bluuebunny/crossref_metadata_2025 huggingface dataset:
Circadian variation of EEG power spectra in NREM and REM sleep in humans: Dissociation from body temperature
Authors: Dijk Derk‐Jan
Published: March 2003

In humans, EEG power spectra in REM and NREM sleep, as well as characteristics of sleep spindles such as their duration, amplitude, frequency and incidence, vary with circadian phase. Recently it has been hypothesized that circadian variations in EEG spectra in humans are caused by variations in brain or body temperature and may not represent phenomena relevant to sleep regulatory processes. To test this directly, a further analysis of EEG power spectra – collected in a forced desynchrony protocol in which sleep episodes were scheduled to a 28‐h period while the rhythms of body temperature and plasma melatonin were oscillating at their near 24‐h period – was carried out.

EEG power spectra were computed for NREM and REM sleep occurring between 90–120 and 270–300 degrees of the circadian melatonin rhythm, i.e. just after the clearance of melatonin from plasma in the ‘morning’ and just after the ‘evening’ increase in melatonin secretion. Average body temperatures during scheduled sleep at these two circadian phases were identical (36.72°C). Despite identical body temperatures, the power spectra in NREM sleep were very different at these two circadian phases. EEG activity in the low frequency spindle range was significantly and markedly enhanced after the evening increase in plasma melatonin as compared to the morning phase. For REM sleep, significant differences in power spectra during these two circadian phases, in particular in the alpha range, were also observed.

The results confirm that EEG power spectra in NREM and REM sleep vary with circadian phase, suggesting that the direct contribution of temperature to the circadian variation in EEG power spectra is absent or only minor, and are at variance with the hypothesis that circadian variations in EEG power spectra are caused by variations in temperature.
Gender‐related sleep differences in neonates in thermoneutral and cool environments
Authors: Bach Véronique , Telliez Frédéric , Leke André , Libert Jean‐Pierre
Published: March 2003

Although thermoregulation and sleep exhibit gender differences in adults, the question is still debated in neonates. The aim of this study was to examine the relationship between gender‐related sleep differences and cool defence mechanisms in neonates. Sleep and thermoregulation were recorded in healthy preterm neonates (21 boys and 17 girls, 37 ± 2 weeks post‐conceptional age) exposed to thermoneutral and cool conditions. Sleep was analysed for continuity and structure. Although the cool exposure did not strongly impair body homeothermia, sleep was altered but without any significant gender difference. However, when data recorded under each of the thermal conditions were pooled, some gender differences emerged: boys slept less, with more wakefulness after sleep onset, more active sleep and less quiet sleep than girls. In contrast to sleep architecture, most of the sleep continuity parameters exhibited greater variability in boys than in girls. This variability may bias the statistical analyses and probably explains the varying conclusions reported in the literature regarding gender‐specific sleep‐related differences.
Optimization of sigma amplitude threshold in sleep spindle detection
Authors: Huupponen E. , Värri A. , Himanen S.‐L. , Hasan J. , Lehtokangas M. , Saarinen J.
Published: March 2003

Sleep spindles are transient EEG waveforms of non‐rapid eye movement sleep. There is considerable intersubject variability in spindle amplitudes. The problem in automatic spindle detection has been that, despite this fact, a fixed amplitude threshold has been used. Selection of the spindle detection threshold value is critical with respect to the sensitivity of spindle detection. In this study a method was developed to estimate the optimal recording‐specific threshold value for each all‐night recording without any visual scorings. The performance of the proposed method was validated using four test recordings each having a very different number of visually scored spindles. The optimal threshold values for the test recordings could be estimated well. The presented method seems very promising in providing information about sleep spindle amplitudes of individual all‐night recordings.
Effect of temperature on the pharmacokinetics of benzocaine in rainbow trout (Oncorhynchus mykiss) after bath exposures
Authors: Stehly G.R. , Meinertz J.R. , Gingerich W.H.
Published: March 2003

