Rapid and simultaneous typing of hemoglobin S, hemoglobin C, and seven Mediterranean beta-thalassemia mutations by covalent reverse dot-blot analysis: application to prenatal diagnosis in Sicily

The molecular lesions causing beta-thalassemia in Sicily can be subdivided into two groups. One that occurs at a 71% frequency and consists of the beta 39, IVS 1,110 and IVS 1,6 mutations and the other group at a 20% frequency comprising the -87, beta s, IVS 1,1 and IVS 2,745 mutations. The identification of all these mutations by polymerase chain reaction (PCR) and conventional dot-blot hybridization has been time consuming and expensive. In this article, we describe the implementation of the reverse dot-blot (RDB) hybridization as a rapid nonradioactive method for the identification of the nine most frequent molecular lesions in the beta-globin gene (-87, beta s, beta c, IVS 1,1, IVS 1,6, IVS 1,110, beta 39, IVS 2,1, IVS 2,745) in Sicily. Sixty prenatal diagnoses were performed by this RDB assay, each of which was confirmed by dot-blot/ASO hybridization; thus demonstrating the accuracy of the RDB. The main advantage of this assay is the rapid typing of an individual's DNA for many mutations in a single working day. Because the mutations in this assay are representative for the Mediterranean region, this mutational panel can also be extended to the screening of beta-thalassemia from other Mediterranean regions.     

Recommendations for HIV surveillance in Europe

The HIV/AIDS epidemic in Europe is now at an important turning point. In western Europe, AIDS incidence has been declining since 1996, partly as a result of new, more effective treatment of HIV infections. As these treatments delay the progression of HIV     

Microaggregate counts in frozen-preserved erythrocytes: effects of washing in three blood processors and filtration

Since microaggregates have been implicated in posttransfusion pulmonary insufficiency, their elimination has become an active concern in blood transfusion. Various types of filters, as well as frozen-preserved erythrocytes, have been used to provide blood relatively low in microaggregates. We have counted particles in frozen-stored blood before deglycerolization, after washing in each of three cell processing systems, and after filtration through a 40-micrometer filter. Washing frozen erythrocytes reduced the total particle counts by an average of 89%. Slight differences were found among the three blood processors with respect to particle removal. Passing washed blood through a 40-micrometer filter did not result in significant further reduction in particle counts. Hence, the use of such filters in a frozen-preserved blood system is not warranted.     

Measuring the cochlear blood flow and distortion-product otoacoustic emissions during reversible cochlear ischemia: a rabbit model.

Impairment to the cochlear blood flow likely induces many types of sensorineural hearing loss. Models using several small laboratory animals have been described in the literature that permit the simultaneous monitoring of the cochlear blood flow with laser-Doppler flowmetry and cochlear function using evoked responses. However, these models have not permitted a direct application of the resulting knowledge to the human condition, primarily due to differences in the translucence of the otic capsule between species. In the present study, to approximate conditions relevant to the human patient, the rabbit was utilized to develop a procedure in which laser-Doppler flowmetry could be used to measure the cochlear blood flow in an animal with an opaque otic capsule. At the same time, the cochlear function was monitored non-invasively using distortion-product otoacoustic emissions. In this manner, a laser-Doppler probe was positioned in the round window niche and the cochlear function measured using distortion-product otoacoustic emissions during a systematic series of ischemic episodes. Cochlear ischemia was produced by deliberately compressing the eighth nerve complex at the porus of the internal acoustic meatus, for periods lasting from 1-3 min, while cochlear blood flow and distortion-product otoacoustic emission measures were obtained simultaneously before, during and following the occlusion. Results demonstrated that the cochlear blood flow sharply decreased within 1 s after compression onset, whereas distortion-product otoacoustic emissions showed obstruction-induced changes after a delay of several seconds, provided that the blood flow decreased, at least 40%. Similarly, upon release of the compression, the cochlear blood flow began to recover within 1 s, whereas the recovery of the corresponding distortion-product otoacoustic emissions was slightly delayed. Although not apparent in the distortion-product otoacoustic emission recovery time course, the cochlear blood flow consistently overshot its initial baseline value during the recovery process. Thus, although cochlear ischemia produced changes in the distortion-product otoacoustic emission activity that generally followed the resulting alterations in the cochlear blood flow, the detailed relationship between the two measures was complex.     

