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. Election/Restrictions Applicant’s election of Group II, claims 18, 19 and 20-28 and the species of dsRNA and Deformed Wing Virus in the reply filed on 8/26/2025 is acknowledged. Because applicant did not distinctly and specifically point out the supposed errors in the restriction requirement, the election has been treated as an election without traverse (MPEP § 818.01(a)). Applicant has cancelled all non-elected claims. With respect to the species election regarding a target gene product set forth in claims 19 and 23, this election of species requirement is withdrawn and all of the non-elected target gene product species are hereby rejoined. A new Examiner, David A. Montanari, has taken over prosecution of the instant application. Claim 20 has been cancelled. The instant application is a DIV of 16/029,686, now US Pat. No. 11,382,989. Claims 18 , 19 and 21 -28 are examined in the instant application. Claim Objections Claim 18 objected to because of the following informalities: claim 18 is not grammatically correct. The letter "a" should be inserted between the words "is" and "bee". Appropriate correction is required. 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 appl icant regards as his invention. Claims 18, 19 and 21-2 6 are 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 18 is not clear. Claim 18 recites the limitation that “one or more bacteria are native to the microbiome of a host insect to an insect ”, however it is not clear what the metes and bounds are of the bacteria. It is not clear if the if the bacteria are native to a host insect, another insect which is not the host, or both a host insect and another insect which is not the host insect. Claim 22 depends from cancelled claim 20, thus it is not clear which claim, claim 22 depends from. However, for the purpose s of examination, it is interpreted that claim 22 depends from claim 18. To obviate this rejection, Applicant should correct the dependency of claim 22. Claim 25 is not clear. Claim 25 recites the limitation that “one or more bacteria are native to the microbiome of a host bee to hive components”, however it is not clear what the metes and bounds are of the bacteria. It is not clear if the if the bacteria are native to a host bee, the hive components, or both the host bee and hive components. Claim Rejections - 35 USC § 102/ 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 the appropriate paragraphs of 35 U.S.C. 102 that form the basis for the rejections under this section made in this Office action: A person shall be entitled to a patent unless – (a)(1) the claimed invention was patented, described in a printed publication, or in public use, on sale , or otherwise available to the public before the effective filing date of the claimed invention. 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) 18 is/are rejected under 35 U.S.C. 102( a ) (1) as anticipated by or, in the alternative, under 35 U.S.C. 103 as obvious over Whitten et al. (2016, Proc. R. Soc. B, Vol. 283, pgs. 1-9 , cited on IDS filed on 8/27/2025 ) . Regarding claim 18, Whitten et al. teach a method of genetically modifying bacteria that will modify gene expression in insects when fed the bacteria. Specifically, Whitten teaches (emphasis added) “ RNaseIII-deficient, dsRNA-expressing bacterial strains were created from the symbionts of two very diverse pest species: a long-lived blood-sucking bug, Rhodnius prolixus , and a short-lived globally invasive polyphagous agricultural pest, western flower thrips ( Frankliniella occidentalis ). When ingested, the manipulated bacteria colonized the insects, successfully competed with the wild-type microflora, and sustainably mediated systemic knockdown phenotypes that were horizontally transmissible. This represents a significant advance in the ability to deliver RNAi, potentially to a large range of non-model insects . ” (Abstract lines 5-13). Regarding that the host insect is a bee , Whitten teaches that their method can be effective in honeybees (pg. 8 col. 2 parag. 1 lines 1-6). Regarding the first insect species, R. prolixus , Whitten teaches that their genetically modified bacteria successfully knocked down expression of Nitrophorin-1 , Nitrophorin-2 and dsdagA in the insect R. prolixus (pg. 4 col. 1 parag. 2). While Whitten uses the term “knockdown” it is interpreted that this would be the same as downregulating the expression of a gene as instantly recited. Regarding the second insect species, F. occidentalis , Whitten teaches that “ To deliver dsRNA, an RNaseIII-deficient mutant, BFo2 α , was obtained into which plasmids, directing expression of dsRNA, were introduced. To demonstrate symbiont-mediated gene silencing, we targeted alpha-tubulin (Tub) production that could be essential. Stable bacterial expression of dsTub and the off-target control dsdagA, was confirmed by qRT-PCR prior to introducing the bacteria into insects of different ages via an artificial sucrose-based feeding solution. After 4 days, a highly significant mortality phenotype was observed among larvae exposed to dsTub, particularly in the first (L1) larval stage (figure 5; p , 0.0001). ” (pg. 6 col. 2 parag. 