DETAILED ACTION
Notice of Pre-AIA or AIA Status
The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA .
Claim Status and Formal Matters
This action is in response to papers filed 10/30/2025.
Claims 1-7, 33, 42-54 are pending.
Claim 33 has been amended.
Claims 33, 42-54 are being examined.
Applicant's election with traverse of group II in the reply filed on 3/31/2025 is acknowledged. The traversal is on the ground(s) that the response asserts there is no search . This is not found persuasive because searching methods which require fixed samples and sample onto a replica surface will not inherently provide art on methods that do not require it..
Claims 1-7 withdrawn from further consideration pursuant to 37 CFR 1.142(b), as being drawn to a nonelected invention, there being no allowable generic or linking claim. Applicant timely traversed the restriction (election) requirement in the reply filed on 3/31/2025.
The previous objection to the claims has been withdrawn in view of the amendment.
The response provides arguments with respect to a supplemental IDS. However, there is no IDS in the instant file wrapper.
The ODP rejections have been withdrawn as the cited patent/applications do not specifically recite stamping.
Priority
The instant application was filed 06/14/2023 and is a continuation of 15747245 , filed 01/24/2018,which is a National Stage entry of PCT/US16/43487 with an International Filing Date: 07/22/2016 and Claims Priority from Provisional Application 62196725 , filed 07/24/2015.
Claim Rejections - 35 USC § 112
The following is a quotation of 35 U.S.C. 112(b):
(b) CONCLUSION.—The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the inventor or a joint inventor regards as the invention.
The following is a quotation of 35 U.S.C. 112 (pre-AIA ), second paragraph:
The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the applicant regards as his invention.
Claims 33, 42-54 rejected under 35 U.S.C. 112(b) or 35 U.S.C. 112 (pre-AIA ), second paragraph, as being indefinite for failing to particularly point out and distinctly claim the subject matter which the inventor or a joint inventor (or for applications subject to pre-AIA 35 U.S.C. 112, the applicant), regards as the invention.
Claim 33 has been amended to recite, “method of analyzing RNA from a subject” and “thereby analyzing the subject's RNA.” However the claim recites, “target nucleic acid.” Thus the metes and bounds are unclear if the claim requires detection of RNA or any target nucleic acid.
Further claim 33 has been amended to recite, “stamping the ligated probes onto a replica surface coated with a plurality of immobilized PCR primers to spatially amplify and detect the target nucleic acid proxy.” The claim is confusing, unclear and incomplete as stamping on a surface with immobilized primers provide for amplification or spatial amplification. Further the recitation of “spatial amplification” suggests there is non-spatial amplification. Thus the metes and bounds are unclear how spatial amplification is done and/or how it is differentiated from non-spatial amplification.
Response to Arguments
This is new ground of rejection necessitated by amendment.
Claim Rejections - 35 USC § 103
In the event the determination of the status of the application as subject to AIA 35 U.S.C. 102 and 103 (or as subject to pre-AIA 35 U.S.C. 102 and 103) is incorrect, any correction of the statutory basis (i.e., changing from AIA to pre-AIA ) for the rejection will not be considered a new ground of rejection if the prior art relied upon, and the rationale supporting the rejection, would be the same under either status.
The following is a quotation of 35 U.S.C. 103 which forms the basis for all obviousness rejections set forth in this Office action:
A patent for a claimed invention may not be obtained, notwithstanding that the claimed invention is not identically disclosed as set forth in section 102, if the differences between the claimed invention and the prior art are such that the claimed invention as a whole would have been obvious before the effective filing date of the claimed invention to a person having ordinary skill in the art to which the claimed invention pertains. Patentability shall not be negated by the manner in which the invention was made.
Claim(s) 33, 42-54 is/are rejected under 35 U.S.C. 103 as being unpatentable over Comanescu (Recent Patents on DNA & Gene Sequences 2012, 6, 22-32), Zhang (Chemical Communications (2003) volume 49, pages 10013-10015), Wang, F., et al. The Journal of molecular diagnostics: JMD 14, 22-29 (2012), Nilsson (Nature Biotechnology (2000) volume 18, pages 791-793), Koch (USPGPUB 20100047773), Cuppoletti (WO2009039202)..
