Empirical evidence has suggested that drug treatment courts (DTCs) reduce re-arrest rates. However, DTC program completion rates are low and little is known about the effectiveness of lower levels of program participation.We examined how DTC program referral, enrollment without completion, and completion, affected re-arrest rates during a two-year follow-up.We used statewide North Carolina data from criminal courts merged with DTC data. Propensity score matching was used to select comparison groups based on demographic characteristics, criminal histories, and drug of choice (when available). Average treatment effects on the treated were computed.DTC participation levels included referral without enrollment, , enrollment without completion (n = 954), and completion (n = 747). Recidivism measured as re-arrest on a substance-related charge, on a violent offense charge not involving an allegation of substance abuse, and on any charge (excluding infractions) was examined by felony and misdemeanor status during a two-year follow-up period.Re-arrest rates were high, 53–76 percent. In general, re-arrest rates were similar for individuals who were referred but who did not enroll and a matched comparison group consisting of individuals who were not referred. In contrast, enrollees who did not complete had lower re-arrest rates than a matched group of individuals who were referred but did not enroll, for arrests on any charge, on any felony charge, and on substance-related charges (felonies and misdemeanors). Finally, relative to persons who enrolled but did not complete, those who completed had lower re-arrest rates on any charge, any felony charge, any misdemeanor charge, any substance-related charge, any substance-related misdemeanor or felony charge, and any violent felony charge.Enrolling in a DTC, even without completing, reduced re-arrest rates. Given the generally low DTC completion rate, this finding implies that only examining effects of completion underestimates the benefits of DTC programs. The link between substance use and crime is longstanding and well documented. Among state prisoners, estimates of drug use at the time of a criminal offense are as high as 30 percent and more than half had used drugs in the month prior to arrest. The criThere have been numerous evaluations of the effectiveness of DTCs in reducing recidivism and substance use–7. OveraEvaluations of DTCs based on observational data frequently have involved matched comparison groups and have used propensity score matching in particular–21. GiveUsing longitudinal statewide data from North Carolina, this study examined whether being referred to, enrolling in, and/or completing a DTC program changed behavior for a criminal offender with a substance use problem. Our study offered several advantages over previous studies of DTCs. First, we examined differences of re-arrest rates between three groups of DTC participants, those who (1) were referred but did not enroll, (2) enrolled but did not complete and (3) completed. We hypothesized that, holding other factors constant, higher levels of participation in a DTC program would be associated with reduced recidivism. That is, completing a DTC program would be more effective than enrolling in but not completing the program and enrolling in the program would be more effective than just being referred. Simply being referred to or enrolling in a DTC program may prevent future criminal activity by exposing the prosecuted or convicted person to available community resources for individuals with addictions. This is particularly relevant since far greater numbers of persons are referred to DTCs than who actually enroll, and many more persons enroll without completing. Yet, exSecond, compared to previous studies, we enhanced the generalizability of findings by including data from 19 DTCs in a populous state, North Carolina (NC), during 2005–2012. Third, our analysis used linked administrative data rather than self-reports from sample surveys. Fourth, we examined effects of DTC programs on recidivism for all types of crimes, not only those directly related to substance use or trade in illicit substances.DTCs aim to reduce the incidence of drug and alcohol-related crimes (N.C. Gen. Stat. § 7A-791). Judges, prosecutors, defense attorneys, and probation officers may refer participants to a DTC. Enrollment in DTCs in North Carolina is voluntary, and DTC referral occurs post plea. These programs have three phases, even though attendance and treatment requirements for each phase vary across courts. Further, while each individual court has the discretion to determine which type of sanctions and rewards to issue, the types of rewards and sanctions used by the courts are consistent among all DTCs in the state. For example, the most common sanctions in fiscal year 2008–9 were jail for 24–48 hours (35%), individualized sanction (16%), and community service (6%).Multiple factors may encourage or discourage program completion. One factor that may encourage program completion is suspended sentences whereby offenders who complete the program do not serve their original sentence. In North Carolina, if a sentence is suspended pending successful completion of a DTC, failure to complete the DTC program will result in reinstatement of the original sentence. On the other hand, strict court rules may discourage completion. For example, participants who test positive for drugs while in the program, or who fail to attend court as scheduled may be terminated from the program at any time. Because participation is voluntary, participants may choose to discontinue their participation at any time. Completion of a DTC program may take a year or more. Among those that we observed to have completed the program, the mean length of time to completion was 435 days .The most common type of DTC is the adult drug treatment court. In these, referrals are not limited to persons convicted of a substance-related crime. The general eligibility requirements are the same among all DTCs in North Carolina. To be considered for referral, individuals need to be: addicted to a chemical substance; willing to volunteer for the drug treatment court program, and eligible under the structured sentencing system for a community or intermediate punishment as an alternative to active prison time. However, each county drug program may impose additional eligibility criteria.Another type of DTC, the DWI court, focuses on individuals arrested for DWIs. In North Carolina, DWI courts focus on persons convicted of a DWI who have been sentenced to supervised probation and who were found to have had at least one of the following aggravating factors: (1) driving while license revoked as a result of DWI, (2) causing injury to another person as a result of DWI, or (3) DWI with a child under the age of 16 in the vehicle.This study relied on two administrative databases from the NC Administrative Office of the Courts. The first was the Drug Treatment Court Management Information System (DTC-MIS) database, which tracks individuals referred, enrolled, and graduated from the 19 state-funded drug treatment court programs. The second database, the Automated Criminal Infraction System (ACIS), contained information on all arrests for felonies, misdemeanors, infractions, and traffic offenses in the state from 2005 through 2012. Infractions, which in North Carolina are non-criminal offenses not punishable with jail time , were excluded from the analyses.Information from the ACIS was merged with the DTC-MIS data to document an individual’s court involvement. The DTC record was merged with the most recent ACIS record prior to DTC referral. In total, 3,875 of 5,257 DTC-MIS referrals to DTC and ACIS records were merged (73.7%). Failure to merge may reflect recording errors, e.g., name changes, misspelled names, or errors in data entry of birthdates.The observational unit was a person-index arrest. For persons never referred to a DTC, index arrests were defined as the first criminal arrest in a calendar year. An individual could thus have up to four index arrests in the sample, one for each year 2007–2010. For persons referred to a DTC, the index arrest was defined as the arrest corresponding to referral. The analyses examined three types of involvement with the courts: being referred but not enrolling in a DTC , enrolling in a DTC but not completing the program (n = 954), and completing the DTC program (n = 747). The presence of a referral date in the DTC data identified individuals who were referred. Enrollment was determined by the presence of a recorded admission date, treatment phase-one entry date, or a discharge date. Program completion was determined by satisfying one of the following conditions: (1) the person completed Phase 3 treatment, the final phase of active treatment, (2) the person entered Phase 4 treatment, aftercare, or another post-completion program, or (3) the discharge record indicated the person completed the program.Re-arrest during a two-year follow-up period was the outcome measure. Binary variables indicated if the person was arrested for: (1) a substance-related charge; (2) a charge involving a violent offense excluding those that co-occurred with a substance-related crime, which was infrequent—only 2.7 percent of violent offenses co-occurred with a substance-related crime; and (3) on any charge, excluding infractions.Offenses associated with the index offense accounted for the fact that individuals often incurred several charges on the same day. We developed a hierarchy based on the assumption that substance abuse charges were most likely to directly lead to a referral to a