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Total hip arthroplasty (THA) and hemiarthroplasty are common treatments for severe hip joint disease. To predict the probability of re-admission after discharge when patients are hospitalized will support providing appropriate health education and guidance.The research aims to use logistic regression (LR), decision trees (DT), random forests (RF), and artificial neural networks (ANN) to establish predictive models and compare their performances on re-admissions within 30 days after THA or hemiarthroplasty. The data of this study includes patient demographics, physiological measurements, disease history, and clinical laboratory test results.There were 508 and 309 patients in the THA and hemiarthroplasty studies respectively from September 2016 to December 2018. The accuracies of the four models LR, DT, RF, and ANN in the THA experiment are 94.3%, 93.2%, 97.3%, and 93.9%, respectively. In the hemiarthroplasty experiment, the accuracies of the four models are 92.4%, 86.1%, 94.2%, and 94.8%, respectively. Among these, we found that the RF model has the best sensitivity and ANN model has the best area under the receiver operating characteristic (AUROC) score in both experiments.The THA experiment confirmed that the performance of the RF model is better than the other models. The key factors affecting the prognosis after THA surgery are creatinine, sodium, anesthesia duration, and dialysis. In the hemiarthroplasty experiment, the ANN model showed more accurate results. Poor kidney function increases the risk of hospital re-admission. This research highlights that RF and ANN model perform well on the hip replacement surgery outcome prediction.

Submarine canyons are common features of the coastal ocean. Although they are known to be hotspots of turbulence that enhance diapycnal transport in their stratified waters, the dynamics of canyon mixing processes are poorly understood. Most studies of internal wave dynamics within canyons have focused on a handful of canyons with along-axis slopes less steep than semidiurnal (D2) internal wave characteristics (subcritical). Here, we present the first tidally-resolving observations within a canyon with a steeply sloping axis (supercritical). A process study consisting of two 24-hour shipboard stations and a profiling mooring was conducted in the La Jolla Canyon off the coast of La Jolla, CA. Baroclinic energy flux is oriented up-canyon and decreases from 182 ± 18 W m−1 at the canyon mouth to 46 ± 5 W m−1 near the head. The ratio of horizontal kinetic energy to available potential energy and the observed group speed of each mode are lower than expected for freely propagating D2 internal waves at each station, indicating partial reflection. Harmonic analysis reveals that variance is dominated by the D2 tide. Moving up-canyon, the relative importance of D2 decreases and its higher harmonics are needed to account for a majority of the observed variance, indicating steepening. Steep internal tides cause large isopycnal displacements (∼50 m in 100m water depth) and high strain events. These events coincide with enhanced O(10−7 -10−5 m2 s−3) dissipation of turbulent kinetic energy at mid-depths.

The buttress wall is an alternative construction method for protection of adjacent buildings during excavation. It is generally recognised, but not verified, that the method has good effects in reducing wall movement and ground settlement, because buttress walls behave as T-shaped concrete beams, which possess comparatively larger stiffness than two-dimensional diaphragm walls. However, the behaviour of buttress walls in excavations is not fully understood, because of a lack of well-monitored case histories and systematic studies. In this study, the effectiveness of buttress walls in reducing wall deflection and ground settlement is evaluated through observed behaviour of a case history, and through finite element studies. The case study indicates that the use of buttress walls in a deep excavation can significantly reduce the deflection of a diaphragm wall. Effective prediction of the behaviour of the buttress wall can be achieved using the three-dimensional finite element method. Parametric studies reveal that deflection of the diaphragm wall is strongly influenced by the condition of the restraint at the bottom of the buttress walls, and by the number of buttress walls. As long as the buttress wall bottom is well restrained, and reasonable numbers of buttress walls are installed, wall deflection can be considerably reduced.

The collection of dried blood spots (DBS) on filter paper provides a powerful approach for the development of large-scale, population-based screening programs. DBS methods are particularly valuable in developing countries and isolated rural regions where resources are limited. Large numbers of field specimens can be economically collected and shipped to centralized reference laboratories for genetic and (or) serological analysis. Alternatively, the dried blood can be stored and used as an archival resource to rapidly establish the frequency and distribution of newly recognized mutations, confirm patient identity or track the origins and emergence of newly identified pathogens. In this report, we describe how PCR-based technologies are beginning to interface with international screening programmes for the diagnosis and genetic characterization of human immunodeficiency virus type 1 (HIV-1). In particular, we review recent progress using DBS specimens to resolve the HIV-1 infection status of neonates, monitor the genetic evolution of HIV-1 during early infancy and establish a sentinel surveillance system for the systematic monitoring of HIV-1 genetic variation in Asia.