Early sexual initiation: the role of peer norms

OBJECTIVE: To elucidate which components of peer norms influence the process of sexual initiation for young adolescents. Design. Prospective cohort study. Setting. Fourteen elementary and middle schools in an urban public school district. Participants. The 1389 sixth-grade students who completed the questionnaire at the beginning (time 1) and at the end (time 2) of the school year comprise the study sample. Mean age at time 1 was 11.7 years. RESULTS: Of students entering the sixth grade, 30% (n = 416) reported having already initiated sexual intercourse, 5% (n = 74) reported initiating sexual intercourse during the sixth-grade school year (initiated group), and 63% (n = 873) reported not having initiated sexual intercourse by the end of the sixth-grade school year (never group). Demographic comparisons revealed that students in the initiated group were significantly more likely than students in the never group to be older (11.9 years vs 11.6 years), male (58% vs 37%), African-American (70% vs 51%), attending a poorer school (87% vs 85%), and living in an area with a high proportion of single-parent families (45% vs 41%). Self-reports and reports of peers' participation in nonsexual risk behaviors were more common for students in the initiated group. Students in the initiated group were more likely than students in the never group to perceive: 1) a high prevalence of sexual initiation among peers; 2) social gains associated with early sexual intercourse; and 3) younger age of peers' sexual initiation. Students in the never group were more likely to believe that sexually-experienced 12-year-old boys would be negatively stigmatized compared with students in the initiated group. Three predictive models were developed to test the relationship between peer norms and the process of initiation. These models demonstrate that the strongest predictor of sexual initiation in sixth grade is having high intention to do so at the beginning of sixth grade. The strongest predictor of high intention is belief that most friends have already had sexual intercourse. Perceptions of social gain and stigma for sexually-experienced 12-year-old boys act independently of intention to decrease risk of early sexual initiation. CONCLUSION: Early sexual intercourse is not an unplanned experience for many teens. Decisions about initiation are strongly bound to social context with peers playing an important role in creating a sense of normative behavior. Specific components of peer norms impact the process of sexual initiation in both positive and negative ways. Interventions aimed at delaying the onset of sexual initiation need to focus on cohort norms as well as on an individual's perceptions and behaviors.     

Scintigraphic Visualization of Haemostatic and Thromboembolic Processes Using 111Indium-labelled Platelets

ABSTRACT Five cases of scintigraphic imaging of haemostatic and thromboembolic processes after injection of autologous ¹¹¹In-labelled platelets are presented. The site of a liver puncture, atherosclerotic plaques in the abdominal aorta, deep vein thrombi, pulmonary embolism and the tip of an indwelling venous catheter could be visualized. Activity measurements corresponding to atherosclerotic plaques suggest that an increased sensitivity of this method may be achieved by extending the investigation period to 4–5 days after injection of ¹¹¹In platelets.     

Complications of percutaneous nephrostolithotomy

In 224 patients, renal stones were removed from the urinary tract using either direct extraction with a basket or forceps (59 patients), ultrasonic lithotripsy (164 patients), or infusion chemotherapy (one patient). Residual stone fragments were present more frequently in patients treated with ultrasonic lithotripsy (27%) than with direct extraction (5%). Other complications included hemorrhage (eight patients), catheter dislodgement (four patients), large amounts of urine extravasation (three patients), glycine ascites (three patients), infection (two patients), pneumothorax (one patient), and a prolonged ileus (one patient). More complications occurred among the first 50 patients than the last 50 patients, even though more difficult cases, including patients with staghorn calculi, were accepted during the latter period. Although a learning curve exists, complications can be minimized by attempting to treat more favorable cases during the initial experience.     

Lentiginous macules and patches of neurofibromatosis (an approach to better terminology)

BACKGROUND AND OBJECTIVES: Freckles (ephelides) are small, light brown macules of sun-exposed areas in fair-skinned subjects. On the other hand, freckle-like pigmentation of the axilla is a highly characteristic feature of neurofibromatosis. To what extent (clinically and pathologically) are the two pigmentary defects similar? And to what extent are café-au-lait patches and freckle-like lesions similar pathologically? SUBJECTS AND METHODS: Ten cases of neurofibromatosis and 10 cases of freckles were examined clinically. Two biopsies were taken from the former; one obtained from an axillary freckle-like lesion, and the other from a café-au-lait patch. One biopsy only was taken from freckles. The biopsies were processed for haematoxylin and eosin, silver stain and transmission electron microscopy. RESULTS: Obvious differences were detected between freckles (ephelides) and freckle-like lesions in cases of neurofibromatosis. Café-au-lait patches and freckle-like lesions showed the same pathological changes of lentigo simplex, with increased melanin pigment from the basal layer up to the stratum corneum, moderate elongation of the rete ridges and mild inflammatory infiltrate intermingled with melanophages. Electron microscopy revealed an increase in melanocyte number and the presence of giant melanin granules in freckle-like pigmentation as well as café-au-lait patches. These ultrastructural freckles (ephelides) showed an increase in activity but not the number of melanocytes. CONCLUSIONS: The term freckle-like should be changed to lentiginous macules of neurofibromatosis. On the other hand, the café-au-lait patches should be considered as lentiginous patches. Freckles should be restricted to sun-exposed areas in fair-skinned persons only.