DTC. In descending order, the hierarchy consisted of charges for the following offenses: both a drug and a DWI; a DWI only; drug only; violent crime; alcohol (non-DWI); traffic; and other. We further subdivided these categories into arrests on felony or misdemeanor charges, with felony given a higher priority. For example, if a person had both a drug felony and a violent misdemeanor charge for the index arrest, the index arrest was classified as a drug felony. This same hierarchy was used to classify arrests for offenses that occurred in the two-year look-back period.Drug of choice was classified into mutually exclusive variables representing: alcohol; cocaine-powder; crack; heroin; other narcotics; other or no drug of choice; and a binary variable for missing values. This information was not available for individuals who were not referred to a DTC program.Our statistical approach compared differences in criminal recidivism across groups using propensity score matching (PSM). PSM is a statistical approach commonly used in observational studies where it is not feasible to randomly assign individuals to a treatment condition. RandomiThe three treatment groups considered in this study were those who: (1) were referred but did not enroll; (2) enrolled but did not complete; and (3) completed. To help mitigate issues of heterogeneity of characteristics across groups, the comparison group for each treatment group was drawn from individuals who were most likely to match the treatment group. Comparison groups were drawn from individuals with the next lowest level of participation. Specifically, to compare outcomes with those who were referred but did not enroll with individuals who were not referred, a matched comparison sample was drawn from the 19 counties that had a DTC program. This was done in an effort to balance county characteristics that could have affected re-arrest rates such as resources dedicated toward policing. For the analysis of the effects of enrollment, the comparison group consisted of persons who were referred but did not enroll. Finally, to analyze the effects of completion, the comparison group consisted of persons who were enrolled but did not complete the program.The propensity score for the probability of referral to a DTC was modeled using a logistic regression. One-to-one nearest neighbor matching with a caliper of 0.01 was done using psmatch2 in Stata version 11, 31. TreTo assess the quality of the match, we calculated standardized differences of each covariate before and after matching. A standardized difference compares mean differences in units based on the pooled (treatment plus comparison) standard deviation. The stawhereBefore matching, persons referred to a DTC program differed from the larger population of individuals who had been arrested but who were not referred, as indicated by many standardized differences greater than 10 percent .In the matched comparison sample, nearly three-quarters of individuals who were referred to a DTC program but who did not enroll were re-arrested for an offense (other than an infraction) during the two-year follow-up period Table . The onlt = 4.27, d.f. = 1,772, p < 0.001). Nearly all of this reduction was attributable to a reduction in arrests on felony charges, . The ATT for substance-related arrests was −10.7 percentage points , indicating that those who enrolled but did not complete were statistically significantly less likely to be re-arrested for a substance-related offense than individuals who were referred but who did not enroll. These groups were not statistically significantly different on re-arrest rates for violent offenses.Those who enrolled but did not complete were less likely to be re-arrested for any offense (other than an infraction) during the two-year follow-up than the matched sample of individuals who were referred but did not enroll. Two-thirds of individuals who enrolled but did not complete were re-arrested within two years relative to three-quarters of persons referred but who never enrolled . Even so, half of those who completed a DTC program were re-arrested on some charge, not including infractions, within two years. Those who completed a DTC program were also less likely to be re-arrested for a substance-related offense than the matched comparison sample of individuals who enrolled in a DTC program but did not complete . Nearly all of this difference was attributable to a reduction in substance-related felony arrests in the group who completed a DTC program. While rare in both groups, a smaller percentage of those who completed the DTC program were re-arrested for a violent felony offense than the matched comparison group of individuals who enrolled but did not complete; the ATT for this comparison was −1.3 percent .Program completion was also effective in reducing recidivism as measured by any re-arrest, and the difference in the matched sample between completion and enrolling without completion was 11.2 percentage points (We found substantial reductions in rates of re-arrest among both persons who enrolled in a DTC program but did not complete and those who completed a DTC program. In contrast, just referring a person to a DTC did not reduce recidivism rates for any of the outcomes examined in this study. Importantly, while completion was beneficial in terms of reducing re-arrest, even enrolling without completion led to a substantial improvement in re-arrest rates during the two years following the date of the index arrest. Most prior studies focused solely on differences between graduates and dropouts. Enrollment in a treatment court program substantially decreased recidivism rates for many types of crime, particularly for substance-related crimes.Many DTC programs across the country do not accept persons convicted of a violent crime. Nevertheless, we documented a 1.3 percentage point decrease in re-arrest for violent felony offenses when comparing those who completed a DTC program with those who enrolled but did not complete, which implies that reduced violent crime was a positive externality of a completed substance abuse treatment program. Substance use is a risk factor for serious violent crimes such as domestic violence, child abuse, assault, and murder. DocumenDespite the positive impact of treatment courts on recidivism, participation in these courts was low. Fewer than 5,000 individuals were referred to a treatment court program over a four-year window for the entire state as contrasted with over a quarter million people who were arrested for a substance-related offense during that same time period. In our sample of participants, only 44 percent of referred persons enrolled and only 19 percent completed. This was lower than completion rates of 27–66 percent documented in other studies. Our stuOur study had several strengths. First, the analysis applied to all state-funded DTCs, not just to individual treatment courts or to courts that agreed to participate in a research study. Second, administrative data offer advantages over survey data based on self-reports. Not only are the samples generally much larger, but administrative data are not subject to recall bias as are self-reported data on arrests and disposition of the arrests. Third, our analysis considered effects of DTCs separately for various stages in the completion process. We found no effects for just being referred, indicating that some treatment is a prerequisite for program effectiveness.We acknowledge several limitations. First, we relied exclusively on administrative data, which only included a few participant characteristics. This omission potentially adversely affected the quality of the matches between treatment and comparison groups. There may be unobserved differences in such factors as financial circumstances, social support, motivation to change behavior, and underlying comorbid mental health or addiction disorders. Such differences may have persisted across our study groups based on DTC participation levels—for example those who were referred but did not enroll may have been less likely than referred individuals who enrolled to believe that treatment services would benefit them. Similarly those who completed a DTC program may have had better support networks to facilitate completion relative to individuals who enrolled but did not complete. However, complete, reliable, longitudinal information on court participation is rarely available to researchers. Although detailed surveys of alcohol and illicit drug use have been conducted, these surveys lack questions about arrests, arrest disposition, and respondent involvement in DTC programs.Second, we analyzed DTCs in North Carolina as a group rather than explicitly account for differences in organizational characteristics. While several studies have focused on individual factors such as gender and employment that affect completion–38, few A third limitation involved the use of official state records to document recidivism. This measure may underreport criminal recidivism rates as individuals who engage in illegal activities are often not arrested, 41. RelFourth, results from one state may not generalize to the U.S. as a whole. While many DTCs in the U.S. follow the model set out by the National Drug Court Institute and the National Association of Drug Court Professionals, there are important variations in implementation, e.g., in the sanctions imposed on those who fail to complete the program, 43.These findings are relevant in light of ongoing policy debates about specialty court funding. The soc


