ABSTRACT
INTRODUCCIÓN: La infección por Chlamydia trachomatis es la ITS bacteriana más frecuente del mundo. En el cervix se presenta mayormente de forma asintomática y afecta especialmente a mujeres jóvenes y adolescentes. Puede producir daño permanente en el tracto reproductor femenino, se asocia a parto prematuro, infecciones neonatales transmitidas vía vertical y mayor riesgo de adquirir otras ITS como VIH Y VPH. Por estos motivos se han establecido estrategias de tamizaje para detectar y tratar precozmente la infección asintomática por C. trachomatis en diferentes países. En nuestro país no contamos con un programa nacional de tamizaje. OBJETIVO: Determinar la prevalencia de infección asintomática por Chlamydia trachomatis en mujeres entre 12 y 21 años de la Provincia de Osorno, Región de Los Lagos, Chile. PACIENTES Y MÉTODOS: Se desarrolló un estudio de diseño transversal con una cohorte única de mujeres adolescentes y jóvenes consultantes en el Policlínico de Alto Riesgo Obstétrico y de Ginecología Infantil y Adolescente, del Hospital Base de Osorno, entre enero de 2019 y enero 2020. Se determinó el estado de infección asintomática mediante RPC en tiempo real para C. trachomatis. Se realizó una encuesta a fin de determinar características demográficas, hábitos y conductas sexuales de las pacientes estudiadas. RESULTADOS: Fueron reclutadas 124 mujeres entre 12 a 21 años de edad, de las cuales, 36 (29,3%) se encontraban embarazadas al momento del estudio. La prevalencia de infección asintomática por C. trachomatis fue de 14/124 (11,3%). En las mujeres gestantes se encontraron 6/36 (16,7%) casos positivos de infección por C. trachomatis y 8/88 (9,1%) en las no gestantes. Existe una mayor frecuencia de infección asintomática a menor edad de inicio de actividad sexual (33,3% en aquellas que inician entre 11-12 años vs. 16,2% en las que inician entre 1314 años, 7,4% entre 15-16 y 8% entre 17-21 años; p < 0,05). Esta tendencia no fue observada al comparar el estado de infección con el tiempo de vida sexual activa. Sólo 15,7% de las pacientes utilizó preservativo en todas sus relaciones sexuales. DISCUSIÓN: La infección asintomática por C. trachomatis es frecuente en las mujeres adolescentes y jóvenes sexualmente activas. Las pacientes con inicio más temprano de la actividad sexual coital (bajo 13 años de edad) podrían estar en mayor riesgo. Se requiere con urgencia establecer la frecuencia nacional de infección para desarrollar una estrategia sanitaria para su pesquisa y manejo oportuno en nuestro país.
BACKGROUND: Chlamydia trachomatis infection is the world most common bacterial STI. At uterine cervix it presents mostly asymptomatically and especially affects young women and adolescents. It can cause permanent damage to the female reproductive tract and is associated with premature birth, connatal infections and increased risk of acquiring other STIs such as HIV and HPV. For these reasons, other countries have established screening strategies to detect and treat asymptomatic C. trachomatis infection. Our country don't have a national screening program. AIM: To determine the prevalence of C. trachomatis asymptomatic infection in adolescent and young women in Osorno province, Los Lagos Region, Chile. METHODS: A crosssectional study was performed in adolescent and young women who consult at Hospital Base Osorno in the MaternoFetal and PediatricAdolescent Gynecology ambulatory clinics, between January 2019 and January 2020. The status of asymptomatic infection was determined by PCR for C. trachomatis. A survey was carried out to determine the demographic characteristics, habits and sexual behaviors. RESULTS: 124 women between 12 and 21 years of age were recruited, of which 36 (29,3%) were pregnant at the time of the study. The prevalence of asymptomatic infection by C. trachomatis was 11.3.%. In pregnant women, there were 6/36 (16.7%) positive cases for C. trachomatis and 8/88 (9.1%) in nonpregnant women. We found a higher frequency of asymptomatic infection at younger age of first sexual intercourse (33% in adolescents at 11-12 years old vs. 16.2% at 13-14, 7.4% at 15-16 and 8% at 17-21; p<0.05). Only 15.7% of the patients utilized condoms in all their intercourses. DISCUSSION: Asymptomatic C. trachomatis infection is common in adolescent and young women, with a higher risk in those who onset sexual activity at an early age (less than 13 years old). It is urgently required to determine the national frequency of asymptomatic C. trachomatis infection to develop a national strategy for screening and timely treatment.
