1. A wheel axle assembly for a vehicle, including:
an axle for extending across the vehicle, said axle having hollow ends, said axle assembly including at each end:
a mounting collar rigidly supported by the hollow end of said axle,
a spindle received in said mounting collar and extending into the hollow end of said axle,
said hollow end of said axle defining a key hole extending across said axle,
said spindle including therein opposed bearing surfaces alignable with said key hole when said spindle is inserted into the hollow end of said axle,
a cam key extendable through said key hole of said axle to a position in alignment with said bearing surfaces of said spindle when said spindle is inserted in the hollow end of the axle,
said cam key including cam means for engaging said bearing surfaces one at a time such that when said cam key is oriented in one direction and inserted into the keyhole its cam means engages one of said bearing surfaces and urges the spindle in one direction along the axle and when said cam key is oriented in another direction and inserted into the keyhole its cam means engages the other bearing surface and urges the spindle in the opposite direction along the axle.
2. The wheel axle assembly of claim 1, wherein
said bearing surfaces of said spindle are oppositely sloped.
3. The wheel axle assembly of claim 2, wherein
said cam means of said cam key comprises a single sloped surface for wedge engagement with both said bearing surfaces.
4. The wheel axle assembly of claim 3, wherein
said keyhole extends entirely through said axle, and
said cam key has an end with helical threads that protrude from said keyhole, and
a retainer for engaging said threads outside said axle for holding said cam key in said axle.
5. The wheel axle assembly of claim 1, wherein said axle is of square tubular configuration.
6. The wheel axle assembly of claim 1, and further including:
said axle and said spindle including complementary shapes for aligning said bearing surfaces of said spindle with said keyhole.
7. The wheel assembly of claim 6, wherein said complementary shape of said axle comprises a bar extending across said axle, and wherein said complementary shape of said spindle comprises a recess formed in said spindle that receives said bar.
8. A wheel axle assembly for a vehicle, including:
an axle for extending across the vehicle, said axle having hollow ends, said axle assembly including at each end:
a spindle received in and extending into the hollow end of said axle,
said hollow end of said axle defining a key hole extending across said axle,
said spindle including therein a cam slot alignable with said key hole when said spindle is inserted into the hollow end of said axle,
said cam slot having opposed oppositely sloped bearing surfaces,
a cam key extendable through said key hole of said axle and into said cam slot of said spindle when said spindle is inserted in the hollow end of the axle,
said cam key including cam means for alternately engaging said bearing surfaces of said cam slot for urging the spindle in one direction along the axle when engaging one bearing surface and for urging the spindle in the opposed direction when engaging the other bearing surface.
9. The wheel axle assembly of claim 8, wherein
said cam means of said cam key comprises a single sloped surface for wedge engagement with both said bearing surfaces.
10. The wheel axle assembly of claim 9, wherein
said keyhole extends entirely through said axle, and
said cam key has an end with helical threads that protrude from said keyhole, and
a retainer for engaging said threads outside said axle for holding said cam key in said axle.
11. The wheel axle assembly of claim 8, wherein said axle is of substantially square tubular cross sectional configuration.
12. The wheel axle assembly of claim 8, and further including:
said axle and said spindle including complementary shapes for aligning said bearing surfaces of said spindle with said keyhole.
13. The wheel assembly of claim 12, wherein said complementary shape of said axle comprises a bar extending across said axle, and wherein said complementary shape of said spindle comprises a recess formed in said spindle that receives said bar.
14. In combination, a wheel spindle and a cam key,
said spindle including a rectilinear spindle body having a longitudinal axis, a wheel support end for supporting a wheel and a mounting end for mounting to an axle,
said mounting end defining a cam slot extending radially therethrough and having oppositely sloped opposed bearing surfaces,
said cam key configured to fit in said cam slot and including a sloped cam surface for engagement with one of said bearing surfaces at a time for urging said spindle along its longitudinal axis.
15. The combination of claim 14, and further including a retainer for retaining said cam key in said cam slot.
16. The combination of claim 14, and further including a tubular axle of square cross section,
a mounting collar rigidly mounted on each end of said axle,
said mounting end of one of said spindles received in said mounting collar at each end of said axle and extending into said axle,
said axle defining a keyhole at each of its ends, and
said cam key sized and shaped to extend through said keyhole and said cam slot.
17. An axle assembly for a wheeled vehicle comprising:
a tubular axle,
a spindle inserted into said axle,
a keyhole formed in said axle,
a cam slot formed in said spindle and aligned with said keyhole and including opposed bearing surfaces,
a cam key configured for extending through said keyhole and through said cam slot and including a cam surface for engaging one of said opposed bearing surfaces,
such that when the cam key is in a first position and is moved into the cam slot the cam surface of the cam key engages one of the bearing surfaces of the cam slot and the spindle is urged in a first direction along the axle in response to the cam surface engaging one of the bearing surfaces and when the cam key is in a second position and is moved into the cam slot the cam surface of the cam key engages the other the bearing surface of the cam slot and the spindle is urged in the opposite direction along said axle in response to the cam surface engaging the opposed bearing surface.
18. The axle assembly of claim 17, and further including a bar extending transversely through said axle, and said spindle including a recess that engages said bar for orienting the spindle with respect to the keyhole of said axle.