Few clues were found in the literature about the independent risk factors for PTSD among earthquake survivors in Sichuan province three years after the 2008 earthquake. Ours was the first case-control study with matching factors of age and distance from the epicenter among survivors age 16 years or older, three years after the catastrophe.To identify independent risk factors for PTSD among earthquake survivors.We performed a population-based matched case-control study. The cases were drawn from earthquake areas three years after the Wenchuan earthquake, including 113 cases who met positive criteria for PTSD symptoms according to the PCL-C (PTSD Checklist-Civilian Version) score and 452 controls who did not meet the criteria. Cases and controls were matched individually by birth year (+ three years) and the town they lived in when the earthquake occurred.Independent risk factors for PTSD symptoms included two-week disease prevalence , witnessing someone being killed in the earthquake , having no regular income after the earthquake , receiving mental health support only one time after the earthquake  and lower social support (lower PSSS score) .Earthquake experience, suffering from physical illnesses, lack of stable income, and lower social support were associated with PTSD symptoms. The Ms 8.0 Wenchuan earthquake of May 12, 2008—the strongest earthquake since the establishment of People's Republic of China (PRC)—has caused great life and financial losses. Tens of thousands of people lost their homes and families. According to the Ministry of Civil Affairs, as of August 25, 2008, there were 69 226 people killed, 374 643 injured, and 17 923 missing. Immediately after the disaster, the Chinese government took several positive measures, such as providing financial support from the central government and 19 local provinces for reconstruction of the destroyed areas. Post-disaster reconstruction, planned to be complete within three years, was actually finished ahead of schedule in May 2011, according to claims by Wen Jiabao, the Prime Minister of China People who have survived a catastrophe such as an earthquake may develop psychological trauma and psychiatric disease, including post-traumatic stress disorder (PTSD). PTSD is a common mental health problem among earthquake survivors in ChinaWith the completion of post-Wenchuan earthquake reconstruction in 2011, the survivors' houses are equal to or even better than their house before the disaster, but current evidence reveals that there is still a high prevalence of PTSD among survivors. To provide appropriate interventions for reducing the occurrence of PTSD, we must identify the risk factors for the mental impairment. Several previous articles showed some possible contributing factors for disaster related PTSD which could be divided into five categories: (1) demographic variables, such as female, younger age, lower socioeconomic status, minority, educational and intellectual disadvantage Unfortunately, little evidence coming from local communities revealed the contributors of chronic PTSD, and most of the previous PTSD-related studies were cross-sectional, focusing on special populations such as students, injured, soldiers, and doctors and nurses who were involved in earthquake relief efforts. Therefore, we designed this case control study to identify independent risk factors for PTSD. To reveal the personal factors and factors directly related to the earthquake experience and relief, this study focused on demographic, earthquake related and post-earthquake variables.The Institutional Review Board (IRB) of West China Hospital in Sichuan University approved this study. Due to the illiteracy of many of the people in the earthquake-affected areas, written consent is not common practice and may violate confidentiality. Therefore, a consent form to obtain verbal consent from respondents was proposed and approved by the IRB, together with the study protocol. Prior to the interview, each investigator read carefully to the interviewee the consent form, which contains information on the objectives of the study, the selection process, risks, benefits and freedom of the participation, as well as information on confidentiality This was a matched case-control study, and the sample was selected from 22 townships of 6 counties in Sichuan province from May to July 2011, three years after the Wenchuan earthquake. More detailed sampling information is available in Jin's paper The PTSD Checklist-Civilian Version (PCL-C) was used to screen for symptoms of earthquake related PTSD. PCL-C is a 17-item self-report symptom scale that corresponds to DSM-IV criteriaGeneral demographic information and other factors ,having relatives injured or killed during the earthquake, displacement after the earthquake, and the number of mental health support sessions received after the earthquake) were collected using a predefined questionnaire.Chronic disease was defined as at least one of the following illnesses Perceived social support was measured using Perceived Social Support Scale(PSSS) Our investigators consisted of professors and post-graduate students from West China Hospital and West China School of Public Health, Sichuan University. They were well-trained on the questionnaire and investigation techniques, including mutual communication with participants by dialect.Frequencies, percentages, means and standard deviations were calculated for descriptive analysis. Conditional logistic regression analysis was used to compare cases with their matched controls for PTSD risk factors in both univariate and multivariate analysis. Odds ratio (OR) of the variables were assessed and 95% confidence intervals (CI) of ORs were calculated. The level of statistical significance was less than 0.05 in this study. Data were analyzed with SPSS version 17·0  and Stata version 10.0 .Of the 2525 eligible respondents, 113 cases were identified and 452 controls were individually matched to the cases . All of Sixteen factors were tested by univariate analysis as potential risk factors for PTSD. Being female, two-week disease prevalence, having first class relatives who died or went missing during the earthquake, having no regular income after the earthquake, displacement after the earthquake, receiving mental health support only one time after the earthquake, and lower PSSS score were all significantly associated with higher PTSD symptoms prevalence. Multivariable conditional logistic regression analysis was then performed. Two-week disease prevalence, witnessing someone being killed in the earthquake, having no regular income after the earthquake, receiving mental health support only one time after the earthquake, and lower PSSS score  were revealed to be independent risk factors for PTSD symptoms .This population-based case-control study, including 113 cases and 452 controls, demonstrated that a number of risk factors were responsible for PTSD. Two-week disease prevalence, witnessing someone being killed in the earthquake, having no regular income after the earthquake, receiving mental health support only one time after the earthquake, and lower social support were all identified to be independent risk factors for PTSD symptoms.Interestingly, receiving mental health only one time was associated with a 2.43-fold increase in risk for PTSD, while receiving no mental health support or two or more sessions of mental health support were without significant association with PTSD in either univariate analysis or multivariate analysis. This finding differs from other literatures that claim that victims with psychological help are less prone to express PTSDTwo-week disease prevalence was significantly associated with PTSD symptoms, which indicates people with poor physical health might be at risk for developing PTSD. Similar findings were revealed by another study Some studies reported that lower income is associated with poor mental health Perceived social support refers to the respondents' subjective sense of social support. A lower PSSS score means subjects felt less social support from family, friend and special person with whom one can share his or her joys and sorrows. Our study determined lower social support was a significant predictive factor for PTSD symptoms, which is consistent with other studies Some studiesSome recent published articles Certain limitations of the study should be recognized. First of all, we failed to analyze the impact of age and distance from the epicenter of all subjects' home because of matching. Second, although widely used in the world, PCL-C is a screening measurement of PTSD, not a diagnostic tool. PCL-C was used because in such a large population-based survey, it costs too much time and money to use a formal diagnostic tool. Hence, some of the control and cases subjects may be incorrectly grouped, which would lead to a bias in our results. Third, although we collected data on the number of times of mental health support received, many subjects could only give imprecise information because much time had passed since the earthquake. Therefore, we can only divide the times of mental health assistance into three categories instead of treating this variable as a continuous variable.In summary, this paper revealed risk factors for PTSD among adult earthquake survivors and estimated the magnitude of risk associated with each factor. Our study discussed the relationship between receiving mental health support and PTSD prevalence and provided further support for other risk factors. For many of these risk factors, having a regular income after the earthquake is a clear factor for preventing PTSD. There should be more consideration about providing more job opportunities for earthquake survivors in reconstruction efforts.


