Co-suppression of AtMIPS demonstrates cooperation of MIPS1, MIPS2 and MIPS3 in maintaining myo-inositol synthesis.

KEY MESSAGE: Co-suppressed MIPS2 transgenic lines allow bypass of the embryo lethal phenotype of the previously published triple knock-out and demonstrate the effects of MIPS on later stages of development. Regulation of inositol production is of interest broadly for its effects on plant growth and development. The enzyme L-myo-inositol 1-phosphate synthase (MIPS, also known as IPS) isomerizes D-glucose-6-P to D-inositol 3-P, and this is the rate-limiting step in inositol production. In Arabidopsis thaliana, the MIPS enzyme is encoded by three different genes, (AtMIPS1, AtMIPS2 and AtMIPS3), each of which has been shown to produce proteins with biochemically similar properties but differential expression patterns. Here, we report phenotypic and biochemical effects of MIPS co-suppression. We show that some plants engineered to overexpress MIPS2 in fact show reduced expression of AtMIPS1, AtMIPS2 and AtMIPS3, and show altered vegetative phenotype, reduced size and root length, and delayed flowering. Additionally, these plants show reduced inositol, increased glucose levels, and alteration of other metabolites. Our results suggest that the three AtMIPS genes work together to impact the overall synthesis of myo-inositol and overall inositol homeostasis.

Psychiatric comorbidity of internet addiction in college students: an interview study.

OBJECTIVE: This study was aimed to evaluate the association between Internet addiction and depressive disorder, social phobia and adult attention-deficit/hyperactivity disorder (ADHD) in a sample of Taiwanese college students; and examine gender differences in the psychiatric comorbidity of Internet addiction in this student population. METHODS: Two hundred sixteen college students (132 males, 84 females) were recruited. Internet addiction, major depressive disorder, dysthymic disorder, social phobia, and adult ADHD of all participants were diagnosed based on psychiatric diagnostic interview. RESULTS: This study revealed that adult ADHD and depressive disorders were associated with Internet addiction among college students. However, depressive disorders were associated with Internet addiction in the males but not the females. CONCLUSION: With these results, it seems reasonable to suggest that effective evaluation of, and treatment for, adult ADHD and depressive disorders is required for college students with Internet addiction.

Independent 5' and 3'-end determination of multiple dihydrofolate reductase transcripts.

Multiple dihydrofolate reductase (dhfr) mRNAs, differing substantially in abundance, are produced as a result of the utilization of multiple transcription initiation sites and multiple polyadenylation sites. We have shown that dhfr mRNAs initiating from an upstream promoter region utilize the same collection of six polyadenylation sites and generate multiple dhfr mRNAs at the same relative abundance as do the mRNAs initiating from the major transcription promoter region. These results indicate that the 5' and 3' ends of dhfr mRNAs are independently determined. We show that the relative abundance of steady-state dhfr mRNAs was the same in nuclear and cytoplasmic RNA fractions. This finding makes it unlikely that differences in mRNA stability account for differences in the relative abundance of the multiple dhfr mRNAs in the cytoplasm. Our analysis of the dhfr promoter region revealed the existence of stable cytoplasmic polyadenylated transcripts complementary to the first 300 nucleotides of the dhfr transcripts initiating from the upstream promoter region. Therefore, the dhfr locus hosts two divergent and partially overlapping genes which share the same promoter region.

Dual bidirectional promoters at the mouse dhfr locus: cloning and characterization of two mRNA classes of the divergently transcribed Rep-1 gene.

The mouse dihydrofolate reductase gene (dhfr) is a housekeeping gene expressed under the control of a promoter region embedded in a CpG island--a region rich in unmethylated CpG dinucleotides. A divergent transcription unit exists immediately upstream of the dhfr gene which is coamplified with dhfr in some but not all methotrexate-resistant cell lines. We show that the promoter region for this gene pair consists of two bidirectional promoters, a major and minor promoter, which are situated within a 660-base-pair region upstream of the dhfr ATG translation initiation codon. The major promoter controls over 90% of dhfr transcription, while the minor promoter directs the transcription of the remaining dhfr mRNAs. The major promoter functions bidirectionally, transcribing a divergent 4.0-kilobase poly(A) mRNA (class A) in the direction opposite that of dhfr transcription. The predicted protein product of this mRNA is 105 kilodaltons. The minor promoter also functions bidirectionally, directing the transcription of at least two divergent RNAs (class B). These RNAs, present in quantities approximately 1/10 to 1/50 that of the class A mRNAs, are 4.4- and 1.6-kilobase poly(A) mRNAs. cDNAs representing both class A and class B mRNAs have been cloned from a mouse fibroblast cell line which has amplified the dhfr locus (3T3R500). DNA sequence analysis of these cDNAs reveals that the class A and class B mRNAs share, for the most part, the same exons. On the basis of S1 nuclease protection analysis of RNA preparations from several mouse tissues, both dhfr and divergent genes showed similar levels of expression but did show some specificity in start site utilization. Computer homology searches have revealed sequence similarity of the divergent transcripts with bacterial genes involved in DNA mismatch repair, and we therefore have named the divergently transcribed gene Rep-1.