Subject(s)
Humans , Female , Child , Adolescent , Young Adult , Sexual Behavior , Chlamydia Infections/epidemiology , Sexually Transmitted Diseases/epidemiology , Chile/epidemiology , Chlamydia trachomatis , Prevalence , Cross-Sectional Studies , Asymptomatic InfectionsABSTRACT
Previous work demonstrated that lysine homopeptides adopt a polyproline II (PPII) structure. Lysine homopeptides with odd number of residues, especially with 11 residues (K11), were capable of inhibiting the growth of a broader spectrum of bacteria than those with an even number. Confocal studies also determined that K11 was able to localize exclusively in the bacterial membrane, leading to cell death. In this work, the mechanism of action of this peptide was further analyzed focused on examining the structural changes in bacterial membrane induced by K11, and in K11 itself when interacting with bacterial membrane lipids. Moreover, alanine and proline scans were performed for K11 to identify relevant positions in structure conformation and antibacterial activity. To do so, circular dichroism spectroscopy (CD) was conducted in saline phosphate buffer (PBS) and in lipidic vesicles, using large unilamellar vesicles (LUV), composed of 2-dimyristoyl-sn-glycero-3-phosphoglycerol (DMPG) or bacterial membrane lipid. Antimicrobial activity of K11 and their analogs was evaluated in Gram-positive and Gram-negative bacterial strains. The scanning electron microscopy (SEM) micrographs of Staphylococcus aureus ATCC 25923 exposed to the Lys homopeptide at MIC concentration showed blisters and bubbles formed on the bacterial surface, suggesting that K11 exerts its action by destabilizing the bacterial membrane. CD analysis revealed a remarkably enhanced PPII structure of K11 when replacing some of its central residues by proline in PBS. However, when such peptide analogs were confronted with either DMPG-LUV or membrane lipid extract-LUV, the tendency to form PPII structure was severely weakened. On the contrary, K11 peptide showed a remarkably enhanced PPII structure in the presence of DMPG-LUV. Antibacterial tests revealed that K11 was able to inhibit all tested bacteria with an MIC value of 5 µM, while proline and alanine analogs have a reduced activity on Listeria monocytogenes. Besides, the activity against Vibrio parahaemolyticus was affected in most of the alanine-substituted analogs. However, lysine substitutions by alanine or proline at position 7 did not alter the activity against all tested bacterial strains, suggesting that this position can be screened to find a substitute amino acid yielding a peptide with increased antibacterial activity. These results also indicate that the PPII secondary structure of K11 is stabilized by the interaction of the peptide with negatively charged phospholipids in the bacterial membrane, though not being the sole determinant for its antimicrobial activity.
