Daily adjunctive therapy with vitamin D3 and phenylbutyrate supports clinical recovery from pulmonary tuberculosis: a randomized controlled trial in Ethiopia.

OBJECTIVE: Immunotherapy using vitamin D (vitD3 ) and phenylbutyrate (PBA) may support standard drug regimens used to treat infectious diseases. We investigated if vitD3 + PBA enhanced clinical recovery from pulmonary tuberculosis (TB). METHODS: A randomized controlled trial was conducted in Addis Ababa, Ethiopia. Patients with smear-positive or smear-negative TB received daily oral supplementation with 5000 IU vitD3 and 2 × 500 mg PBA or placebo for 16 weeks, together with 6-month chemotherapy. Primary end-point: reduction of a clinical composite TB score at week 8 compared with baseline using modified intention-to-treat (mITT, n = 348) and per-protocol (n = 296) analyses. Secondary end-points: primary and modified TB scores (week 0, 4, 8, 16, 24), sputum conversion, radiological findings and plasma 25(OH)D3 concentrations. RESULTS: Most subjects had low baseline plasma 25(OH)D3 levels that increased gradually in the vitD3 + PBA group compared with placebo (P < 0.0001) from week 0 to 16 (mean 34.7 vs. 127.4 nmol L-1 ). In the adjusted mITT analysis, the primary TB score was significantly reduced in the intervention group at week 8 (-0.52, 95% CI -0.93, -0.10; P = 0.015) while the modified TB score was reduced at week 8 (-0.58, 95% CI -1.02, -0.14; P = 0.01) and 16 (-0.34, 95% CI -0.64, -0.03; P = 0.03). VitD3 + PBA had no effect on longitudinal sputum-smear conversion (P = 0.98). Clinical adverse events were more common in the placebo group (24.3%) compared with the vitD3 + PBA group (12.6%). CONCLUSION: Daily supplementation with vitD3 + PBA may ameliorate clinical TB symptoms and disease-specific complications, while the intervention had no effect on bacterial clearance in sputum.

High plasma efavirenz level and CYP2B6*6 are associated with efavirenz-based HAART-induced liver injury in the treatment of naïve HIV patients from Ethiopia: a prospective cohort study.

The objective of this study was to assess the incidence, timing and identify pharmacogenetic, efavirenz (EFV) pharmacokinetic and biochemical predictors of EFV-based antiretroviral therapy (ART) drug-induced liver injury (DILI). ART-naïve HIV patients (n = 285) were prospectively enrolled. Pretreatment laboratory evaluations included hepatitis B surface antigen and C antibody, CD4 count and viral load. Liver tests were done at baseline, 1st, 2nd, 4th, 8th, 12th, 24th and 48th weeks during ART. Plasma EFV and 8-hydroxyefvairenz concentration was determined at week 4 using liquid chromatography-mass spectrometry. CYP2B6, CYP3A5, ABCB1 3435C/T and UGT2B7*2 genotyping was done using Taqman genotyping assay. Data were analyzed using survival analysis and Cox proportional hazards model. The incidence of DILI was 15.7% or 27.9 per 100 person-years and that of severe injury was 3.4% or 6.13 per 100 person-years. The median time for the development of DILI and severe injury was 2 and 4 weeks after initiation of ART, respectively. There was significant association of DILI with lower baseline platelet, albumin, log plasma viral load and CD4 count (P = 0.031, 0.037, 0.06 and 0.019, respectively). Elevated baseline alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, plasma EFV level and CYP2B6*6 were good predictors for the development of DILI (P = 0.03, 0.01, 0.016, 0.017 and 0.04, respectively). We report for the first time CYP2B6*6 as a putative genetic marker and high plasma EFV concentration as intermediate biomarker for vulnerability to EFV-induced liver injury in HIV patients. CYP2B6 genotyping and/or regular monitoring of EFV and lever enzymes level during early therapy is advised for early diagnosis and management of DILI.

Nine diiron(II) complexes of three bis-tetradentate pyrimidine based ligands with NCE (E = S, Se, BH3) coligands.

