piecewise linear trend as follows:
* BEDS 0 to one hundred = the value of the variable if BEDS < one hundred, = one hundred if BEDS [greater than or equal to] one hundred.
* BEDS a hundred to 500 = 0 if BEDS < 100, = the price of the variable minus one hundred if a hundred [less than or equal to] BEDS < 500, = 500 if BEDS [greater than or equal to] 500
* BEDS over 500 = 0 if BEDS < 500, = the value of the variable minus 500 if BEDS [greater than or equal to] 500.
DEBT = debt per bed in provider. Debt is defined as bonds issued plus loans.
OCCUPANCY = percent of beds in provider occupied, entering the regression equation as follows:
* OCCUPANCY 0 to 0.10 = the price of the variable if OCCUPANCY < 0.10, = 0.10 if OCCUPANCY [greater than or equal to] 0.10.
* OCCUPANCY 0.10 to 0.50 = 0 if OCCUPANCY<0.10, = the price of the variable minus 0.10 if 0.10 [less than or equal to] OCCUPANCY < 0.50, = 0.50 if OCCUPANCY [greater than or equal to] 0.50.
* OCCUPANCY over 0.50 = 0 if OCCUPANCY < 0.50, = the value of the variable minus 0.50 if OCCUPANCY [greater than or equal to] 0.5.
CONVERT = dummy variable, taking the value of 1 if a health center transformed from NP fame to FP fame between 1991 and 1995, and zero in any other case.
We make use of a piecewise linear regression permitting for two adjustments in the slope coefficients for BEDS and OCCUPANCY. This relaxes the belief of linearity within the relationship, which we discovered changed into no longer supported by means of the information, and constitutes a comparatively parsimonious way to describe the patterns within the records. The estimation methodology used is typical least squares (OLS) with heteroscedasticity adjustment for general blunders in line with White (1980).
display 1 studies descriptive data for the independent and dependent variables of the sample hospitals for the years 1991 and 1995. The profitability of our sample hospitals increased between years 1991 and 1995. any such sample turned into exhibited by means of both FP and NP hospitals. The more desirable financial efficiency of hospitals relates to improvements in collection and digital funds, as well as hospital administration study DC0-260ing to function conveniently beneath the new regime of working can charge PPS, via shortening patient LOS and elevating inpatient case combine.
Hospitals reveal an increase of their measurement between years 1991 and 1995. Occupancy costs declined for the years 1991 to 1995. Declines in occupancy prices were accompanied for each FP and NP hospitals; besides the fact that children, the occupancy rate for NP hospitals changed into greater in both the 1991 and 1995 years. despite the fact, hospitals changing from NP to FP repute confirmed steeper declines in occupancy fees than non-changing hospitals.
EMPIRICAL facts
show 2 reviews the empirical effects for our regression model. A tremendous controversy in the healthcare container centers on the results of possession manage on the economic efficiency of hospitals. The indicator variable of ownership status enters drastically within the regression equation. The terrible signal of its coefficient estimate means that FP hospitals display superior economic performance in comparison to NP hospitals, in response to the variables examined. subsequently, the speculation that the category of ownership has no impact on profitability is rejected. FP hospitals have a stronger ROA than NP hospitals. this sort of finding is consistent with the outcomes in Sear (1991, 1992), Forgione (1987), Herzlinger & Krasker (1987), Forgione, Schiff & Crumbley (1996), and others.
The educating status variable has a poor coefficient suggesting inferior financial performance of educating hospitals as compared to non-educating hospitals. instructing hospitals supply practicing for interns and residents. always instructing hospitals have an affiliation with scientific faculties and keep a charitable role within the community. The significant difference between educating and non-instructing hospitals is the scope of functions supplied by the teaching hospitals. teaching hospitals tend to be greater and discovered in urban, economically depressed areas with a high charity care burden (HCIA, 1997). as a result, instructing hospitals deliver access for the indigent inhabitants in the surrounding enviornment, with limited, or no, compensation. instructing hospitals therefore exhibit inferior financial performance relative to non-teaching hospitals, which emanates from the costs associated with training, teaching, and charitable functions.
In the same method, hospitals with below one hundred beds in service are extra ecocnomic than hospitals with more than 500 beds. Small hospitals, always discovered in rural areas had been characterized with no, or low, penetration of managed care. additionally, rural hospitals usually face less competition and provide a wide array of capabilities. better hospitals continually operate in city places with more native competitors and a far better penetration of managed care--which puts big downward drive on revenues, and shifts a superior uncompensated charity care burden onto the hospitals (ProPAC, 1997).