Subject(s)
Anti-Bacterial Agents , Antimicrobial Cationic Peptides , Bacteria/growth & development , Polylysine , Alanine/chemistry , Anti-Bacterial Agents/chemistry , Anti-Bacterial Agents/pharmacology , Antimicrobial Cationic Peptides/chemistry , Antimicrobial Cationic Peptides/pharmacology , Polylysine/chemistry , Polylysine/pharmacology , Proline/chemistryABSTRACT
Small RNA and chaperone proteins form synergistic duos that play pivotal roles in controlling gene expression in bacteria. This is the case for Hfq, a highly pleiotropic pretranslational modulator of general protein expression, which responds to harsh environmental conditions and influences fitness and virulence in a wide range of pathogenic Enterobacteria. Given this relevancy, we evaluated the presence and potential role of Hfq in the fish pathogen Piscirickettsia salmonis, a Gram-negative bacterium that threatens the sustainability of Chilean salmon production. Using bioinformatics tools were identified and characterized two variants of Hfq, which share the consensus RNA-binding domains and the active sites described functional Hfq other bacteria. Additionally, we demonstrated that hfq-1 and hfq-2 were transcriptionally active when growing in cell-free media and in infected susceptible fish cell line. Expression of both genes differed under different growth conditions and under stress, suggesting that their roles might be independent and different, depending on the bacterial physiological status. In conclusion, we demonstrate the existence of two different and functional ORF coding for the hfq marker in marine bacteria and a preliminary analysis indicating that these two novel proteins might have relevant roles in the biology and pathogenic potential of P. salmonis.
Subject(s)
Host Factor 1 Protein/genetics , Oncorhynchus mykiss , Piscirickettsia/isolation & purification , Piscirickettsiaceae Infections/veterinary , Salmo salar , Amino Acid Sequence , Animals , Cell Line , Chile , Fish Diseases/microbiology , Host Factor 1 Protein/metabolism , Piscirickettsiaceae Infections/microbiology , Sequence AlignmentABSTRACT
Bronchiectasis in adults is a chronic disorder associated with poor quality of life and frequent exacerbations in many patients. There have been no previous international guidelines.The European Respiratory Society guidelines for the management of adult bronchiectasis describe the appropriate investigation and treatment strategies determined by a systematic review of the literature.A multidisciplinary group representing respiratory medicine, microbiology, physiotherapy, thoracic surgery, primary care, methodology and patients considered the most relevant clinical questions (for both clinicians and patients) related to management of bronchiectasis. Nine key clinical questions were generated and a systematic review was conducted to identify published systematic reviews, randomised clinical trials and observational studies that answered these questions. We used the GRADE approach to define the quality of the evidence and the level of recommendations. The resulting guideline addresses the investigation of underlying causes of bronchiectasis, treatment of exacerbations, pathogen eradication, long term antibiotic treatment, anti-inflammatories, mucoactive drugs, bronchodilators, surgical treatment and respiratory physiotherapy.These recommendations can be used to benchmark quality of care for people with bronchiectasis across Europe and to improve outcomes.
Subject(s)
Humans , Anti-Bacterial Agents/therapeutic use , Anti-Inflammatory Agents/therapeutic use , Bronchiectasis/drug therapy , Bronchiectasis/surgery , Bronchodilator Agents/therapeutic use , Physical Therapy ModalitiesABSTRACT
OBJECTIVES: To evaluate the accuracy of magnetic resonance imaging measurements of cartilage tissue-mimicking phantoms and to determine a combination of magnetic resonance imaging parameters to optimize accuracy while minimizing scan time. METHOD: Edge dimensions from 4 rectangular agar phantoms ranging from 10.5 to 14.5 mm in length and 1.25 to 5.5 mm in width were independently measured by two readers using a steel ruler. Coronal T1 spin echo (T1 SE), fast spoiled gradient-recalled echo (FSPGR) and multiplanar gradient-recalled echo (GRE MPGR) sequences were used to obtain phantom images on a 1.5-T scanner. RESULTS: Inter- and intra-reader reliability were high for both direct measurements and for magnetic resonance imaging measurements of phantoms. Statistically significant differences were noted between the mean direct measurements and the mean magnetic resonance imaging measurements for phantom 1 when using a GRE MPGR sequence (512x512 pixels, 1.5-mm slice thickness, 5:49 min scan time), while borderline differences were noted for T1 SE sequences with the following parameters: 320x320 pixels, 1.5-mm slice thickness, 6:11 min scan time; 320x320 pixels, 4-mm slice thickness, 6:11 min scan time; and 512x512 pixels, 1.5-mm slice thickness, 9:48 min scan time. Borderline differences were also noted when using a FSPGR sequence with 512x512 pixels, a 1.5-mm slice thickness and a 3:36 min scan time. CONCLUSIONS: FSPGR sequences, regardless of the magnetic resonance imaging parameter combination used, provided accurate measurements. The GRE MPGR sequence using 512x512 pixels, a 1.5-mm slice thickness and a 5:49 min scan time and, to a lesser degree, all tested T1 SE sequences produced suboptimal accuracy when measuring the widest phantom.