The adaptive decision-making, risky decision, and decision-making style of Internet gaming disorder.

BACKGROUND: Persistent gaming, despite acknowledgment of its negative consequences, is a major criterion for individuals with Internet gaming disorder (IGD). This study evaluated the adaptive decision-making, risky decision, and decision-making style of individuals with IGD. METHODS: We recruited 87 individuals with IGD and 87 without IGD (matched controls). All participants underwent an interview based on the Diagnostic and Statistical Manual of Mental Disorders (5th Edition) diagnostic criteria for IGD and completed an adaptive decision-making task; the Preference for Intuition and Deliberation Scale, Chen Internet Addiction Scale, and Barratt Impulsivity Scale were also assessed on the basis of the information from the diagnostic interviews. RESULTS: The results demonstrated that the participants in both groups tend to make more risky choices in advantage trials where their expected value (EV) was more favorable than those of the riskless choice. The tendency to make a risky choice in advantage trials was stronger among IGD group than that among controls. Participants of both groups made more risky choices in the loss domain, a risky option to loss more versus sure loss option, than they did in the gain domain, a risky option to gain more versus sure gain. Furthermore, the participants with IGD made more risky choices in the gain domain than did the controls. Participants with IGD showed higher and lower preferences for intuitive and deliberative decision-making styles, respectively, than controls and their preferences for intuition and deliberation were positively and negatively associated with IGD severity, respectively. CONCLUSIONS: These results suggested that individuals with IGD have elevated EV sensitivity for decision-making. However, they demonstrated risky preferences in the gain domain and preferred an intuitive rather than deliberative decision-making style. This might explain why they continue Internet gaming despite negative consequences. Thus, therapists should focus more on decision-making styles and promote deliberative thinking processes to mitigate the long-term negative consequences of IGD.

N-nitrosophenacetin: its synthesis, characterization, mutagenicity, and teratogenicity.

Reaction of phenacetin (CAS: 62-44-2; p-acetophenetidide) with nitrous fumes (N2O3) in glacial acetic acid at 0-5 degrees C yields N-nitrosophenacetin (NP), 2-nitrophenacetin, N-nitroso-2-nitrophenacetin (NNP), and other compounds. Both NP and NNP are fairly stable at low temperature (-30 degrees C) but extremely labile at ambient temperature. NP (median lethal dose to Sprague-Dawley rat: 21 mg/kg body wt) is 80 times more toxic than its parent compound phenacetin and is directly mutagenic to bacterial cells including Salmonella typhimurium and Sarcina lutea. The mutagenicity of NP is comparable to that of N-methyl-N'-nitro-N-nitrosoguanidine [(MNNG) CAS: 70-25-7; 1-methyl-3-nitro-1-nitrosoguanidine] and requires no microsomal metabolic activation. The teratogenic potential of NP was studied in White Leghorn chick embryos given a single dose of 5-15 micrograms/egg on day 6 of incubation. A low incidence of exencephaly and eyelid defect and a high incidence of feather and claw malformations were found in the treated group; no such malformed embryos were found in the control group. The teratogenicity of NP was found to be weaker than that of MNNG, but stronger than that of N-methyl-N-nitrosourea (CAS: 684-93-5), dimethylnitrosamine (CAS: 62-75-9; N-nitrosodimethylamine), and diethylnitrosamine (CAS: 15-18-5; N-nitrosodiethylamine).

Treatable domain and optimal frequency for brain tumors during ultrasound hyperthermia.

