Distribution of Human Papillomavirus (HPV) Genotypes in HIV-Negative and HIV-Positive Women with Cervical Intraepithelial Lesions in the Eastern Cape Province,South Africa

South African women have a high rate of cervical cancer cases, but there are limited data on human papillomavirus (HPV) genotypes in cervical intraepithelial neoplasia (CIN) in the Eastern Cape province, South Africa. A total of 193 cervical specimens with confirmed CIN from women aged 18 years or older, recruited from a referral hospital, were tested for HPV infection. The cervical specimens, smeared onto FTA cards, were screened for 36 HPV types using an HPV direct flow kit. HPV prevalence was 93.5% (43/46) in CIN2 and 96.6% (142/147) in CIN3. HIV-positive women had a significantly higher HPV prevalence than HIV-negative women (98.0% vs. 89.1%, p = 0.012). The prevalence of multiple types was significantly higher in HIV-positive than HIV-negative women (p = 0.034). The frequently detected genotypes were HPV35 (23.9%), HPV58 (23.9%), HPV45 (19.6%), and HPV16 (17.3%) in CIN2 cases, while in CIN3, HPV35 (22.5%), HPV16 (21.8%), HPV33 (15.6%), and HPV58 (14.3%) were the most common identified HPV types, independent of HIV status. The prevalence of HPV types targeted by the nonavalent HPV vaccine was 60.9% and 68.7% among women with CIN2 and CIN3, respectively, indicating that vaccination would have an impact both in HIV-negative and HIV-positive South African women, although it will not provide full protection in preventing HPV infection and cervical cancer lesions.     

An Eco-Epidemiological Study of Contamination of Soil with Infective Forms of Intestinal Parasites

The objectives of the present work were to screen topsoil samples collected from public squares in two cities within the Argentine Patagonia for the presence of infective forms of intestinal parasites and to examine the possible relationship between positive findings and the environmental, socioeconomic, and cultural conditions of that region. For this purpose we studied 13 public squares, their 13 custodians, and 44 family groups within their respective surrounding areas. Of the 226 topsoil samples analyzed, 44.3% proved positive for infective forms of intestinal parasites, with 17.3% of these containing more than one species. The frequency of appearance of positive samples was dependent on the season of the year (p < 0.001), while presence of the parasites was related to the soil pH (p < 0.05) but independent of the soil relative humidity (p > 0.05). Some of the organisms detected are associated with zoonoses. We observed the presence of Capillaria spp. and Spirocerca spp. under cool desert climatic conditions. Within the group of custodians we detected hematologic alterations one positive serology for toxoplasmosis and documented behavior conducive to risk of infection with the parasites found in those squares. Within the family group an acquaintance with parasitic zoonoses and their prevention was an inconsistent finding, with toxocarosis and toxoplasmosis being the diseases associated with the greatest degree of ignorance. Furthermore, we consider the failure to de-parasitize pets and the practice of feeding them with raw meat, as typically found in our family survey, to be factors contributing to a greater likelihood of public square contamination. From the results obtained here, we propose a spatial organization approach for the purpose of detecting zones at risk of contracting zoonotic parasitoses within urban environments.     

Synthesis, characterization, and preliminary in vivo tests of new poly(ethylene glycol) conjugates of the antitumor agent 10-amino-7-ethylcamptothecin

Despite the high antitumor activity of camptothecins, few derivatives have been developed and tested for human treatment of solid tumors, due to unpredictable toxicity mainly connected to their poor water solubility. We report the conjugation of the antitumor agent 10-amino-7-hydroxy camptothecin (SN-392) to linear or branched poly(ethylene glycol)s (PEGs) of different loading capacity through a tri- or tetrapeptide spacer selectively cleaved by lysosomal enzymes (cathepsins). A synthetic strategy based on the chemoselective acylation of the aromatic amino group in the presence of the unprotected C20 tertiary alcohol allowed high overall yields. Two conjugates demonstrated good stability at physiological pH and in mouse plasma (nonspecific proteases) but slowly released the drug payload in the presence of the lysosomal enzyme cathepsin B1. Compound 3, selected for in vivo experiments, was very active against P388, P388/ADM leukaemia, and Meth A fibrosarcoma cell lines, scoring T/C% values comparable with the camptothecin derivative CPT-11. Pharmacokinetic studies indicated that 3 acts as a reservoir of 10-amino-7-ethylcamptothecin, as the mean residence time (MRT) is about 3-fold higher than that of the free drug.     

Signaling capacity of FcgammaRII isoforms in B-CLL cells

Two main isoforms of Fcgamma receptor II (CD 32) have been described in humans: activatory FcgammaRIIA and inhibitory FcgammaRIIB. We have previously reported that B cells from a subset of chronic lymphocytic leukemia (B-CLL) patients express not only FcgammaRIIB, as normal B lymphocytes, but also the myeloid FcgammaRIIA. The aim of this study was to evaluate the signaling capacity of both FcgammaRII isoforms in B-CLL cells. We found that FcgammaRIIA expressed by leukemic cells failed to induce Ca(2+) mobilization or protein tyrosine phosphorylation, suggesting that the receptor is not functional. By contrast, FcgammaRIIB effectively diminished BCR-triggered ERK 1 phosphorylation, which indicates that it is able to transduce inhibitory signals in B-CLL cells. Moreover, we found that FcgammaRIIB homoaggregation in either B-CLL or non-malignant tonsillar B cells did not result in apoptosis as was reported for murine B splenocytes. Together, these results show that FcgammaRIIB, but not FcgammaRIIA is biologically active in B-CLL cells and might influence leukemic cell physiology in vivo.     