Subject(s)
Cartilage/diagnostic imaging , Dimensional Measurement Accuracy , Magnetic Resonance Imaging/methods , Phantoms, Imaging , Observer Variation , Radiographic Image Interpretation, Computer-Assisted , Reference Values , Reproducibility of Results , Signal-To-Noise Ratio , Time FactorsABSTRACT
The previously monotypic genus Pseudeurybata Hennig is revised to include seven species from Central America and Mexico and one species from South America. Pseudeurybata rufilabris (Enderlein) and Pseudeurybata compeditus (Hennig) are given as new combinations and a lectotype is designated for P. rufilabris (Enderlein). Pseudeurybata browni and P. alces from Costa Rica, P. guatemalensis from Guatemala, P. dasypogon from Mexico, and P. zeta from Colombia and Ecuador are described as new. All species are keyed and illustrated.
Subject(s)
Diptera/classification , Animal Distribution , Animal Structures/anatomy & histology , Animal Structures/growth & development , Animals , Body Size , Diptera/anatomy & histology , Diptera/growth & development , Female , Male , Organ Size , South AmericaABSTRACT
OBJECTIVES: To evaluate the accuracy of magnetic resonance imaging measurements of cartilage tissue-mimicking phantoms and to determine a combination of magnetic resonance imaging parameters to optimize accuracy while minimizing scan time. METHOD: Edge dimensions from 4 rectangular agar phantoms ranging from 10.5 to 14.5 mm in length and 1.25 to 5.5 mm in width were independently measured by two readers using a steel ruler. Coronal T1 spin echo (T1 SE), fast spoiled gradient-recalled echo (FSPGR) and multiplanar gradient-recalled echo (GRE MPGR) sequences were used to obtain phantom images on a 1.5-T scanner. RESULTS: Inter- and intra-reader reliability were high for both direct measurements and for magnetic resonance imaging measurements of phantoms. Statistically significant differences were noted between the mean direct measurements and the mean magnetic resonance imaging measurements for phantom 1 when using a GRE MPGR sequence (512x512 pixels, 1.5-mm slice thickness, 5:49 min scan time), while borderline differences were noted for T1 SE sequences with the following parameters: 320x320 pixels, 1.5-mm slice thickness, 6:11 min scan time; 320x320 pixels, 4-mm slice thickness, 6:11 min scan time; and 512x512 pixels, 1.5-mm slice thickness, 9:48 min scan time. Borderline differences were also noted when using a FSPGR sequence with 512x512 pixels, a 1.5-mm slice thickness and a 3:36 min scan time. CONCLUSIONS: FSPGR sequences, regardless of the magnetic resonance imaging parameter combination used, provided accurate measurements. The GRE MPGR sequence using 512x512 pixels, a 1.5-mm slice thickness and a 5:49 min scan time and, to a lesser degree, all tested T1 SE sequences produced suboptimal accuracy when measuring the widest phantom.
Subject(s)
Cartilage/diagnostic imaging , Dimensional Measurement Accuracy , Magnetic Resonance Imaging/methods , Phantoms, Imaging , Observer Variation , Radiographic Image Interpretation, Computer-Assisted , Reference Values , Reproducibility of Results , Signal-To-Noise Ratio , Time FactorsABSTRACT
Mesoconius Enderlein, including Zelatractodes Enderlein new synonym, is revised for Central America with one described species (Mesoconius hemithorax Frey) and five new species (M. bicolor, M. dianthus, M. nigrihumeralis, M. tigrinus, and M. zadbi). The relatively large species of this entirely Neotropical genus, most of which are restricted to narrow high elevation ranges, lack the male genital fork that characterizes all other Taeniapterinae.