1 lines 1-11). Whitten concludes by teaching (emphasis added) “ symbiont-mediated RNAi is a powerful means to interrogate gene function in invertebrates. We have demonstrated this transformative technology in two evolutionarily distinct insect species. Based on this success, we believe the technology can potentially be translated to address gene function in many diverse arthropod species , dependent on exploiting facultative gut symbiotic bacteria specific to these species. In addition, the technology provides a potential means of highly targeted biocontrol against tropical disease vectors and agricultural pest species. ” (pg. 8 col. 2 parag. 2). Thus, while Whitten does not teach the active step of administering an effective amount of genetically engineered bacteria to a bee, it would have been obvious in view of Whitten teaching that honeybees can be a target for their method of knocking down gene expression via genetically modified bacteria expressing dsRNA. Accordingly, the teachings of Whitten anticipate or render obvious the invention of claim 18. 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) 18 , 19 and 21-24 is/are rejected under 35 U.S.C. 103 as being unpatentable over Whitten et al. (2016, Proc. R. Soc. B, Vol. 283, pgs. 1-9 , cited on IDS filed on 8/27/2025 ) in view of Paldi et al. (US Pat. No. 8,822,426, issued 9/2/2014) . Regarding claim 18, Whitten et al. teach a method of genetically modifying bacteria that will modify gene expression in insects when fed the bacteria. Specifically, Whitten teaches (emphasis added) “ RNaseIII-deficient, dsRNA-expressing bacterial strains were created from the symbionts of two very diverse pest species: a long-lived blood-sucking bug, Rhodnius prolixus , and a short-lived globally invasive polyphagous agricultural pest, western flower thrips ( Frankliniella occidentalis ). When ingested, the manipulated bacteria colonized the insects, successfully competed with the wild-type microflora, and sustainably mediated systemic knockdown phenotypes that were horizontally transmissible. This represents a significant advance in the ability to deliver RNAi, potentially to a large range of non-model insects . ” (Abstract lines 5-13). Regarding the host insect is a bee, Whitten teaches that their method can be effective in honeybees (pg. 8 col. 2 parag. 1 lines 1-6). Regarding the first insect species, R. prolixus , Whitten teaches that their genetically modified bacteria successfully knocked down expression of Nitrophorin-1 , Nitrophorin-2 and dsdagA in the insect R. prolixus (pg. 4 col. 1 parag. 2). While Whitten uses the term “knockdown” it is interpreted that this would be the same as downregulating the expression of a gene as instantly recited. Regarding the second insect species, F. occidentalis , Whitten teaches that “ To deliver dsRNA, an RNaseIII-deficient mutant, BFo2 α , was obtained into which plasmids, directing expression of dsRNA, were introduced. To demonstrate symbiont-mediated gene silencing, we targeted alpha-tubulin (Tub) production that could be essential. Stable bacterial expression of dsTub and the off-target control dsdagA, was confirmed by qRT-PCR prior to introducing the bacteria into insects of different ages via an artificial sucrose-based feeding solution. After 4 days, a highly significant mortality phenotype was observed among larvae exposed to dsTub, particularly in the first (L1) larval stage (figure 5; p , 0.0001). ” (pg. 6 col. 2 parag. 1 lines 1-11). Whitten concludes by teaching “ symbiont-mediated RNAi is a powerful means to interrogate gene function in invertebrates. We have demonstrated this transformative technology in two evolutionarily distinct insect species. Based on this success, we believe the technology can potentially be translated to address gene function in many diverse arthropod species, dependent on exploiting facultative gut symbiotic bacteria specific to these species. In addition, the technology provides a potential means of highly targeted biocontrol against tropical disease vectors and agricultural pest species. ” (pg. 8 col. 2 parag. 2). Whitten does not teach: a gene product from Nosema ceranae (claim s 19 and 23 ) a target gene as TOM70, TIM22 and TOM40 (claim 21) , reducing the susceptibility to CCD or infection (claim 22); and at least two noncontiguous dsRNAs (claim 24) . Regarding a gene product from N. ceranae and colony collapse disorder in claims 19, 22 and 23 , Paldi et al. teach (emphasis added): “ The importance of honeybees and other pollinating insects to the global world economy far s urpasses their contribution in terms of honey production. The United States Department of Agriculture (USDA) estimates that every third bite we consume in our diet is dependent on a honeybee to pollinate that food. The total contribution of pollination in terms of added value to fruit crops exceeds S15 billion per annum, with indirect potential consequence