The pharmacokinetics of benzocaine during bath exposures at 1 mg/L were determined in rainbow trout acclimated at 6 °C, 12 °C or 18 °C for at least 1 month. Individual fish were exposed to benzocaine in a recirculating system for 4 h and pharmacokinetic parameters were estimated in a unique manner from the concentration of benzocaine in the bath water vs. time curve. Elimination from plasma was also determined after the 4 h exposure. The uptake clearance and metabolic clearance increased with increased acclimatization temperatures (uptake clearance 581 ± 179 mL/min/kg at 6 °C and 1154 ± 447 mL/min/kg at 18 °C; metabolic clearance 15.2 ± 4.1 mL/min/kg at 6 °C and 22.3 ± 4.2 mL/min/kg at 18 °C). The apparent volume of distribution had a trend for increasing with temperature that was not significant at the 5% level (2369 ± 678 mL/kg at 6 °C to 3260 ± 1182 mL/kg at 18 °C). The elimination half‐life of benzocaine in plasma was variable and did not differ significantly with temperature (60.8 ± 30.3 min at 6 °C to 35.9 ± 13.0 min at 12 °C). Elimination of benzocaine from rainbow trout is relatively rapid and even more rapid at higher acclimatization temperatures based on calculated metabolic clearances and measured plasma concentrations, but was not evident by measurement of terminal plasma half‐lifes.
Comparison of three doses of dexmedetomidine with medetomidine in cats following intramuscular administration
Authors: Ansah  , Raekallio  , Vainio
Published: March 2003

Cats (n = 6) were administered dexmedetomidine (DEX) and medetomidine (MED) at three different dose levels in a randomized, blinded, cross‐over study. DEX was administered at 25, 50 and 75 μg/kg (D25, D50 and D75), corresponding to MED 50, 100 and 150 μg/kg (M50, M100 and M150). Sedation, analgesia and muscular relaxation were scored subjectively. Heart and respiratory rates and rectal temperature were measured. Corresponding doses of DEX and MED were compared. Effects were also compared between dose levels for each compound. At dose level 2 (D50‐M100), the duration of effective clinical sedation was significantly shorter after DEX (202.5±16.0 min) than after MED (230.0±41.2 min). Proceeding from D50‐M100 to D75‐M150, the duration of effective clinical sedation was increased more after DEX (by 57.5±38.4 min) than after MED (by 14.2±41.9 min) Increasing from D50‐M100 to D75‐M150, heart rate was further decreased after DEX (by 8.1±13.4%) but not after MED. There was no statistically significant difference between corresponding doses of DEX and MED for any of the other parameters studied. Changes in sedation, analgesia and muscular relaxation were dose‐dependent. It was concluded that anaesthetic effects of medetomidine in cats are probably due entirely to its d‐isomer and that dexmedetomidine at 25, 50 and 75 μg/kg induces dose‐dependent sedation, analgesia and muscular relaxation of clinical significance in cats.
Characterization of urinary metabolites of testosterone, methyltestosterone, mibolerone and boldenone in greyhound dogs
Authors: Williams T. M. , Kind A. J. , Hyde W. G. , Hill D. W.
Published: March 2003

Androgenic steroids are used in female greyhound dogs to prevent the onset of estrus; moreover, these steroids also have potent anabolic activity. As anabolic steroids increase muscle mass and aggression in animals, the excessive use of these agents in racing greyhounds gives an unfair performance advantage to treated dogs. The biotransformation of most anabolic steroids has not been determined in greyhound dogs. The objective of the present study was to identify the urinary metabolites of testosterone, methyltestosterone, mibolerone, and boldenone in greyhound dogs. These steroids were administered orally (1 mg/kg) to either male or female greyhound dogs and urine samples were collected pre‐administration and at 2, 4, 8, 12, 24, 72, and 96 h post‐administration. Urine extracts were analyzed by high‐performance liquid chromatography/mass spectrometry (HPLC/MS) to identify major metabolites and to determine their urinary excretion profiles. Major urinary metabolites, primarily glucuronide, conjugated and free, were detected for the selected steroids. Sulfate conjugation did not appear to be a major pathway for steroid metabolism and excretion in the greyhound dog. Phase I biotransformation was also evaluated using greyhound dog liver microsomes from untreated dogs. The identification of several in vivo steroid metabolites generated in this study will be useful in detecting these steroids in urine samples submitted for drug screening.
Pharmacokinetics of selamectin following intravenous, oral and topical administration in cats and dogs
Authors: Sarasola P. , Jernigan A. D. , Walker D. K. , Castledine J. , Smith D. G. , Rowan T. G.
Published: March 2003

Pharmacokinetics of selamectin following intravenous, oral and topical administration in cats and dogs. J. vet. Pharmacol. Therap.24, 265–272.