Subject(s)
Diptera/classification , Animal Distribution , Animal Structures/anatomy & histology , Animal Structures/growth & development , Animals , Body Size , Central America , Diptera/anatomy & histology , Diptera/growth & development , Ecosystem , Female , Male , Organ SizeABSTRACT
The Caribbean and North American species of the large, otherwise neotropical genus Grallipeza Rondani (Diptera: Micropezidae: Taeniapterinae) are revised and keyed. One species, G. nebulosa (Loew), is endemic to the southeastern United States and eighteen species are endemic to single Caribbean islands. Of these, the following nine are described as new: Grallipeza abeja, G. albiterga, G. cliffi, G. grenada, G marleyi, G. mellea, G. paraplacida, G. perezi and G. turba.
Subject(s)
Diptera/anatomy & histology , Diptera/classification , Animal Distribution , Animals , Female , Male , North America , Phylogeny , Species Specificity , West IndiesABSTRACT
Abstract A highly sensitive real-time PCR procedure to detect and quantify the number of Pisciricketsia salmonis units in seawater samples from affected farm sites has been developed. The purpose was to determine a fallowing period that would allow safe restocking of the target farm with new fish. Bacterial load was determined in water samples by comparing the obtained amplification values against a standard curve generated by the amplification of known concentrations of the ITS-ribosomal component of P. salmonis DNA, cloned in a suitable vector. The standard curve was linear over the range of 10(1)-10(10) log units. Target samples were taken every 10 days over a 40-day period, at 5 m depth and at the surface. In a highly affected area of southern Chile, the number of bacterial units in farm water decreased to zero at day 50. Therefore, a fallowing period of 50 days post-removal of cages of affected fish appears to be appropriate before restocking. This procedure could be adapted to control disease problems because of other pathogens in fish farm waters.
Subject(s)
Fish Diseases/microbiology , Fisheries , Piscirickettsia/physiology , Piscirickettsiaceae Infections/veterinary , Seawater/microbiology , Water Microbiology , Animals , DNA, Bacterial/analysis , Piscirickettsiaceae Infections/microbiology , Reference Standards , Salmon , Seawater/chemistry , Time FactorsABSTRACT
Because of the continued importance of correct condom-use in controlling the HIV epidemic and the limited availability of tools for assessing correct condom-use, methods for assessing condom-application skills, especially when direct observation is not feasible, are needed. Accordingly, in the context of a high-risk population (The Bahamas) for HIV, a 17-item scale--the Condom-use Skills Checklist (CUSC)--was developed for use among young adolescents and adults. The rationale and approach to developing the scale and some measures of internal consistency, construct validity, and criterion-related validity have been described. It is concluded that the scale offers a reasonable alternative to direct observation among older subjects and that further development may make it more useful among pre-adolescents.
Subject(s)
Condoms/statistics & numerical data , Contraception Behavior/statistics & numerical data , HIV Infections/prevention & control , Health Knowledge, Attitudes, Practice , Adult , Bahamas , Child , Female , Humans , Male , Psychometrics/methods , Psychometrics/statistics & numerical data , Reproducibility of Results , Surveys and QuestionnairesABSTRACT
Neutropenia associated with race/ethnicity has essentially been unexplained and, although thought to be benign, may affect therapy for cancer or other illnesses. A recent study linked a single nucleotide polymorphism (SNP) (rs2814778) in the Duffy antigen/receptor chemokine gene (DARC) with white blood cell count. We therefore analysed the association of the rs2814778 CC, TC and TT genotypes with absolute neutrophil count (ANC) among asymptomatic women from the Caribbean, Europe and the United States. Among 261 study participants, 33/47 women from Barbados/Trinidad-Tobago, 34/49 from Haiti, 26/37 from Jamaica, and 29/38 US-born black women, but only 4/50 from the Dominican Republic and 0/40 US- or European-born whites (P = 0.0001) had the CC genotype. In a linear regression model that included percentage African ancestry, national origin, cytokines, socio-economic factors and the ELA2 rs57834246 SNP, only the DARC rs2814778 genotype and C-reactive protein were associated with ANC (P < 0.0001). Women with the CC genotype had lower ANC than other women. Further research is needed on the associations of rs2814778 genotype with neutropenia and treatment delay in the setting of cancer. A better understanding of these associations may help to improve cancer outcomes among individuals of African ancestry.