The prevalence of allergic diseases has increased in the past decades. It is unknown whether expression of certain microRNAs (miRNAs) in neonatal leucocytes is correlated to IgE production and/or allergic diseases.This study investigated the association of miRNA expression in neonatal leucocytes with cord blood IgE (CBIgE) elevation and development of allergic disease.We screened for the expression of a panel of 157 miRNAs in mononuclear leucocytes from human umbilical cord blood (CB) samples with elevated CBIgE and tracked the association of down-regulated miRNA expression to the miRNA-targeted gene expression and to children with allergic rhinitis (AR).Among the initial screen of 10 CB samples with elevated CBIgE, expression of eight of the 157 miRNAs was low. Of these eight down-expressed miRNAs, three remained down-regulation in a validation with other 20 CB samples, and two of the three miRNAs, miR-21 and miR-126, were significantly lower in monocytes from AR children. Further analysis of mRNA expression of the miR-21-targeted genes identified that TGFBR2 expression on monocytes was significantly up-regulated in CB with elevated CBIgE, and in AR patients. Transfection of miR-21 precursor into monocytes from patients with AR increased miR-21 expression and decreased TGFBR2 expression.This study demonstrated the first in the literature that lower miR-21 expression in CB and increased TGFBR2 expression is associated with antenatal IgE production and development of AR.

Prevalence of allergic diseases in children has increased worldwide over the past decades. Allergy sensitization may occur in fetal life. This study investigated whether gene-gene and gene-environment interactions affected cord blood IgE (CBIgE) levels.A total of 575 cord blood DNA samples were subjected to a multiplex microarray for 384 single nucleotide polymorphisms (SNPs) in 159 allergy candidate genes. Genetic association was initially assessed by univariate and multivariate analyses. Multifactor dimensionality reduction (MDR) was used to identify gene-gene and gene-environment interactions. Environmental factors for analysis included maternal atopy, paternal atopy, parental smoking, gender, and prematurity.Twenty-one SNPs in 14 genes were associated with CBIgE elevation (or =0.5 KU/l) in univariate analysis. Multivariate analysis identified eleven genes (IL13, IL17A, IL2RA, CCL17, CXCL1, PDGFRA, FGF1, HAVCR1, GNAQ, C11orf72, and ADAM33) which were significantly associated with CBIgE elevation. MDR analyses of gene-gene interactions identified IL13 interacted with IL17A and/or redox genes on CBIgE elevation with the prediction accuracy of 62.52%. Analyses of gene-environment interactions identified that maternal atopy combined with IL13, rs1800925 and CCL22, rs170359 SNPs had the highest prediction accuracy of 67.15%. All the high and low risk classifications on gene-gene and gene-environment interactions by MDR analyses could be validated by Chi-square test.Gene-gene (e.g. immune and redox genes) and gene-environment (e.g. maternal atopy and FGF1or redox genes) interactions on IgE production begin in prenatal stage, suggesting that prevention of IgE-mediated diseases may be made possible by control of maternal atopy and redox responses in prenatal stage.

A two day meeting hosted by the World Health Organization (WHO) and the U.S. Centers for Disease Control and Prevention (CDC) was held in May 2006 in Entebbe, Uganda to review the laboratory performance of virologic molecular methods, particularly the Roche Amplicor DNA PCR version 1.5 assay, in the diagnosis of HIV-1 infection in infants. The meeting was attended by approximately 60 participants from 17 countries. Data on the performance and limitations of the HIV-1 DNA PCR assay from 9 African countries with high-burdens of HIV/AIDS were shared with respect to different settings and HIV- subtypes. A consensus statement on the use of the assay for early infant diagnosis was developed and areas of needed operational research were identified. In addition, consensus was reached on the usefulness of dried blood spot (DBS) specimens in childhood as a means for ensuring greater accessibility to serologic and virologic HIV testing for the paediatric population.

We attempted to establish normal Doppler flow velocity waveform patterns in the human fetal ductus venosus (DV), and also to establish a standardized measurement technique.Ductus venosus blood flow was measured in a prospective study involving 545 fetuses aged between 8 and 38 weeks in utero, the mothers of whom received prenatal care in Kaohsiung Chang Gung Memorial Hospital in a 12-month period in 1998-1999. Several DV hemodynamic parameters were assessed, including peak systolic velocity (DVP), peak systolic/diastolic (S/D) ratio, time-averaged velocity (TAMX), maximum velocity during atrial contraction (DVM), pulsatility index (PI), Pourcelot's resistance index (RI), and fetal heartbeat (FHB).Technically acceptable ductus venosus blood flow velocity waveform patterns were collected from 490 of 545 pregnant women (89.9%). The mean +/- SD value for the peak systolic DV velocity during the time period of 8 to 38 weeks in utero was 0.33 +/- 0.11 meters/sec (m/s), the TAMX being 0.24 +/- 0.09 m/s. The maximum velocity during atrial contraction was 0.15 +/- 0.09 m/s, and the peak S/D velocity ratio was 2.5 +/- 1.01. The PI, Pourcelot's RI and fetal heart beat were, 0.67(+/- 0.21), 0.64 (+/- 0.11), and 163.3 (+/- 18.82 bpm), respectively. Significant increases in DVP, TAMX, and DVM with advancing gestational age were established, and decreases in PI, RI, S/D, and FHB with advancing gestational age were also observed.Further investigation of DV hemodynamics throughout pregnancy may enable a greater understanding of normal placental perfusion, the fetal venous return to the heart and associated cardiac function.