Viewing the body prepares the brain for touch: effects of TMS over somatosensory cortex

Viewing the body can improve tactile perception. We investigated whether this could be due to a remodeling of somatosensory cortical areas during vision of the body. Single-pulse transcranial magnetic stimulation (TMS) was delivered over the primary and secondary somatosensory areas of subjects who showed clear visual-tactile enhancement while they performed a tactile grating discrimination task. Before the tactile stimulus, subjects viewed either their right index finger through a semisilvered mirror or an object reflected by the mirror and positioned to appear in the same location as the finger. In a first experiment we observed that TMS over primary somatosensory cortex significantly reduced subjects' accuracy whilst viewing the hand. No such reduction was found when subjects viewed a neutral object. In a second experiment, we disrupted the activity of primary and secondary somatosensory areas in different sessions. When stimulating the primary somatosensory cortex, a reduction in accuracy was again found while viewing the hand, but not a neutral object. TMS over secondary somatosensory cortex had no effect in any condition. Our results show that vision of the body may act at an early stage in stimulus elaboration and perception, allowing an anticipatory tuning of the neural circuits in primary somatosensory cortex that underlie tactile acuity.     

Classical and nonclassical 21-hydroxylase deficiency: a molecular study of Argentine patients

OBJECTIVE: To characterize the molecular basis of the 21-hydroxylase deficiency in a group of Argentine patients presenting the classical and nonclassical forms of the disease. DESIGN: To analyse the frequency of point mutations in the CYP21 gene by DNA amplification and mutation detection. PATIENTS: Forty-one patients from 36 nonrelated families: 25 nonclassical (NC), 11 salt-wasting (SW) and five simple virilizing (SV). A total of 27 parents and 13 nonaffected siblings were also analysed. MEASUREMENTS: Basal steroid hormones and 17-hydroxyprogesterone levels following adrenal stimulation with adrenocorticotrophic hormone were measured, together with an analysis of 10 point mutations in the CYP21 gene. RESULTS: A total of 83% and 74.4% classical and nonclassical chromosomes, respectively, were characterized. The intron 2 mutation was the most prevalent among classical alleles. In addition, a high frequency for R356W was observed in both groups (13.3 and 6.9%, respectively), while V281L was the most frequent mutation among the nonclassical patients with a frequency of 39.5%. No alleles containing P30L were observed, and one de novo mutation (R356W) was found. A total of 68.3% patients were fully genotyped, and all but one showed no genotype/phenotype discrepancy. Though the cut-off value for post-ACTH 17-hydroxyprogesterone stimulation was 30.25 nmol/l (10.00 microg/l), the lowest value observed in the fully genotyped nonclassical group was 42.35 nmol/l (14.00 microg/l). CONCLUSIONS: The high number of unidentified alleles in the nonclassical group suggests that less frequent mutations, or the presence of new ones, might be the cause of the disease in the Argentine population. Alternatively, the cut-off value in the ACTH-stimulated 17-hydroxyprogesterone test might overestimate the diagnosis of the nonclassical form by including some patients with heterozygous status.     

Junin virus-induced astrocytosis is impaired by iNOS inhibition

Because Junin virus (JV) experimental encephalitis of mice and rats is characterized by mild histopathological changes that do not seem to justify per se lethality after intracerebral infection, such a murine model seems adequate to investigate the potential role of inducible nitric oxide synthase (iNOS) as a pathogenic factor. Concomitant with a predominant astrocyte reaction, increased immunoperoxidase expression of iNOS, mitochondrial superoxide dismutase (SODm) and glutathione peroxidase (GPX) was disclosed in brain of mice infected with JV strain #44. When specific inhibition of iNOS was achieved by intraperitoneal administration of amino guanidine (AG), significantly greater mortality was observed in treated animals (70% vs. 40%), together with similar infective titers ( approximately 10(7) PFU/g) but lower astrocytosis, as shown by glial fibrillary acidic (GFAP) labeling. As regards SODm and GPX immunochemical expression in neurons, no differences were found between mice with or without AG treatment. The present results suggest that the apparent protective role of nitric oxide (NO), when synthesized by iNOS, is unrelated to reduced viral replication but rather to enhanced astrocyte activation behaving as a beneficial cell response to virus-induced CNS damage.     

Thyroglobulin reference values in a pediatric infant population.

Few reports have addressed normal serum thyroglobulin (Tg) values in newborns and infants. In the present study, serum Tg was measured in 228 normal children (110 females and 118 males) aged from 3 to 180 days of life, all presenting normal age-related serum thyrotropin (thyroid-stimulating hormone [TSH]) values and negative anti-Tg and antithyroperoxidase antibodies. Serum Tg was measured by Radioimmunoassay (RIA) (two methods) and Immunometricassay (IMA) (three methods). Mean Tg values measured by the five methods exhibited among-method biases, although a significant positive correlation was observed. Serum Tg levels measured by the five methods showed a correlation with age, but not with TSH or gender. During the first days of life, relatively high mean Tg levels were observed, which progressively decreased until they reached a plateau. Therefore, with the aim of establishing reference values, the population was separated into two age groups: from 3 to 15 days of life (group A) and from 16 to 180 days of life (group B). Mean Tg concentration in group A was statistically higher than in group B. Tg centile distributions were calculated with the aim of establishing the normal levels of serum Tg for each method. We conclude that for a correct interpretation of serum Tg levels, the age and the methods used should be considered.