Subject(s)
Humans , Female , Neutropenia , Ethnicity , Chemokines , Genotype , Trinidad and Tobago , Caribbean RegionABSTRACT
BACKGROUND: Low white blood cell counts (WBC) or absolute neutrophil counts (ANC) may delay or prevent the completion of appropriate chemotherapy, especially among women receiving adjuvant therapy for breast and colon cancer, and affect cancer survival. Because race/ethnicity is also associated with survival, the authors compared WBC and ANC in healthy American-born women of African descent and European descent, and women from Barbados/Trinidad-Tobago, the Dominican Republic, Haiti, and Jamaica. METHODS: Blood samples from 261 healthy women ages 20 to 70 years were tested for WBC with differential, cytokine and growth factor levels, and ancestry informative and neutrophil elastase polymorphisms. The authors analyzed the association between neutropenia and serum WBC growth factor levels, cytokine levels, and neutrophil elastase c199a polymorphism. RESULTS: The median WBC and ANC differed among the 6 groups (P < .01 for WBC and P < .0001 for ANC). Dominicans were found to have higher median WBC and ANC than all other groups (P < .03). Neutropenia (ANC < 1500 cu/mm) was observed among 2.7% to 12.5% of the groups of predominantly African descent; no other groups were found to have neutropenia (P < .05). Granulocyte-colony-stimulating factor was found to be lower in white women, but tumor necrosis factor-alpha and C-reactive protein were not found to be correlated with ethnicity. Women of African origin were more likely to have polymorphisms of African ancestry (P < .001) and c199a alleles (P < .0001), which were also associated with low ANC levels. CONCLUSIONS: In the current study, the authors observed a strong association between neutropenia and African descent among asymptomatic women from the U.S. and the Caribbean. Among women of African descent who develop a malignancy, this association may contribute to racial disparities in treatment and outcomes.
Subject(s)
Black or African American , Neutropenia/ethnology , Neutropenia/epidemiology , Adult , Aged , Ethnicity , Female , Hispanic or Latino , Humans , Leukocyte Count , Middle Aged , Neutrophils , United States/epidemiology , United States/ethnology , West Indies/epidemiology , White PeopleABSTRACT
BACKGROUND: Low white blood cell counts (WBC) or absolute neutrophil counts (ANC) may delay or prevent the completion of appropriate chemotherapy, especially among women receiving adjuvant therapy for breast and colon cancer, and affect cancer survival. Because race/ethnicity is also associated with survival, the authors compared WBC and ANC in healthy American-born women of African descent and European descent, and women from Barbados/Trinidad-Tobago, the Dominican Republic, Haiti, and Jamaica. METHODS: Blood samples from 261 healthy women ages 20 to 70 years were tested for WBC with differential, cytokine and growth factor levels, and ancestry informative and neutrophil elastase polymorphisms. The authors analyzed the association between neutropenia and serum WBC growth factor levels, cytokine levels, and neutrophil elastase c199a polymorphism. RESULTS: The median WBC and ANC differed among the 6 groups (P < .01 for WBC and P < .0001 for ANC). Dominicans were found to have higher median WBC and ANC than all other groups (P < .03). Neutropenia (ANC < 1500 cu/mm) was observed among 2.7% to 12.5% of the groups of predominantly African descent; no other groups were found to have neutropenia (P < .05). Granulocyte-colony-stimulating factor was found to be lower in white women, but tumor necrosis factor-alpha and C-reactive protein were not found to be correlated with ethnicity. Women of African origin were more likely to have polymorphisms of African ancestry (P < .001) and c199a alleles (P < .0001), which were also associated with low ANC levels. CONCLUSIONS: In the current study, the authors observed a strong association between neutropenia and African descent among asymptomatic women from the U.S. and the Caribbean. Among women of African descent who develop a malignancy, this association may contribute to racial disparities in treatment and outcomes.