Microsatellite instability was first reported in hereditary nonpolyposis colorectal cancer (HNPCC) as well as other cancers, including endometrial and ovarian cancers. Single base repeat markers of human MSH3 and MSH6 genes were found to precipitate the action of human MSH2. The marker BAT-26 was reported to be a simple, low-cost, and rapid marker for detection replication errors (RER) and the status of colorectal cancers. We analyzed di-nucleotide repeats of the microsatellite markers (D2 S123, D5 S82, D5S299, D10S197, D17S791, D18S34), single base repeat markers (DeltaP3, hMSH3, hMSH6, and TGFbeta-RII), and BAT-26 to evaluate microsatellite instability in cervical cancer. Altogether 80 paired cervical cancers were studied. Our results showed that microsatellite instability is not common in cervical cancer, and the mutation of the single base repeat of mismatch repair (MMR) genes (hMSH3 and hMSH6) is also uncommon. The BAT-26 is not a good marker to detect the RER status of cervical cancer.

The purpose of our study was to assess the influence of intra-uterine insemination (IUI) on the results of maternal serum Down syndrome screening. 43 women with IUI pregnancies and 4507 healthy women who conceived were studied. Ovulation in IUI pregnancies was induced by clomiphene and/or human menopausal gonadotrophin (hMG). Maternal serum levels of free beta-human chorionic gonadotrophin (hCG) and alpha-fetoprotein (AFP) were measured for Down syndrome screening. It was considered screen-positive when the risk of Down syndrome was 1 in 270 or greater in the second trimester. The value of maternal serum AFP was significantly lower in the IUI group (median=0.760 MoM) than in the control group (median=1.050 MoM). However, the value of free beta-hCG was not significantly different between the two groups. The positive rate of maternal serum Down syndrome in IUI pregnancies was similar to that of the control group. Our results indicate that IUI pregnancy may be associated with a lower level of AFP, although the mechanism for this difference remains unknown.

Retinal detachment is an unusual complication of hypertensive disorder in pregnancy. It has been reported in 1% to 2% of patients with severe preeclampsia and in 10% of patients with eclampsia. Choroidal ischemia may be the cause of retinal detachment. We know that mild arteriolar spasm involving the bulbar conjunctival vessels has been observed in the normal pregnancy, but in pregnancy-induced hypertension the vasospasm may be severe and result in choroidal ischemia. Most patients with retinal detachment in pregnancy-induced hypertension have had full spontaneous resolution within a few weeks, and they did not have any sequelae. Medical treatment with antihypertensive drugs and steroids may be helpful. We report two rare cases of retinal detachment and persistent hypertension in association with postpartum eclampsia and post-cesarean section preeclampsia. These patients had normotension throughout pregnancy. Preeclampsia or eclampsia developed after delivery, and blurred vision, headache, and reduced vision accompanied serous retinal detachment. The serous retinal detachment disappeared within 3 weeks. Good outcomes were found in the follow-up examinations in both of these cases. For women who had been normotensive at the time of delivery and then complained in the postpartum period of blurred vision, headaches, nausea and vomiting, we should consider the possibility of retinal detachment and perform fundoscopy.

Deletion of the proximal portion of chromosome 4q is apparently rare. To our knowledge, prenatal diagnosis of the interstitial deletion of 4q12-21.1 has never been reported. We present a prenatal case of 4q deletion in association with a positive Down syndrome screening test of an elevated maternal serum free beta human chorionic gonadotrophin (beta-hCG) level. The prenatal sonogram revealed intra-uterine growth retardation (IUGR) and shortening of the femur. Facial dysmorphism included micrognathia, depressed nasal bridge and low-set ears, these anomalies were evident at the postnatal examination. All of the anomalies were consistent with those described in proximal 4q deletion syndrome. Our case suggests that chromosome studies may be indicated for patients with high maternal serum free beta-hCG and IUGR in the early second trimester.

A polymorphism associated with a thermolabile variant (C677T) of the enzyme methylenetetrahydrofolate reductase has been associated with both elevated total homocysteine (tHcy) levels and risk for cardiovascular disease. Data from the Stroke Prevention in Young Women Study were used to determine the prevalence of the C677T genotype and to assess whether environmental factors modified the association between genotype and tHcy concentration. The C677T genotype prevalence was 80% -/-, 20% +/-, and 0% +/+ among 46 African-American women; and 39% -/-, 53% +/-, and 8% +/+ among 77 white women (P0.01). There was a trend toward higher tHcy levels in African-American women with the +/- genotype when compared with the -/- genotype (6.9 mumol/L vs 5.3 mumol/L respectively, p = 0.10); no association was found among the white women (6.0 mumol/L, -/-; 4.5 mumol/L, +/-; and 6.2 mumol/L, +/+; p = 0.67). Among African American women, those who smoked and were +/- genotype had the highest tHcy levels (8.0 mumol/L); while among white women, those who smoked and were -/- had the highest tHcy levels (8.1 mumol/L). Despite being hampered by a limited sample size, the thermolabile allele is significantly less common among African-American than white women. The association between genotype and tHcy concentration is influenced by smoking and multivitamin use.