The pharmacokinetics of selamectin were evaluated in cats and dogs, following intravenous (0.05, 0.1 and 0.2 mg/kg), topical (24 mg/kg) and oral (24 mg/kg) administration. Following selamectin administration, serial blood samples were collected and plasma concentrations were determined by high performance liquid chromatography (HPLC). After intravenous administration of selamectin to cats and dogs, the mean maximum plasma concentrations and area under the concentration–time curve (AUC) were linearly related to the dose, and mean systemic clearance (Clb) and steady‐state volume of distribution (Vd(ss)) were independent of dose. Plasma concentrations after intravenous administration declined polyexponentially in cats and biphasically in dogs, with mean terminal phase half‐lives (t½) of approximately 69 h in cats and 14 h in dogs. In cats, overall Clb was 0.470 ± 0.039 mL/min/kg (±SD) and overall Vd(ss) was 2.19 ± 0.05 L/kg, compared with values of 1.18 ± 0.31 mL/min/kg and 1.24 ± 0.26 L/kg, respectively, in dogs. After topical administration, the mean Cmax in cats was 5513 ± 2173 ng/mL reached at a time (Tmax) of 15 ± 12 h postadministration; in dogs, Cmax was 86.5 ± 34.0 ng/mL at Tmax of 72 ± 48 h. Bioavailability was 74% in cats and 4.4% in dogs. Following oral administration to cats, mean Cmax was 11929 ± 5922 ng/mL at Tmax of 7 ± 6 h and bioavailability was 109%. In dogs, mean Cmax was 7630 ± 3140 ng/mL at Tmax of 8 ± 5 h and bioavailability was 62%. There were no selamectin‐related adverse effects and no sex differences in pharmacokinetic parameters. Linearity was established in cats and dogs for plasma concentrations up to 874 and 636 ng/mL, respectively. Pharmacokinetic evaluations for selamectin following intravenous administration indicated a slower elimination from the central compartment in cats than in dogs. This was reflected in slower clearance and longer t½ in cats, probably as a result of species‐related differences in metabolism and excretion. Inter‐species differences in pharmacokinetic profiles were also observed following topical administration where differences in transdermal flux rates may have contributed to the overall differences in systemic bioavailability.
Systemic manifestations and liver disease in patients with chronic hepatitis C and type II or III mixed cryoglobulinaemia
Authors: Donada C. , Crucitti A. , Donadon V. , Tommasi L. , Zanette G. , Crovatto M. , Santini G. F. , Chemello L. , Alberti A.
Published: March 2003

The aim of this study was to evaluate the prevalence of cryoglobulins in patients with chronic hepatitis B and C virus infection and to investigate the association of type II and type III mixed cryoglobulinaemia with systemic manifestations and liver disease stage and outcome in hepatitis C virus (HCV)‐positive patients. We analysed the prevalence of cryoglobulinaemia in a cohort of patients with chronic liver disease and compared the systemic manifestations and liver involvement in HCV‐positive patients with type II or type III mixed cryoglobulinaemia. The prevalence of serum cryoglobulins was significantly higher in HCV‐positive patients than in hepatitis B surface antigen (HBsAg)‐positive patients (55.4 vs 20.6%). In HCV‐positive patients, stage of liver disease correlated with the prevalence of cryoglobulinaemia. Patients with type II cryoglobulins showed a significantly higher risk of cirrhosis and of extrahepatic manifestations while patients with type III cryoglobulins had a significantly higher prevalence of hepatocellular carcinoma. During follow‐up the former had an odds ratio of 11.9 of death from extrahepatic complications while the latter had an odds ratio of 3.4 of dying from hepatic disease. Our study confirms the high frequency of mixed cryoglobulinaemia in patients with chronic hepatitis C virus infection. The presence and type of cryoglobulins seem to be associated with different clinical manifestations and outcome.
Visualization and characterization of GB virus‐C particles: evidence for a nucleocapsid
Authors: Xiang J. , Daniels K.J. , Soll D.R. , Schmidt W.N. , LaBrecque D.R. , Stapleton J.T.
Published: March 2003