The larger the number of full-time personnel adjusted for case combine, the lessen the profitability, maintaining all different variables regular. Case combine index (CMI) is similar to product mix in a manufacturing context. it's a measure of the mixture of patient disorder forms handled within the medical institution, relative to the countrywide normal, and proxies for relative resource consumption. consequently, a health facility with an above-typical CMI is expected to eat extra components than a medical institution with a reduce CMI. employee full-time-equivalents (FTEs) are divided by means of the CMI to provide an adjusted (standardized) FTE measure. An FTE worker is a fine proxy for variable charge. besides the fact that children, employees has a low coefficient cost with a low importance stage. This means that hospitals could be operating on an superior variety of employees, and any reduction in the variety of employees would now not cause tremendous development in profitability, besides the fact that children, the level of value does not warrant potent conclusions.
traditionally, the measure of performance of the sanatorium trade has relied on monetary ratios derived from the hospitals' fiscal statements (salary statement and balance sheet). The economic ratios measure the hospital's historic performance. Banks, creditors and rating companies use these ratios to assist predict the hospital's future efficiency and credit score worthiness. besides the fact that children, this look at demonstrates that additional, vital measures should be utilized in evaluating health facility efficiency, reminiscent of occupancy cost, staffing ratio, and total expense per adjusted discharge. These measures are likely to explain one of the underlying elements that produce favorable or destructive financial efficiency.
for instance, considering the implementation of operating can charge PPS, and in the era of declining use of inpatient functions vis a vis outpatient cures, occupancy price is a key predictor of economic efficiency. A declining trend in occupancy expense has an hostile impact on standard cost, profitability, and liquidity. At a reduce expense of occupancy, working rate per adjusted discharge is enhanced, so that they can prevent means to operate with financial effectivity.
The efficiency of the health center trade is multidimensional. main ameliorations exist across hospitals when it comes to their vicinity, scope of services supplied, size, ownership, organizational constitution, and volume of graduate scientific training offered. in addition, linked to these structural and placement differences, are variables akin to inpatient and payer mix, executive law, and a number of different non-fiscal elements, over which a clinic may additionally have tremendously little manage, and which may still be considered carefully. furthermore, the use of the regression model ought to be considered in comparison with different quantitative and qualitative displays. as an instance, in model (l), the classification of possession did have a significant impact on health facility profitability measured by the ROA. youngsters, the median of ROA of NP hospitals extended from 4.075 percent in 1991 to four.995 percent in 1995 (a 20 percent increase over a 5 yr duration); meanwhile, the ratio for FP hospitals increased from 5.535 p.c to 11.80 % for a similar period (a 60 percent raise). The empirical work showed that the affiliation with a college of medicine and site in city versus suburban, or city versus rural areas had considerable affect on health center profitability, services, and access for indigent populations (HCIA, Inc., 1997). The sample of hospitals that transformed from NP to FP fame was extraordinarily small (133 hospitals). The conversion in ownership popularity did not show an immense impact on a health facility's conduct, might be because of antecedent lifestyle of the administration, and/or because of constraining regulation in the healthcare trade.
These findings lift here two issues: (1) whether the reimbursement price beneath PPS should equitably contain other traits and native factors; and (2) because of government legislation and different components beside the type of ownership, clinic managers in widespread DC0-260 might also no longer be capable of make dramatic alterations in hospital stages of profitability.
CONCLUSIONS AND FUTURE directions
The results of this research show that operating effectivity isn't uniform across possession handle classes. additionally, the impact of the category of possession on healthcare functions for the indigent populations needs additional examination. as a result of rapid changes in the healthcare device, existing fashions can also not produce stable consequences over time.
As discussed in Morck et al. (1988), insider ownership may additionally reduce the likelihood of mergers and takeovers in non-healthcare industries. analysis related to the prediction of hospitals' mergers and takeovers is therefore advised. Prediction of medical institution bankruptcy is a different enviornment for future research. There might possibly be a powerful correlation between bankruptcy and charge equipment, debt provider load, entry to fitness care, mergers, and acquisitions. The effect of effectivity on first-rate of service, the top rate for receiving health services and organizational structure of hospitals also warrant future research. The style in NP conversions to FP fame is attracting the attention and opposition of the regulators and public citizen businesses (Forgione, 1999). additional research on this phenomenon is also informed.