The prior art as exemplified below demonstrates that the use of fixed samples was known for nucleic acid analysis, including FFPE samples with in situ hybridization. Further the art recognizes RNA is poorly preserved in fixed samples. The art recognizes the use of Rnl2 to ligate two probes of DNA molecules in which the 3’ of a probe having 2 RNA bases and the 5’ end of another probe is phosphorylated for multiplex detection of RNA. Further the prior art demonstrates that detection of RNA in FFPE samples was known.
While claim 33 has been amended to recite, “to spatially amplify and detect the target nucleic acid proxy, thereby analyzing the subject's RNA.” However the amendment is unclear how the stamping results in spatial amplification. Thus the broadest reasonable interpretation is the amplification somehow an intended outcome of stamping and not a positive active step.
Comanescu teaches, “ RNA is notoriously poorly preserved in FFPE tissue.” (abstract). Comanescu teaches, “RNA has been found to be heavily degraded and fragmented so that only short sequences, approximately 100–200 nucleotides long, can be recognized and amplified.” (page 27 top) Comanescu teaches numerous methods of analyzing RNA and DNA from fixed samples including DASL (DNA- mediated Annealing, extension, selection ligation which provides for amplification and detection of nucleic acids.(page 28)
Comanescu does not specifically teach applying a sub-probe with two RNA bases at the 3’ end and a second sub probe to anneal within a fixed permeabilized sample
Zhang teaches multiplex ligation-dependent probe amplification for ultrasensitive multiplexed miRNA detection using ribonucleotide modified RNA (title). Zhang teaches that the method can be used to detect nucleic acids at low concentrations (10013, 2nd column, top).
Zhang teaches
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Zhang teaches, “Our strategy for multiplexed miRNA detection based on the
MLPA technique is illustrated in Fig. 1. For detection of each miRNA target (such as miRNA 1 and miRNA 2), the probe A and probe B (1A, 1B and 2A, 2B corresponding to miRNA 1 and miRNA 2, respectively) are designed. universal primer-specific sequence used for PCR amplification (green), a target-specific sequence (blue and sky blue) and a stuffer sequence in between (red). The target-specific sequences in probe A and probe B are simply designed to be complementary to the half-sequence of each miRNA target at the 3’- and the 5’-terminal, respectively. T4 RNA ligase 2 is employed in this work to ligate DNA probes due to its high specificity.’ J. Nandakumar and S. Shuman have documented that T4 RNA ligase 2 can effectively catalyze the ligation reactions of 3’-OH RNA and 5'-PO, RNA or DNA, which does not discriminate between RNA
and DNA on the 5/-PO, side of the ligation nick. Moreover, they have also proved that the ligation efficiency achieved by using a DNA probe modified only with 3 or 2 ribonucleotides at the 3'-OH side is similar to that achieved by using an RNA probe, but the ligation efficiency will become much lower upon using a DNA probe modified with only a single 3’-OH ribonucleotide.”” To save on costs, probe A is modified with two ribonucleotides (brown) at its 3’-terminus and probe B is modified with a phosphate group at its 5’-terminus. In the presence of miRNA targets, probe A and probe B hybridize immediately to the adjacent sites of the target sequences and therefore, can be ligated with T4 RNA ligase 2. The ligated DNA probes are then amplified by PCR with fluorescein-labeled primers. The universal primer-specific sequences in probe A and probe B permit simultaneous PCR amplification of different miRNA targets using only one primer pair. For different miRNA targets, the stuffer sequences in probe A and probe B have different length, which are used to modulate the size of PCR products. Therefore, each miRNA target can give rise to an amplification product of unique size, permitting that the PCR products produced by different miRNA targets can be well separated using capillary electrophoresis (CE) and sensitively detected using laser-induced fluorescence.” (10013 bottom 2nd column-10014). Zhang teaches, “we rationally design one of the DNA probes in MLPA modified with two ribonucleotides at its 3'-terminus, which can greatly improve the ligation efficiency of DNA probes templated by miRNAs.” (10013 bottom 2nd column)
While Zhang teaches the applying, annealing, ligating of multipartite DNA probes with 2 RNA bases at the 3’ end to 5’ phosphate for multiple detection of miRNA, Zhang does not specifically teach fixed samples.