of S75 billion dollars. Microsporidia are basal fungi and obligate intracellular parasites of other eukaryotes characterized by extreme genomic and cellular reduction. Two described species of micro s poridia, Nosema apis and Nosema ceranae , cause a widespread and destructive disease in adult honey bee. Nosema disease is widespread across the world, and it has been observed that nosema pathogenesis, together with increased viral load, are the best predictors of weak and collapsing colonies. In Europe, disappearing colony Syndrome has been directly attributed to Nosema ceranae , and the risk of colony depopulation is six times higher in colonies infected with N. ceranae than in uninfected ones. Recently, it was shown that natural Nosema ceranae infection can cause the sudden collapse of bee colonies. ” Specifically, Paldi teaches (emphasis added): “ According to yet another aspect of some embodiments of the present invention there is provided a bee-ingestible composition comprising an isolated nucleic acid agent comprising a nucleic acid sequence downregulating expression of a gene product of a Nosema parasite. ” (col. 4 lines 1 6 -20). Regarding claim 21 and the down regulated expression of a gene product from N. ceranae , Paldi teaches that they engineered dsRNA targeting the genes TOM70, TIM22, TOM40, Imp2, mitochondrial Hsp70, ATM1-ABC transporter proteins, Frataxin, Ferredoxin, ERV1, ferredoxin, NADPH oxido-reductase FNR, pyruvate dehydrogenase C. Subunit, pyruvate dehydrogenase β subunit, mitochondrial glycerol-3-phosphate dehydroge nase (mtG3PDH), manganese-containing Superoxide dismutase (MnSOD), DNAJ (Hsp70 interacting), Iron Sulfur cluster ISU1 and Cystein desulfurase Nsf 1 (col. 5 lines 12-19, col. 7 lines 24-44 and Fig. 6). Regarding claim 24, Paldi teaches that they can use two noncontiguous dsRNA ( col. 5 lines 31-35 and col. 17 lines 59-67). Significantly, Paldi teaches in Example 5 that feeding dsRNA targeting Nosema proteins led to significant increases in survival of bees infected with Nosema (starts at col. 30). Thus at the time of filing the ordinary artisan would have found it prima facie obvious to modify the teachings of Whitten regarding a method of downregulating gene expression in honeybees using genetically modified bacteria expressing dsRNA with the teachings of Paldi regarding downregulating expression of genes involved in CCD in honeybees using dsRNA to arrive at the claimed invention. One of ordinary skill in the art would have been motivated to make such a modification since Whitten teaches that symbiotic bacteria or bacteria that are native to the microbiome of an insect are a successful vector for expressing dsRNA to downregulate expression of target gene product in an insect. Specific motivation is provided by Paldi to modify the teachings of Whitten to target genes of N. ceranae since they are a parasitic organism known to contribute to colony collapse of honey bees. There would have been a reasonable expectation of success that the genetically modified bacteria of Whitten could be modified to target the genes of N. ceranae since Paldi teaches successful targeting of genes such as TOM70 using dsRNA fed to honeybees. Thus the cited art provides the requisite teachings and motivations to make and use the invention as claimed. Claim (s) 25 and 26 is/are rejected under 35 U.S.C. 103 as being unpatentable over Whitten et al. (2016, Proc. R. Soc. B, Vol. 283, pgs. 1-9 , cited on IDS filed on 8/27/2025 ) in view of P owell et al. ( 2016, Pest Management Science, Vol. 73, pgs. 53-63 ) and Paldi et al. (US Pat. No. 8,822,426, issued 9/2/2014) . Regarding claim 18, Whitten et al. teach a method of genetically modifying bacteria that will modify gene expression in insects when fed the bacteria. Specifically, Whitten teaches (emphasis added) “ RNaseIII-deficient, dsRNA-expressing bacterial strains were created from the symbionts of two very diverse pest species: a long-lived blood-sucking bug, Rhodnius prolixus , and a short-lived globally invasive polyphagous agricultural pest, western flower thrips ( Frankliniella occidentalis ). When ingested, the manipulated bacteria colonized the insects, successfully competed with the wild-type microflora, and sustainably mediated systemic knockdown phenotypes that were horizontally transmissible. This represents a significant advance in the ability to deliver RNAi, potentially to a large range of non-model insects . ” (Abstract lines 5-13). Regarding the host insect is a bee, Whitten teaches that their method can be effective in honeybees (pg. 8 col. 2 parag. 1 lines 1-6). Regarding the first insect species, R. prolixus , Whitten teaches that their genetically modified bacteria successfully knocked down expression of Nitrophorin-1 , Nitrophorin-2 and dsdagA in the insect R. prolixus (pg. 4 col. 1 parag. 