19. A method of mounting and de-mounting a spindle to an axle, the axle having a hollow end, comprising:
when the spindle is to be attached to the axle:
inserting a spindle longitudinally into a hollow end of the axle,
orienting a cam key in a first position and inserting the cam key transversely through the axle and through a cam slot of the spindle with a cam surface on the cam key engaging a first bearing surface of the cam slot and urging the spindle into the axle, and
when the spindle is to be removed from the axle:
orienting the cam key in a second position and inserting the cam key transversely through the axle and through the cam slot of the spindle with the cam surface on the cam key engaging a second bearing surface on the cam slot and urging the spindle out of the axle.
The claims below are in addition to those above.
All refrences to claim(s) which appear below refer to the numbering after this setence.
1. A method for detecting in a biological sample comprising microorganisms the presence of microorganisms resistant to a predetermined therapeutic agent, comprising the following steps:
a. inoculating the sample, on a first tube with, and on a second tube without, at least one such therapeutic agent and incubating it;
b. adding to both tubes a fluorescent marker;
c. performing a fluorescence analysis to obtain one or more fluorescence parameters for each of the two tubes;
wherein the presence of microorganisms in the biological sample having the phenotype of resistance to said therapeutic agent is obtained by comparing the one or more fluorescence parameters between the two tubes.
2. A method for detecting in a biological sample comprising microorganisms the presence of microorganisms resistant to a predetermined therapeutic agent, comprising the following steps:
a. inoculating the sample, on a first tube with, and on a second tube without, at least one such therapeutic agent and incubating it;
b. adding to both tubes a fluorescent marker;
c. performing a flow cytometric analysis to obtain one or more flow cytometric parameters for each of the two tubes;
wherein the presence of microorganisms in the biological sample having the phenotype of resistance to said therapeutic agent is obtained by comparing the one or more flow cytometric parameters between the two tubes.
3. The method according to claim 2 wherein the one or more flow cytometric parameters comprise forward scatter andor side scatter andor fluorescence scatter parameters.
4. The method according to claim 3 wherein the parameters of the fluorescence scatter signal comprise intensity, spectral profile andor cell count.
5. The method according to claim 1 further comprising one or more of the preparatory steps:
i. identifying the type of microorganism present in the biological sample and according to such identification selecting the therapeutic agent or agents, for example whether the type of microorganism is a gram-positive or gram-negative bacteria or fungi;
ii. concentrating the biological sample and remove, if present, any detritus.
6. The method according to claim 1 wherein the biological sample is an uncultured sample of urine, a blood culture, water, or a suspension of microorganisms obtained from pure cultures.
7. The method according to claim 1 further comprising a centrifugation step or filtration step.
8. The method according to claim 1 wherein step a) further comprises including at least one lysing agent in the inoculate.
9. (canceled)
10. The method according to claim 1 wherein in the step a) the sample is inoculated in a suitable culture medium.
11. The method according to claim 1 wherein the incubation time in the step a) is between 30 minutes and 2 hours.
12. The method according to claim 1 wherein the incubation temperature is between 25 and 35\xb0 C.
13. The method according to claim 1 wherein in step b) the fluorescent marker is present for a reaction time between 5 and 60 minutes.
14. The method according to claim 1 wherein the biological sample in step c) further comprises an enzymatic inhibitor in the case of detection of extended spectrum beta-lactamases (ESBLs) presence.
15. (canceled)
16. The method according to claim 1 which comprises the following steps:
i. inoculating the sample with a carbapenem and incubating it;
ii. removing the microorganism in the biological sample, preferably by filtration;
iii. adding a strain with known behaviour;
iv. adding to both samples a fluorescent marker;
v. performing a flow cytometric analysis for obtaining one or more flow cytometric parameters for each of the two tubes;
vi. comparing the profile generated with the control strain.
17. The method according to claim 1 wherein the said microorganism is a bacteria, a fungi or a protists, preferably wherein the bacteria is a gram positive or negative; gram variable or bacteria unable to Gram stain like Mollicutes; or an alcohol-acid resistant bacteria.
18. (canceled)
19. The method according to claim 17 wherein the microorganism is a Staphylococcus spp, Streptococcus spp, Enterococcus spp; Enterobacteriacea, Escherichia coli, Klebsiella pneumonia, Proteus spp; Pseudomonas aeruginosa, Acinetobacter baummanii, Burkloderia cepaciae; Neisseria spp, Haemophilus spp; Mycobacteria spp, Nocardia spp; Legionella spp, anaerobic bacteria; Candida spp, Cryptococcus neoformans, Pneumocystis jirovecii, or moulds like Aspergillus spp; Giardia spp.
20. The method according to claim 1 wherein the therapeutic agent is an antibacterial drug selected from the group consisting of penicillins, cephalosporins, carbepenems, aztreonam, macrolides, quinolones, aminoglycosides, glycopeptides, a lipopeptide, tetracyclines colistin, chloramphenicol, clindamycin, fosfomycin, linezolid, nitrofurantoin, sulphonamide, rifampin, trimethoprim, trimethoprim-sulfamethoxazole or tigecycline, polyenes, 5-fluorocytosine, azoles or echinocandins.
21. The method according to claim 1 wherein the fluorescent marker is a nucleic acid stain, a metabolic stain, a membrane potential stain, a probe for organelles, a fluorescent tracer of cell morphology and fluid flow, a probe for cell viability, proliferation and function or a probe for reactive oxygen species.
22. The method according to claim 21 wherein the fluorescent marker is an acridine dye, cyanine dye, fluorone dye, oxazin dye, phenanthridine dye, or a rhodamine dye.
23. A kit for carrying out the method of claim 1.
24-25. (canceled)
26. A fluorescence device for carrying out the method of claim 1.
27. (canceled)