Saccharomyces cerevisiae and has been implicated in binding and catalytic enhancement of Top3 in the dissolution of double Holliday junctions. Deletion of RMI1 results in a severe growth defect resembling that of top3Δ. Despite the importance of Rmi1 for cell viability, little is known about its functional domains, particularly in Rmi1 of S. cerevisiae, which does not have a resolved crystal structure and the primary sequence is poorly conserved. Here, we rationally designed point mutations based on bioinformatics analysis of order/disorder and helical propensity to define three functionally important motifs in yeast Rmi1 outside of the proposed OB-fold core. Replacing residues F63, Y218 and E220 with proline, designed to break predicted N-terminal and C-terminal α-helices, or with lysine, designed to eliminate hydrophobic residues at positions 63 and 218, while maintaining α-helical structure, caused hypersensitivity to hydroxyurea. Further, Y218P and E220P mutations, but not F63P and F63K mutations, led to reduced Rmi1 levels compared to wild type Rmi1, suggesting a role of the C-terminal α-helix in Rmi1 stabilization, most likely by protecting the integrity of the OB-fold core. Our bioinformatics analysis also suggests the presence of a disordered linker between the N-terminal α-helix and the OB fold core; a P88A mutation, designed to increase helicity in this linker, also impaired Rmi1 function in vivo. In conclusion, we propose a model that maps all functionally important structural features for yeast Rmi1 based on biological findings in yeast and structure-prediction-based alignment with the recently established crystal structure of the N-terminus of human Rmi1.Rmi1 is a member of the Sgs1/Top3/Rmi1 (STR) complex of  In yeast the RecQ-like Sgs1 helicase forms a complex with the topoisomerase Top3 and the recQ-mediated genome instability 1 protein Rmi1 (STR) and facilitates both early and late steps of DNA double-strand break (DSB) repair –7. This ing Sgs1 ,11. Simiing Sgs1 .rmi1Δ mutant, the contribution of Rmi1 to the function of the STR complex and the regions of Rmi1 that are critical for these functional contributions are still poorly understood. Rmi1, was first discovered in S. cerevisiae in a screen for components of the Sgs1/Top3 pathway [rmi1Δ phenotypes, Rmi1 has no known catalytic function. It has been shown to stimulate the final decatenation step of dHJ dissolution by Sgs1/Top3, while inhibiting the relaxation activity of the topoisomerase on negatively supercoiled DNA, possibly by affecting the conformation of the topoisomerase gate [Despite the growth defect of the ase gate –15. Thisase gate –18.S. cerevisiae Rmi1, is capable of binding BLM and Topo IIIα, and contains an oligonucleotide-binding (OB) fold that is similar in structure to that of the replication protein A subunit RPA70, though it is suggested that it is incapable of binding DNA like RPA [The N-terminal 219 residues of the 625-residue long human Rmi1 have been crystallized, providing some clues to its role in catalytic enhancement and stability of the BTR complex . The N-tlike RPA ,21. Humalike RPA . It has like RPA .rmi1 mutations through genetic screens has proved difficult; one rmi1 mutant, the temperature-sensitive E69K, was identified through this conventional approach [in vivo mutational analysis of RMI1 function in yeast. This approach has identified structural motifs that are essential for Rmi1 function and stability and we propose hypotheses for how these motifs contribute to Rmi1’s role in maintaining the functional integrity of the STR complex. As an alternative to primary sequence alignments, which have been only minimally informative because of the poor sequence conservation among Rmi1 homologues, we present a structure-based alignment between yeast and human Rmi1 that maps the location of conserved motifs and suggests differences in the size and structure of the DUF1767 domain.Because of the severe growth retardation, low viability and rapid accumulation of suppressor mutations, the identification and functional analysis of deleterious approach . In an eS. cerevisiae Rmi1 and the N-terminal 240 residues of the 625-residue human Rmi1 were analyzed with algorithms for helical propensity [The 241 residues of opensity ,27, and opensity .RMI1 plus 500 bp up- and downstream was amplified by PCR from the endogenous RMI1 locus of KHSY1338 . The fragment was inserted into XbaI-digested pRS415 by gap-repair cloning using the non-homologous-endjoining deficient yeast strain KHSY2331  and standard lithium-acetate transformation [RMI1 and the promoter region in the resulting plasmid pKHS621 was verified by sequencing. Point mutations were introduced into the RMI1 ORF in pKHS621 by QuikChange site-directed mutagenesis (Agilent Technologies). To construct pKHS621 derivatives that express myc-epitope-tagged Rmi1 and rmi1 mutant proteins, pKHS621 was linearized with BoxI and the HIS3-linked myc-epitope-coding sequence from pFA6a-13MYC-HIS3MX6 [The open reading frame of ormation . The int-HIS3MX6  was inseMATα, ura3Δ0, leu2Δ0, his3Δ1, lys2Δ0, rmi1::HIS3, TOP3.V5.VSV.KANMX6), grown to OD600 = 0.5 in synthetic complete media lacking leucine (SC-Leu), and spotted in 10-fold dilutions on yeast extract/peptone/dextrose (YPD) and on YPD supplemented with 150 mM hydroxyurea. Growth was documented after 3 to 5 days of incubation at 30°C.Derivatives of pKHS621 were transformed into KHSY4695 (rmi1Δ) was transformed with plasmids pKHS621, pKHS624, and pKHS626. Independent cultures were set up from 9 to 12 transformants for each plasmid and grown to approximately 2 x 107 cells/ml. Actual cell counts were determined using a hemocytometer and cultures were diluted to plate ~ 400 cells. Plates were incubated for 3 days at 30°C and colonies counted. Viability was calculated by dividing the number of colony forming units by the number of cells plated based on the hemocytometer count.Yeast strain KHSY4695  from the Yeast Cross and Capture Collection (GE Dharmacon) was transformed with pKHS630 or its derivatives  and lengths range from 241 residues in S. cerevisiae to 625 residues in humans. The crystal structure of the N-terminal 219 residues of human Rmi1 was recently resolved [Determining functionally important residues in resolved . It reveresolved ,33 couldin vivo. We found that rmi1Δ cells expressing rmi1-A128P and rmi1-A139P exhibited the same HU sensitivity as the rmi1Δ mutant complemented with wildtype RMI1, whereas the rmi1-P88A allele was only able to partially suppress the HU hypersensitivity of rmi1Δ , and to ~18% when compared to the N-terminal 241 residues of human Rmi1. Because of the low level of sequence identity between human and yeast Rmi1 we decided to analyze the alignment of the N-termini of twelve closely related Rmi1 sequences from fully sequenced Saccharomyces and non-Saccharomyces yeast species in PhylomeDB v4 [S.c. Rmi1) that contain hydrophobic residues in the i,i+4 pattern typical of an α-helix and are separated from each other by residues with the lowest helical propensity, proline and glycine. To test the possibility that these two regions could become helical upon binding to another protein, possibly Sgs1, or could be analogous to α1 and α2 in human Rmi1, we replaced L7 and Y35 with proline  or increasing its hydrophilicity (rmi1-Y218K) abolished Rmi1 function  and the yeast phylome indicates two regions of similarity in the N-terminal 57 residues, we tested if binding-induced helix formation could also be a function of the extended unstructured N-terminus of yeast Rmi1, but found that introducing proline residues where prospective helices might form , did not impair Rmi1 function. This limited analysis, however, cannot exclude the possibility that shorter helices fully capable of supporting Rmi1 function in vivo can still be induced with the L7P and Y35P mutations present.The N-terminal region flanking the OB-fold in the crystal structure of human Rmi1 forms a three-helix bundle that has been designated a DUF1767 domain . The thrch other . In humach other . The F63ch other ; interesIn contrast to the N-terminal α-helix, our findings suggest that the C-terminal α-helix defined by the Y218P/K and E220P mutations plays a major role in stabilizing Rmi1 as seen in other proteins containing an OB-fold . Our strFinally, our bioinformatics analysis suggests a disordered loop in yeast Rmi1 linking the OB-fold core to an N-terminal α-helix. The structural disorder in this linker appears to depend largely on a single proline, whereas multiple prolines are present in human Rmi1, suggesting a more flexible linkage of the three-helix bundle to the OB-fold core . A mutatS1 Table(PDF)Click here for additional data file.