display 1 DESCRIPTIVE facts FOR stylish AND independent VARIABLES Conversion Variable fame 1991 1995 Return on belongings (ROA) NP to FP 4.34 5.32 FP -0.85 10.55 NP -7.24 -4.seventy six possession popularity NP to FP 29 29 (possession) FP seventy nine seventy nine (variety of hospitals) NP 70 70 teaching repute (educating) NP to FP 5 6 (variety of hospitals) FP 13 12 NP 15 15 Conversion of ownership From NP to FP 29 29 popularity right through 1991-1995 (CONVERT) (number of hospitals) Full-time equivalent NP to FP 5.sixty three 4.seventy nine employees per a hundred FP 5.02 4.43 discharges, adjusted for NP 5.65 5.60 case mix (employees) length of stay per adjusted NP to FP 4.seventy five three.77 acute case mix (LOS) FP four.50 3.58 NP four.25 three.495 Debt per mattress (DEBT) NP to FP 134,095.75 156,6695 FP 112,808.seventy five eighty one,562.09 NP 178,088.36 226,665.34 Sole neighborhood issuer NP to FP 0 1 (SOLE) FP 2 1 (number of hospitals) NP 3 3 number of beds in carrier NP to FP 259 263 (BEDS) FP 208 213 NP 324 335 Occupancy expense (OCCUPANCY) NP to FP 50.eighty forty eight.12 FP 50.23 46.forty three NP 55.75 50.forty six notice: FP denotes for-earnings hospitals and NP denotes not-for-gainhospitals. supply: Medicare cost report statistics, provided via HCIA, Inc., Baltimore, Maryland. reveal 2 PIECEWISE LINEAR average LEAST SQUARES REGRESSION OF RETURN ON property ON health center features FOR YEARS 1991 AND 1995 independent Variables Coefficient t-values estimates regular four.8274 0.80 possession 5.7153 4.54 *** teaching -three.8813 -2.seventy six*** CONVERT -12.33613 -5.39*** employees -0.0503 -0.25 LOS -1.3847 -2.10 ** DEBT -0.0002 -four.79 *** SOLE three.1271 0.86 yr 7.2830 5.seventy one *** BEDS 0 to 100 -0.1447 -2.82*** BEDS a hundred to 500 0.0038 0.ninety two BEDS over 500 -0.0018 -0.28 OCCUPANCY 0 to 10 0.6640 0.28 OCCUPANCY 10 to 50 0.9650 1.35 OCCUPANCY over 50 1.1125 1.62 F statistic (marginal significance degree) 10.ninety seven 0.0001 *** Adjusted [R.sup.2] 0.32 Notes: The pattern contains 178 hospitals in 1991 (seventy nine FP and ninety nine NP) and 178 hospitals in 1995 (seventy nine FP, 29 FP that were NP in 1991 [converted from NP to FP] and 70 NP). The common mistakes are adjusted for heteroscedasticity in accordance with White (1980). The F statistic corresponds to the null hypothesis that the coefficient estimates for all variables blanketed in the regression equation are jointly zero. *** shows statistical value at the 0.01 stage ** shows statistical significance on the 0.05 degree REFERENCES
Arrington, B., & Haddock, C. (1990). Who definitely earnings from not-for-gains? health carrier analysis, 25(2), 291-304.
Fitzgerald, J., & Jacobson, B. (1987). examine fails to prove for-earnings superiority. health progress, 32-37.
Forgione, D.A. (1987). Incentives and efficiency in the fitness Care industry: The Case of For/Non-income Multihospital programs, Doctoral Dissertation, school of management, school of Massachusetts at Amherst, Amherst, MA.
Forgione, D.A. (1999). these conversion blues. Journal of fitness Care Finance, 25(4), 38-forty six.
Forgione, D.A., Schiff, A.D., & Crumbley, D.L. (1996). Assessing health facility performance: a listing of economic and nonfinancial metrics. international affiliation of administration Journal, discussion board on research in Healthcare financial management, 8(2), 65-eighty three.
Haddock, C., Arrington, B., & Skelton, A. (1989). Who cash in on now not-for-income: A reconsideration. fitness service management research, 2(2), eighty two-103.
HCIA, Inc. (1997). The Comparative efficiency of U.S. Hospitals: The Sourcebook. Deloitte & Touche LLP, Chicago, IL.
Herzlinger, R., & Krasker, W. (1987). Who earnings from non-gains? Harvard business evaluation, 93-105.
Morck, R., Shleifer, A., & Vishny, R.W. (1988). management possession and market valuation: an empirical analysis. Journal of economic Economics, 20(1), 293-316.
Needleman, J., Choilet, D., & Lamphere, J. (1997). health facility conversion trends. fitness Affairs, 16(2), 187-195.
ProPAC (1997). Medicare and the American health Care gadget, report to The Congress, June 1997. prospective payment assessment commission. Washington, DC, ninety four.
Sear, A.M. (1992). working features and comparative performance of investor owned multihospital methods. clinic and fitness functions Administration, 37(three), 403-415.
Sear, A.M. (1991). assessment of effectivity and profitability of investor-owned multihospital methods with now not-for-profit hospitals. fitness Care administration overview, 16(2),31-37.
Sloan, F.A., & Vraciu, R.A. (1983). Investor-owned and never-for-profit hospitals: addressing some issues. health Affairs, 2(1), 25-37.
Walker, C. (1993). A cross-sectional evaluation of medical institution profitability. Journal of sanatorium advertising and marketing, 7(2), 121-138.
Watt, J., Renn, S., Hahn, J., Derzon, R., & Schramm, C. (1986). The outcomes of possession and multihospital gadget membership on medical institution functional ideas and economic efficiency. For-profit commercial enterprise in fitness Care, countrywide Academy Press, Washington, DC, 260-290.
White, H. (1980). A heteroscedasticity-consistent covariance matrix estimator and an immediate test for heteroscedasticity. Econometrica, 48, 817-838.
Mustafa Younis Jackson State school (usa) Dana A. Forgione Florida overseas university (united states) Mahmud Khan Tulane school (country) John Barkoulas school of Tennessee (u . s . a .) The authors would like to thank Mr. George Pillari, Chairman and CEO, CBCA Inc. Oakland, CA for his beneficiant support for this analysis, and Jacqueline Pugh, MD, Director, Veteran evidence-based mostly research Dissemination and Implementation middle, VA fitness capabilities analysis center of Excellence, and Professor of medicine on the school of Texas health Science center at San Antonio, for her valuable feedback.
address for correspondence: Mustafa Younis, school of Allied health, The clinical Mall, 350 West Woodrow Wilson Avenue, Suite 3450, Jackson State school, Jackson, MS 39213 u . s . a ., myounis@mail1.jsums.edu.