Three bis-tetradentate acyclic amine ligands differing only in the arm length of the pyridine pendant arms attached to the 4,6-positions of the pyrimidine ring, namely, 4,6-bis[N,N-bis(2'-pyridylethyl)aminomethyl]-2-phenylpyrimidine (L(Et)), 4,6-bis[N,N-bis(2'-pyridylmethyl)aminomethyl]-2-phenylpyrimidine (L(Me)), and 4,6-[(2'-pyridylmethyl)-2'-pyridylethyl)aminomethyl]-2-phenylpyrimidine (L(Mix)) have been used to synthesize nine air-sensitive diiron(II) complexes: [Fe(II)(2)L(Et)(NCS)(4)]·MeOH·¾H(2)O (1·MeOH·¾H(2)O), [Fe(II)(2)L(Et)(NCSe)(4)]·H(2)O (2·H(2)O), [Fe(II)(2)L(Et)(NCBH(3))(4)]·(5/2)H(2)O (3·(5/2)H(2)O), [Fe(II)(2)L(Me)(NCS)(4)]·½H(2)O (4·½H(2)O), [Fe(II)(2)L(Me)(NCSe)(4)] (5), [Fe(II)(2)L(Me)(NCBH(3))(4)]·(3/2)H(2)O (6·(3/2)H(2)O), [Fe(II)(2)L(Mix)(NCS)(4)]·½H(2)O (7·½H(2)O), [Fe(II)(2)L(Mix)(NCSe)(4)]·(3/2)H(2)O (8·(3/2)H(2)O), and [Fe(II)(2)L(Mix)(NCBH(3))(4)]·(3/2)H(2)O (9·(3/2)H(2)O). Complexes 3·(5/2)H(2)O, 4·½H(2)O, 5, 6·(3/2)H(2)O, and 8·(3/2)H(2)O were structurally characterized by X-ray crystallography, revealing, in all cases, both of the iron(II) centers in an octahedral environment with two NCE (E = S, Se, or BH(3)) anions in a cis-position relative to one another. Variable temperature magnetic susceptibility measurements showed that all nine diiron(II) complexes are stabilized in the [HS-HS] state from 300 K to 4 K, and exhibit weak antiferromagnetic coupling. Mössbauer spectroscopy confirmed the spin and oxidation states of eight of the nine complexes (the synthesis of air-sensitive complex 3 was not readily reproduced).

Alcohol use and its associated factors among Ethiopian military personnel.

BACKGROUND: Studies indicate that alcohol use is more prevalent in the military and that such use is related to coping mechanisms for stress. Alcohol use could result in health and social problems. However, little is known about alcohol use in Ethiopian military personnel. OBJECTIVE: To assess the level of alcohol use and its associated factors in the Ethiopian military. METHODS: A cross-sectional study including participants from the Ground and Air Forces of the Ethiopian National Defence Forces was conducted from February to June 2021. A representative sample of 502 military personnel from the two forces participated in the study. A structured questionnaire was developed to assess the individual level of alcohol use and its associated factors. The Alcohol Use Disorder Identification Test (AUDIT) was used to identify likely problematic alcohol use (AUDIT score ≥8). Hierarchical multivariable logistic regression models were run to identify associated factors with alcohol consumption. RESULTS: Approximately half of the respondents (49.8%, 95% CI 45.4% to 54.0%) were alcohol drinkers. Of the current alcohol users, 142 (63.1 %) were infrequent users; 60 (26.7 %) were moderate drinkers; and 23 (10.2 %) were heavy drinkers. Based on the AUDIT composite score, 71 (33.0%) of male participants were classified as having a score indicative of hazardous and harmful drinking and possible alcohol dependence behaviours. After adjusting for covariates, alcohol drinking was statistically significantly associated with higher odds of being: male, younger age, part of the Ground Force, smoker and high risk-taker. CONCLUSIONS: This study provides an initial step to addressing patterns of harmful and hazardous alcohol use in the Ethiopian National Defence Forces. Findings indicate the need to integrate alcohol abuse prevention into existing health education and behaviour change efforts of the Ethiopian National Defence Forces.

