ABSTRACT
Though generally safe, research continues to demonstrate negative side effects of antibiotic administration on the gastrointestinal (GIT) microbiota across species. In horses, antibiotic associated diarrhea (AAD) is a life-threatening condition linked to the GIT microbiota. This study tested the hypothesis that short term antibiotic administration to healthy horses would negatively impact the fecal microbiota as measured by their ability to digest nutrients and through fecal shedding of disease-associated-bacteria. Twenty-four horses were assigned to one of four treatment groups: control (CO); potassium penicillin/gentamicin sulfate (KPG); ceftiofur crystalline free acid (EX); trimethoprim/sulfamethoxazole (SMZ); and treated for 4 days. Fecal samples were collected before treatment began (S0), the day after treatment conclusion (S5), and at 10, 14, 21, and 28 days after initiating treatment. Horses had highly individualized responses to antibiotic administration. All horses receiving antibiotics experienced significantly softer stool compared to controls. Lactobacillus spp. were dramatically reduced in all antibiotic treated S5 samples. Horses receiving antibiotics were significantly more likely to test positive for C. difficile or C. perfringens on fecal qPCR. In conclusion, response to antibiotic administration displays high inter-individual variability, but shows changes to the functions of fecal microbiota that may depend on the antibiotic used.
Subject(s)
Clostridioides difficile , Microbiota , Animals , Horses , Anti-Bacterial Agents/adverse effects , Feces/microbiology , BacteriaABSTRACT
We have previously localized a locus causing familial nonspecific dementia to the centromeric region of chromosome 3 in a pedigree from the Jutland area of Denmark. This pedigree shows anticipation. Here we present further analysis of these anticipation data which are suggestive of trinucleotide repeat expansion involvement. We also outline our strategies to clone the mutant gene via its putative associated trinucleotide repeat sequence.
Subject(s)
Chromosomes, Human, Pair 3/physiology , Dementia/genetics , Frontal Lobe/metabolism , Temporal Lobe/metabolism , Adult , Child , Cosmids/genetics , DNA Fingerprinting , Dementia/metabolism , Denmark , Disease Progression , Genetic Linkage/genetics , Humans , Immunohistochemistry , Pedigree , Trinucleotide RepeatsABSTRACT
Most drugs for cancer therapy are targeted to relative differences in the biological characteristics of cancer cells and normal cells. The therapeutic index of such drugs is theoretically limited by the magnitude of such differences, and most anticancer drugs have considerable toxicity to normal cells. Here we describe a new approach for developing anticancer drugs. This approach, termed variagenic targeting, exploits the absolute difference in the genotype of normal cells and cancer cells arising from normal gene sequence variation in essential genes and loss of heterozygosity (LOH) occurring during oncogenesis. The technology involves identifying genes that are: 1) essential for cell survival; 2) are expressed as multiple alleles in the normal population because of the presence of one or more nucleotide polymorphisms; and 3) are frequently subject to LOH in several common cancers. An allele-specific drug inhibiting the essential gene remaining in cancer cells would be lethal to the malignant cell and would have minimal toxicity to the normal heterozygous cell that retains the drug-insensitive allele. With antisense oligonucleotides designed to target two alternative alleles of replication protein A, 70-kDa subunit (RPA70) we demonstrate in vitro selective killing of cancer cells that contain only the sensitive allele of the target gene without killing cells expressing the alternative RPA70 allele. Additionally, we identify several other candidate genes for variagenic targeting. This technology represents a new approach for the discovery of agents with high therapeutics indices for treating cancer and other proliferative disorders.