Estrogen Inhibits Bone Resorption via without delay Inducing Apoptosis of the Bone-resorbing Osteoclasts summary

Estrogen deficiency explanations bone loss, which will also be prevented by way of estrogen alternative therapy. using a lately developed approach for isolation of highly purified mammalian osteoclasts, we showed that 17 β-estradiol (E2) turned into able to directly inhibit osteoclastic bone resorption. At concentrations beneficial for inhibiting bone resorption, E2 also without delay brought on osteoclast apoptosis in a dose- and time-dependent manner. ICI164,384 and tamoxifen, as pure and partial antagonists, respectively, completely or partially blocked the impact of E2 on both inhibition of osteoclastic bone resorption and induction of osteoclast apoptosis. These facts imply that the defensive results of estrogen in opposition t postmenopausal osteoporosis are mediated partially via the direct induction of apoptosis of the bone-resorbing osteoclasts by an estrogen receptor– mediated mechanism.

Estrogen deficiency, caused by means of either menopause or ovariectomy, results in pathological bone loss, which may also be avoided by estrogen substitute therapy ((1), (2)). although it is believed that estrogen's main motion in preventing bone loss is thru inhibition of osteoclastic bone resorption, the specific mechanism of such consequences isn't clear, mostly due to technical difficulties in acquiring purified useful osteoclasts ((three), (4)). Osteoclasts are terminally differentiated multinucleate cells the leading function of which is to dissolve bone matrix and minerals in the resorption section of bone home improvement ((5)). Recruitment, differentiation, and activity of osteoclasts are tightly managed by systemic and local elements. as an example, nutrition D3, prostaglandins, TGF-β, IL-1, IL-6, and TNF-α stimulate osteoclast differentiation and activity via direct or oblique mechanisms, whereas calcitonin directly inhibits osteoclast pastime ((6)–(10)). The destiny of osteoclasts after bone resorption is largely unknown. certain factors, reminiscent of calcitonin, inactivate osteoclasts with out induction of phone death, whereas other elements, reminiscent of bisphosphonates and nutrition K2, are recommended to induce apoptosis of osteoclasts ((eleven)–(13)). Apoptosis, or programmed telephone loss of life, is characterised by way of nucleosomal DNA fragmentation and grossly changed morphology of the nuclei with out a transformation in the morphology of the intracellular organelles ((14), (15)).

Estrogen consequences on osteoclasts are notion to be mediated ultimately via nonosteoclastic cells. as an example, loss of estrogen at menopause or by way of ovariectomy is linked to increased secretion of IL-1, IL-6, and TNF-α from the peripheral blood monocytes, bone marrow stromal cells, or osteoblasts, and decreased expression of TGF-β in bone ((sixteen)–(19)). multiplied ranges of those factors influence in increased osteoclastogenesis ((20)). in addition, Hughes et al. ((21)) these days suggested that estrogen promoted apoptosis of murine osteoclast–like cells mediated by means of TGF-β in a combined cellphone inhabitants in culture. using a recently developed approach for isolation of particularly purified genuine osteoclasts, we confirmed in this study DC0-260 the direct outcomes of estrogen on osteoclastic bone-resorbing pastime and osteoclast apoptosis. These estrogen effects on osteoclasts had been blocked by means of remedy with antiestrogens. additionally, Northern blot evaluation verified plentiful estrogen receptor α (ERα)1 mRNA expression in isolated osteoclasts, suggesting that estrogen may also also have a direct have an effect on on osteoclasts (22– 24). The facts presented here point out that estrogen inhibits osteoclastic bone resorption endeavor in part by means of focused on osteoclasts at once to undergo apoptosis through ER-mediated mechanisms.

materials and methods Osteoclast preparation.

Purified rabbit osteoclasts had been organized by using the formulation of Kakudo et al. ((25)) from unfractionated bone cells got based on the technique described by Takada et al. ((26)). in short, mobilephone suspensions received from minced lengthy bones of 10-d-old rabbits (Japan White; Saitama Experimental Animals provide Co., Saitama, Japan) were agitated by means of vortexing and plated in 10-cm tissue culture dishes (Becton Dickinson Labware, Lincoln Park, NJ) coated with 0.24% collagen gel (Nitta Gelatin Co., Tokyo, Japan). After a 3-h incubation, adherent nonosteoclast cells were removed from the collagen gel with the aid of sequential remedy with 0.001% pronase E and nil.01% collagenase (Wako Pure Chemical Industries, Osaka, Japan). The remaining osteoclasts have been then accrued via 0.1% collagenase answer medication and replated. When these telephone suspensions were seeded onto tissue culture dishes, osteoclasts connected and opened up on the dishes. via staining these cells for tartrate-resistant acid phosphatase (TRAcP, a marker of osteoclasts) recreation the usage of a leukocyte acid phosphatase equipment (Sigma Chemical Co., St. Louis, MO) after a 2-h incubation, we estimated that the purity of the TRAcP-fine multinucleate cells (>three nuclei) became >ninety nine% (Fig. 1). When the cells harvested from collagen gels were cultured on dentine slices, they shaped resorption pits as judged by using scanning electron microscopic remark ((25)). We used these pure osteoclast mobile suspensions for all experiments.