We report a fetus with partial trisomy 13 [46,XY,dup(13)(q14-31)] which was found during an amniocentesis performed due to the age of the mother. This duplication segment (13q 14-31) has not been reported in literature. The clinical features of our patient included cleft lip/palate, low set ears, depressed nasal bridge, hypertelorism, and epicanthal fold. After termination of the pregnancy, the fetus was sent for an autopsy. The autopsy report was compatible with the gross findings and no other abnormalities, including central nervous, cardiac, and renal system, were found. The level of maternal alpha-fetoprotein, 2.67 MoM, was opposite with the common phenomenon in complete trisomy 13.

Advances in genetic technology, such as the polymerase chain reaction (PCR) (1) and the work of the Human Genome Project to map and sequence the human genome (2-4), are expediting our understanding of the genetic determinants of common disease. For example, genetic factors are proving to be important in determining who is susceptible to infectious agents such as the human immunodeficiency virus (5), as well as in identifying populations who are at risk for cancer because of exposure to environmental hazards such as cigarette smoke and aniline dyes (6-9). DNA is now being used in epidemiologic investigations to study genetic risk factors (10). Genetic risk factors include rare gene mutations, such as the BRCA1 mutation which confers high risk for breast or ovarian cancer (11, 12), as well as genetic polymorphisms, such as the polymorphic genes that code for carcinogen-metabolizing enzymes that affect the way people process carcinogens, thereby increasing or decreasing their risk for sporadic forms of cancer (13). Because of new technologies that have become a part of routine molecular methods in the last decade, especially PCR, specimens other than fresh whole blood can be used, thereby, in many cases, making specimen collection in the field possible for the first time (14). All nucleated cells, including cells from hair follicles, buccal swabs, and urine specimens, are suitable specimens for DNA analysis (15-20).

Peripheral blood mononuclear cell specimens were collected from 13 HIV-1-infected IV drug users in Kuala Lumpur, Malaysia, as well as one HIV-infected baby, between 1992 and 1993. DNA was then amplified by nested polymerase chain reaction and a 345-bp fragment of the C2V3 region of the env gene was sequenced. 11 of the 14 Malaysian sequences clustered with the B' subtype, one different from the typical subtype B US strains HIVMN and HIVSF2. Two sequences grouped in the C subtype and had sister taxa closer to the Indian C subtype sequences than those from Zambia. The sequence from the infant was identified as a subtype E virus, grouped more closely with subtype E strains from Thailand than subtype E viruses from the Central African Republic.

Global surveillance of HIV-1 subtypes for genetic characterization is hampered by the biohazard of processing and the difficulties of shipping whole blood or cells from many developing country regions. We developed a technique for the direct automated sequencing of viral DNA from dried blood spot (DBS) specimens collected on absorbent paper, which can be mailed unrefrigerated in sturdy paper envelopes with low biohazard risk. DBS were collected nonrandomly from HIV-1-infected, mostly asymptomatic, patients in five Asian countries in 1991, and shipped via airmail or hand carried without refrigeration to Bangkok, and then transshipped to North America for processing. After more than 2 years of storage, including 6 months at ambient temperatures, proviral DNA in the DBS was amplified by nested PCR, and a 389-nucleotide segment of the C2-V3 env gene region was sequenced, from which 287 base pairs were aligned and subtyped by phylogenetic analysis with neighbor-joining and other methods. From southern India, there were 25 infections with subtype C and 2 with subtype A. From Myanmar (Burma), we identified the first subtype E infection, as well as six subtype BB, a distinct cluster within subtype B that was first discovered in Thailand and that has now appeared in China, Malaysia, and Japan. From southwest China, one BB was identified, while a "classical" B typical of North American and European strains was found in Indonesia. From Thailand, five DBS of ambiguous serotype were identified as three B, one BB, and one E. A blinded control serotype E specimen was correctly identified, but a serotype BB control was not tested. Most HIV-1 in southern India appears to be env subtype C, with rare A, as others have reported in western and northern India. The subtypes BB and E in Myanmar, and the BB in China, suggest epidemiological linkage with these subtypes in neighboring Thailand. DBS are a practical, economical technique for conducting large-scale molecular epidemiological surveillance to track the global distribution and spread of HIV-1 variants.The global surveillance of HIV-1 subtypes for genetic characterization is frustrated by the danger of processing and the difficulties of shipping whole blood or cells from many developing country regions. The authors therefore developed a technique for the direct automated sequencing of viral DNA from dried blood spot (DBS) specimens collected on absorbent paper. Such specimens can be mailed unrefrigerated in paper envelopes with low biohazard risk. 51 DBS specimens were collected nonrandomly from HIV-1-infected, mostly asymptomatic individuals in India, Myanmar, China, Indonesia, and Thailand in 1991, then shipped via airmail or hand carried without refrigeration to Bangkok from where they were forwarded to North America for processing. After more than 2 years in storage, including 6 months at ambient temperatures, proviral DNA in 42 of the DBS was successfully amplified by nested polymerase chain reaction, and a 389-nucleotide segment of the C2-V3 env gene region was sequenced, from which 287 base pairs were aligned and subtyped by phylogenetic analysis with neighbor-joining and other methods. From southern India, there were 25 infections with subtype C and two with subtype A; the first subtype E infection was identified from Myanmar, as well as six subtype B(B); one B(B) was identified from southwest China; subtype B was identified from Indonesia; and five DBS of ambiguous serotype classified as three B, one B(B), and one E were identified from Thailand. DBS can be used as a practical, cost-effective way of tracking the global distribution and spread of HIV-1 variants.