Subject(s)
Adult , Middle Aged , Aged , Humans , Female , Comparative Study , Research Support, Non-U.S. Gov't , Blood Cell Count , Black or African American , Hispanic or Latino , Leukocyte Count , Neutropenia/ethnology , Neutropenia/epidemiology , Neutrophils , United States/epidemiology , United States/ethnology , West Indies/epidemiology , Caribbean Region , Trinidad and TobagoABSTRACT
Although anal intercourse carries great risk for HIV transmission, little research has focused on it among the general population, particularly pre- and early adolescents. This study describes the prevalence of anal and vaginal intercourse among Bahamian pre- and early adolescents and associations with other risk behaviours, family interactions and intrapersonal correlates. Data were from 1274 sixth-grade students aged 9-14 years who completed self-administered questionnaires at baseline of a larger school-based behavioural intervention study. Youth who reported having had anal intercourse engaged in significantly higher rates of several risk behaviours and were significantly more likely to engage in risk behaviours over the next six months, compared with youth with a history of vaginal intercourse only, who in turn were more likely than virgin adolescents. Youth indulging in anal intercourse also perceived significantly lower levels of parental monitoring. Multivariate analyses revealed that anal intercourse, vaginal intercourse, reduced parental monitoring, depression and perceived friend high-risk involvement were associated with both past involvement and future intention to engage in other risk behaviours. Anal intercourse poses a direct threat to the health of these children and is a flag for a constellation of other risks.
Subject(s)
Risk-Taking , Sexual Behavior/statistics & numerical data , Sexually Transmitted Diseases/transmission , Adolescent , Bahamas , Child , Coitus , Depression , Female , HIV , HIV Infections/transmission , Humans , Male , Parent-Child RelationsABSTRACT
Lylmphocyte subsets/CD4 T Helper cell enumeration in HIV care and treatment in resource constrained settings can be difficult to ascertain as a result of the lack of the necessary instrumentation, capacity and infrastructure. However. it is imperative to gain such information for patient monitoring in HIV. The Total Lymphocyte Count (TLC) is useful as a surrogate marker for CD4 count as recommended by the World Health Organisation (WHO) and to calculate CD4% for pacdiatric use. This study therefore sets out to determine and compare the accuracy of the total lymphocyte counts obtained from three haematology analysers designated A. B and C. that are in regular use for routine haemnatological parameters at the main referral hospital in Barbados. West Indies. The TLC of 263 HIV treatment naive individuals attending the HIV Reference Unit in Barbados were enumnerated on the three haematology analysers. The lymphosumn (Sum of lymphocyte subsets: T-helper cell. T-cytotoxic cells. B lymphocytes and Natural killer cells) should be equal to the TLC. and is derived by immunophenotypic analysis on a 4-colour flowcytometer. Machine C had the highest positive correlation between the TLC and the lymphosumn with and R' of 0.9031 compared to machine A with an R values of 0.7119 and Machine B with R(2) values of 0.637. These results show that there can be dramatic inaccuracies when using routine haematology analysers for both routine use. as a surrogate marker of CD4 or for derivation of CD4% in HIV management. It further demonstrates that all haematology analyzers require some form of Quality control. The possible lack of accuracy of the TLC by haematology analysers should be taken into consideration when following the recommendations of the WHO in resource poor settings or using it as a denominator for calculating CD4%.