Subject(s)
Antineoplastic Agents/therapeutic use , Loss of Heterozygosity , Neoplasms/drug therapy , Oligoribonucleotides, Antisense/therapeutic use , Antineoplastic Agents/pharmacology , Cell Survival/drug effects , DNA-Binding Proteins/antagonists & inhibitors , DNA-Binding Proteins/genetics , Drug Design , Feasibility Studies , Gene Targeting , Genetic Variation , Genome, Human , HeLa Cells , Humans , Neoplasms/genetics , Oligoribonucleotides, Antisense/pharmacology , Replication Protein A , Suppression, Genetic , Tumor Cells, CulturedABSTRACT
Single-nucleotide polymorphisms (SNPs) are the most common type of genetic variation in man. Genes containing one or more SNPs can give rise to two or more allelic forms of mRNAs. These mRNA variants may possess different biological functions as a result of differences in primary or higher order structures that interact with other cellular components. Here we report the observation of marked differences in mRNA secondary structure associated with SNPs in the coding regions of two human mRNAs: alanyl tRNA synthetase and replication protein A, 70-kDa subunit (RPA70). Enzymatic probing of SNP-containing allelic fragments of the mRNAs revealed pronounced allelic differences in cleavage pattern at sites 14 or 18 nt away from the SNP, suggesting that a single-nucleotide variation can give rise to different mRNA folds. By using phosphorothioate oligodeoxyribonucleotides complementary to the region of different allelic structures in the RPA70 mRNA, but not extending to the SNP itself, we find that the SNP exerts an allele-specific effect on the accessibility of its flanking site in the endogenous human RPA70 mRNA. This further supports the allele-specific structural features identified by enzymatic probing. These results demonstrate the contribution of common genetic variation to structural diversity of mRNA and suggest a broader role than previously thought for the effects of SNPs on mRNA structure and, ultimately, biological function.
Subject(s)
Alanine-tRNA Ligase/genetics , DNA-Binding Proteins/genetics , Genetic Variation , Polymorphism, Genetic , RNA, Messenger/chemistry , RNA, Messenger/genetics , Alleles , Base Sequence , DNA Helicases/genetics , Escherichia coli/enzymology , Humans , Macromolecular Substances , Molecular Sequence Data , Mutagenesis, Site-Directed , Nucleic Acid Conformation , Polymerase Chain Reaction , Replication Protein A , Ribonuclease H/metabolismABSTRACT
Folate derivatives are essential for DNA synthesis and methylation. A large proportion of the Caucasian population is heterozygous for a common substitution, 677C-->T (alanine-->valine), in methylenetetrahydrofolate reductase (MTHFR), an enzyme of folate interconversion. Homozygous mutant individuals, approximately 10-15% of North Americans, have been reported to have a reduced risk of colorectal cancer. We examined lymphocyte and tumor tissue DNA from colorectal carcinoma patients from two different populations to assess loss of heterozygosity (LOH) of MTHFR. We observed LOH in approximately 16% of colorectal tumors; in 8 of the 11 tumors with LOH, the mutant valine allele was lost. Additional studies are required to determine if preferential loss of the mutant allele is a common finding that could contribute to colorectal tumorigenesis.
Subject(s)
Colorectal Neoplasms/enzymology , Colorectal Neoplasms/genetics , Loss of Heterozygosity , Oxidoreductases Acting on CH-NH Group Donors/genetics , Alleles , DNA, Neoplasm/genetics , Female , Genotype , Humans , Methylenetetrahydrofolate Reductase (NADPH2) , Ovarian Neoplasms/enzymology , Ovarian Neoplasms/genetics , Polymerase Chain ReactionABSTRACT
This paper reports the treatment progress of methadone maintenance clients who were discharged or withdrew from treatment and then were readmitted for a second episode of treatment. Thirty-nine clients in a contingency contract condition remained in treatment long enough (6 months) during both the initial and a second treatment episode, to be exposed to discharge sanctions that were part of the contingency contract. Of these clients 34 failed treatment during the initial treatment episode. Nine (26%) of these initial treatment failures improved their performance in the second episode compared to the first, and only one (20%) of five initial treatment successes who left treatment during their first treatment episode for non-contract reasons showed a poorer performance (failing the second after succeeding in the first episode). Of 17 clients in a condition that applied no contingencies for positive urines, three of 14 (21%) who failed during the initial treatment episode improved their performance, and two of three (67%) who succeeded during the initial treatment episode failed in the second episode. For a subset of clients the efficacy of contingency contracting may not be realized until it is reapplied during a subsequent admission.
Subject(s)
Methadone/therapeutic use , Substance-Related Disorders/rehabilitation , Adult , Female , Humans , Male , Methadone/urine , Random Allocation , Treatment OutcomeABSTRACT
The recurrent translocation t(8;16)(p11;p13) is a cytogenetic hallmark for the M4/M5 subtype of acute myeloid leukaemia. Here we identify the breakpoint-associated genes. Positional cloning on chromosome 16 implicates the CREB-binding protein (CBP), a transcriptional adaptor/coactivator protein. At the chromosome 8 breakpoint we identify a novel gene, MOZ, which encodes a 2,004-amino-acid protein characterized by two C4HC3 zinc fingers and a single C2HC zinc finger in conjunction with a putative acetyltransferase signature. In-frame MOZ-CBP fusion transcripts combine the MOZ finger motifs and putative acetyltransferase domain with a largely intact CBP. We suggest that MOZ may represent a chromatin-associated acetyltransferase, and raise the possibility that a dominant MOZ-CBP fusion protein could mediate leukaemogenesis via aberrant chromatin acetylation.