Persons with a thermolabile form of the enzyme 5,10 methylenetetrahydrofolate reductase (MTHFR) have reduced enzyme activity and increased plasma homocysteine which can be lowered by supplemental folic acid. Thermolability of the enzyme has recently been shown to be caused by a common mutation (677C--T) in the MTHFR gene. We studied 41 fibroblast cultures from NTD-affected fetuses and compared their genotypes with those of 109 blood specimens from individuals in the general population. 677C--T homozygosity was associated with a 7.2 fold increased risk for NTDs (95% confidence interval: 1.8-30.3; p value: 0.001). These preliminary data suggest that the 677C--T polymorphism of the MTHFR gene is a risk factor for spina bifida and anencephaly that may provide a partial biologic explanation for why folic acid prevents these types of NTD.

The human immunodeficiency virus (HIV) was introduced readily into Asia and has quickly spread between Asian states through both parenteral and sexual modes of transmission. Only 1 year after Thailand's epidemic wave among intravenous drug users (IDUs) in 1988, the virus spread to the adjacent Myanmar and Malaysia, and another year later IDUs were infected in parts of India and China bordering Myanmar. Several methods can be used to quantify the genetic diversity, divergence, or variation within or between subtypes, genotypes, or isolates. Consensus sequences, representing the most common nucleotide in the genome, are often generated for comparison. 8 subtypes A through F, H, and O have been described for HIV-1 based on the genetic similarities and differences in the env gene or viral envelope. Subtype A and D have been found primarily in central and western Africa. Subtype B is predominant in Europe, the Western hemisphere, Japan, and Australia. Subtype C has been found mostly in southern Africa, the Central African Republic, and India. Subtype E was first identified in Thailand and recently in the Central African Republic. Subtype F has been found in Romania and is a rare variant in Brazil. Isolates from Gabon and the Russian Federation were designated subtype H. An "outlier" subtype O containing 2 human and 2 chimpanzee isolates has been identified in Cameroon and Gabon. Sequencing of the relatively conserved gag gene of geographically diverse HIV-1 isolates yielded a classification with 7 subtypes A-D and F-H. Other topics discussed include genome characterization, comparison with foreign isolates, segregation by mode of transmission, and biologic properties of HIV-1 variants in Thailand; regional diversity of HIV-1 subtypes and substantial spread of HIV-2 in India; as well as HIV transmission and infections in Japan, Australia, Cambodia, China, Taiwan, Philippines, Malaysia, Myanmar, and in states created out of the former Soviet Union.

Maternal antibodies against the V3 loop principal neutralizing domain (PND) have been reported to protect against perinatal HIV-1 transmission. To study this association in an African city with a long-standing HIV epidemic and no established "consensus sequence" for the V3 loop region of gp120, we determined the DNA sequence for the V3 region of HIV-1 from 13 HIV-1-infected residents of Kinshasa, Zaire, and developed peptide enzyme immunoassays (EIAs) reflecting the V3 loop PND for those HIV-1 strains. Using the most broadly reactive locally derived V3 loop peptide in a limited-antigen EIA, there was no significant difference in the perinatal HIV-1 transmission risk between 64 women with anti-V3 loop antibody (transmission risk, 30%) and 104 women without anti-V3 loop antibody (transmission risk, 25%; p = 0.5); this finding was unchanged after we controlled for maternal AIDS and low birth weight. Although we used assays for V3 loop antibody based on local HIV-1 strains and evaluated a large number of mother-child pairs, we found no evidence that maternal anti-V3 loop PND antibody protects against perinatal HIV-1 transmission.

A nonradioactive, colorimetric microplate hybridization procedure was used to assay human immunodeficiency virus (HIV) DNA, amplified by the polymerase chain reaction (PCR). Under the PCR conditions used, four proviral copies per 150,000 cells were detected by amplifying a series of DNA mixtures that contained various copy numbers of HIV. Assays of PCR-amplified DNA from peripheral blood mononuclear cells of seronegative individuals yielded negative results (104 of 104), whereas samples from seropositive individuals yielded99% positive results (141 of 142). Similar results were obtained in a chemiluminescent assay with an acridinium ester-labeled probe and in a solution hybridization assay in which a 32P-labeled probe was used.