PURPOSE: To examine the optimal ultrasound frequency and the treatable domain determined by the tumor size and tumor depth when an external ultrasound heating system is employed for the brain tumor hyperthermia. METHODS AND MATERIALS: This work employs a simplified model of a scanned ultrasound transducer power deposition (a cone with convergent/divergent shape) and a search algorithm to investigate the optimal frequency and the treatable domain. The distributions of temperature and SAR (specific absorption rate) ratio are used to determine the appropriateness of the acoustic window size and the input power level for a yielded set of tumor conditions. The factors considered are the acoustic window size, tumor size and depth, ultrasound frequency, and the acoustic absorption of the post-target bone behind the tumor. RESULTS: Simulation results demonstrate that the optimal frequency depends on the tumor depth and the acoustic absorption of the post-target bone. However, it is almost independent of the acoustic window size. The optimal frequency shifts to a higher level for a deeper tumor heating to reduce the effect of the high acoustic absorption of post-target bone. Moreover, the treatable domain is proportional to the acoustic window size and related to the ultrasound frequency. CONCLUSION: It may not be possible to deliver appropriate ultrasonic energy to heat a brain tumor without overheating the normal brain tissue and/or the post-target bone under the constraints of the available acoustic window size for the ultrasonic beam, ultrasonic attenuation of brain tissue, high absorption of post-target bone, and high blood perfusion rate. The results of this study can be a guideline for designing an optimal ultrasound heating system, arranging the transducers, and implementing further treatment planning for the brain tumor hyperthermia.

A theoretical study of cylindrical ultrasound transducers for intracavitary hyperthermia.

PURPOSE: The purpose of this paper was to examine the heating patterns and penetration depth when a cylindrical ultrasound transducer is employed for intracavitary hyperthermia treatments. METHODS AND MATERIALS: The present study employs a simulation program based on a simplified power deposition model for infinitely long cylindrical ultrasound transducers. The ultrasound power in the tissue is assumed to be exponentially attenuated according to the penetration depth of the ultrasound beam, and a uniform attenuation for the entire treatment region is also assumed. The distribution of specific absorption rate (SAR) ratio (the ratio of SAR for a point within the tissue to that for a specific point on the cavity surface) is used to determine the heating pattern for a set of given parameters. The parameters considered are the ultrasound attenuation in the tissue, the cavity size, and the transducer eccentricity. RESULTS: Simulation results show that the ultrasound attenuation in the tissue, the cavity size, and the transducer eccentricity are the most influential parameters for the distribution of SAR ratio. A low frequency transducer located in a large cavity can produce a much better penetration. The cavity size is the major parameter affecting the penetration depth for a small cavity size, such as interstitial hyperthermia. The heating pattern can also be dramatically changed by the transducer eccentricity and radiating sector. In addition, for a finite length of cylindrical transducer, lower SAR ratio appears in the regions near the applicator's edges. CONCLUSION: The distribution of SAR ratio indicates the relationship between the treatable region and the parameters if an appropriate threshold of SAR ratio is taken. The findings of the present study comprehend whether or not a tumor is treatable, as well as select the optimal driving frequency, the appropriate cavity size, and the eccentricity of a cylindrical transducer for a specific treatment.

Optimization of power deposition and a heating strategy for external ultrasound thermal therapy.

The purpose of this paper is to examine the thermal dose distribution, to configure the optimal absorbed power deposition, and to design an appropriate heating strategy for ultrasound thermal therapy. This work employs simulation programs, which are based on the transient bio-heat transfer equation and an ideal absorbed power deposition or an ideal temperature elevation within a cube of tissue, to study the optimal absorbed power deposition. Meanwhile, a simplified model of a scanned ultrasound transducer power deposition (a cone with convergent/divergent shape) is used to investigate the heating strategy for a large tumor with a sequence of heating pulses. The distribution of thermal dose equivalence defined by Sapareto and Dewey is used to evaluate the heating result for a set of given parameters. The parameters considered are the absorbed power density, heating duration, temperature elevation, blood perfusion, and the size of heating cube. The results demonstrate that the peak temperature is the key factor determining the thermal dose for this short-duration heating. Heat conduction has a very strong influence on the responses of temperature and thermal dose for a small heating cube and the boundary portion of a large heating cube. Hence, for obtaining the same therapeutic result, a higher power density is required for these two conditions to compensate the great temperature difference between the heating cube and the surrounding tissue. The influence of blood perfusion on the thermal dose is negligible on the boundary portion of the heating cube, while in the central portion it may become a crucial factor as a lower power density is used in this portion to save the delivered energy. When using external ultrasound heating method to treat a large tumor, the size of heating unit, the sequence of heating pulses, and the cooling-time interval between the consecutive heating pulses are the important factors to be determined to have an appropriate treatment within a reasonable overall treatment time.