Impact of the Mechanical Properties of a Functionalized Cross-Linked Binder on the Longevity of Li-S Batteries.

One of the very challenging aspects of Li-S battery development is the fabrication of a sulfur electrode with high areal loading using conventional Li-ion binders. Herein, we report a new multifunctional polymeric binder, synthesized by the free-radical cross-linking polymerization of [2-(acryloyloxy)ethyl]trimethylammonium chloride (AETMAC) and ethylene glycol diacrylate (EGDA) to form poly(AETMAC- co-EGDA), that not only helps to confine the soluble polysulfide species but also has the desired mechanical properties to allow stable cycling of high-sulfur loading cathodes. Through a combination of spectroscopic and electrochemical studies, we elucidate the chemical interactions that inhibit polysulfide shuttling. We also show that extensive cross-linkage enables this polymeric binder to exhibit a low degree of swelling as well as high tensile modulus and toughness. These attributes are essential to maintain the architectural integrity of the sulfur cathode during extended cycling. Using this material, Li-S cells with a high-sulfur loading (6.0 mg cm-2) and a low-intermediate electrolyte/sulfur ratio (7 µL:1 mg) achieve an areal capacity of 5.4 mA h cm-2 and can be (dis)charged for 300 cycles with stable reversible redox behavior after the initial cycles.

Conceptualizations of subjective recovery from recent onset psychosis and its associated factors: A systematic review.

AIM: There is no standard definition of "subjective recovery" from psychosis, its nature is currently contested and debated among service-users and professionals. Individual studies have explored conceptualizations of subjective recovery from recent onset psychosis, but there have been no previously published systematic reviews on the topic. The aim of this review was to examine and synthesize quantitative and qualitative studies examining the concept of subjective recovery from recent onset psychosis and identify common factors associated with this recovery process. METHODS: Relevant electronic databases (Medline, CINAHL, PsychInfo and ProQuest Dissertations and Theses) were searched and hand searches were also carried out. Publications in each database from the inceptions of the databases to April 12, 2017 were included. Data from selected articles were extracted using a piloted extraction form and thematic integrative analysis was performed. RESULTS: Ten studies with different study designs were included in this review. Subjective recovery was conceptualized into 3 main themes: "recovery as outcome", "recovery as process" and "endeavours during recovery". Factors contributing to subjective recovery were categorized into 4 main themes; "treatment related", "illness related", "individual related" and "social environment" related. Non-linear and subjective nature of the process of recovery were reinforced by the review findings. CONCLUSIONS: Studies in subjective recovery from recent onset psychosis are limited to developed countries. Acquiring hope and self-confidence, overcoming symptoms and stigma through mobilizing all resources available were accentuated in conceptualizing subjective recovery and related factors. Recovery-oriented health care services should acknowledge individual differences and involve service users in their care decisions.

Genotyping and molecular detection of multidrug-resistant Mycobacterium tuberculosis among tuberculosis lymphadenitis cases in Addis Ababa, Ethiopia.

Multidrug-resistant tuberculosis (MDR-TB) has emerged as a major public health problem. Drug-resistance surveillance data show that 3.9% of new and 21% of previously treated TB cases were estimated to have had rifampicin/ multidrug-resistant tuberculosis (MDR/RR-TB) in 2015. This implies that the MDR-TB is increasing alarmingly. Hence, a better understanding of drug resistance mechanisms and genotypes associated with multidrug resistance in M. tuberculosis is crucial for improving diagnostic and therapeutic methods to treat individuals with MDR-TB. The aim of this study was to analyze molecular drug resistance mutations of MDR-TB isolates from the cases of TB-lymphadenitis in relation to its genetic lineages. A cross-sectional study was conducted on culture positive cases from July to October, 2014 in Addis Ababa, Ethiopia. Sixty isolates were included to analyze drug resistance mutated gene responsible for MDR-TB in relation to its molecular genotyping. Mycobacterial culture, GenoTypeMTBDR plus and Spoligotyping were used to undertake the study. Of 60 TBLN isolates, 8.3% were identified MDR-TB cases and one isolate was isoniazid mono-resistant. Eleven isolates in T3-ETH genetic sub lineage were sensitive to both RMP and INH, while only 2 isolates were MDR-TB. Most of the RMP- resistant isolates showed mutation in codon S531L and all isolates mutated in the katG gene conferring INH resistant strains had mutations in codon of S315T1. Screening for the rpoB and katG gene mutation of tuberculosis lymphadenitis is useful in Ethiopia for an early detection and treatment of MDR-TB. Besides, there is a drug resistance variation among different lineages of Tuberculosis lymphadenitis which has important consequences for the development of efficient control strategies.