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However, Wang teaches in situ hybridization analysis in formal fixed paraffin embedded tissues.(title) Wang teaches, “cells were placed on slides and fixed in 4% formaldehyde for 60 minutes, followed by protease digestion (2.5 g/mL) at 23°C to 25°C. The cells were then incubated in order at 40°C with the following solutions: target probes in hybridization buffer A [6SSC (1SSC is 0.15 mol/L NaCl, 0.015 mol/L Na-citrate), 25% formamide, 0.2% lithium dodecyl sulfate, blocking reagents] for 3 hours; preamplifier (2 nmol/L) in hybridization buffer B (20% formamide, 5SSC, 0.3% lithium dodecyl sulfate, 10% dextran sulfate, blocking reagents) for 30 minutes; amplifier (2 nmol/L) in hybridization buffer B at 40°C for 15 minutes; and label probe (2 nmol/L) in hybridization buffer C (5SSC, 0.3% lithium dodecyl sulfate, blocking reagents) for 15 minutes. After each hybridization step, slides were washed with wash buffer (0.1 SSC, 0.03% lithium dodecyl sulfate) three times at room temperature. For multiplex detection, equimolar amounts of target probes, preamplifier, amplifier, and label probe of each amplification system were used. Chromogenic detection was performed using DAB followed by counterstaining with hematoxylin (AmericanMasterTech Scientific, Lodi, CA.”
Nilsson teaches enhanced detection and distinction of RNA by enzymatic probe ligation (title). Nilsson teaches, “It was recognized early on that the T4 DNA ligase can ligate DNA oligonucleotides hybridizing to RNA strands5,6. RNA-templated ligation of DNA probes has been used to generate molecules, amplifiable by polymerase chain reaction (PCR) by means of general sequences present at the remote ends of a pair of ligation probes7. The method has been applied to detect viral RNA extracted from clinical and archival specimens with increased sensitivity compared to nested reverse transcription (RT)-PCR8,9. RNA-templated ligation of RNA probes has been used for detection of transcripts in experiments where ligation products were amplified by the Qβ replicase10. It is thus possible to substitute RNA-templated ligation of either DNA or RNA probes for an RT step before amplification.”(791, 2nd column, 2nd paragraph). In figure 1, Nilsson teaches two probes that are enzymatically ligated together to detect RNA.
Nilsson teaches, “Ligase-mediated gene detection could therefore provide highly sensitive and accurate ligase-mediated detection and distinction of RNA sequence variants in solution, on DNA microarrays, and in situ.” 791, 2nd column, 1st paragraph).
Nilsson teaches, “RNA-templated ligation of RNA probes has been used for detection of transcripts in experiments where ligation products were amplified by the QB replicase!”. It is thus possible to substitute RNA-templated ligation of either DNA or RNA probes for an RT step before amplification. However, no analysis has been presented of optimal reaction conditions for RNA-templated DNA ligation, and it is not known how well probe ligation reactions can discriminate among variants of RNA target sequences. We found that under low-salt and low-ATP conditions, high concentrations of T4 DNA ligase efficiently joined DNA oligonucleotides, hybridized in juxtaposition on RNA target strands (M.N., manuscript in preparation). In order to determine if probe ligation reactions can be used to distinguish RNA sequence variants, a set of four in vitro transcripts of amplified synthetic oligonucleotides were prepared that differed in one centrally located position.” (page 791, last paragraph).
Nilsson teaches the use of nM concentrations of reactants for the analysis. Thus Nilsson teaches the use of two or more copies of multipartite probes (experimental protocol).