Although depression is known to affect millions of people worldwide, individuals seeking aid from qualified health care professionals are faced with a number of barriers to treatment including a lack of treatment resources, limited number of qualified service providers, stigma associated with diagnosis and treatment, prolonged wait times, cost, and barriers to accessibility such as transportation and clinic locations. The delivery of depression interventions through the Internet may provide a practical solution to addressing some of these barriers.The purpose of this scoping review was to answer the following questions: (1) What Web-delivered programs are currently available that offer an interactive treatment component for depression?, (2) What are the contents, accessibility, and usability of each identified program?, and (3) What tools, supports, and research evidence are available for each identified program?Using the popular search engines Google, Yahoo, and Bing (Canadian platforms), two reviewers independently searched for interactive Web-based interventions targeting the treatment of depression. The Beacon website, an information portal for online health applications, was also consulted. For each identified program, accessibility, usability, tools, support, and research evidence were evaluated and programs were categorized as evidence-based versus non-evidence-based if they had been the subject of at least one randomized controlled trial. Programs were scored using a 28-point rating system, and evidence- versus non-evidence-based programs were compared and contrasted. Although this review included all programs meeting exclusion and inclusion criteria found using the described search method, only English language Web-delivered depression programs were awarded an evaluation score.The review identified 32 programs meeting inclusion criteria. There was a great deal of variability among the programs captured in this evaluation. Many of the programs were developed for general adolescent or adult audiences, with few (n=2) focusing on special populations . Cognitive behavioral therapy was the most common therapeutic approach used in the programs described. Program interactive components included mood assessments and supplementary homework sheets such as activity planning and goal setting. Only 12 of the programs had published evidence in support of their efficacy and treatment of depressive symptoms.There are a number of interactive depression interventions available through the Internet. Recommendations for future programs, or the adaptation of existing programs include offering a greater selection of alternative languages, removing registration restrictions, free trial periods for programs requiring user fees, and amending programs to meet the needs of special populations . Furthermore, discussion of specific and relevant topics to the target audience while also enhancing overall user control would contribute to a more accessible intervention tool. In October 2012, the World Health Organization estimated that 350 million people worldwide suffer from depression [Not all individuals face the same risk. Certain populations are more susceptible to depression relative to others. For example, it is estimated that 14%-77% of traumatic brain injury survivors suffer from depression post injury, which is a much higher than the general population -11. The There are a number of depression treatment options available including medication, lifestyle, and psychological interventions. Unfortunately, many therapies are unavailable to patients due to a lack of qualified service providers. In Canada, it is estimated that only 33% of individuals seeking mental health services actually receive treatment . PatientNot only are individuals hindered by accessibility issues but also by the stigma associated with a mental illness diagnosis such as depression. Only 42% of Canadians said that they would socialize with a friend suffering from a significant mental health issue , and 46%Recent advances in computer technology offer potential alternative treatment options. The percentage of the global population that had access to the Internet nearly doubled from 18% in 2006 to 35% in 2011 . As the Regarding Web-based intervention treatments for depression specifically, the Canadian Network for Mood and Anxiety Treatments suggested computer-assisted cognitive behavioral therapy (CBT) programs as a second line treatment in cases “where first line treatments are not indicated or cannot be used or when first line treatments have not worked” . HoweverReviews of existing Web-based programs for the treatment of depression are available on the Beacon website , as wellWe conducted a scoping review of interactive Web-based treatment programs for depression to answer the following questions: (1) What Web-based programs are currently available that offer an interactive treatment component for depression?, (2) What are the contents, accessibility, and usability of each identified program?, and (3) What tools, supports, and research evidence are available for each identified program? Unlike the systematic reviews that have been published to date, our review included publicly available Web-based programs with and without supporting research trials. In addition, we have compared and contrasted those with and without supporting evidence on predetermined evaluation criteria.A search for interactive programs targeting depression was conducted in April 2014 using Google, Yahoo, and Bing (Canadian versions)—popular and comprehensive search engines easily accessible to many individuals. A list of the search terms used is included in For the purposes of this scoping review, interactive was defined as a program requiring user engagement and input . If a program provided only reading materials on the symptoms of depression, treatments, and relapse prevention, it was classified as psychoeducational and did not meet the “interactive” inclusion criterion.Online depression treatmentCBT depression online treatmentDepression and CBT online programsOnline methods of depression treatmentComputerized cognitive behavior therapy programs for treatment of depressionOnline cognitive behavior therapy programs treating depressionComputer based depression treatment programsInternet delivered depression treatment programsE-therapy for depressionEach of the terms in The Beacon website  was addiIn order to systematically evaluate each identified program, we used criteria adapted from previously published guidelines  to genera in the case that the point could not be evaluated using the program of interest and/or attempts to contact program developers. A program evaluation scoring system was also created based on the above mentioned article  to proviTwo reviewers independently examined each program and scored them using the evaluation criteria outlined in An identified program was categorized as an evidence-based program (EBP) if it was the intervention in at least one published randomized controlled trial (RCT). All other programs were categorized as non-evidence-based programs (NBP).We identified 27 websites collectively offering 32 programs in accordance with inclusion and exclusion criteria  included in A summary of the countries where programs were developed is provided in Four of the EBP were offered in alternative languages . Two programs were available only in Dutch (Interapy and Kleur je Leven). Only one NBP was offered in another language and was not available in English (Internetpsykiatri: Swedish only). The remaining 19 programs were offered exclusively in English.The registration processes and fee structures varied by program for both the EBP and NBP and included those that were freely available (n=12), those with a fee required (n=8), those with an access code required (n=4), those for which an application to the course was required (n=4), those with multiple registration criteria (n=1), and those with unknown registration criteria n=3). See . See 3.Eight programs had an associated fee ranging from AUD $55  for five/six lessons to US $400 for eight sessions . Most programs offered a free trial period, allowing users to interact with the program prior to paying the fee.Most programs (n=21) required the input of personal information to access course material. Registration was restricted to certain countries for five programs . Registration was not required for three programs, and registration requirements were unknown for three programs.The programs that did not mandate registration allowed users to access program content without entering any personal information. However, users could not track their progress through these programs without registration. Programs with