Mycobacterium tuberculosis in central Ethiopia: drug sensitivity patterns and association with genotype.

Drug resistance tuberculosis (TB) and the emergence of multidrug resistant (MDR) isolates are significant concerns regarding TB control programs in several countries. This study was undertaken to evaluate the drug sensitivity of Mycobacterium tuberculosis and to assess its association with strains and lineages of M. tuberculosis. A total of 279 M. tuberculosis strains isolated from Central Ethiopia were tested for their drug sensitivity patterns to first line TB drugs using the conventional proportion method on Löwenstein Jensen media. The association between drug sensitivity and strain type was assessed on 263 isolates of the 279 isolates. Of the 268 M. tuberculosis isolates obtained from new cases, 209 (78%) were susceptible to first line TB drugs, and 59 (22.2%) bacterial isolates were resistant to at least one of the first line drugs. The highest mono-resistance (7.5%) pertained to streptomycin (STM). Remarkably, seven of eleven isolates (63.6%) previous treatment for TB were resistant to at least one of the first line drugs. The prevalence of MDR-TB was 1.5% (4/268) for newly identified TB cases, all of which were members of the Euro-American Lineage. There was no statistically significant association (P > 0.05) between drug sensitivity, and either strains, sub-lineages or main lineages of M. tuberculosis. A significant proportion of M. tuberculosis was resistant to at least one first line anti-TB drug. Moreover, the frequencies of resistance to either isoniazid or rifampicin were high compared to data that were previously reported in some part of the country.

Sharp Injury and Exposure to Blood and Body Fluids among Health Care Workers in Health Care Centers of Eastern Ethiopia.

BACKGROUND: Health care workers are facing certain occupational hazards because of sharp injury and exposure to human blood and body fluids as a result of handling wastes. Though much attention is paid for the protection of these workers, the number of exposures and injuries do not show a sign of decline from time to time. OBJECTIVE: To examine the occurrence of sharp injury and exposure to blood and body fluids in health care workers in health care centers in Ethiopia. METHODS: In a case-control study, a randomly selected sample of 65 health facilities with 391 cases and 429 controls were studied. Data were collected through a self-administered questionnaire. Detailed analysis of exposure among the health care workers was done by logistic regression analysis with generalized estimating equations model to control correlation effects of responses within the cluster of health facilities. RESULTS: The number of health care workers who got sharp injury was 217 (26.5%). 296 (36.1%) had exposure to blood and body fluids. Working at Harari region (adjusted OR 0.44, 95% CI 0.26 to 0.75) and East Hararghea (adjusted OR 0.61, 95% CI 0.40 to 0.94), being male (adjusted OR 0.56, 95% CI 0.44 to 0.91), and a being nurse (adjusted OR 0.188, 95% CI 0.06 to 0.63) were independent risk factors of the exposure. CONCLUSION: Regardless of the anticipated low self-reporting for exposure status, the number of health care workers reported having sharp injury and exposure to blood and body fluids was high. Such high exposures indicate that health care workers are at high risk of acquiring blood-borne viral infections such as hepatitis B, hepatitis C, and HIV.

Molecular typing of mycobacteria isolated from tuberculous lymphadenitis cases in Addis Ababa, Ethiopia.