GB virus type C (GBV‐C) is a member of the hepacivirus genus within the Flaviviradae. Persistent GBV‐C infection is common in humans, yet it remains unclear if GBV‐C causes any disease. Although GBV‐C infection has been associated with acute non‐A to non‐E post‐transfusion hepatitis, it does not appear to cause chronic hepatitis. GBV‐C is closely related to hepatitis C virus (HCV), but indirect evidence suggests that it does not encode a core protein at the amino terminus of the open reading frame (ORF). This has led to speculation that GBV‐C does not have a nucleocapsid. We evaluated the buoyant density of GBV‐C, and found very low density particles consistent with virions, and intermediate density particles consistent with nucleocapsids in GBV‐C‐infected people. In addition, electron microscopy demonstrated an apparent nucleocapsid within an enveloped particle. Although these biophysical data strongly suggest that GBV‐C utilizes a nucleocapsid, they do not indicate the origin of the protein content of this particle. To assess this, we evaluated patient plasma for reactivity with a synthetic oligopeptide representing a conserved region near the amino terminus of the predicted ORF. Specific antibody was detected in some individuals, similar to data of Feucht et al. who identified antibody against a recombinant core protein in GBV‐C‐infected people. These data indicate that GBV‐C particles contain nucleocapsids. At least in some patients, the region upstream of the GBV‐C E1 protein coding region appears to be expressed, and this region may represent the structural protein of the nucleocapsid.
Interim analysis of a randomized controlled trial of combination of ribavirin and high dose interferon‐α in interferon nonresponders with chronic hepatitis C
Authors: Ferenci P. , Stauber R. , Steindl‐Munda P. , Gschwantler M. , Fickert P. , Datz C. , Müller C. , Hackl F. , Rainer W. , Watkins‐Riedel T. , Lin W.
Published: March 2003

This trial investigated the efficacy of a combination of high‐dose interferon‐α (IFN‐α) with ribavirin in IFN nonresponders. Study protocol: 304 patients with chronic hepatitis C were treated with 5 MU IFN‐α2b (IntronA®, Schering‐Plough) per TIW for 3 months. Nonresponders (defined by HCV‐RNA positivity in serum after the 3 months of therapy) were randomized either to continue with IFN (5 MU IFN per TIW followed by 10 MU per TIW for each 3 months) alone (group A) or in combination with ribavirin (1–1.2 g per day) (group B). ALT was measured in monthly intervals, HCV‐RNA in 3 monthly intervals. Pretreatment characteristics of the randomized patients were as follows: group A, n = 76; m/f, 54/22; 16% cirrhosis, age, 45.7 ± 12 years; ALT (U per litre), 66 ± 35; group B, n = 81; m/f, 57/24; 17% cirrhosis, age, 48.2 ± 12; ALT, 71 ± 40. After 9 months of treatment, nine (11.6%) and 27 (32.5%, P = 0.0066) patients were HCV‐RNA negative and 51 and 39 were HCV‐RNA positive, in groups A and B, respectively. There were 17 drop‐outs in group A and 15 in group B. Six months after treatment only two patients in group A (2.5%) and five (6%, P = 0.06) in group B had normal ALT and no detectable HCV‐RNA in serum. In addition to the well‐known side‐effects of IFN the mean haemoglobin concentration dropped by 2 g per litre in group B. These data indicate that a combination of high‐dose IFN with ribavirin is effective in inducing a short‐lasting complete response in one‐third of IFN nonresponders. Prolonged treatment with IFN/ribavirin may be necessary to obtain a sustained response.
Comparison of thrice weekly vs daily human leucocyte interferon‐α therapy for chronic hepatitis C
Authors: Chemello L. , Cavalletto L. , Bernardinello E. , Boccato S. , Casarin P. , Cavinato F. , Urban F. , Pontisso P. , Cecchetto A. , Gatta A. , Alberti A. ,   the TVVH Study Group
Published: March 2003