Specific absorption rate ratio patterns of cylindrical ultrasound transducers for breast tumors.

The purpose of this paper is to examine the optimal driving frequency and to configure the ultrasound energy deposition schema for a various size and location of breast tissues when a portion or the entire cylindrical ultrasound transducer is employed for breast hyperthermia treatments. This work employs a computer simulation program based on an ideal ultrasound power deposition from a cylindrical transducer. The ultrasound power within the breast is assumed to be exponentially attenuated according to the penetration depth of the ultrasound beam and a uniform absorption for the entire breast is also assumed. The distribution of the specific absorption rate (SAR) ratio is employed to determine the heating pattern of a set of given parameters. The control parameters considered are the ultrasound frequency in the breast tissue, the active portion of cylindrical transducer, and the shifting distance between the central axes of the breast and the transducer. The effect of the breast size on the SAR ratio is also considered. Simulation results demonstrate that the breast size, the ultrasound frequency in breast tissue, the shifting distance, and the active portion of the cylindrical transducer are the potential parameters for influencing the distribution of the SAR ratio. High frequencies should be used for the superficial heating treatments and the active portion of the transducer can be changed to obtain a region with an appropriate SAR ratio to cover the treatment region. Low frequencies are used for deep heating treatments and the region of the high SAR ratio can be moved by shifting the transducer and its pattern is varied with the transducer's active portion. The distribution of the SAR ratio indicates the domain of treatable tumor size and tumor depth for a given set of parameters (driving frequency, shifting distance and active portion of the transducer, as well as breast diameter). Findings of this study can be used to know whether or not the tumor is treatable as well as to select the optimal driving frequency and the appropriate active portion of the cylindrical transducer for a treatment, and hopefully to design an appropriate cylindrical ultrasound heating system for breast tumors.

Relationship between acoustic aperture size and tumor conditions for external ultrasound hyperthermia.

External ultrasound hyperthermia is a very flexible modality for heating deep-seated tumors due to its deep penetration and focusing ability. However, under the constraints of the available acoustic aperture size for the ultrasonic beam, ultrasonic attenuation, as well as other anatomic properties, it may not be able to deliver sufficient ultrasonic energy to heat a large tumor located in a deep region without overheating the normal tissue between the tumor and the aperture. In this work, we employ a simulation program based on the steady-state bioheat transfer equation and an ideal ultrasound power deposition (a cone with convergent/divergent shape) to examine the relationship between the minimal diameter of the acoustic aperture and the tumor conditions. Tissue temperatures are used to determine the appropriate aperture diameter and the input power level for a given set of tumor conditions. Due to the assumed central axis symmetry of the power intensity deposition and anatomic properties, a two-dimensional (r-z) simulation program is utilized. Factors determining the acoustic aperture diameter and the input power level considered here are the tumor size, tumor depth, ultrasonic attenuation in tissue, blood perfusion, and temperature of the surface cooling water. Simulation results demonstrate that tumor size, tumor depth, and ultrasonic attenuation are major factors affecting the aperture diameter of the ultrasonic beam to obtain an appropriate temperature distribution, while blood perfusion and the temperature of the surface cooling water are the minor factors. Plots of the effects of these factors can be used as the guideline for designing an optimal ultrasound heating system, arranging the transducers, and planning further treatments.

Self-tuning fuzzy logic control for ultrasound hyperthermia with reference temperature based on objective functions.