mandatory registration required users to input basic personal information before gaining access to program content, allowing for sessional data storage. For example, registration allowed users to create personal profiles through which mood assessments, worksheets, and module progression were recorded.The identified programs predominantly targeted adult audiences (n=19), with a few programs developed for adolescents or students (n=3), combined populations , a specified special population (n=2), or unknown audiences (n=3) see .The majority of programs contained content that was specific to the adult end-user. Sessions varied by program and offered an array of materials targeting adult concerns surrounding depression. Topics included but were not limited to problem solving, goal setting and planning, tackling financial issues, workplace stress, in addition to challenging negative thoughts. Programs targeting adolescents were mainly focused on academic life and the stressors associated with the learning environment. Topics included time management, stress associated with exams, relationships, confidence and self-esteem, as well as other social issues faced by youth.Two NBP were developed for specific target populations: After Deployment (discharged military personnel) and eCentre Clinic Wellbeing Plus Course (older adults). Sessions covered many issues faced by these individuals including post-traumatic stress syndrome, insomnia, depression, and anxiety.Evaluation scores for EBP ranged from 68%-92%, with an average score of 80%. Evaluation scores for NBP ranged from 39%-92%, with an average score of 73%. A detailed breakdown of scores per program is provided in A number of programs (n=17) provided user statistics and/or user reviews. Varying by program, statistics aimed to demonstrate overall program efficacy and effectiveness using program completion rates, number of registered users, number of users completing the course, and reductions in validated measure scores from pre- to post-intervention. See Programs delivered their interventions through various therapeutic techniques. The majority of the EBP (n=6) delivered CBT-focused treatments, four offered integrated therapies , and two did not define their therapeutic approach. Nine NBP provided CBT-based materials, seven offered combination therapy models, and four NBP were categorized as unknown as they did not define their therapeutic approach. See Treatment interventions were delivered in a number of ways for both EBP and NBP. Nine EBP contained a multimodal format  with only one delivered in a text-only format. Two EBP were unknown . The majority of the NBP (n=12) were offered in a multimodal format. Six NBP were offered in a text-only format, and two programs were categorized as unknown. See Many programs included additional features such as email reminders, calendar applications, journal space, progress tracking reports as well as mood tracking. EBP and NBP both offered a number of support tool options, with most programs offering multiple features . See Many programs provided lesson reinforcement activities and worksheets . Worksheets covered topics discussed in current or previous modules and encouraged user engagement. Many worksheets incorporated activity planning, goal setting, problem solving, and thought evaluation. Self-reflection activities further identified troublesome areas and encouraged corrective action.Most programs incorporated mood or depression assessment tools . Assessments were delivered prior to user registration, integrated into module content and/or independent of session programming. Programs administering assessments provided feedback and results immediately upon user completion. When evaluating some programs , it was unclear whether assessments were administered. In total, 17 programs used validated measures: Patient Health Questionnaire (PHQ)-9, Beck Depression Inventory (BDI), and Center for Epidemiologic Studies Depression Scale (CES-D).None of the EBP offered a peer-support forum (three programs were categorized as unknown). The majority (n=11) of NBP did not include a peer-support forum (one program was categorized as unknown). Only eight NBP were found to have this service available to its users. See Six EBP offered clinician support, four offered no clinician support, and two programs were unknown. Eight NBP offered clinician support, 11 programs did not offer clinician support, and one program was not evaluated. See Crisis links were defined as email addresses, phone numbers, and/or hotlines connected to distress centers providing counseling services to at-risk users. If present, telephone numbers and distress centers were from within the program’s country of origin. In total, 20 programs (both EBP and NBP) had a crisis link with contact information and phone numbers. Most EBP provided this service (five programs were unknown); however, six NBP did not (one program unknown). See Only 12 of the 32 programs had at least one published RCT evaluating their efficacy. It was beyond the scope of the current review to summarize the results of these trials and the reader is directed to recently published systematic reviews and meta-analyses  see Mul.Use and wide spread dissemination of Web-based mental health care interventions is expanding and reflected by the number of currently available depression treatment programs captured in this scoping review. Web-based approaches may have several benefits beyond those of conventional psychotherapy -38. Sucha) . EBP had a smaller range in scores (68%-92%) and a higher overall average score (80%) relative to the wider range for NBP (39%-92%) and a lower overall average score (73%). While generally comparable, there were some subtle differences between EBP and NBP programs on the evaluation criteria of accessibility, program structure, use of validated assessment tools, additional features, peer support forums, safeguards, and the provision of user-statistics as further discussed below.We have identified 32 existing interactive Web-based programs and have found varying degrees of accessibility, quality, and evidence supporting their efficacy. Only 12 of the 32 programs had at least one peer-reviewed, published article describing the results of an efficacy study. In examining the programs, authors noted a number of similarities and differences when comparing EBP to NBP by each evaluation point see . Using tWhile Web-based programs can generally improve access to mental health care, some aspects of existing programs may present users with alternative access barriers. For example, some programs were available only in one language , with the majority of NBP available only in English. The addition of alternative languages could promote open accessibility to any user seeking treatment. Furthermore, some programs had accessibility restrictions based on country of residence see , which ma; 47%) were freely available  as compared to the EBP , a higher proportion of the NBP  than the EBP  had a fee for accessing the program. Requirements such as therapist referral, administrator acceptance for registration, and/or user fees may act as deterrents to use as they necessitate additional motivation and resources on the part of the user. The need to obtain a referral, enter personal information to register, and/or wait for access negates the benefits of anonymity and convenience afforded by Web-based tools. Conversely, registration requirements enable the user to track their progress and build on previously completed modules. In addition, the registration of personal information would allow the program deliverers to contact the user when in need, such as when increases in depression symptoms or suicide risk  are reported. Also, referral-based programs often allowed for integrated therapist contact. For fee-based programs, free demonstration/trial modules could be provided to allow users to assess the program prior to making a financial commitment.While a higher proportion of the NBP (9/19a) of EBP offered the guided approach relative to 63% (12/19a) of the NBP. Future studies should evaluate the relative effectiveness of guided versus unguided interventions.In examining the programs captured in the review, CBT was the most commonly incorporated therapeutic approach . Future programs could be geared toward the needs of special populations such as individuals with cognitive impairments or persons in a caregiving role. Accommodations for cognitively impaired individuals may include larger text sizing, multimodal delivery (audio and video files), in addition to programming specific to their impairments . Caregivers of chronically ill patients also demonstrate increased psychological distress and burden ,41. Apara) and NBP (4/18a). Personalized treatment plans may enhance user engagement by appealing to their specific treatment needs and offering relevant treatment information. Take for example a user suffering from only a mild form of depression; they may not have found additional anxiety information useful causing them to lose interest in continuing with the program despite its potential benefit. In programs offering a personalized treatment plan, program suggestions were based on an initial assessment. All but a few EBP and NBP offered assessments; however, not all of the assessment tools used were validated . Of EBP, 75% (6/8a) relative to 67% (12/18a) of NBP employed validated assessment tools . Programs should strive to offer validated assessment tools to provide users with accurate feedback in regards to their depressive and anxiety symptoms. During this emotionally sensitive time period, individuals could be heavily influenced by program feedback and results, necessitating accurate and valid depictions of depression symptoms over time.Although a target audience was identified for each of the programs , course content was often generic for all users within the targeted population. Personalized treatment plans  were offered in only a few programs: 22% of EBP  and NBP  offered three or more of these additional features  of EBP and 50% (8/19a) of NBP offered therapist support. Similarly, 60% (6/10a) of EBP and 50% (8/19a) of NBP provided a therapist name and their credentials . Providing users with this information may provide them peace of mind that they are being cared for and monitored by an accredited individual capable of intervening if required. A recent study using MoodGYM plus brief face-to-face therapist support indicated positive results in the reduction of symptoms of depression in a primary care setting [Many of the programs included additional integrated therapist contact, peer support discussion forums, and crisis links. Programs offering therapist support  were delivered via telephone, video conference, or live chat : 60%  of NBP offered this feature. However, only 24% (4/17a) of NBP offered forums that were monitored by an overseeing authority, facilitating safe user interaction and positive constructive topics of conversation . Peer support offers a level of familiarity not offered with clinician support. The need for relatedness to others enduring similar emotional issues can be both comforting and motivating; however, the effectiveness of peer support upon symptom resolution has yet to be evaluated in this context.Other avenues of support offered within the evaluated programs included peer discussion forums, blogs, and shared user spaces . Unlike EBP, none of which provided peer-support forums, 44% (8/18a) of NBP offered this feature. Due to the sensitive nature of the treatment and topics discussed, all programs should offer or provide information for available crisis links.In addition to peer and clinician support, some programs offered crisis links via telephone hotlines, email, or chat functions. Hotlines provided support to users under distress when therapists or other social support options were unavailable. Safeguards  were available in all the evaluated EBP; however, only 68% , statistics varied greatly. Examining the existing literature, completion rates in RCTs of Web-delivered treatments for depression have primarily ranged from 55%-67% [To evaluate program usability, we contacted each of the program’s administrators. Many were unable or did not wish to disclose user statistics in regards to registration, attrition, and program completion. Those that responded to inquiry emails or posted statistics on their program websites are listed in  55%-67% -54 with Although not included in this review (as they did not meet the inclusion and exclusion criteria), three novel and noteworthy Web-based treatment programs for depression were identified: Depression Quest , MoodbusIn summary, many interactive treatment programs for depression are available on the Web; however, the efficacy and validity of most of these programs  have not been evaluated using RCTs. When comparing those programs that are evidence-based to those that have not been empirically evaluated, more of the EBP programs seemed to use a guided approach, employ validated assessment tools, offer additional features, incorporate safeguards, and provide user statistics. More of the NBP programs were available without fees or referrals  and offered peer-support forums. Based on our review, several programs emerged that are easily accessible, free to use, and have supporting evidence for their efficacy including E Couch, MoodGYM, and This Way Up  to identify programs conforming to evaluation inclusion criteria. Searches were repeated until results became redundant; however, it is possible that despite our efforts to be inclusive, some programs were missed. It is also important to note that searches conducted in other countries (using the above mentioned search engines) may not yield the same search results. Provided that many search engines suggest local websites and may also receive funding from local advertisers, our search results may be highly specific to our proximity . Moreover, the rapidly evolving nature of the Internet means that the same search conducted several months from now could yield a different set of results and conclusions. New programs may be developed, existing programs may be discontinued, adapted, or amalgamated, and new research trials could support or refute the efficacy of these programs.It should also be noted that programs were evaluated from the perspective of a researcher and not of an end-user. A person with depression, for example, may have different opinions regarding the usability and quality of the identified programs. The intended end-user would likely provide developers with a more subjective opinion. Programs may also be biased to the type of user they capture, that is, individuals with less severe forms of depression. Individuals suffering from more severe forms of depression may face greater decreases in motivation and are less likely to access and participate in an intervention program. Consequently, users with severe forms of depression may be underrepresented in RCTs and in collected user feedback and data. Feedback is necessary to facilitate change as well as outline likes and dislikes for various features. Overall, end-user input should be sought as it is crucial to improving treatment delivery and program functionality.At this time, there are few programs available for special populations . It is important that program functionality accommodate accessibility of varying populations to ensure adequate treatment is delivered. Studies investigating the needs of these special populations could inform the development of new programs and the adaptation of existing ones. Future programs could also aim for increased accessibility. This could entail multilanguage delivery, elimination of residency restrictions, elimination of registration fees or referrals, flexibility in module timing, and minimization of mandatory user response .With respect to research, although the minority of the identified programs were evidence-based as defined by the presence of at least one evaluative RCT, the trials that have been completed to date have been generally of strong quality with adequate sample sizes -34. In aIn this review, we identified 27 websites offering 32 programs with interactive components aimed at reducing symptoms of depression among users. The programs varied widely in terms of content, accessibility, usability, method of delivery , and supplementary tools. A minority of the programs identified had empiric evidence to support their efficacy for the treatment of depressive symptoms.In choosing to use or refer a Web-based treatment program for depression, the user may wish to consider the following factors: ease of use and accessibility, availability of additional features and support needs, and most critically, programs that have been validated with good quality research. Users are encouraged to critically evaluate their program choice and should investigate research supporting program claims.Web-delivered interventions afford many potential advantages to individuals. Users can log on to their preferred program in the comfort of their own home 24 hours a day, 365 days a year. This may help to increase accessibility, reduce prolonged wait times, and address privacy concerns. Furthermore, it is potentially cost-efficient and convenient, allowing users to seek treatment when desired. Developers should continue to create such programs and tailor additional sites to the needs of specialty groups.