Standard treatment for chronic hepatitis C currently consists of 3–6 million units (MU) of interferon‐α (IFN‐α) given thrice weekly (t.i.w.) for 12 months, obtaining rates of sustained response (SR) that usually do not exceed 15–25%. Some recent reports have suggested that daily administration of IFN‐α may be more efficacious. More than 7 years ago, when standard therapy for hepatitis C was usually given for 6 months, we conducted a randomized clinical trial comparing daily vs t.i.w. treatment. In this study, 149 patients with chronic hepatitis C were randomized to received 3 MU of IFN‐α either t.i.w. for 6 months or daily for 3 months followed by t.i.w. for 3 months. All patients were treated with human leucocyte IFN‐α and were followed‐up for up to 72 months after inclusion. Overall, patients treated daily or t.i.w. had similar rates of virological response after 3 months of induction [24/49 (50%) vs 40/100 (40%)], at the end of therapy [15/49 (31%) vs 36/100 (36%)] and at the end of follow‐up [6/49 (12%) vs 9/100 (9%)]. However, when patients infected with HCV types other than HCV‐1 were studied, there was a trend favouring the daily schedule that was associated with a higher [5/20 (25%) vs 5/48 (10%)] rate of long‐term SR. All patients with a virological response – hepatitis C virus (HCV) RNA negative in serum as determined using the polymerase chain reaction – at 6 months after therapy remained in biochemical and virological remission at long‐term follow‐up, while seven of eight subjects who had normal alanine aminotransferase (ALT) levels but were serum positive for HCV RNA at 6 months, relapsed later, indicating that serum HCV RNA is better than ALT at predicting long‐term cure after IFN‐α therapy in chronic hepatitis C.
Acute hepatitis A and acquired immunity to hepatitis A virus in hepatitis B virus (HBV) carriers and in HBV‐ or hepatitis C virus‐related chronic liver diseases in Thailand
Authors: Pramoolsinsap C.
Published: March 2003

A number of studies have suggested that the clinical course of hepatitis A virus (HAV) infection is more severe in patients with chronic liver disease (CLD). A study was undertaken to determine the impact of acute HAV in asymptomatic hepatitis B surface antigen (HBsAg) carriers (n = 20) and patients with hepatitis B virus (HBV)‐(n = 8) or hepatitis C virus (HCV)‐related (n = 4) CLD. Disease progression was compared with that in 100 patients with isolated HAV infection. No patient with HAV infection alone developed complications, and all recovered fully. Fulminant or submassive hepatitis occurred in 55% of HBsAg carriers and 33% of patients with HBV‐ or HCV‐related CLD. The mortality rate in HBsAg carriers (25%) was not significantly different from that in the patients with CLD (33%). The seroprevalence of anti‐HAV immunoglobulin G in 820 individuals was also determined. Approximately 50% of the individuals had acquired HAV infection between the ages of 21 and 30 years. It was demonstrated that HAV infection may have a more severe clinical course in patients with underlying CLD, particularly among older individuals. Vaccination for such patients should be considered.
Prevention of hepatitis C in Italy: lessons from surveillance of type‐specific acute viral hepatitis
Authors: Mele  , Tosti  , Marzolini  , Moiraghi  , Ragni  , Gallo  , Balocchini  , Santonastasi  , Stroffolini  ,   the SEIEVA collaborating group
Published: March 2003