determine 1

Purified osteoclasts harvested from collagen gels. 2,000 osteoclasts have been plated on a dentine slice and stained for TRAcP exercise after a 2-h incubation. (×a hundred).

Assay for Osteoclastic Bone-resorbing pastime.

In an assay to consider osteoclastic bone-resorbing recreation, resorption pits formed by way of osteoclasts on dentine slices were measured for area, and their number changed into counted. briefly, purified osteoclasts have been replated onto dentine slices (round, 6 mm in diameter) in each smartly of 96-smartly plates (Becton Dickinson) at one hundred fifty cells/smartly and cultured for 24 h in phenol purple–free α-MEM (life technologies, Inc., Grand Island, big apple) supplemented with 0.1% BSA (Sigma Chemical Co.). Dentine slices have been then brushed with a rubber policeman to eradicate cells after observation beneath a fluorescence microscope, and stained with acid hematoxylin (Sigma Chemical Co.) for 5 min. total number of pits (reflecting active osteoclast quantity) became counted under a lightweight microscope, and complete area of resorption pits (reflecting bone-resorbing activity) turned into quantified by using densitometric evaluation of photographs of the entire enviornment of dentine slices put into a laptop. 17β-Estradiol (E2) and ICI164,384 (ICI) were provided through Shionogi prescribed drugs Inc. (Osaka, Japan) and Dr. A.E. Wakeling (ZENECA pharmaceuticals, Cheshire, UK), respectively. Tamoxifen (TAM) was obtained from Sigma Chemical Co. on the point of replating, agents had been introduced to the cultures on the desired concentrations. on the end of the culture length, osteoclasts have been fastened with formalin–acetone solution (37% formalin: acetone: 18 mM citric acid/19 mM Na Citrate/ 12 mM NaCl; 7:65:25) and stained for TRAcP pastime to assess the osteoclast number on the dentine slices.

Osteoclast Apoptosis.

The method used to realize osteoclast apoptosis through fluorescence microscopy changed into described up to now ((27)). After remedy with E2, recombinant human TGF-β1 (R&D programs, Minneapolis, MN) or salmon calcitonin (CT; 28), cells had been fastened with 2% glutaraldehyde answer (Wako Pure Chemical Co.) for 10 min, and stained with 0.2 mM Hoechst 33258 for visualization of chromatin condensation beneath a fluorescence microscope (VANOX AHB-LB, Olympus Co., Tokyo, Japan). Transmission electron microscopy changed into performed as follows. After fixation of osteoclasts on dentine slices with 2% glutaraldehyde solution for 1 h, the cells were immersed in 0.sixteen M EDTA (pH 7.2) and zero.2 M sucrose at 4°C for 2 wk to decalcify the dentine slices ((29)), and postfixed in 1% osmium tetraoxide answer (Electron Microscopy Science, castle Washington, PA) for 1 h. After dehydration in graded ethanol solutions, the cells have been embedded in epoxy resin. Sections had been reduce, stained with four% uranyl acetate, and examined below a transmission electron microscope (JEM-100CX, JEOL).

Northern Blot evaluation.

For evaluation of the mRNA expression of osteoclast-specific genes (cathepsin okay [Cat K] and carbonic anhydrase II [ECA II]), total RNA was extracted by the AGPC components ((30)) from osteoclasts cultured on dentine slices in phenol purple–free α-MEM supplemented with 0.1% BSA with or devoid of 0.1 nM E2 for 6 or 24 h, and for examination of the mRNA expression of ERs, osteoclasts isolated by way of the medicine with 0.001% pronase E answer ((31)) had been cultured on tissue lifestyle dishes for three h in phenol pink–free α-MEM supplemented with 0.1% BSA, and then complete RNA turned into extracted through the identical formula. The RNA changed into blotted onto a nitrocellulose membrane after formaldehyde agarose gel electrophoresis, and northern blotting became performed. 32P-labeled radioactive probes have been organized by the random primer labeling technique, and the blot changed into hybridized in 50% formamide/5× SSPE/5× Denhart's reagent/0.2 mg/ml salmon sperm DNA/labeled probe. After hybridization, the membrane changed into washed beneath stringent washing circumstances (0.1× SSPE/0.1% NaPPi/1% SDS) at sixty five°C earlier than autoradiography. The hybridization probes have been rabbit Cat k cDNA ((30)), rabbit CA II cDNA, and human ERα cDNA, all of which were bought from American class culture collection (Rockville, MD). A rat ERβ cDNA changed into cloned via RT-PCR system from rat testes. A human β-actin cDNA probe changed into used as a reference.