BACKGROUND: In Ethiopia, one of the world's 22 high tuberculosis (TB) burden countries, one third of the tuberculosis (TB) cases are attributed to tuberculosis lymphadenitis (TBLN). However, information on the molecular type of the mycobacterial species and strains that cause TBLN in the country is scarce. OBJECTIVE: To identify the species and strains of mycobacteria that cause TBLN in Ethiopia. METHODS AND RESULTS: A cross-sectional study was conducted among 206 presumed TBLN cases to characterise positive culture isolates. Mycobacterium tuberculosis complex species and strains were identified using region of difference 9 deletion and SITVIT WEB, respectively. Of the 206 fine-needle aspirate samples collected, 74 (36%) were culture-positive: 73 (98.6%) of the isolates were M. tuberculosis, and the remaining 1.4% were M. bovis. Further characterisation of the 73 M. tuberculosis isolates led to 26 distinct spoligotype international types (SITs) and 13 newly identified patterns. The most prevalent strains were SIT149, SIT53, SIT26 and SIT37 of sublineages T3-ETH, T1, CASI-DELHI and T3, respectively; these accounted for 52.6% of the total number of strains. CONCLUSION: TBLN was mainly caused by M. tuberculosis and highly clustered strains SIT149, SIT53, SIT26 and SIT37 of sublineages T3-ETH, T1, CASI-DELHI and T3, respectively.

Characteristics of trichiasis patients presenting for surgery in rural Ethiopia.

AIMS: To determine the characteristics of trichiasis patients presenting for surgery in Wolayta Zone of Ethiopia. METHODS: Patients referred for trichiasis surgery by community health agents were evaluated by trained integrated eye care workers (IECWs) for the presence of trichiasis, locations of inturned lashes, severity of trichiasis, corneal opacity, and visual acuity. RESULTS: 1635 individuals with trichiasis presented for surgery. 82% had bilateral trichiasis; 91% of patients reported trichiasis duration of >2 years. Epilation was practised by over three fourths of the study subjects. A high proportion of patients tested positive for ocular Chlamydia trachomatis at presentation. 17% had monocular blindness and 8% were binocularly blind. Corneal opacity was highly associated with the trichiasis duration and severity and visual loss was associated with corneal opacity. CONCLUSION: Severe trichiasis reflects the magnitude of the trachoma problem in Ethiopia. Visual impairment due to trichiasis is highly associated with disease severity and duration. Early intervention to correct trichiasis before it become severe is recommended to prevent visual impairment.

Low vision and blindness in adults in Gurage Zone, central Ethiopia.

AIM: To determine the magnitude and causes of low vision and blindness in the Gurage zone, central Ethiopia. METHODS: A cross sectional study using a multistage cluster sampling technique was used to identify the study subjects. Visual acuity was recorded for all adults 40 years and older. Subjects who had a visual acuity of <6/18 were examined by an ophthalmologist to determine the cause of low vision or blindness. RESULTS: From the enumerated population, 2693 (90.8%) were examined. The prevalence of blindness (<3/60 better eye presenting vision) was 7.9% (95% CI 6.9 to 8.9) and of low vision (6/24-3/60 better eye presenting vision) was 12.1% (95% CI 10.9 to 13.3). Monocular blindness was recorded in 16.3% of the population. Blindness and low vision increased with age. The odds of low vision and blindness in women were 1.8 times that of the men. The leading causes of blindness were cataract (46.1%), trachoma (22.9%), and glaucoma (7.6%). While the prevalence of vision reducing cataract increased with age, the prevalence of trachoma related vision loss did not increase with age, suggesting that trichiasis related vision loss in this population might not be cumulative. CONCLUSION: The magnitude of low vision and blindness is high in this zone and requires urgent intervention, particularly for women. Further investigation of the pattern of vision loss, particularly as a result of trachomatous trichiasis, is warranted.

Sexual violence among female high school students in Debark, north west Ethiopia.