Using data from the surveillance system for type‐specific acute viral hepatitis, the temporal incidence trend of non‐A, non‐B acute hepatitis and risk factors for acute hepatitis C have been evaluated in Italy. The association between hepatitis C and the potential risk factors (odds ratios, OR) was estimated using hepatitis A patients as controls. The independent roles of the different risk factors were estimated by multiple logistic regression analysis. The incidence of non‐A, non‐B acute hepatitis declined from 5 per 100 000 to 1 per 100 000 between 1985 and 1996. Anti‐HCV data collected by SEIEVA since 1991 showed that 60% of patients with non‐A, non‐B acute hepatitis were positive for antibodies to the hepatitis C virus (anti‐HCV) at the time of hospitalization. During the 6 months prior to the disease onset, the most frequently reported risk factors were multiple sexual partners, other parenteral exposure and intravenous drug use; transmission by blood transfusion declined from 20% in 1985 to 2% in 1996. On multivariate analysis, intravenous drug use (OR=35.5; 95% CI=23.1–54.4), surgical intervention (OR=4.6; 95% CI=3.3–6.5), dental treatment (OR=1.5; 95% CI=1.1–1.9) and two or more sexual partners (OR=2.2; 95% CI=1.6–3.0) were all independent predictors of hepatitis C. These findings indicate that HCV infection is decreasing in Italy. Intravenous drug use, multiple sexual partners, surgical intervention and dental therapy are the main modes of transmission.
Pretreatment symptoms and dosing regimen predict side‐effects of interferon therapy for hepatitis C
Authors: Cotler  , Wartelle  , Larson  , Gretch  , Jensen  , Carithers Jr
Published: March 2003

We analysed data from a multicentre interferon (IFN) treatment trial to evaluate symptoms in patients with chronic hepatitis C and to identify factors that might predict development of debilitating IFN side‐effects. Two hundred and twenty‐two patients (120 US, 102 French) received 3 or 5 million units (MU) of IFN‐α three times weekly (t.i.w.) for 3 months. Those who had detectable hepatitis C virus (HCV) RNA, as detected by the branched DNA signal amplification (bDNA) assay, at 3 months were intensified to daily therapy, while patients who were bDNA negative continued t.i.w. dosing for the subsequent 3 months of treatment. Symptoms were assessed at baseline, and adverse effects were evaluated at 6 months of therapy. Prior to treatment, the most common symptom that interfered with daily functioning was fatigue, occurring in 25% of patients. The frequency of debilitating fatigue, myalgia, arthralgia, headache, the presence of dry eyes and dry mouth, and use of antidepressant medication increased significantly from baseline to 6 months of IFN therapy (all P < 0.01). In multivariate analysis, the development of a debilitating side‐effect at 6 months of treatment was associated with the presence of that symptom prior to therapy in all cases. Symptoms and adverse effects varied by gender and country. Compared with patients maintained on t.i.w. dosing, those who were dose intensified to daily IFN reported more debilitating fatigue, malaise, myalgia, arthralgia, fever, nausea, and headache, and the presence of dry mouth (all P < 0.05). In conclusion, patient characteristics, including pretreatment symptoms, gender and nationality, as well as daily IFN dosing are associated with the development of debilitating adverse effects on IFN therapy.
Long‐term follow‐up of sustained responders to interferon therapy, in patients with chronic hepatitis C
Authors: Toyoda H. , Kumada T. , Tokuda A. , Horiguchi Y. , Nakano H. , Honda T. , Nakano S. , Hayashi K. , Katano Y. , Nakano I. , Hayakawa T. , Nishimura D. , Kato K. , Imada K. , Imoto M. , Fukuda Y. ,   for the Yon‐Ken HCV‐HCC Follow‐up Study Group
Published: March 2003