The purpose of this paper is to develop and evaluate a self-tuning fuzzy logic controller for a scanned focused ultrasound hyperthermia system with the reference temperature (Tr) determined from objective functions. This work employs simulation programs to develop the power deposition for the scanned focused ultrasound system and to solve the responses of temperature profiles based on the transient bioheat transfer equation. A fuzzy logic control algorithm is employed to determine the output power level for the heating system and an observer for blood perfusion variation is used to enhance the capability of the controller to adjust the required output power level for the treatment due to the drastic change of the blood perfusion. The reference temperature (Tr) for the controller is based on objective functions to tune its value during the heating process, while a control temperature (Tc) from the thermosensors located in the tumor region is used as the input for the controller. The objective function based on the entire temperature profile is used to evaluate the appropriateness of the heating temperature distribution for a time-variational blood perfusion. Simulation results demonstrate that the tumor region can be rapidly heated to the desired temperature level and maintained at that level despite blood perfusion variation. The resulting temperature profile, the objective function, and the output power level are related to the magnitude of blood perfusion, but are almost independent of the Tc location and the initial setting value of Tr. The fuzzy logic control algorithm with Tr determined from objective functions can be used for controlling the entire temperature distribution through a single control temperature, and the combination of control and optimization allows appropriate temperature fields to be created during the entire heating process. The control algorithm does not require the accurate prior knowledge of the locations of the thermosensors and the appropriate setting value for Tr.

Optimal configuration of multiple-focused ultrasound transducers for external hyperthermia.

External ultrasound hyperthermia is considered to be a very flexible modality for heating deep-seated tumors owing to its penetration and focusing ability. However, using this flexibility requires that many complicated, interacting decisions be made to obtain optimal treatment. This paper presents the feasibility of arranging multiple-focused ultrasound transducers to produce an appropriate heating pattern for a specific treatment, based on the optimal scan parameters obtained from an optimization algorithm. The variable scan parameters of the heating system optimized are the transducer tilt and rotation angles, focal depth, scan radius, and output acoustical power. After obtaining the optimal scan parameters, multiple transducers are systematically arranged according to these scan parameters. Three-dimensional ultrasound power deposition and temperature distribution for a specific treatment are calculated for this multiple ultrasound transducer system. A more uniform temperature distribution in the treatment region for a large, highly perfused tumor can be achieved by scanning the system with respect to the central scan axis and/or swinging the transducers inwards and outwards. The maximum heating depth of focused ultrasound transducers used in this heating system is also studied. Simulation results demonstrate that the optimal arrangement of this multiple-focused ultrasound transducer system is highly promising for heating deep, large, and highly perfused tumors.

Inverse error-diffusion using classified vector quantization.

This correspondence extends and modifies classified vector quantization (CVQ) to solve the problem of inverse halftoning. The proposed process consists of two phases: the encoding phase and decoding phase. The encoding procedure needs a codebook for the encoder which transforms a halftoned image to a set of codeword-indices. The decoding process also requires a different codebook for the decoder which reconstructs a gray-scale image from a set of codeword-indices. Using CVQ, the reconstructed gray-scale image is stored in compressed form and no further compression may be required. This is different from the existing algorithms, which reconstructed a halftoned image in an uncompressed form. The bit rate of encoding a reconstructed image is about 0.51 b/pixel.

Pilot point temperature regulation for thermal lesion control during ultrasound thermal therapy.

The fundamental goal of ultrasound thermal therapy is to provide proper thermal lesion formations for effective tumour treatment. The quality of the therapy depends mostly on its positional precision. To date, most ultrasound thermal therapy treatments have focused on the formation of power or temperature patterns. The non-linear and time-delay effects of thermal dose formation prohibit direct control of the thermal dose distribution. In the paper, the control of thermal lesions by regulation of the temperature of a pilot point is proposed. This scheme utilises the high correlation between temperature elevation and thermal dose at the forward boundary of thermal lesions. To verify the feasibility, a 2D ultrasound phased array system was used to generate thermal lesions of various sizes, and the temperature elevation required to generate a thermal dose threshold was investigated. Results showed that the required temperature elevation was found to be a reasonably constant value of 52.5 degrees C under differing conditions when the focal area was small. When the focal area under consideration was large, the required temperature elevation became a monotonic function of blood perfusion rate, ranging from 49.2 to 52.5 degrees C. When the reference temperature of the pilot point was set at a conservative value (52.5 degrees C), the thermal lesions were controlled precisely under a wide range of blood perfusion and power pattern changes, tested by using a more realistic model that takes into account thermal-induced attenuation and blood perfusion changes. This changed the complex thermal dose control problem into a simple temperature regulation problem, which makes implementation of thermal lesion control easier, giving the scheme a high potential for application to current ultrasound thermal therapy systems.

The association between Internet addiction and psychiatric disorder: a review of the literature.