TUNEL Assay.

To become aware of in situ DNA fragmentation of apoptotic osteoclasts, we employed the TUNEL (TdT-mediated dUTP-biotin nick conclusion-labeling) assay through the use of a TACS Blue Label™ equipment (Trevigen, Inc., Gaithersburg, MD) in accordance with the manner counseled through the company.

effects And discussion Estrogen at once affects Osteoclast feature.

Direct discount of osteoclastic bone resorption with the aid of E2 begun inside 6 h of treatment (facts no longer shown), and fantastic reduction changed into accompanied by way of 24 h (Fig. 2, A–C). The ratio of pit enviornment per pit number was reduced by means of the medication with E2, indicating that E2 inhibition of bone resorption occurred at the degree of individual osteoclast activity (Fig. 2 C, inset). The level of inhibition of osteoclastic bone resorption with the aid of E2 turned into similar to that received for calcitonin (Fig. 2 C).

determine 2

Estrogen inhibition of bone resorption via purified rabbit osteoclasts. Purified osteoclasts had been incubated with medium (A) missing or (B) containing 0.1 nM E2 for twenty-four h on dentine slices (×40). After removing of osteoclasts, resorption pits formed through osteoclasts have been stained with acid hematoxylin and followed under a lightweight microscope. (C) Inhibitory effects of E2 on osteoclast-mediated bone resorption. Purified osteoclasts were cultured on dentine slices (150 cells/slice) in medium (Con) or in medium containing 0.001–1 nM E2 (E2) or 1 nM salmon calcitonin (CT). Osteoclastic bone resorption undertaking changed into measured in terms of pit enviornment shaped by purified osteoclasts after 24 h of incubation. Pit area per pit quantity is additionally indicated (inset). Values are skill ± SD, n = 4. *P <0.05, **P <0.01, ***P <0.005 compared with handle organizations. facts are representatives of those received in three further unbiased experiments.

To check the molecular mechanisms of estrogen-inhibited osteoclastic bone-resorbing recreation, we examined even if E2 would have an effect on the mRNA of Cat okay and CA II, enzymes worried in the bone-resorbing recreation of osteoclasts. E2 decreased mRNA levels of each Cat k and CA II in a time-dependent manner (Fig. 3, A and B). Such reduction become correlated with E2 inhibition of formation of resorption pits through the osteoclasts (facts now not proven). These records corresponded to the consequences that E2 inhibition of bone resorption occurred on the degree of particular person osteoclast activity (Fig. 2 C, inset).

figure 3

negative legislation of mRNA degrees for Cat okay and CA II with the aid of estrogen. (A) Northern blot evaluation. total RNAs (3 μg) from ∼25,000 purified osteoclasts cultured on dentine slices (one hundred fifty cells/dentine slices) with (E2) or devoid of 0.1 nM E2 (Con) for six or 24 h were used as samples. (B) Relative abundance of Cat ok and CA II mRNAs. The relative abundance of Cat ok and CA II mRNAs on Northern blotting changed into evaluated from the values of densitometric scanning. The values shown were normalized with recognize to the abundance of β-actin (ACT), and expressed as potential ± SD of samples from three different impartial experiments.

Estrogen at once Promotes Osteoclast Apoptosis.

To take into account the mechanism of estrogen-brought about osteoclast inactivation, we evaluated the morphological alterations in osteoclasts upon estrogen remedy at the subcellular level. Apoptosis is quite distinct from necrosis, which is accompanied by mobilephone disintegration, and gives an geared up potential of phone loss of life to minimize hurt to the encompassing cells ((32)). To examine direct results of estrogen on osteoclast apoptosis, we followed pure genuine osteoclasts attached to dentine slices during this analyze, as >ninety% of the osteoclasts plated remained connected to the dentine slices after 24 h of cultures. furthermore, we established by video monitoring that osteoclasts didn't detach conveniently even all through the morphological exchange to the apoptotic appearance, notably up to 24 h (facts now not proven). After 24 h of E2 treatment, osteoclasts displayed qualities of apoptosis, including chromatin condensation and grossly modified mobile and nuclear morphology as detected with the aid of fluorescence microscopy (Fig. 4 A). These observations had been additional tested through transmission electron microscopy (Fig. 4 C). E2 medicine did not alternate the morphology of intracellular organelles of osteoclasts, and cell membranes remained intact (Fig. 4 C). Such facets are attribute of apoptosis, and obviously indicated that the reason for cellphone dying become not necrosis, which is accompanied through mobile disintegration ((14), (15)). furthermore, in situ DNA fragmentation in these cells became detected by way of the TUNEL components (Fig. 4 B). The variety of DNA fragment–bearing osteoclasts (TUNEL-high-quality cells) become correlated with the variety of morphological apoptotic osteoclasts detected by way of Hoechst 33258 staining. In unfractionated bone cells, E2 also induced osteoclast apoptosis with out causing alterations in nonosteoclastic cells (data now not proven).