Subject(s)
Acetyltransferases/genetics , Chromosomes, Human, Pair 16 , Chromosomes, Human, Pair 8 , Leukemia, Monocytic, Acute/genetics , Leukemia, Myelomonocytic, Acute/genetics , Nuclear Proteins/genetics , Trans-Activators , Transcription Factors/genetics , Translocation, Genetic , Amino Acid Sequence , Animals , Base Sequence , CREB-Binding Protein , Chromosome Mapping , Cloning, Molecular , Cricetinae , Gene Expression , Histone Acetyltransferases , Humans , Molecular Sequence Data , Polymerase Chain Reaction , Sequence Homology, Amino Acid , Zinc Fingers/geneticsABSTRACT
As part of an effort to identify the gene responsible for the predominant form of polycystic kidney disease (PKD1), we used a gridded human P1 library for contig assembly. The interval of interest, a 700-kb segment on chromosome 16p13.3, can be physically delineated by the genetic markers D16S125 and D16S84 and chromosomally characterized as a GC-rich isochore enriched for CpG islands, genes, and Alu-like repeats. Our attempts to recover CEPH YACs that encode this region of chromosome 16 were unsuccessful. However, we screened an arrayed P1 library using 15 distinct probes from the D16S125-D16S84 interval and identified 56 independent P1 clones. Only one probe from the interval was unsuccessful in identifying a P1 clone. Forty-four P1 clones were determined to be unique based on restriction enzyme analysis, and 42 of these were found to originate from chromosome 16p13.3, based on FISH to metaphase chromosomes. The 700-kb interval could be defined by a single sequence-ready contig comprised of 12 P1 clones and 1 cosmid clone. Our studies support the use of multiple libraries to generate the requisite physical reagents for positional cloning and encourage the use of Escherichia coli-based large-insert cloning systems to recover clones from YAC-deficient chromosomal intervals.
Subject(s)
Genetic Diseases, Inborn/genetics , Polycystic Kidney, Autosomal Dominant/genetics , Bacteriophage P1/genetics , Chromosome Mapping , Chromosomes, Human, Pair 16/genetics , Cloning, Molecular , Cosmids/genetics , Gene Library , Genetic Markers/genetics , Humans , In Situ Hybridization, Fluorescence , Sequence Tagged SitesABSTRACT
We describe an integrated approach to large-scale physical mapping using an Alu-PCR hybridization screening strategy in conjunction with direct PCR-based screening to construct a continuous yeast artificial chromosome map covering >20 mb in human chromosome 3, bands p14-p21, composed of 205 loci, connected by 480 yeast artificial chromosomes, with average interlocus distance of approximately equal to 100 kb. We observe an inverse distribution of Alu-PCR and (CA)n markers. These results suggest that the two screening methods may be complementary and demonstrate the utility of Alu-PCR hybridization screening in the closure of high-resolution human physical maps.
Subject(s)
Chromosomes, Artificial, Yeast , Chromosomes, Human, Pair 3 , Repetitive Sequences, Nucleic Acid , Animals , Chromosome Mapping , Cricetinae , DNA Probes , Genetic Markers , Humans , Hybrid Cells , Mice , Nucleic Acid Hybridization , Polymerase Chain ReactionABSTRACT
The t(7;11)(p15;p15) translocation is a recurrent chromosomal abnormality associated primarily with acute myeloid leukaemia (FAB M2 and M4). We present here the molecular definition of this translocation. On chromosome 7 positional cloning revealed the consistent rearrangement of the HOXA9 gene, which encodes a class I homeodomain protein potentially involved in myeloid differentiation. On chromosome 11 the translocation targets the human homologue of NUP98, a member of the GLFG nucleoporin family. Chimaeric messages spliced over the breakpoint fuse the GLFG repeat domains of NUP98 in-frame to the HOXA9 homeobox. The predicted NUP98-HOXA9 fusion protein may promote leukaemogenesis through inhibition of HOXA9-mediated terminal differentiation and/or aberrant nucleocytoplasmic transport.