Koch teaches, “ 0347] In Situ Detection of RNA Using a Turtle Probe [0348] In situ detection of EBER1 (Epstein-Barr Early Region) RNA in paraffin embedded formalin fixed human tonsil tissue infected with Epstein-Barr virus (EBV) (see FIG. 5). Pretreatment: The formalin fixed paraffin embedded tissue was deparaffinized with xylene for 2.times.10 minutes and then washed in an ethanol series of 99%, 85%, 70% to remove residual xylene. The tissue was then dehydrated and air dried at room temperature. The tissue was treated with 0.05% pepsin (Sigma) dissolved in 0.1 M HCl for 15 minutes at 37.degree. C. The pepsin treatment was terminated by submerging the slide in wash buffer (0.1 M Tris-HCl, 0.15 M NaCl and 0.05% Tween-20). The tissue was refixed in 0.4% paraformaldehyde in 1.times.PBS for 20 minutes, and washed in wash buffer for 5 minutes at 37.degree. C. and dehydrated and air dried at room temperature. [0349] Probe Hybridization : A Hybridization mixture containing: 0.1 .mu.M EB1-turtle probe, 20% formamide, 2.times.SSC, 0.2 .mu.g/.mu.l BSA, 5% glycerol, and 0.2 .mu.g/.mu.l carrier DNA was added to the slide and covered with a cover glass. The cover glass was sealed to the slide with heat resistant glue. The slide was heated for 2 minutes at 95.degree. C., cooled to 37.degree. C. and incubated at that temperature for 30 minutes. After hybridization, the slide was washed in 2.times.SSC with 0.05% tween-20 for 5 minutes at 37.degree. C., in wash buffer for 5 minutes at 37.degree. C., and finally dehydrated and air dried at room temperature. Hybridization could be performed at 37.degree. C. without first heating to 95.degree. C., but heating to 95.degree. C. has been found to increase the number of signals. Carrier DNA or RNA may not always be required, but often seems to increase the number of signals. [0350] Probe ligation: An advantage of using a turtle probe is that this probe contains its own ligation template, so that probe-ligation is performed on this self-contained DNA template, rather than using the target RNA as template. This probe design will be preferred is most cases since DNA ligation using an RNA template is much less efficient than DNA ligation using a DNA template (compare FIGS. 5 and 7). [0351] Ligation of the probe was performed in a mixture containing: 1.times.T4 DNA ligase buffer (Fermentas), 0.2 .mu.g/.mu.l BSA, and 0.1 U/.mu.l T4 DNA ligase (Fermentas) for 30 minutes at 37.degree. C. After incubation with the ligase mixture, the slide was washed in wash buffer for 5 minutes at 37.degree. C. “
Thus the art demonstrates the use of MLPA to detect nucleic acids in samples and their ability to detect short nucleic acids at low concentrations. The art teaches RNA and DNA in fixed samples are often fragmented and degraded. The art demonstrates rnl2 increases ligation efficiency relative to other ligases by use of two ribonucleotide bases at 3’ end of one probe to the 5’ end of another probe. Further the prior art suggests and teaches the use of ligases in situ to detect nucleic acid based on context in cells. Further the art teaches in situ hybridization to RNA. Thus the claims are nothing the obvious combination of known art.
Therefore it would have been prima facie obvious to one of ordinary skill in the art prior to the effective filing date of the claims to perform the method of Zhang in situ in fixed cells by applying the probes and rnl2 ligase in solution to the permeabilized sample, and amplifying in solution, which causes dissociation of the nucleic acid proxy form the target RNA which is in solution and amplification. The artisan would be motivated to apply Zhang’s method to a fixed sample to target short sequence and minimizes issues associated with degradation of nucleic acids in fixed samples. The artisan would further be motivated to use the method of Zhang as the art demonstrates it detects low concentrations of nucleic acids in samples due to the increased efficiency of rnl2 relative to other ligases. The artisan would have a reasonable expectation of success as Wang, Koch and Nilsson demonstrates fixes cells for analysis in situ by probe ligation was known.
While Comanescu, Zhang, Nilsson, Wang, and Koch teach the use of fixed samples. Comanescu, Zhang, Nilsson, Wang, and Koch do not specifically teach stamping of the ligated target nucleic acid proxy onto a target surface and amplifying the target nucleic acid proxy, and hybridizing the amplified product with multicolored fluorescent oligonucleotides.
However Cuppoletti teaches, “[0002] The present application relates generally to polymer microarrays and methods for fabricating polymer microarrays. In particular, the application relates to hydrogel based microarrays fabricated by a nanostamping process.”
Cuppoletti teaches, “0014] Specific molecules can spontaneously arrange on various surfaces forming two-dimensional mono-molecular layers called self-assembled monolayers (SAMs). Patterned DNA SAMs can be used as templates for a novel printing technique for organic materials called Supramolecular NanoStamping (SuNS). Supramolecular NanoStamping (SuNS) is a newly developed stamping technique that enables the transfer of spatial together with chemical information from a template containing DNA features to a secondary substrate. (Yu AA et al. J. Mater. Chem., (2006) 16, 2868 - 2870). This method, like the DNA/RNA information transfer, uses the reversible assembly of DNA double strands as a way of transferring patterns from a surface to another. The method relies on the biochemical ability of DNA to replicate and avoids the reproducibility problems associated with traditional monomer-by monomer chemical synthesis of nucleic acids to generate microarrays. One of the main advantages of SuNS is that multiple DNA strands each encoding different information can be printed at the same time in parallel. (Yu AA et al. Nano Lett. 2005 Jun;5(6):1061-1064).”
