Twelve clarithromycin-resistant (MIC, >1 g/ml) Helicobacter pylori isolates were analyzed for point mutations in the 23S rRNA gene. Sequence analysis of all of the resistant isolates revealed a T-to-C transition mutation at position 2182. Transformation experiments confirmed that a single T-to-C transition mutation at position 2182 is associated with clarithromycin resistance. Eradication ofHelicobacter pylori infection by treatment with two antimicrobials (clarithromycin and amoxicillin or metronidazole) and a proton pump inhibitor is recommended by various consensus groups (5, 13). The prevalence of antimicrobial susceptibility of H. pylori varies with geographical regions, and clarithromycin resistance is the major cause of treatment failure (3,5,20). Alteration in either one or both copies of the H. pylori 23S rRNA gene is associated with resistance to clarithromycin, and the mechanism is a point mutation (an adenine-to-guanine transition at either position 2142 or 2143 or an adenine-to-cytosine transversion at position 2142) of the 23S rRNA (2,8,15,18,19,23). However, a T2182C mutation has also been proposed to be associated with clarithromycin resistance (10). In the present study we have examined clarithromycin-resistant (Cla r ) H. pylori (CRHP) isolates and characterized a T2182C mutation involved in clarithromycin resistance.Twelve CRHP isolates and three clarithromycin-susceptible H. pylori (CSHP) strains isolated from individual pretreatment patients attending the Dhaka Medical College hospital during 1999 to 2001 for routine endoscopy were examined. The bacteria were grown in brain heart infusion (BHIB) agar with 7% sheep blood and incubated at 37°C in 5% O 2 -10% CO 2 -85% N 2 for 3 to 6 days. A pure culture from a single colony was stored at Ϫ70°C until further study.The MICs of clarithromycin for the isolates were determined by the agar dilution method (MIC breakpoint, Ն1 g/ ml) and by the E-test strip (AB Biodisk, Solna, Sweden) as described elsewhere (11,12,16). All tests were repeated twice, and CSHP strain 26695 (MIC, 0.032 g/ml) was used as a control. Chromosomal DNA of the isolates was extracted by the cetyltrimethylammonium bromide method. A 1,300-bp fragment of the 23S rRNA (bp 1627 to 2926; GenBank accession number U27270) was amplified with primers Cla1 (5Ј-G GCTCTTTGAGTCCTTTTAGGAC-3Ј) and Cla4 (5Ј GCAT TACTGCGCTCACACAT-3Ј) as described previously (21). The PCR product was purified with a Microcon centrifugal filter device (Millipore Corporation, Bedford, Mass.), and a cycle sequencing reaction was performed with the same primers. DNA sequencing was performed under standard conditions with an ABI PRISM 310 automated sequencer (PerkinElmer Applied Biosystems, Foster City, Calif.). DNA sequence editing and analysis were performed by DNASTAR package 5.06 (DNASTAR Inc.) software. In order to confirm the role of the T2182C mutation, the 1,300-bp PCR fragment from CRHP isolate Gj50 (MIC, 4 g/ml) was transformed to CSHP strain DM26B as described earlier (19). Briefly, exponentially growing CSH...
The prevalence of STIs/RTIs among SWs in brothels in Bangladesh is high. An intervention strategy addressing both symptomatic and asymptomatic infections and periodic screening of SWs for RTIs/STIs is essential for successful HIV and STIs prevention programs.
Supporting caregivers’ mental wellbeing and ability to provide psychosocial stimulation may promote early childhood development. This paper describes the systematic approach of developing an integrated stimulation intervention, identifying the feasibility and challenges faced throughout the period. We developed an integrated curriculum by culturally adapting three interventions (Reach Up, Thinking Healthy, and general nutrition advice) and piloted this curriculum (Mar–April 2017) in courtyard groups sessions and individual home visits with pregnant women (n = 11) and lactating mothers (of children <24 months) (n = 29). We conducted qualitative interviews with the participants (n = 8) and the community health workers who delivered the intervention (n = 2). Most participants reported willingness to attend the sessions if extended for 1 year, and recommended additional visual cues and interactive role-play activities to make the sessions more engaging. Participants and community health workers found it difficult to understand the concept of “unhealthy thoughts” in the curriculum. This component was then revised to include a simplified behavior-focused story. Community health workers reported difficulty balancing the required content of the integrated curriculum but were able to manage after the contents were reduced. The revised intervention is likely feasible to deliver to a group of pregnant and lactating mothers in a low-resource setting.
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