Subject(s)
Chromosomes, Human, Pair 11 , Chromosomes, Human, Pair 7 , Homeodomain Proteins/genetics , Leukemia, Myelomonocytic, Acute/genetics , Membrane Proteins/genetics , Nuclear Pore Complex Proteins , Nuclear Proteins/genetics , Translocation, Genetic , Amino Acid Sequence , Base Sequence , Chromosome Mapping , Cloning, Molecular , Homeodomain Proteins/physiology , Humans , Molecular Sequence Data , RNA, Messenger/genetics , RNA, Neoplasm/genetics , Repetitive Sequences, Nucleic Acid/genetics , Sequence Analysis, DNAABSTRACT
The initial step of mapping STSs to chromosome 3 has been by mapping to a reference panel of 21 somatic cell hybrids containing fragments of chromosome 3. In this study we map 638 STSs to 23 bins on chromosome 3. The bin information greatly facilitates further mapping by radiation hybrids and YAC clones.
Subject(s)
Chromosomes, Human, Pair 3 , Sequence Tagged Sites , Chromosome Mapping , Humans , Polymerase Chain ReactionABSTRACT
We have developed a surface mounting technology for the rapid construction of ordered restriction maps from individual DNA molecules. Optical restriction maps constructed from yeast artificial chromosome DNA molecules mounted on specially derivatized glass surfaces are accurate and reproducible, and the technology is amenable to automation. The mounting procedures described here should also be useful for fluorescence in situ hybridization studies. We believe these improvements to optical mapping will further stimulate the development of nonelectrophoretic approaches to genome analysis.
Subject(s)
Chromosomes, Artificial, Yeast , Restriction Mapping , Saccharomyces cerevisiae/genetics , Automation , DNA, Fungal/chemistry , DNA, Fungal/genetics , DNA, Fungal/isolation & purification , Microscopy, Fluorescence , Molecular WeightABSTRACT
We report here an efficient approach to the establishment of extended YAC contigs on human chromosome 2 by using an interspersed repetitive sequences (IRS)-PCR-based screening strategy for YAC DNA pools. Genomic DNA was extracted from 1152 YAC pools comprised of 55,296 YACs mostly derived from the CEPH Mark I library. Alu-element-mediated PCR was performed for each pool, and amplification products were spotted on hybridization membranes (IRS filters). IRS probes for the screening of the IRS filters were obtained by Alu-element-mediated PCR. Of 708 distinct probes obtained from chromosome 2-specific somatic cell hybrids, 85% were successfully used for library screening. Similarly, 80% of 80 YAC walking probes were successfully used for library screening. Each probe detected an average of 6.6 YACs, which is in good agreement with the 7- to 7.5-fold genome coverage provided by the library. In a preliminary analysis, we have identified 188 YAC groups that are the basis for building contigs for chromosome 2. The coverage of the telomeric half of chromosome 2q was considered to be good since 31 of 34 microsatellites and 22 of 23 expressed sequence tags that were chosen from chromosome region 2q13-q37 were contained in a chromosome 2 YAC sublibrary generated by our experiments. We have identified a minimum of 1610 distinct chromosome 2-specific YACs, which will be a valuable asset for the physical mapping of the second largest human chromosome.
Subject(s)
Chromosomes, Artificial, Yeast , Chromosomes, Human, Pair 2 , Cloning, Molecular/methods , Genome, Human , Repetitive Sequences, Nucleic Acid , Animals , Base Sequence , Cricetinae , DNA, Satellite/genetics , Humans , Hybrid Cells , Molecular Sequence Data , Polymerase Chain ReactionABSTRACT
We have developed a new technique for the generation of YAC contigs in the mouse genome that is based on the ability to detect overlapping clones by hybridization of shared IRS-PCR products. As a demonstration of the technique, a 5-cM, > 5 megabase contig was developed on the distal half of mouse Chromosome (Chr) 1, spanning the region from Lamb2 to At3. The contig covers roughly 5% of the genetic distance of the chromosome and is comprised of more than 80 clones; 71 probes were assigned physical order to the chromosome, of which 59 were new markers generated in this study. Eight of the new probes were shown to be polymorphic between C3H/HeJ-gld and M. spretus. Three probes were mapped on a [(C3H/HeJ-gld x M. spretus) x C3H/HeJ-gld] interspecific backcross to integrate the physical map with a high-resolution genetic map of the region.