figure 4

Estrogen-precipitated osteoclast apoptosis. Osteoclasts have been cultured on dentine slices with 0.1 nM E2 for 24 h. (A) A fluorescence micrograph shows common and apoptotic osteoclasts (arrowhead) connected to a dentine slice in a 24-h subculture. ×250. (B) The TUNEL assay indicates DNA fragmentation in an apoptotic osteoclast. ×250. (C) This micrograph, acquired by means of electron transmission microscopy, demonstrates the gross morphological alterations in an apoptotic osteoclast. Nuclear fragments are indicated by means of the arrowheads. ×2,000.

Quantification of E2-brought about osteoclast apoptosis confirmed a dose-based increase (Fig. 5 A), which correlated with the dose latitude of E2 for inhibition of bone resorption (Fig. 2 B). In distinction to estrogen, calcitonin, which inhibits osteoclast recreation at once through its receptors on osteoclasts, did not trigger osteoclasts to undergo apoptosis, suggesting that inhibition of osteoclastic bone resorption via E2 is mediated through a distinctively distinctive mechanism than that used with the aid of calcitonin. As for indifferent cells, big mobile disintegration prevented us from detecting osteoclast apoptosis in them in 24-h and older cultures. E2-brought on osteoclast apoptosis was additionally time based (Fig. 5 B). E2 (0.1 nM) inhibition of formation of resorption pits via osteoclasts was correlated with a reduction within the expression of mRNAs for Ca ok and CA II and with an extended induction of osteoclast apoptosis in a time direction look at. These findings indicate that E2 at once acts on osteoclasts and inhibits osteoclastic bone resorption by way of causing osteoclast inactivation partially due to apoptosis. A recent document indicated that estrogen promoted TGF-β–mediated apoptosis of in vitro–developed murine osteoclast-like cells in combined cell cultures ((21)). TGF-β1 (10 ng/ml) induces apoptosis of our pure genuine osteoclasts. although, the incidence was decrease than that via E2 treatment in our mature cells or that of TGF-β–precipitated apoptosis in murine combined cellphone cultures (Fig. 5 A, reference (21)). The causes of this discrepancy should be would becould very well be in part as a result of their distinctive species; despite the fact, these results suggest that estrogen-more suitable osteoclast apoptosis mediated by means of TGF-β might turn up in an indirect manner. The direct induction of osteoclast apoptosis by way of estrogen anyway oblique induction may exist.

figure 5

Dose- and time-elegant method of estrogen-brought on osteoclast apoptosis. (A) Purified osteoclasts had been cultured on dentine slices (a hundred and fifty cells/slice) for twenty-four h in medium (Con) or in medium containing 0.001–1 nM E2, 1–20 ng/ml TGF-β1, or 1 nM CT. Apoptotic osteoclasts had been quantified beneath a fluorescence microscope. (B) Time-elegant consequences of E2 on osteoclast apoptosis and osteoclast quantity. under the identical way of life conditions, purified osteoclasts had been incubated in medium without 0.1 nM E2 (apoptosis: ○; cell quantity: □) or with 0.1 nM E2 (apoptosis: •, cell number: ▪) for 6, 12, or 24 h. Apoptotic osteoclasts are expressed as a percentage of total number of adherent osteoclasts. Values are means ± SD, n = four. *P <0.05, **P <0.005 compared with time = 0 groups. records are representative of these of three additional unbiased experiments.

Estrogen Inhibits Osteoclastic Bone-resorbing undertaking and Promotes Osteoclast Apoptosis through ER-mediated Mechanisms.

using a pure or a partial antagonist which can block estrogen results by way of binding to the ER, we examined even if these estrogen results on osteoclasts were ER mediated or no longer. ICI, a pure antiestrogen, had no impact on pit formation when expressed when it comes to pit area, whereas TAM, a partial estrogen antagonist, partly inhibited the resorption undertaking of purified osteoclasts (Fig. 6 A). When combined with E2, both ICI and TAM reversed the inhibitory impact of E2, indicating that E2 inhibited osteoclastic bone-resorbing exercise via ER. In distinction, the inhibitory effect of calcitonin changed into no longer littered with both ICI or TAM (Fig. 6 A). in keeping with their lack of, or partial, capacity to inhibit pit formation, ICI (0.001–a hundred nM) did not set off apoptosis of osteoclasts, whereas TAM brought on osteoclast apoptosis partially at 1 nM , however no longer in a statistically enormous method, at 0.001–0.1 nM or at 10–a hundred nM (Fig. 6 B, data no longer shown). These contradictory effects of antiestrogens should be would becould very well be because of diversity of ERs and coregulators ((33), (34)). When combined with E2, ICI and TAM blocked E2-prompted osteoclast apoptosis as a pure or partial antagonist, respectively (Fig. 6 B, precise variety of attached osteoclasts, plated at 150 cells, onto dentine slice after 24 h of way of life: manage, 143.5 ± 5.2; 1 nM ICI, 139.5 ± 7.three; 1 nM TAM, 141.0 ± 7.eight; 0.1 nM E2, 137.0 ± 10.9; 0.1 nM E2 + 1 nM ICI, 141.0 ± 7.2; 0.1 nM E2 + 1 nM TAM, 136.9 ± 7.8). These consequences imply that estrogen consequences on osteoclasts should be would becould very well be mediated via the ER and not be a outcome of toxicity. most effective a excessive dose of TAM became said to be in a position to induce osteoclast apoptosis in rat osteoclasts as well as E2 medicine ((35)). during this tradition device, besides the fact that children, the best attention of TAM for both inducing osteoclast apoptosis and rescuing osteoclasts from undergoing apoptosis by the medication with E2 was 1 nM, which should be would becould very well be as a result of partial mimicking of estrogen effects. TAM can also act on osteoclasts as a partial agonist rather than as a partial antagonist.