OBJECTIVES: To assess the prevalence, outcome and awareness of sexual violence among high school female students. DESIGN: A school-based cross-sectional survey. SETTING: Debark Town, north-west Ethiopia. SUBJECTS: Two hundred and sixteen female high school students, grade 9-11 were included for the quantitative study. For the qualitative data, 16 individuals for the focus group discussion (10 well-recognised female figures in the town and six high school students) and head of the police department for in-depth interview were enrolled. RESULTS: Sixty two per cent of the respondents had heard of sexual violence committed on young females. Sexual violence was reported by 65.3% of the respondents. The prevalence of performed and attempted rape were 8.8% and 11.5%, respectively. The age range of performed rape victims was between 12 and 21 years. Of the 19 (8.8%) who reported rape being performed on them, unwanted pregnancy, suicide attempt, vaginal discharge and abortion were the consequences in 21%, 15.8%, 10.5% and 5.3%, respectively. CONCLUSION: Sexual violence is a major public health problem with high rates of underreporting. Sex education should be given on a regular basis and policy making bodies and the police be well aware of this high magnitude and take strong measures to reduce it.

Impact of child mortality and fertility preferences on fertility status in rural Ethiopia.

OBJECTIVES: To determine the effects of child mortality and fertility preference on fertility status in rural Ethiopia. DESIGN: Case-referent where the cases were women with number of children ever born alive was less than five and controls were women with number of children ever born alive greater or equal to five. SETTING: Butajira rural health project study base which is a field epidemiologic laboratory found 35 Kms south of the capital city of Ethiopia, Addis Ababa run by the Department of Community Health of Tikur Anbessa Medical Faculty. MAIN OUTCOME MEASURES: Fertility status measured by number of children ever born alive; death of a child or an infant is a predictor. SUBJECTS: A total of 219 women with number of children ever born alive less than five and 899 women with number of children ever born alive greater or equal to five were included in the study. RESULTS: Child mortality affected number of children ever born alive significantly (OR= 7.39, 95% CI: 4.62, 9.08). As the number of children died increased proportionally, there is a proportional increase in the risk of higher fertility (X2 for trend 4.92, d.f =4, p-value 0.02). Number of children desired before marriage, desire for more children and sex preference were not associated with increased fertility in this study. Of all the socio-demographic and reproductive variables, later age at first marriage and first birth showed lower number of children ever born alive with (OR= 1.82, 95 % CI: 1.24, 2.83) and (OR= 3.08, 95 % CI: 2.03, 4.68) respectively. Breast-feeding duration of more than six months showed association with less number of children ever born alive (OR= 1.92, 95% CI: 1.30,2.80). CONCLUSION: The study finding implies high fertility status is strongly associated with child death and hence measures that curb child mortality are believed to decrease fertility status besides promoting child survival.

Solvent control: dinuclear versus tetranuclear complexes of a bis-tetradentate pyrimidine-based ligand.

A new bis-tetradentate acyclic amine ligand L(Et) has been synthesized from 4,6-bis(aminomethyl)-2-phenylpyrimidine and 2-vinylpyridine. Dinuclear complexes, Mn(II)(2)L(Et)(MeCN)(H(2)O)(3)(ClO(4))(4) (1), Fe(II)(2)L(Et)(H(2)O)(4)(BF(4))(4) (2), Co(II)(2)L(Et)(H(2)O)(3)(MeCN)(2)(BF(4))(4) (3), Ni(II)(2)L(Et)(H(2)O)(4)(BF(4))(4) (4), Ni(II)(2)L(Et)(H(2)O)(4)(ClO(4))(4)·8H(2)O (4'), Cu(II)(2)L(Et)(BF(4))(4)·MeCN (5), Zn(II)(2)L(Et)(BF(4))(2)(BF(4))(2)·½MeCN (6), were obtained from 1 : 2 reactions of L(Et) and the appropriate metal salts in MeCN, whereas in MeOH tetranuclear complexes, Mn(II)(4)(L(Et))(2)(OH)(4)(ClO(4))(4) (7), Fe(II)(4)(L(Et))(2)(F)(4)(BF(4))(4)·5/2H(2)O (8), Co(II)(4)(L(Et))(2)(F)(4)(BF(4))(4)·3H(2)O (9), Ni(II)(4)(L(Et))(2)(F)(4)(BF(4))(4)·4H(2)O (10), Cu(II)(4)(L(Et))(2)(F)(4)(BF(4))(4)·3H(2)O (11) and Zn(II)(4)(L(Et))(2)(F)(4)(BF(4))(4) (12), result. Six complexes have been structurally characterized: in all cases each L(Et) is bis-tetradentate and provides a pyrimidine bridge between two metal centres. As originally anticipated, complexes 1, 4' and 6 are dinuclear, while 9, 10 and 12 are revealed to be tetranuclear, with two M(2)(L(Et))(4+) moieties bridged by two pairs of fluoride anions. Weak to moderate antiferromagnetic coupling between the metal centres is a feature of complexes 2, 3, 4, 8, 9 and 10. The dinuclear complexes 1-6 undergo multiple, mostly irreversible, redox processes. However, the pyrimidine-based dicopper(II) complex 5 undergoes a two electron quasi-reversible reduction, Cu(II)(2)→ Cu(I)(2), and this occurs at a more positive potential [E(m) = +0.11 V (E(pc) = -0.03 and E(pa) = +0.26 V) vs. 0.01 M AgNO(3)/Ag] than for either of the dicopper(II) complexes of the analogous pyrazine-based ligands.