Subject(s)
Chromosomes, Artificial, Yeast/genetics , Repetitive Sequences, Nucleic Acid , Animals , Base Sequence , Chromosome Mapping , Cloning, Molecular , Crosses, Genetic , DNA Primers/genetics , Electrophoresis, Gel, Pulsed-Field , Female , Genetic Linkage , Genetic Markers , Genome , Male , Mice , Mice, Inbred C3H , Molecular Sequence Data , Muridae , Polymerase Chain ReactionABSTRACT
We report here the construction of a genetic linkage map and an overlapping set of clones containing DNA markers linked to the causative locus for facioscapulohumeral muscular dystrophy (FSHD) on 4q35. Multipoint linkage analysis placed eight loci in the following order with odds greater than 1000:1: cen-D4S171-FXI-D4S426-D4S187-D4S130-D4S 163-D4S139-D4F35S1-qter. The most likely position of D4S809 was distal to D4F35S1. Thirty-four yeast artificial chromosomes (YACs) were isolated by PCR-based assays for STSs derived from DNA markers with known genetic and physical order. Walking from the insert ends of 2 YACs identified 7 additional YACs, bridging the gaps between three of the markers. Two new YACs were found by hybridization of a cosmid inter-Alu PCR product to dot blots of inter-Alu PCR products of YAC DNA pools. All YAC clones were positioned using the genetic and physical order of the STSs and inter-Alu PCR fingerprint data. Eleven of the YACs and two cosmids were mapped by fluorescence in situ hybridization to confirm the location of the clones and to detect chimerism. The 43 YACs were assembled into two contigs. The larger contig spans approximately 2.4 Mb and contains markers closest to the FSHD gene.
Subject(s)
Chromosomes, Human, Pair 4 , Genes , Muscular Dystrophies/genetics , Base Sequence , Chromosome Mapping , Chromosome Walking , Chromosomes, Artificial, Yeast , Gene Library , Genetic Linkage , Genetic Markers , Humans , Molecular Sequence DataABSTRACT
In a 3 x 2 factorial design, 360 new admissions to methadone maintenance were randomly assigned to one of three levels of counseling: (1) "medication only," (2) "standard" counseling, and (3) "enhanced" services; and one of two contingency contracting conditions: (1) no contingencies (NC), and (2) contingency contracting (CC). Contingency contracting included discharge for continuous positive urines; subsequently CC subjects were discharged at a greater rate than the NC group. However, CC subjects were more likely to be readmitted. NC subjects provided more urines positive for any illicit drug use than did CC subjects. For opiate positives a significant level of counseling by contingency contracting interaction was found with medication only/CC subjects obtaining fewer opiate positives than medication only/NC subjects. The impact of reduced or enhanced services and of contingency contracting will not be fully understood until longer term follow-up (18 and 24 month) is completed. Results suggest that contingency management procedures could be utilized in settings offering minimum services (e.g., "interim methadone") to achieve treatment outcomes similar to programs offering standard counseling services.
Subject(s)
Behavior Therapy/methods , Counseling , Methadone/therapeutic use , Opioid-Related Disorders/rehabilitation , Substance Abuse Detection , Adolescent , Adult , Combined Modality Therapy , Feedback , Female , Follow-Up Studies , Humans , Male , Middle Aged , Treatment OutcomeABSTRACT
An overlapping set of 21 yeast artificial chromosomes (YACs) spanning the RET proto-oncogene [Takahashi et al., Oncogene 3 (1988) 571-578] and D10S102 markers on human chromosome 10 was isolated in a series of hybridization-based chromosomal walks in a YAC library. Genetic linkage analyses implicate this chromosomal region as the location of the gene (MEN2A) responsible for multiple endocrine neoplasia type 2A. Four YACs carrying a RET sequence-tagged site (STS) and two YACs carrying a D10S102 STS were used to initiate chromosome walks. These were based on hybridization of Alu element-mediated polymerase chain reaction (Alu-PCR) products from YACs to dot blots of Alu-PCR products from complex pools of YAC clones. The hybridization anchor content of YACs identified in the walks was confirmed by probing blots of Alu-PCR products from individual YACs and by comparing Alu-PCR fingerprints of each YAC. Ten hybridization-based Alu-PCR anchors and three STS anchors were ordered within eleven intervals created by the 21 overlapping YACs. The order of anchors requiring the fewest gaps in the YACs is consistent with the walking results and establishes the STS anchor order as D10S102-D10S94-RET. The overlapping set of YACs represents about 1.55 Mb of the human genome according to restriction mapping of four representative YACs in the contig. These results demonstrate the power of Alu-PCR hybridization for chromosomal walking and provide a rich source of overlapping YACs which can be used to identify candidate MEN2A genes.