We have investigated the cause of dual serological reactivity to human immunodeficiency virus (HIV) types 1 and 2, a common occurrence in West Africa. Serum specimens from 111 individuals from Cote d'Ivoire classified by commercial western blot as HIV-1 (n = 15), HIV-2 (32), and dually reactive (64) were further tested by more specific serological tests (a synthetic peptide enzyme immunoassay [Pepti-LAV 1/2] and western blots prepared from antigen in which oligomeric forms of the transmembrane protein were disrupted by trichloroacetic acid [WB-TCA]). Peripheral blood mononuclear cells were tested for HIV-1 and HIV-2 with the polymerase chain reaction (PCR) and virus culture. Of 104 samples that were concordant by both WB-TCA and Pepti-LAV, 82 (79%) were confirmed by PCR results. Virus culture was concordant with serology for specimens (35/38) in which any virus was detected. Our findings indicate that mixed HIV-1/HIV-2 infections are common in Cote d'Ivoire, and suggest that natural infection by one HIV type does not prevent heterologous infection.

A chitobiase gene from Vibrio parahemolyticus was cloned into plasmid pUC18 in Escherichia coli strain DH5 alpha. The plasmid construct, pC120, contained a 6.4 kb Vibrio DNA insert. The recombinant gene expressed chitobiase [EC 3.2.1.30] activity similar to that found in the native Vibrio. The enzyme was purified by ion exchange, hydroxylapatite and gel permeation chromatographies, and exhibited an apparent molecular weight of 80 kDa on SDS-polyacrylamide gel electrophoresis. Chitobiose and 6 more substrates, including beta-N-acetyl galactosamine glycosides, were hydrolyzed by the recombinant chitobiase, indicating its putative classification as an hexosaminidase [EC 3.2.1.52]. The enzyme was resistant to denaturation by 2 M NaCl, thermostable at 45 degrees C and active over a very unusual (for glycosyl hydrolases) pH range, from 4 to 10. The purified cloned chitobiase gave 4 closely focussed bands on an isoelectric focusing gel, at pH 4 to 6.5. The N-terminal 43 amino acid sequence shows no homology with other proteins in commercial databanks or in the literature, and from its N-terminal sequence, appears to be a novel protein, unrelated in sequence to chitobiases from other Vibrios reported and unrelated to hexosaminidases from other organisms.

A prospective study was conducted in the Bronx, New York, of 70 infants of human immunodeficiency virus (HIV)-infected (n = 33) and uninfected (n = 37) mothers who had a history of intravenous drug use or of intravenous drug-using sex partners. Infants were observed from birth to a median age of 23 months (range 3 to 54 months). HIV infection was confirmed in seven infants (21%) of seropositive mothers; six developed HIV disease, with symptoms observed in the first year. Of these, three died (3, 9, and 36 months) of HIV-related causes; 3 of 4 survivors were greater than 25 months of age. HIV symptoms preceded or were concurrent with abnormalities in T-lymphocyte subsets; postneonatal polymerase chain reaction confirmed HIV infection in five infants with symptoms and one without symptoms. Among infants of seropositive mothers, seven without laboratory evidence of HIV (including polymerase chain reaction) had findings suggestive of HIV infection, including persistent generalized lymphadenopathy, hepatosplenomegaly, oral candidiasis, parotitis, and inverted T-lymphocyte ratios. These findings were not observed in infants of seronegative mothers. Although the presence of HIV proviral sequences was associated with HIV disease, the observation of indeterminate symptoms in at-risk infants indicates the importance of long-term clinical follow-up to exclude HIV infection. Disease manifestations in comparable infants of seronegative mothers are important for assessment of the impact of maternal drug use, development of specific clinical criteria for early diagnosis of HIV and eligibility for antiretroviral therapy.

UV irradiation provides a simple and efficient way to minimize contamination or false positivity which often occurs in laboratories performing routine PCR tests. Here, we characterize several parameters of the effect of UV irradiation on DNA template, primers, deoxynucleoside triphosphate and Taq polymerase. UV irradiation of DNA results in the formation of pyrimidine dimers and thus prevents them from being effective templates in subsequent PCR. Reduction of the HIV DNA templates in polypropylene microcentrifuge tubes by more than 1000-fold can be achieved by UV irradiation. The sensitivity of the primers is sequence- and concentration-dependent. Oligonucleotides with neighboring thymine bases are more susceptible to UV than those without. Taq polymerase is highly UV sensitive, whereas deoxynucleotide triphosphate is relatively UV resistant.

Six months after receiving 58 units of blood components, a 65-year-old white man from New York City, with no other risk factors for human T-lymphotropic virus type I (HTLV-I) infection, developed HTLV-I-associated myelopathy/ tropical spastic paraparesis (HAM/TSP). Investigation of blood donors identified a 25-year-old white Hispanic woman from Florida whose platelets had been given to the patient and who was seropositive for the virus on a serum specimen obtained 2 years after the donation. She was born in Cuba and had had 2 sexual relationships with men who either had been born in or had resided in the Caribbean. Polymerase chain reaction (PCR) studies of peripheral blood mononuclear cells indicated that both donor and recipient were infected with HTLV-I. Molecular studies of a 595-nucleotide sequence in the 5' envelope region of HTLV-I indicated that the viruses from donor and recipient were identical in each of 32 positions in which published HTLV-I sequences demonstrate molecular heterogeneity; the donor and recipient viruses were also identical in 2 additional positions in which they differed from all published sequences. Transfusion-associated HAM/TSP has occurred in the United States, but additional cases should be prevented by screening blood donations for HTLV-I. Molecular studies of HTLV-I may prove useful in defining the genetic heterogeneity of HTLV-I isolates in the United States and in studying transmission of this virus.