Evaluation of Toluidine Blue O staining for the diagnosis of Pneumocystis jiroveci in expectorated sputum sample and bronchoalveolar lavage from HIV-infected patients in a tertiary care referral center in Ethiopia.

BACKGROUND: Pneumocystis pneumonia (PCP) is becoming increasingly recognized in sub-Saharan Africa. The currently recommended diagnostic methods using induced sputum (IS) and bronchoalveolar lavage (BAL) are neither technically feasible nor affordable for a wider clinical use in developing countries. Therefore, there is a need for a simple and affordable diagnostic test. METHODS: The yield of Toluidine Blue O (TBO) stain, immunofluorescence (IF), and polymerase chain reaction (PCR) for the diagnosis of Pneumocystis jiroveci were compared in 78 expectorated sputum and 118 BAL samples of 131 HIV-infected patients presenting with atypical chest X-ray and sputum smear-negative for acid-fast bacilli. RESULTS: A total of 56 (42.7%) patients tested positive for P. jiroveci by PCR, 39 (29.4%) by IF, and 28 (21.4%) by TBO stain. The sensitivity of TBO as compared to IF and PCR was 71.4% and 34.5% in expectorated sputum and 68% and 41.5% in BAL samples, respectively, with a specificity approaching 100% in both. The sputum PCR showed high concordance rate with BAL PCR. The sensitivity and specificity of sputum PCR as compared to BALPCR was 78.9% and 89%, respectively. In both TBO and IF positive BAL samples, majority were from patients who could not produce sputum (p<0.001). The density of P. jiroveci clusters in BAL samples did not correlate with prior co-trimoxazole use, immunologic status of the patient or overall mortality. CONCLUSION: Compared to IF, TBO staining has an acceptable sensitivity and very high specificity both in expectorated sputum and BAL samples. Expectorated sputum is, therefore, the most practical specimen and TBO staining an inexpensive diagnostic method to be recommended for high-HIV, resource-constrained settings. Bronchoscopy for the diagnosis of PCP is often not required for patients who can produce sputum. For patients who cannot produce sputum, however, the cost and efficacy of TBO in IS sample needs to be investigated in resource-poor countries.

Application of hexafluoroacetone as protecting and activating reagent in amino acid and peptide chemistry.

Using hexafluoroacetone as protecting and activating reagent, multifunctional amino acids like aspartic acid can be functionalized regioselectively. This strategy offers i.a. a two-step synthesis for aspartame and preparatively simple access to multifunctional natural and unnatural amino acids, like 4-oxo-L-amino acids, 5-diazo-4-oxo-L-amino acids, 4-substituted L-proline derivatives and various heterocyclic L-amino acids. On application of this strategy to amino diacetic acid N-substituted glycines become readily available.