Subject(s)
Chromosomes, Artificial, Yeast , Chromosomes, Human, Pair 10 , Multiple Endocrine Neoplasia/genetics , Repetitive Sequences, Nucleic Acid , Base Sequence , Chromosome Walking , DNA Primers , Genetic Linkage , Genetic Markers , Humans , Molecular Sequence Data , Polymerase Chain Reaction , Proto-Oncogene Mas , Restriction MappingABSTRACT
We have isolated and mapped by fluorescence in situ hybridization 80 new cosmids on the short arm of chromosome 3. These markers were isolated from a radiation-reduced hybrid, DM1, made from a cell line that was monochromosomal for human chromosome 3. Selected cosmids were used in double-label cohybridization experiments in which polymerase chain reaction products, generated by an Alu oligonucleotide primer from genomic DNA, were used for chromosome banding. Fifty-six of the cosmid probes map between 3p14.3 and 3p22 while 24 other probes cluster around bands 3p23-3p25. Three probes that appeared to map close to the chromosome 3 region bearing a t(3,8)p14.2; q24.1 translocation associated with renal cell carcinoma were analyzed by interphase mapping techniques and hybridized to metaphase spreads from the translocation cell line. These 80 probes will be useful in the elucidation of genetic alterations associated with diseases such as small cell lung carcinoma, renal cell carcinoma, and von Hippel-Lindau disease.
Subject(s)
Chromosome Mapping , Chromosomes, Human, Pair 3 , Cosmids/genetics , Animals , Base Sequence , Cloning, Molecular , Cricetinae , DNA/genetics , DNA Probes , Genetic Markers , Humans , Hybrid Cells , In Situ Hybridization, Fluorescence , Molecular Sequence Data , Polymerase Chain ReactionABSTRACT
OBJECTIVE: To describe the clinical course and genetic studies of renal carcinoma in members of a family with the constitutional chromosome translocation, t(3;8) (p14;q24). DESIGN: A follow-up study that updates our 1979 report of renal carcinoma in 10 of these relatives. SETTING: A cancer center and university hospital. PATIENTS: Members of the family, including five carriers of the 3;8 translocation who were in remission of renal cancer. MEASUREMENTS: Clinical follow-up of the family and genetic analyses of the renal cancer specimens of three patients. RESULTS: Renal carcinoma recurred in all five patients in the family at 1 to 16 years of follow-up. Three patients have died of renal cancer, and two are in a second remission. The renal cancers from three family members consistently reveal loss of the entire derivative chromosome 8, which bears the chromosome 3p segment spanning band p14 to the telomere. In contrast, no genetic change was detected in the derivative chromosome 3 or in normal chromosomes 3 and 8. CONCLUSIONS: This family illustrates the importance of clinical follow-up of patients with a hereditary cancer that can develop at multiple foci and recur over time. The inherited 3;8 translocation and loss of the translocated distal chromosome 3p in tumor specimens of family members may help localize the gene or genes involved in the pathogenesis of both familial and sporadic renal carcinoma.
Subject(s)
Carcinoma, Renal Cell/genetics , Chromosomes, Human, Pair 3 , Chromosomes, Human, Pair 8 , Kidney Neoplasms/genetics , Translocation, Genetic , Adult , Alleles , DNA, Neoplasm/genetics , Female , Follow-Up Studies , Humans , Karyotyping , Male , Middle Aged , Neoplasm Recurrence, Local/genetics , PedigreeABSTRACT
Just before and 4 months after initiation of a condom giveaway program, a questionnaire regarding sexual behavior and condom acquisition was administered to 103 men attending an outpatient drug abuse treatment clinic. Jars filled with a variety of condoms were placed in every clinic room. Condom taking varied as a function of room. Sixty percent of the subjects reported taking condoms. At follow-up, clients reported increases in condom possession and in use of condoms for vaginal intercourse.