Background: Beare first described a 2-year-old boy who had abnormal physical feature including hypertelorism, cutis gyratum, acanthosis nigrican, cleft palate, functional pyloric obstruction and abnormal genitalia. Ten cases of Beare–Stevenson cutis gyrata syndrome have been reported. Here, we present, the first case of antenatal ultrasound image of this rare syndrome. A 31-year-old Taiwanese, gravida 2, para 1, was referred for fetal evaluation at 32 weeks gestation because of polyhydramnios and fetal head anomaly. Conventional sonography revealed a single fetus with amniotic index of 27.3 cm, cover leaf skull, and a skin fold-like umbilical cord. Three-dimensional sonography of the face of the fetus showed, high wide forehead, ocular proptosis, and depressed nasal bridge. The patient delivered a male infant weighing 2980 g at 39 weeks gestation. Physical examination of the infant showed furrows of corrugated appearance of the skin, craniofacial anomalies, particularly craniosynosis, displaced ears, prominent umbilical stump and hypospadia. Computer tomography and magnetic resonance image were performed and revealed fronto-ethmoid-nasal, cephalocele, pansynostosis and frontal base defect. Conclusion: A total of 10 cases of Beare–Stevenson cutis gyrata syndrome have been previously reported. All of the previously reported cases of the syndrome were diagnosed postnatally. Our patient presented with ocular hypertelorism, low-set and creased ears, ocular proptosis, choanal atresia, cloverleaf skull, craniosynosis, cutis gyrata, acanthosis nirican, prominent umbilical stump and bifida scrotum. Here we present the first case of antenatal ultrasound image of this rare syndrome.

We molecularly cloned unintegrated viral DNA of the BALB/c endogenous N-tropic and B-tropic murine leukemia retroviruses and in vitro passaged N-tropic Gross (passage A) murine leukemia retroviruses. Recombinant genomes were constructed in vitro by exchanging homologous restriction enzyme fragments from N- or B-tropic parents and subsequent recloning. Infectious virus was recovered after transfection of these recombinant genomes into NIH-3T3 cells and cocultivation with the Fv-1 nonrestrictive SC-1 cells. XC plaque assays of recombinant virus progeny on Fv-ln and Fv-lb cells indicated that the Fv-l host range was determined by sequences located between the BamHI site in the p30 region of the gag gene (1.6 kilobase pairs from the left end of the map) and the HindIII site located in the pol gene (2.9 kilobase pairs from the left end of the map).

Nucleotide sequences of the region that corresponds to the site of tRNA primer binding for a functional retrovirus were determined in five murine leukemia virus-related sequence clones from mouse chromosomal DNA, which contain a unique 170 to 200-base-pair additional internal segment in the long terminal repeats. The 3'-terminal 18-nucleotide sequence of a major glutamine tRNA isoacceptor was found to match well with the putative primer binding site: 18 of 18 in three clones, 17 of 18 in one clone, and 16 of 18 in one clone. This implies that most of these endogenous proviral sequences of the mouse genome, if replicated as retroviruses, will be different from ecotropic murine leukemia viruses and most mammalian type C retroviruses in using glutamine tRNA, rather than proline tRNA, as a primer.

Polymerase chain reaction (PCR) was used to detect human immunodeficiency virus (HIV)-1 DNA in peripheral blood mononuclear cells to assess in hemophilic men whether any were HIV-seropositive but uninfected or seronegative but infected and in seronegative sex partners of seropositive hemophilic men whether any were infected. Of 40 seropositive men, 38 (95%) were PCR-positive; one was PCR-indeterminate and one PCR-negative. None of 41 seronegative men who used only donor-screened, virus-inactivated coagulation factor products were PCR-positive. However, two of six who received noninactivated products were PCR-positive; one had low T-helper cell counts and died of unrelated causes and the other had seroconverted 11 mo later. PCR with a second primer pair also detected HIV-1 DNA in these two men. None of 25 seronegative female sex partners of seropositive men, including six men with AIDS and seven with AIDS-related symptoms, were PCR-positive. These data suggest that most seropositive hemophilic men are HIV-infected; whether some are infected with defective virus remains to be resolved as does the infection status of seropositive PCR-negative men. Identification of two seronegative PCR-positive men supports the possibility that HIV-1 DNA can be detected before seroconversion.