Interferon (IFN) therapy has been proven to induce the normalization of serum alanine aminotransferase (ALT) levels and to eradicate the hepatitis C virus (HCV) in some patients with chronic hepatitis C, and these patients are usually defined as ‘sustained responders’. However, there have been some reports of hepatocellular carcinoma (HCC) in these patients, and the development of HCC remains life‐threatening in patients who clear HCV. We analysed the long‐term prognoses of patients with chronic hepatitis C in whom HCV was eradicated with IFN. We investigated 392 sustained responders to IFN therapy, from 1277 patients with chronic HCV infection who received IFN treatment at one of our institutions between April 1989 and March 1999. We analysed the medical records and looked for the development of HCC. About 30% of the sustained responders had been lost to follow‐up 3 years after the end of IFN therapy, and the follow‐up rate of sustained responders was significantly lower than that of non‐sustained responders (P < 0.0001). HCC were found in eight patients: in seven patients HCC developed within 5 years after completion of IFN therapy; but in one patient, a single HCC less than 3 cm in diameter was detected between 7 and 8 years after completion of IFN. Of the five patients who had regular medical follow‐up, the HCC was solitary, and the patients survived without any evidence of recurrence. Of the three patients who had not been followed‐up, two died from HCC and HCC recurred in the third. These results suggest that HCC can develop in sustained responders and that sustained responders should be followed‐up closely after completion of IFN so that HCC may be detected at an early stage. The optimal duration of the follow‐up period of the sustained responders remains unclear. Additional prospective studies are required in order to establish an appropriate follow‐up protocol for sustained responders to IFN.
Prevalence of anti‐hepatitis E virus antibodies in different Indian animal species
Authors: Arankalle V. A. , Joshi M. V. , Kulkarni A. M. , Gandhe S. S. , Chobe L. P. , Rautmare S. S. , Mishra A. C. , Padbidri V. S.
Published: March 2003

Prevalence of IgG antibodies to hepatitis E virus (IgG‐anti‐HEV) was determined among different animal species from India. Seropositivity varied from 4.4% to 6.9% in cattle, 54.6–74.4% in pigs and 2.1–21.5% in rodents. Of the 44 dogs screened, 10 were positive (22.7%). None of the 250 goat sera tested were found to be anti‐HEV positive. Among rodents, over 50% serum samples collected in 1985 from Bandicota bengalensis were positive for anti‐HEV antibodies. No evidence of HEV infection was obtained following experimental inoculation of an Indian strain (AKL‐90) of HEV into anti‐HEV negative pigs and goats. The results document varied prevalence of anti‐HEV antibodies in different animal species from India and of inability of Indian pigs and goats to support replication of at least one human strain of HEV.
Biochemical response to interferon therapy correlates with interferon sensitivity‐determining region in hepatitis C virus genotype 1b infection
Authors: Yoshioka K. , Kobayashi M. , Orito E. , Watanabe K. , Yano M. , Sameshima Y. , Kusakabe A. , Hirofuji H. , Fuji A. , Kuriki J. , Arao M. , Murase K. , Mizokami M. , Kakumu S. ,   the IFN Treatment Group of Affiliated Hospitals of the Third Department of Internal Medicine at Nagoya University School of Medicine
Published: March 2003

Biochemical responders maintain normal alanine aminotransferase levels after interferon (IFN) therapy despite persistent presence of hepatitis C virus (HCV) RNA in their sera. There have been few reports on predictive factors for biochemical response. A region associated with sensitivity to IFN was identified in the nonstructural protein 5 A of genotype 1b [aa 2209–2248; IFN sensitivity‐determining region (ISDR)]. The substitutions in ISDR correlate with sustained response to IFN. In this report, we assessed the association of ISDR with biochemical response. The sequences of ISDR were determined in 62 patients with HCV genotype 1b treated by IFN in two randomized controlled trials. 30 patients had wild ISDR (identical to HCV‐J), 20 intermediate ISDR (1–3 amino acid substitutions compared with HCV‐J), and 12 mutant ISDR (four or more amino acid substitutions). All 12 patients with mutant ISDR had a sustained response, while only one of those with wild or intermediate ISDR had a sustained response (P < 0.0001). In the 49 patients other than sustained responders, the patients with intermediate ISDR obtained biochemical response significantly more frequently (52.6%, 10/19) than those with wild‐type ISDR (20.0%, 6/30) (P < 0.05). Multivariate analysis indicated the number of substitutions in ISDR as the most important predictor for biochemical response (discriminant coefficient=1.08, P < 0.05) and sustained response (discriminant coefficient=6.13, P < 0.0001). In phylogenetic analysis, clustering of sustained responders and biochemical responders was observed. These results demonstrate that the substitutions in ISDR are the most important predictor for biochemical response to IFN in patients infected with genotype 1b as well as for sustained response.
In vitro infection of human peripheral blood mononuclear cells by a defective hepatitis B virus with a deletion in the PreS1 region of the viral genome
Authors: Cabrerizo M. , Bartolomé J. , Carreño V.
Published: March 2003