BCR-ABL1 fusions is required for the development of personalized medicine approach for minimal residual disease monitoring at the DNA level. Next generation sequencing (NGS) of amplicons larger than 1000 bp simplified and accelerated a process of characterization of patient-specific BCR-ABL1 genomic fusions. NGS of large regions upstream and downstream the individual breakpoints in BCR and ABL1 genes, respectively, also provided information about the sequence variants such are single nucleotide polymorphisms.In chronic myeloid leukemia, the identification of individual  BCR-ABL1 transcript level monitoring is a crucial part of therapy response evaluation in patients with chronic myeloid leukemia (CML). Molecular response (MR) in CML is expressed in terms of log reduction in BCR-ABL1 transcript levels from a standardized baseline on an international scale [BCR-ABL1 transcripts [BCR-ABL1 gene would be useful for MRD monitoring in patients with therapy cessation.al scale ,3. Althonscripts ,5. It isBCR-ABL1 fusion is unique to each CML patient, since breakpoints are scattered within intron 13 (718 bp) or 14 (2128 bp) of BCR and within intron 1 of ABL1 (140 Kbp). The exact genomic sequence of the BCR-ABL1 fusion must therefore be characterized to design a personalized real-time polymerase chain reaction (PCR) assay. The sensitivity of personalized real-time PCR assay quantifying clone-specific fusions was reported to be down to less than 10−5 (depending on the amount of DNA entering the reaction) when nested PCR was used [BCR-ABL1 genomic fusions using conventional sequencing has been a challenging task [BCR-ABL1 fusions, we set up a strategy based on the generation of large amplicons by multiplex Long-Range PCR (mLR-PCR) and on subsequent ultra-wide sequencing using NGS technology. NGS provided sequences of large regions upstream and downstream the BCR-ABL1 fusions, allowing us to assess the frequency of annotated single nucleotide polymorphisms (SNPs), which could be associated with the fusion gene formation.At the genomic level the was used ,7. As amron 13 71 bp or 14BCR-ABL1 positive cell lines K562 and JURL-MK were included as controls of BCR-ABL1 fusion identification. Twelve healthy individuals were used as controls for SNP detection in the BCR region from exon 13 to exon 15.The patient cohort consisted of 24 males and 22 females with CML. Two BCR-ABL1 DNA present in the sample. Two-round mLR-PCR was performed with HiFi Accuprime Taq DNA polymerase (Invitrogen) enabling amplification of large regions. The amplification strategy is presented in Figure BCR exon 13 and 10 primers in ABL1 intron 1 [BCR exon 13 and 20 primers in ABL1 intron 1 [BCR) and NG_012034.1 (ABL1) were used for sequence analysis.DNA was isolated from total leukocytes of peripheral blood at the time of diagnosis ensuring high amount of intron 1 , the lonintron 1  productsBCR were located in intron 13 in 8/46 patients, carrying the e13a2 mRNA fusion, and in intron 14 in 38/46 patients and in both cell lines, carrying the e14a2 mRNA fusion . Only 10 of them recurred in two patients, whereas the remaining 46 were detected only once. This heterogeneity in SNP detection was caused by the extreme length of intron 1  , where significant differences between MRD levels measured by BCR-ABL1 expression and Ig/TCR DNA-based approach were described [TEL/ABL1  [TEL/AML1 (acute lymphoid leukemia) [TMPRSS2/ERG (prostate cancer) [EML4/ALK  [Although increasing number of studies has been published in last few years -16, analescribed . Moreove or ALL) , TEL/AMLeukemia) , TMPRSS2 cancer)  or EML4/ cancer) .


To the Editor: As in many West Africa nations, vectorborne diseases represent a substantial health burden in Mali; however, beyond malaria, the incidence and etiology of many of these diseases is poorly understood. Of the estimated 14.1 million persons living in sub-Saharan Mali, ≈70% live in remote rural settings with an ecologic landscape that puts inhabitants at an increased risk for contact with rodent and arthropodborne diseases. We retrospectively analyzed serum samples for evidence of recent (IgM+) and previous (IgG+) infection with chikungunya (CHIKV), dengue (DENV), West Nile (WNV), Lassa (LASV), Crimean-Congo hemorrhagic fever (CCHFV), and Ebola (EBOV) virus, as well as Old World hantaviruses (OW-HANV) and Leptospira spp., which is regularly misdiagnosed as an acute viral infection.http://www.hhs.gov/ohrp/humansubjects/guidance/belmont.html). This research was conducted under an institutional review board–approved document.We tested 376 deidentified serum samples collected from acutely ill patients who had a history of fever and hemorrhagic, diarrheal, or icteric syndromes  Figure. Plasmodium falciparum malaria and yellow fever virus infections. Commercially available IgM capture and conventional IgG ELISAs were used for serologic testing for CHIKV ; DENV  and WNV ; OW-HANVs ; and Leptospira spp. . Conventional IgM/IgG ELISAs were used for LASV  and CCHFV , and reagents for the EBOV IgM/IgG ELISA (infected/uninfected cell lysates) were prepared at the Rocky Mountain Laboratories  and validated with serum from experimentally infected monkeys. With the exception of the CHIKV, Leptospira spp., and in-house EBOV assays, the tests conducted in this study are under preclinical development for human diagnostic assays.Samples had previously tested negative for acute Leptospira spp.) or in-house quality-control assessments (EBOV), in a blinded fashion. Serologic reactivity was assessed according to manufacturer recommendations. For the EBOV ELISA, samples were deemed positive if optical density at 405 nm was >3 SD above that of the average of known negative samples.Samples were tested at a 1:100 dilution according to manufacturer specifications  and WNV (0.27%). Of rodentborne pathogens, OW-HANVs had a seroprevalence of 7.2%, whereas LASV was considerably lower (0.27%). CCHFV IgM was documented in 4.8% of samples. Overall, little annual variation in the IgM seroprevalence was noted, except for Leptospira spp., for which 2 obvious peaks in seroprevalence were observed (Serologic evidence suggestive of acute infection (IgM+) with 1 of the pathogens tested for was observed for 39.9% of samples . At 14.4observed .Leptospira spp., an acute dual infection that might be underrecognized . No samples were reactive with EBOV, and the low incidence of LASV infection is not surprising because the samples analyzed here were collected outside of the 1 documented LASV-endemic region in Mali (The relatively high IgG seroprevalence for most of the pathogens tested supports the findings of the IgM assays and further suggest the circulation of and potential for human exposure to these agents in Mali . GeograpWe used commercially available diagnostic platforms, primarily IgM capture and conventional IgG ELISAs, many of which are validated for human diagnostics. Ideally, diagnostics for zoonotic diseases would not rely on IgM/IgG serologic analysis because of caveats including IgM persistence and cross-reactivity between closely related pathogens (Leptospira spp., are contributing to human illness in Mali. These results add to those recently documented in studies conducted in Sierra Leone, implying that several of these zoonotic pathogens are widely distributed yet underreported throughout West Africa (Despite these limitations, these serologic findings indicate that flaviviruses, bunyaviruses, and togaviruses, as well as Technical Appendix. Distribution of serum samples tested, by region and year. IgM and IgG seroprevalence rates of selected vectorborne pathogens, by region of sample collection.