Human T-lymphotropic virus type 2 (HTLV-2) provirus in circulating cells of the monocyte/macrophage lineage in patients dually infected with human immunodeficiency virus type 1 and HTLV-2 and having predominantly sensory polyneuropathy.

We investigated the presence of human T-lymphotropic virus type 2 (HTLV-2) DNA in the peripheral blood mononuclear cell subsets obtained from 18 patients coinfected with human immunodeficiency virus type 1 and HTLV-2, 6 of whom also had predominantly sensory polyneuropathy (PSP). HTLV-2 DNA and RNA were found in CD8- and CD19-positive cells, and, for patients with PSP, in CD14-positive cells as well. Furthermore, the patients with PSP had higher proviral loads than those without PSP.

Prevalence of hepatitis G virus RNA in human immunodeficiency virus type 1-positive intravenous drug users.

OBJECTIVE: The aim of this study was to evaluate the prevalence of the new human flavivirus hepatitis G virus (HGV) in Italian intravenous drug users (IDUs) and its interaction with human immunodeficiency virus type 1 (HIV-1) and hepatitis C virus (HCV). STUDY DESIGN/METHODS: Seventy-nine IDUs with different clinical stages of HIV-1 infection and 20 non-IDU patients with chronic HCV infection were included in the study. HGV RNA was detected by means of reverse transcription-polymerase chain reaction (RT-PCR) used for the amplification of two HGV-related sequences included in the 5'-noncoding (NCR) and NS5a regions. RESULTS: Eighteen (22.8%) of the 79 IDUs were positive for plasma HGV RNA; there was no difference in mean serum alanine aminotransferase (ALT) levels between the HGV-positive and HGV-negative patients. No significant correlation was observed between HGV and other viral markers (hepatitis B virus [HBV], HCV, human T-cell lymphotropic virus type II [HTLV-II]) or HCV genotype. The number of patients with symptomatic HIV-1 infection in whom HGV RNA was detected was significantly lower than the number of those who were asymptomatic (6 of 49 [12.2%] versus 12 of 30 [40%]; P = 0.004). The mean plasma HGV RNA titer was higher in the asymptomatic than in the symptomatic patients (4.6 versus 3.2 log PCR-amplified units in 1 mL of plasma sample [PU/mL]; P = 0.03). CONCLUSIONS: Our results show a considerable spread of HGV levels among Italian HIV-1-positive IDUs and do not indicate that HGV infection enhances liver impairment. We suggest that the greater prevalence of HGV RNA in IDUs with asymptomatic HIV-1 infection may reflect the relatively recent HGV infection in this population.

Quantitation of glycated hemoglobins in human adult blood by capillary isoelectric focusing.

A precise and reproducible method for assessment of glycated hemoglobin in human adult red blood cells is reported, based on capillary isoelectric focusing (IEF). In order to obtain baseline resolution between adult hemoglobin (Hb A) and its glycated form (Hb A1c), two species which differ by minute delta pI values, < 0.03 pH units, the following procedure was adopted: the focusing mixture consisted of 5% Ampholine, pH 6-8, 0.5% Pharmalyte, pH 3-10, 3% short-chain liquid polyacrylamide and an equimolar mixture of two "separators", 0.33 M beta-alanine and 0.33 M 6-aminocaproic acid. The last two compounds flatten the pH gradient in the pI region of the two Hbs, thus allowing full separation. Additionally, the Hb samples, instead of being pulse-loaded, are uniformly distributed in the background electrolyte. A longer capillary life-time is obtained if all nonbuffering ions are eliminated; thus, as catholyte, 50 mM Lys (pH 9.7) is utilized and as anolyte 50 mM acetic acid (pH 3.5) is adopted. The percentages of Hb A1c, as obtained by capillary IEF, are in good agreement (+/- 6%) with data obtained by one of the standard zone electrophoretic methods in clinical chemistry, i.e., the Helena REP Glyco gel system.

An isoelectric focusing study of seasonal variation in rattlesnake venom proteins.

Adult specimens of seven southern Pacific rattlesnakes (Crotalus viridis helleri), four northern black-tailed rattlesnakes (Crotalus molossus molossus), and six western diamondback rattlesnakes (Crotalus atrox) were housed under controlled light and temperature and milked of venom monthly for 20 months. Ambient conditions were modelled to simulate seasonal change. Weighed amounts of lyophilized venom from each snake were compared chronologically for variation in isoelectric focusing patterns, using natural and immobilized gradients. No variation in patterns was evident over this time period for any individual snake. However, intraspecific differences were obvious in the venom samples. The pattern seems indicative of a species, however, concentration of various protein constituents seems individual and genetically "fingerprinted'. Unlike other physiological functions that demonstrate cyclicity in response to temperature and photoperiod, concentration ratios of venom components appear to be constant regardless of external cues. These findings may further emphasize the medical importance of treating snakebite victims symptomatically as individuals. A variation exists in the components of venoms of any given species, as well as in the physiological sensitivities of humans to a venom.

Electrophoretic and chromatographic techniques for the differential diagnosis of a haemoglobin abnormality: Hb E heterozygosity.

A method is described for separating haemoglobin (Hb) E (beta 26 Gly----Lys) from Hb A2 (a normal minor Hb component in adult blood). The technique allows the distinction between subjects carrying beta-thalassaemia trait and patients who are simultaneously alpha-thalassaemic and heterozygous for Hb E, the standard electrophoretic pattern often being similar in these two circumstances. Complete separation between Hb E and Hb A2 (2 mm empty space in between) is obtained by isoelectric focusing in immobilized pH gradients in an ultra-narrow pH 7.55-7.65 gradient. The apparent pI values of the two species (at 10 degrees C and at an average ionic strength of 5.6 mequiv. l-1) have been calculated to be 7.603 for Hb E and 7.607 for Hb A2. Thus, the system reported here affords a resolution of at least 0.004 pH unit.

Immobilized pH gradients and reversed-phase high-performance liquid chromatography: a strategy for characterization of haemoglobin variants with electrophoretic mobility identical to that of Hb A. The case of Hb San Diego.

The preparative aspects of immobilized pH gradients applied to abnormal haemoglobins (Hb) is described. As shown with the example of Hb San Diego, this method is successful even with as small a difference in pHi as 0.01 pH unit. For characterization of such neutral variants, reversed-phase high-performance liquid chromatography is demonstrated to be a very efficient tool.

Neonatal screening of beta-thalassemias by thin layer isoelectric focusing.

As an alternative to the recently proposed screening for beta-thalassemias by isoelectric focusing (IEF) of denatured globin chains in urea-detergent gels [6], IEF of intact hemoglobins, obtained from umbilical cord blood in neonatal units, is reported here. For maximum separation, IEF should be performed in nonlinear pH gradients, in gels containing 0.2 M beta-alanine and 0.2 M 6-amino caproic acid, which flatten the pH gradient around pH 7, thus increasing the resolution between HbA and HbFac. The method can unambiguously detect homozygous and heterozygous beta-thalassemic conditions. A bimodal distribution of HbA at birth has been found: In heterozygous patients, HbA values of 9.02% (range 6.8-9.98%) have been found while in normal newborns HbA levels of 20.34% (range 11.02-30.6%) have been demonstrated.