2). While Whitten uses the term “knockdown” it is interpreted that this would be the same as downregulating the expression of a gene as instantly recited. Regarding the second insect species, F. occidentalis , Whitten teaches that “ To deliver dsRNA, an RNaseIII-deficient mutant, BFo2 α , was obtained into which plasmids, directing expression of dsRNA, were introduced. To demonstrate symbiont-mediated gene silencing, we targeted alpha-tubulin (Tub) production that could be essential. Stable bacterial expression of dsTub and the off-target control dsdagA, was confirmed by qRT-PCR prior to introducing the bacteria into insects of different ages via an artificial sucrose-based feeding solution. After 4 days, a highly significant mortality phenotype was observed among larvae exposed to dsTub, particularly in the first (L1) larval stage (figure 5; p , 0.0001). ” (pg. 6 col. 2 parag. 1 lines 1-11). Whitten concludes by teaching “ symbiont-mediated RNAi is a powerful means to interrogate gene function in invertebrates. We have demonstrated this transformative technology in two evolutionarily distinct insect species. Based on this success, we believe the technology can potentially be translated to address gene function in many diverse arthropod species, dependent on exploiting facultative gut symbiotic bacteria specific to these species. In addition, the technology provides a potential means of highly targeted biocontrol against tropical disease vectors and agricultural pest species. ” (pg. 8 col. 2 parag. 2). Whitten does not teach: Reducing the susceptibility of a bee colony to infestation by the small hive beetle; and at least two noncontiguous dsRNAs. Regarding reducing the susceptibility of a bee colony to infestation by the small hive beetle ( A. tumida ) , Powell et al. teach using a dsRNA to increase the mortality of the small hive beetle (see Abstract). Specifically, Powell teaches “ The small hive beetle ( A. tumida ), a scavenger and predator of the European honey bee, has already spread from Africa to countries including the United States, Australia, Canada and Mexico and has potential to establish in Europe and the United Kingdom. Current pest control measures are challenged by the need for target specificity and high efficacy. RNAi, able to cause the destruction of target-specific mRNAs, offers possibilities for the development of a new approach to combat this economically significant pest without jeopardising the health of honey bee populations. Here we report significant dose-dependent mortality of A. tumida following the injections of 2–12.5 ng doses of dsRNAs targeting Laccase 2 and V-ATPase subunit A mRNAs. Analysis of relative mRNA levels by qPCR confirmed target gene knockdown, and significantly enhanced levels of gene suppression over time demonstrated that the RNAi effect was persistent and systemic. ” (pg. 60 col. 1 Discussion bridge pg. 61 col. 1 parag. 1). (ii) Regarding claim 26, Paldi teaches that at the time of filing the ordinary artisan can use two noncontiguous dsRNAs when downregulating genes expressed by a parasitic organism (col. 5 lines 31-35 and col. 17 lines 59-67). Thus at the time of filing the ordinary artisan would have found it prima facie obvious to modify the teachings of Whitten regarding a method of downregulating gene expression in honeybees using genetically modified bacteria expressing dsRNA with the teachings of Powell that the small hive battle can be controlled via dsRNA and with the teachings of Paldi regarding using two noncontiguous dsRNA to arrive at the claimed invention. One of ordinary skill in the art would have been motivated to make such a modification since Whitten teaches that symbiotic bacteria or bacteria that are native to the microbiome of an insect are a successful vector for expressing dsRNA to downregulate expression of target gene product in an insect. Specific motivation is provided by Powell to modify the teachings of Whitten to target genes of A. tumida since they are a n organism that is known to contribute to the death of honey bee hives . There would have been a reasonable expectation of success that the genetically modified bacteria of Whitten could be modified to target the genes of A. tumida since P owell teaches successful targeting of genes via dsRNA that increase the mortality of A. tumida . Thus the cited art provides the requisite teachings and motivations to make and use the invention as claimed. Claim (s) 27 and 28 is/are rejected under 35 U.S.C. 103 as being unpatentable over Whitten et al. (2016, Proc. R. Soc. B, Vol. 283, pgs. 1-9 , cited on IDS filed on 8/27/2025 ) in view of Kwong et al. (2013, Int. J. Systematic and Evol. Micro., Vol. 63, pgs. 2008-2018) . Regarding claim 18, Whitten et al. teach a method of genetically modifying bacteria that will modify gene expression in insects when fed the bacteria. Specifically, Whitten teaches (emphasis added) “ RNaseIII-deficient, dsRNA-expressing bacterial strains were created from the symbionts of two very diverse pest species: a long-lived blood-sucking bug, Rhodnius prolixus , and a short-lived globally invasive polyphagous agricultural pest, western flower thrips ( Frankliniella occidentalis ). When ingested, the manipulated bacteria colonized the insects, successfully competed with the wild-type microflora, and sustainably mediated systemic knockdown phenotypes that were horizontally transmissible. This represents a significant advance in the ability to deliver RNAi, potentially to a large range of non-model insects . ” (Abstract lines 5-13). Regarding the host insect is a bee, Whitten teaches that their method can be effective in honeybees (pg. 8 col. 2 parag. 