Subject(s)
HIV Infections/diagnosis , Lymphocyte Count/instrumentation , Biomarkers/analysis , CD4 Antigens/blood , CD4 Antigens/immunology , CD4 Lymphocyte Count/instrumentation , CD4 Lymphocyte Count/methods , CD4 Lymphocyte Count/standards , Equipment and Supplies/economics , Equipment and Supplies/standards , HIV Infections/therapy , Humans , Lymphocyte Count/methods , Lymphocyte Count/standards , Quality ControlABSTRACT
Reported strains of Piscirickettsia salmonis, a pathogen of salmonid fishes, were analyzed by amplifying part of the internal transcribed spacer (ITS) of the ribosomal RNA (rRNA) operon followed by denaturing gradient gel electrophoresis (DGGE) of the amplicons. All amplified fragments differing in sequence were distinguished by migration during DGGE. A simpler format, constant denaturant gel electrophoresis (CDGE), allowed the same diagnostic distinctions among strains. Sampling during 1997 and 1998 of salmonids from 5 different sites on and near Chiloé Island in southern Chile displaying piscirickettsiosis revealed only P. salmonis resembling LF-89, the type strain first isolated in 1989. These observations are encouraging for control strategies, which might otherwise be compromised by unpredictable shifts of P. salmonis types in salmon farms. A competitive PCR assay offered insight about the power of PCR for quantification and about specific tissue invasiveness by this intracellular pathogen. This approach revealed that the PCR could amplify approximately 1 to 10 P. salmonis genome equivalents against a background of > 99.9% salmonid DNA. It also raised the possibility that the salmonid brain is an important site for P. salmonis survival, with its bacterial load in 1 individual having been about 100 times the loads observed in liver and kidney. Pathogen detection by competitive PCR in a surface seawater sample from a netpen in use indicated a density of about 3000 to 4000 P. salmonis cells (or their DNA remnants) 1(-1). Such quantitative estimates should aid decisions about disease prevention and management as, for example, choice of netpen sites following fallow periods and certification of ova, which are known conduits of infection.
Subject(s)
Bacterial Infections/veterinary , DNA, Bacterial/chemistry , Fish Diseases/microbiology , Gammaproteobacteria/genetics , Genetic Variation , Salmonidae , Animals , Aquaculture , Bacterial Infections/microbiology , Base Sequence , Chile , DNA, Bacterial/isolation & purification , Molecular Sequence Data , Oncorhynchus kisutch , Oncorhynchus mykiss , Polymerase Chain Reaction/veterinary , Polymorphism, Genetic , RNA, Bacterial/genetics , RNA, Ribosomal/genetics , Sequence Analysis, DNAABSTRACT
The attributes of the PCR allowed implementation of an assay for specific detection of Piscirickettsia salmonis from a few microliters of fish serum. This opens the way to less invasive modes of sampling for this microbial pathogen in salmonids.
Subject(s)
Fish Diseases/diagnosis , Gram-Negative Bacteria/isolation & purification , Gram-Negative Bacterial Infections/veterinary , Polymerase Chain Reaction/methods , Salmonidae/microbiology , Animals , Aquaculture , Bacteremia/diagnosis , Bacteremia/veterinary , DNA, Bacterial/analysis , DNA, Bacterial/isolation & purification , DNA, Ribosomal/analysis , DNA, Ribosomal/isolation & purification , Gram-Negative Bacteria/genetics , Gram-Negative Bacterial Infections/diagnosis , Gram-Negative Bacterial Infections/microbiology , Muscles/microbiology , Operon/genetics , RNA, Ribosomal, 23S/geneticsABSTRACT
One form of a group of enzymes known as aspartate kinases, primarily reported in prokaryotes and plants, might also exist in animal cells. Here we report the immunodetection of an aspartate kinase-like activity in human platelets using antibodies against the pure form of the enzyme purified from Escherichia coli. Moreover, the enrichment of platelet extracts with the bacterial kinase results in the phosphorylation of discrete forms mainly of membrane-bound endogenous polypeptides.