Internet addiction is a newly emergent disorder. It has been found to be associated with a variety of psychiatric disorders. Information about such coexisting psychiatric disorders is essential to understand the mechanism of Internet addiction. In this review, we have recruited articles mentioning coexisting psychiatric disorders of Internet addiction from the PubMed database as at November 3, 2009. We describe the updated results for such disorders of Internet addiction, which include substance use disorder, attention-deficit hyperactivity disorder, depression, hostility, and social anxiety disorder. We also provide discussion for possible mechanisms accounting for the coexistence of psychiatric disorders and Internet addiction. The review might suggest that combined psychiatric disorders mentioned above should be evaluated and treated to prevent their deteriorating effect on the prognosis of Internet addiction. On the other hand, Internet addiction should be paid more attention to when treating people with these coexisting psychiatric disorders of Internet addiction. Additionally, we also suggest future necessary research directions that could provide further important information for the understanding of this issue.

Areca quid chewing and methamphetamine use in Taiwanese adolescents.

The association between areca quid chewing and methamphetamine (MAMP) use in Taiwanese adolescents was examined in 200 MAMP users and 400 non-users. Subjects were interviewed individually regarding the experience of chewing areca quid and the evidence of lifetime areca quid use disorder. The ages at which they initially chewed areca quid and used MAMP were also evaluated. The results revealed that areca quid chewing and areca quid use disorders were more common in both male and female MAMP users than in non-users. Among those who chewed areca quid and used MAMP, the mean age at initial MAMP use was older than the mean age at initial chewing of areca quid in males but not in females. MAMP users who chewed areca quid were subdivided into three groups according to the sequence of initial MAMP use and chewing areca quid. Although most males and females started chewing areca quid before using MAMP, this pattern was more prevalent in males. Adolescent MAMP users need to be educated about deleterious health sequelae caused by chewing areca quid. Adolescent areca quid chewers need to be monitored for possible progression to MAMP use.

Breast cancer education program based in Asian grocery stores.

Culturally and linguistically compatible university students were trained as community health educators to provide breast cancer education and screening information to shoppers at Asian grocery stores. Information about early detection of breast cancer was shared with 8,877 women, who reported speaking 40 different languages. Baseline surveys were completed by 1,202 women; 779 took part in the follow-up survey. The survey questions assessed baseline knowledge, attitudes, and screening behaviors regarding breast cancer, tested the efficacy of the intervention, and sought barriers to accessing screening services. Screening adherence at baseline was low, but reported screening compliance had increased by follow-up. This study confirms the cost-effectiveness of student health educators and Asian grocery store sites as venues to reach the diverse age, ethnic, and socioeconomic segments of the Asian community, while demonstrating the community's receptiveness to the dissemination of health information and introducing bilingual students to health education and research careers.

Identification of functional murine adenosine deaminase cDNA clones by complementation in Escherichia coli.

Total poly(A+) RNA derived from a mouse cell line with amplified adenosine deaminase genes was used as template to synthesize double-stranded cDNA. The cDNAs were inserted into the PstI site of the beta-lactamase gene in plasmid pBR322 following G-C tailing. After transformation into adenosine deaminase-deficient Escherichia coli hosts, recombinant plasmids containing functional murine adenosine deaminase cDNAs were identified by selecting for functional complementation. Analysis of plasmids containing functional adenosine deaminase cDNA sequences strongly suggested that adenosine deaminase expression resulted mainly from beta-lactamase/adenosine deaminase fusion proteins even when the adenosine deaminase codons were out-of-frame with respect to the beta-lactamase gene codons upstream. The nucleotide sequence of a 1.65-kilobase pair cDNA insert in one of the functional recombinant clones was determined and found to contain a 1056-nucleotide open reading frame. When this 1056-nucleotide open reading frame was inserted into a mammalian expression vector and introduced into monkey kidney cells, a high level of authentic mouse adenosine deaminase was produced. Nucleic acid blot analysis using a full-length adenosine deaminase cDNA clone as probe revealed that the mouse adenosine deaminase structural gene was at least 21 kilobase pairs in size and encoded three polyadenylated mRNAs. Analysis of the cDNA library from which the functional clones were isolated suggested that this approach of cloning functional mammalian adenosine deaminase cDNA clones by genetic complementation of enzyme-deficient bacteria could be accomplished even if the abundance of the adenosine deaminase mRNA sequences were as low as approximately 0.001%.