1 lines 1-6). Regarding the first insect species, R. prolixus , Whitten teaches that their genetically modified bacteria successfully knocked down expression of Nitrophorin-1 , Nitrophorin-2 and dsdagA in the insect R. prolixus (pg. 4 col. 1 parag. 2). While Whitten uses the term “knockdown” it is interpreted that this would be the same as downregulating the expression of a gene as instantly recited. Regarding the second insect species, F. occidentalis , Whitten teaches that “ To deliver dsRNA, an RNaseIII-deficient mutant, BFo2 α , was obtained into which plasmids, directing expression of dsRNA, were introduced. To demonstrate symbiont-mediated gene silencing, we targeted alpha-tubulin (Tub) production that could be essential. Stable bacterial expression of dsTub and the off-target control dsdagA, was confirmed by qRT-PCR prior to introducing the bacteria into insects of different ages via an artificial sucrose-based feeding solution. After 4 days, a highly significant mortality phenotype was observed among larvae exposed to dsTub, particularly in the first (L1) larval stage (figure 5; p , 0.0001). ” (pg. 6 col. 2 parag. 1 lines 1-11). Whitten concludes by teaching “ symbiont-mediated RNAi is a powerful means to interrogate gene function in invertebrates. We have demonstrated this transformative technology in two evolutionarily distinct insect species. Based on this success, we believe the technology can potentially be translated to address gene function in many diverse arthropod species, dependent on exploiting facultative gut symbiotic bacteria specific to these species. In addition, the technology provides a potential means of highly targeted biocontrol against tropical disease vectors and agricultural pest species. ” (pg. 8 col. 2 parag. 2). Whitten does not teach: Using a S. alvi to express a heterologous nucleic acid in a bee. Regarding using S. alvi to express a heterologous nucleic acid in a bee, Kwong e t al. teaches the isolation, identification and characterization of a novel bacterium, S. alvi from the gut of the Western honey bee, A. mellifera and the bumble bee s , B. bo mbus and B. vagans (see Abstract and Methods). Kwong continues to teach that “ Bacteria that live in close, symbiotic association with insects can perform beneficial functions including nutrient synthesis, digestion and provision of disease resistance (Dillon & Dillon, 2004; Koch & Schmid-Hempel, 2011b). Honey bees (Apis spp.) and bumble bees (Bombus spp.) have recently been found to possess a highly specific gut microbial composition, suggesting they too may participate in an intimate host–microbe symbiosis (Martinson et al., 2011; Koch & Schmid-Hempel, 2011a). ” (pg. 2008 col. 2 last two lines bridge pg. 2009 col. 1 parag. 1). Thus, at the time of filing the ordinary artisan would have found it prima facie obvious to modify the teachings of Whitten regarding a method of downregulating gene expression in honeybees using genetically modified bacteria expressing dsRNA with the teachings of Kwong regarding a novel bacterium expressed in bees to arrive at the claimed invention. One of ordinary skill in the art would have been motivated to make such a modification since Kwong teaches a novel bacterium that is present in the gut of at least three species of bee. One of ordinary skill in the art would have been motivated to use S. alvi to express a heterologous nucleic acid since S. alvi is naturally present in several species of bee . There would have been a reasonable expectation of success that the S. alvi of Kwong could be modified to express a heterologous nucleic acid since Whitten teaches successful genetic modification of several strain of bacteria from insects. Thus, the cited art provides the requisite teachings and motivations to make and use the invention as claimed. Conclusion No claims are allowed. Any inquiry concerning this communication or earlier communications from the examiner should be directed to FILLIN "Examiner name" \* MERGEFORMAT DAVID A MONTANARI whose telephone number is FILLIN "Phone number" \* MERGEFORMAT (571)272-3108 . The examiner can normally be reached FILLIN "Work Schedule?" \* MERGEFORMAT M-Tr 8-6 . 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Visit https://www.uspto.gov/patents/apply/patent-center for more information about Patent Center and https://www.uspto.gov/patents/docx for information about filing in DOCX format. For additional questions, contact the Electronic Business Center (EBC) at 866-217-9197 (toll-free). If you would like assistance from a USPTO Customer Service Representative, call 800-786-9199 (IN USA OR CANADA) or 571-272-1000. /DAVID A MONTANARI/ Examiner, Art Unit 1632