The pancreatic polypeptide response to a meal depends on various mechanisms, which are only partly understood. The aim of this study was to define whether humoral factors and nutrients which modulate postprandial gastric acid secretion play a role in the regulation of pancreatic polypeptide secretion. Our study was performed in the cat, a species in which pancreatic polypeptide release has never been explored. The animals were provided with a gastric fistula and a Heidenhain pouch and received, in a random order, mixed liver (50 g) per os or different nutriments which were introduced directly into the gastric fistula in identical final volumes: 50 g mixed liver; 1.5 to 12 g oligopeptides; or 2 g of triglycerides or glycogen. Acid output and pancreatic polypeptide secretion were measured over 150 min. In the cat as in dog and man, a mixed meal induced a five to ten-fold increase of plasma pancreatic polypeptide. The protein fraction of the meal was the most potent stimulus for release of this peptide and the pancreatic polypeptide response to protein seemed to be dose-related. The lipid and carbohydrate components of the meal were only weak stimulants. In the cat, a central vagal stimulation is effective on pancreatic polypeptide release, as on gastric acid secretion, since 2-deoxyglucose stimulated both secretions (about 15 p. 100 of maximal response to a meal). The integrated pancreatic polypeptide release and the Heidenhain pouch acid secretion in response to nutriments were correlated during the 30-120 min period.(ABSTRACT TRUNCATED AT 250 WORDS)

The early diagnosis of infection with human immunodeficiency virus (HIV) in infants born to infected mothers is essential for early treatment, but current tests cannot detect HIV infection in newborns because of the presence of maternal antibodies. We used the polymerase chain reaction, a new technique that amplifies proviral sequences of HIV within DNA, to detect HIV infection in peripheral-blood mononuclear cells obtained from infants of seropositive women during the neonatal (age less than 28 days) and postneonatal periods. In blood obtained during the neonatal period, the polymerase chain reaction was positive in five of seven infants in whom the acquired immunodeficiency syndrome (AIDS) later developed (a mean of 9.8 months after the test). The test was also positive in one of eight newborns who later had nonspecific signs and symptoms suggestive of HIV infection (mean follow-up, 12 months). No proviral sequences were detected in neonatal samples from nine infants who remained well (mean follow-up, 16 months). HIV proviral sequences were detected in samples obtained during the postneonatal period (median age, five months) in all of 6 infants tested who later had AIDS and in 4 of 14 infants with nonspecific findings suggestive of HIV infection. No proviral sequences were detected in 25 infants who remained well (mean follow-up, 17 months) after being born to HIV-seropositive mothers, or in 15 infants born to HIV-seronegative mothers. We conclude that the polymerase chain reaction will be a useful technique to diagnose HIV infection in newborns and to predict the subsequent development of AIDS. However, larger studies will be required to determine the sensitivity and specificity of the test.

The variability and discordance of human immunodeficiency virus type 1 (HIV-1) antibody enzyme immunoassay determinations on serial specimens derived, to our knowledge, from the first documented case of HIV-2 infection in North America are described. The initial specimen was weakly reactive, but two subsequent serum specimens were both nonreactive by enzyme immunoassay. All specimens were indeterminate for HIV-1 antibody by HIV-1 Western blot analysis. Serum HIV-2 antibody was demonstrated by enzyme immunoassay using whole virus lysate, HIV-2-specific synthetic peptide assays, and HIV-2 Western blot analysis. Human immunodeficiency virus type 2 genomic sequences were demonstrated in peripheral blood mononuclear cells using gene amplification technology. Human immunodeficiency virus type 2, isolated from peripheral blood lymphocytes, had typical morphologic features of lenti-virus by electron microscopy. Western blot analysis and other specific assays should be considered in individuals with clinical evidence suggesting HIV infection who are nonreactive for HIV-1 antibody by enzyme immunoassay or who have atypical reactivity patterns.

Previously, in vitro recombinant DNA studies demonstrated that genetic determinants of N-tropism and B-tropism, or Fv-1-related host range properties of murine leukemia viruses, were located in a BamHI-HindIII DNA segment derived from the 5' portion of the cloned viral genome. We sequenced this segment and its immediate 5' region from cloned DNA of two BALB/c mouse C-type viruses (WN1802N and WN1802B) and found base differences at 12 positions out of the otherwise identical 1,390-base-pair sequences. Analysis of the most likely reading frame showed that 6 of the 12 base differences would result in four encoded amino acid changes, three of which occur at positions 109 (glutamine in WN1802N versus threonine in WN1802B), 110 (arginine in WN1802N versus glutamic acid in WN1802B), and 159 (glutamic acid in WN1802N versus glycine in WN1802B) of the p30 protein. The remaining one is located at position 36 (threonine in WN1802N versus isoleucine in WN1802B) of the viral polymerase protein. Significant conformational alteration of the p30 protein could be predicted from these amino acid changes.

A colorimetric method of in situ hybridization has been developed for the rapid detection of human immunodeficiency virus (HIV) in formalin-fixed paraffin-embedded material. Following optimization of digestion conditions, biotin-labeled DNA probes are detected with an alkaline phosphatase conjugate. The method is verified using fixed paraffin-embedded cell blocks of HIV-infected and uninfected lymphocyte cell cultures. Hybridization specifically detects both viral RNA and proviral DNA. Formalin fixation for intervals up to 21 d did not significantly hamper the signal under the appropriate digestion conditions; however, Trump's fixation for even 12 h greatly reduced the intensity of the hybridization. This technique for in situ hybridization is amenable to automation, provides results within 6 h, and results in good morphologic preservation. A key feature of the technique is the use of human placental DNA as an endogenous positive control to optimize the empirically determined conditions for protein digestion.