figure 6

Estrogen influences osteoclasts via ER-mediated mechanisms. (A) Antiestrogens recovered E2-inhibited osteoclastic bone resorption. ICI at 1 nM or 1 nM TAM was brought to osteoclast cultures in the presence or absence of 0.1 nM E2 or 1 nM salmon CT. 24 h later, osteoclastic bone resorption pastime become measured in terms of pit area. (B) Antiestrogens blocked E2-triggered osteoclast apoptosis. under the same lifestyle conditions, purified osteoclasts have been incubated in medium with or with out 1 nM ICI or 1 nM TAM in the presence or absence of 0.1 nM E2, and osteoclast apoptosis was measured after a 24-h remedy. Apoptotic osteoclasts are expressed as a percentage of total number of adherent osteoclasts. (C) Northern blot analysis of ERα and ERβ mRNA expression in osteoclasts. complete RNA (50 μg) from ∼5 × one hundred and five purified osteoclasts cultured on tissue tradition dishes with 0.1 nM E2 for 6 h had been used as samples. Values are means ± SD, n = 4. *P <0.05, **P <0.01, ***P <0.005 compared with manage or estrogen-treated organizations for A and B. information are consultant of those of three extra unbiased experiments.

several isoforms of ERα and one subtype of ER, called ERβ, had been suggested ((36)–(38)). In rabbit osteoclasts, we up to now established the expression of mRNAs (∼1.5, 2, and 6 kb with the 6-kb mRNA regarded to be putative ERα mRNA) which hybridized with ERα cDNA in Northern blots ((22)). in this look at, Northern blot evaluation published that osteoclasts expressed ERα mRNA, but ERβ mRNA changed into undetectable (Fig. 6 C). ER cDNA-hybridizing mRNAs anyway putative ERα in pure osteoclasts might be isoforms of ERα mRNA or another mRNA but no longer ERβ mRNA. These statistics imply that rabbit osteoclasts may be managed via estrogen through ERαs.

In conclusion, the facts offered in this study DC0-260 show that moreover the oblique outcomes of estrogen on bone resorption mediated through soluble factors secreted from nonosteoclastic cells and cellphone-to-mobile contact with nonosteoclastic cells, estrogen can feature at once on osteoclasts to inhibit their bone-resorbing recreation. The ability of estrogen to induce apoptosis of osteoclasts at correlative concentrations valuable for inhibition of bone resorption could be the mechanism underlying such results. The consequences of the antiestrogens additionally imply that osteoclast apoptosis caused via estrogen is mediated by the ER. These findings, therefore, may additionally shed new light on our figuring out of the cellular mechanism in which estrogen offers shielding outcomes on the skeleton, and guide the validity of using estrogen substitute therapy for treating postmenopausal osteoporosis.

Acknowledgments

We thank Ms. Yoko Katagiri and Ms. Sachiko Ishii for technical suggestions and guidance of the manuscript, respectively. we're also grateful to Ms. Yumiko Kanda for her assistance with the transmission electron microscopy.

Footnotes

Dr. T. Kameda and Dr. H. Mano contributed equally to this work.

1 Abbreviations used in this paper: CA II, carbonic anhydrase II; Cat k, cathepsin k; CT, calcitonin; E2, 17β-estradiol; ER, estrogen receptor; ICI, ICI164,384; TAM, tamoxifen; TRAcP, tartrate-resistant acid phosphatase; TUNEL, TdT-mediated dUTP-biotin nick conclusion-labeling.

Submitted: 9 September 1996

Revision received eleven June 1997

References:

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DC0-260 Questions and Answers

Answer: B, C, E

QUESTION: 57

Answer: B

QUESTION: 58

Answer: B, D

QUESTION: 59

Answer: B, D

QUESTION: 60

Answer: B, D

QUESTION: 61

Answer: B, C, D

QUESTION: 62

Answer: B, C, E

QUESTION: 63

Answer:

Reference:

DELL DC0-260 Exam (Dell Certified Storage Networking(R) Professional) Detailed Information

DELL DC0-260

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