Cuppoletti teaches amplifying of target with primers in the hydrogel (0105).
Cuppoletti teaches multiple labels for labeling oligonucleotides (0124)
Therefore it would have been prima facie obvious to one of ordinary skill in the art prior to the effective filing date of the claims to stamp the ligated target nucleic acid proxies produced by the method of Comanescu, Zhang, Nilsson, Wang, and Koch to a target surface and amplifying the target nucleic acid proxy. The artisan would be motivated to provide replicates of the target nucleic proxies for further assays. The artisan would have a reasonable expectation of success as the artisan is merely using known method to stamp, amplify and hybridize.
With regards to claim 42-44, Zhang teaches, “Our strategy for multiplexed miRNA detection based on the MLPA technique is illustrated in Fig. 1. For detection of each miRNA target (such as miRNA 1 and miRNA 2), the probe A and probe B (1A, 1B and 2A, 2B corresponding to miRNA 1 and miRNA 2, respectively) are designed. universal primer-specific sequence used for PCR amplification (green), a target-specific sequence (blue and sky blue) and a stuffer sequence in between (red). The target-specific sequences in probe A and probe B are simply designed to be complementary to the half-sequence of each miRNA target at the 3’- and the 5’-terminal, respectively. T4 RNA ligase 2 is employed in this work to ligate DNA probes due to its high specificity.’ J. Nandakumar and S. Shuman have documented that T4 RNA ligase 2.” ” (10013 bottom 2nd column-10014).
With regards to claim 45, Comanescu teaches, “ RNA is notoriously poorly preserved in FFPE tissue.” (abstract)
The specification teaches, “"Bridge PCR" is a method of localized, solid-phase PCR amplification, in which DNA25 fragment are amplified by primers attached to a surface (rather than in solution)”
Thus the broadest reasonable interpretation of BRIGE PCR or solid phase amplification is attached to a solid support.
With regards to claim 46-48, Cupploetti teaches, “[0106] Once the hydrogel is in contact with the surface of the template microarray, the primers are allowed to anneal with the template strands. The hydrogel is wetted with solution containing nucleic acid polymerase and dNTPs, allowing the primers to extended along the template strands as shown in Figure 1C.” Thus Cupploetti teaches solid phase PCR or Bridge PCR.
With regards to claim 49, Wang teaches, “FFPE tumor tissues” (page 23, tissue specimens). Thus Wang teaches biopsies or tumor tissue.
With regards to claims 50-54, Zhang teaches, “Our strategy for multiplexed miRNA detection based on the MLPA technique is illustrated in Fig. 1. For detection of each miRNA target (such as miRNA 1 and miRNA 2), the probe A and probe B (1A, 1B and 2A, 2B corresponding to miRNA 1 and miRNA 2, respectively) are designed. universal primer-specific sequence used for PCR amplification (green), a target-specific sequence (blue and sky blue) and a stuffer sequence in between (red). The target-specific sequences in probe A and probe B are simply designed to be complementary to the half-sequence of each miRNA target at the 3’- and the 5’-terminal, respectively. T4 RNA ligase 2 is employed in this work to ligate DNA probes due to its high specificity.’ J. Nandakumar and S. Shuman have documented that T4 RNA ligase 2 can effectively catalyze the ligation reactions of 3’-OH RNA and 5'-PO, RNA or DNA, which does not discriminate between RNA and DNA on the 5/-PO, side of the ligation nick. Moreover, they have also proved that the ligation efficiency achieved by using a DNA probe modified only with 3 or 2 ribonucleotides at the 3'-OH side is similar to that achieved by using an RNA probe, but the ligation efficiency will become much lower upon using a DNA probe modified with only a single 3’-OH ribonucleotide.”” To save on costs, probe A is modified with two ribonucleotides (brown) at its 3’-terminus and probe B is modified with a phosphate group at its 5’-terminus. In the presence of miRNA targets, probe A and probe B hybridize immediately to the adjacent sites of the target sequences and therefore, can be ligated with T4 RNA ligase 2. The ligated DNA probes are then amplified by PCR with fluorescein-labeled primers. The universal primer-specific sequences in probe A and probe B permit simultaneous PCR amplification of different miRNA targets using only one primer pair. For different miRNA targets, the stuffer sequences in probe A and probe B have different length, which are used to modulate the size of PCR products. Therefore, each miRNA target can give rise to an amplification product of unique size, permitting that the PCR products produced by different miRNA targets can be well separated using capillary electrophoresis (CE) and sensitively detected using laser-induced fluorescence.” (10013 bottom 2nd column-10014).
Response to Arguments
The response traverses the rejection by providing the representatives interpretation of the claims. This is noted.
The response continues by noting the rejection concedes not specifically teach applying a sub-probe with two RNA bases at the 3’ end and a second sub probe to anneal within a fixed permeabilized sample. This is conceded in the rejection. However, Comanescu teaches DASL (DNA- mediated Annealing, extension, selection ligation which provides for amplification and detection of nucleic acids.(page 28). Fan (Genome Research (2004) pages 878-885) exemplifies DASL
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The response continues by noting the rejection relies on the teachings of Zhang for sub-probe with two RNA bases at the 3’ end and a second sub probe to anneal within a fixed permeabilized sample.
The response continues by asserting Zhang teaches detection of miRNA which are not fragmented. This argument has been thoroughly reviewed but is not considered persuasive as the claims do not require fragmented sequences and the miRNA are of similar size to the fragmented nucleic acids in FFPE samples.
The response asserts there is noting in Zhang to motivate the artisan to use it in FFPE samples. This argument has been thoroughly reviewed but is not considered persuasive as Zhang teaches, “we rationally design one of the DNA probes in MLPA modified with two ribonucleotides at its 3'-terminus,which can greatly improve the ligation efficiency of DNA probes templated by miRNAs” (10013 bottom 2nd column).
The response continues by providing arguments with respect to spatial resolution. This argument has been thoroughly reviewed but is not considered persuasive as the metes and bounds are unclear how or what is required of spatial amplification.
The response continues traversing the rejection by asserting Wang does not disclosed ligation. This argument is not persuasive as both Comanescu and Zhang teach ligation.
The response continues by conceding Nilsson and Koch teach enzymatic ligation, but identify limitations in some aspect of enzymatic ligation. These arguments are not persuasive as the limitations argued are not present in the claims or relied upon in the rejection.
The response continues by asserting, “Koch requires circular nucleic acid probes and rolling circle amplification procedures, both of which are contrary to Applicant's invention.” This argument is confusing as claim 47 recites rolling circle amplification. Thus the argument is inconsistent with the claims.
The response continues by asserting Cuppoletti does not teach the steps of the claimed method. This argument has been thoroughly reviewed but is not considered persuasive as the cited art as whole teaches the other active steps of the claims.
Thus the rejection is maintained.
Summary
No claims are allowed.
Conclusion
Applicant's amendment necessitated the new ground(s) of rejection presented in this Office action. Accordingly, THIS ACTION IS MADE FINAL. See MPEP § 706.07(a). Applicant is reminded of the extension of time policy as set forth in 37 CFR 1.136(a).
A shortened statutory period for reply to this final action is set to expire THREE MONTHS from the mailing date of this action. In the event a first reply is filed within TWO MONTHS of the mailing date of this final action and the advisory action is not mailed until after the end of the THREE-MONTH shortened statutory period, then the shortened statutory period will expire on the date the advisory action is mailed, and any nonprovisional extension fee (37 CFR 1.17(a)) pursuant to 37 CFR 1.136(a) will be calculated from the mailing date of the advisory action. In no event, however, will the statutory period for reply expire later than SIX MONTHS from the mailing date of this final action.
Any inquiry concerning this communication or earlier communications from the examiner should be directed to STEVEN C POHNERT PhD whose telephone number is (571)272-3803. The examiner can normally be reached Monday- Friday about 6:00 AM-5:00 PM, every second Friday off.
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/Steven Pohnert/ Primary Examiner, Art Unit 1683
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