summary. Previously, we identified a defective hepatitis B virus (HBV) which contains a 183 nucleotide deletion in the PreS1 region of the viral genome affecting the S gene promoter in sera from hepatitis B surface antigen (HBsAg)‐negative patients with serum HBV‐DNA. The aim of this study was to analyse the infectivity of this mutant. Peripheral blood mononuclear cells (PBMC) from a healthy donor were incubated with serum samples from 2 HBsAg‐negative patients with serum HBV‐DNA (infected with wild‐type and deletion mutant HBV), from an HBsAg carrier (infected with wild‐type HBV) and from a healthy donor. After 1 week, HBV‐DNA was detected by polymerase chain reaction (PCR) in all supernatants and cells incubated with the HBV‐DNA‐positive inocula. DNase and trypsin pretreatment confirmed intracellular localization of HBV‐DNA in cells. HBV‐RNA and covalently closed circular HBV‐DNA were also detected in PBMC, indicating that the viral DNA infecting these cells was transcriptionally active. Deletion mutant and wild‐type HBV were detected in the supernatants and cells infected with the two HBsAg‐negative sera, while only wild‐type HBV was detected in the supernatant and cells incubated with the serum from the HBsAg‐carrier. In conclusion, this HBV deletion mutant can infect, replicate and release viral particles in in vitro infected PBMC.
The long‐term efficacy of plasma‐derived hepatitis B vaccine in babies born to carrier mothers
Authors: Young B. W. Y. , Lee S. S. , Lim W. L. , Yeoh E. K.
Published: March 2003

summary. The long‐term efficacy of a childhood hepatitis B vaccination programme was evaluated. A total of 112 newborn babies of hepatitis B carrier mothers were given hepatitis B immune globulin (HBIG) and a 10‐μg three‐dose regimen of plasma‐derived vaccine administered at a conventional (0, 1, 6 months), delayed (2, 3, 8 months) or accelerated (0, 1, 2 months) schedule. The vaccinees were followed up to determine their anti‐HBs status over a 16‐year period. Upon completion of the vaccination schedules, 92.6% developed antibody against surface antigen (anti‐HBs) seroconverion, the rate of which fell to 33.3% at year 16. The three schedules were equally effective in preventing chronic infection, with a protective efficacy of 88.9% from hepatitis B surface antigen (HBsAg) carriage, compared with historical control. Vaccinees on the delayed schedule had a slightly higher seroconversion rate over years, and were better able to maintain an anti‐HBs level of ≥ 100 iu/L. Overall, a quarter demonstrated evidence of exposure to the virus, being positive for antibody against core antigen or HBsAg, or mounting a rise in anti‐HBs during the follow‐up period. We conclude that a three‐dose hepatitis B vaccination regimen is generally effective in protecting newborns of hepatitis B carrier mothers from infection and chronic carriage. Booster is not needed even after 16 years of monitoring.
Early viral kinetics on treatment with pegylated interferon‐α‐2a in chronic hepatitis C virus genotype 1 infection*
Authors: Jessner W. , Stauber R. , Hackl F. , Datz C. , Watkins‐Riedel T. , Hofer H. , Gangl A. , Kessler H. , Ferenci P.
Published: March 2003

summary. Even with pegylated (PEG) interferons (IFN), therapy of chronic hepatitis C (genotype 1) remains unsatisfactory. The initial viral response to IFN identifies patients infected with IFN resistant viruses not responding to standard IFN/ribavirin therapy. The impact of primary IFN unresponsiveness for PEG‐IFN‐α‐2a/ribavirin therapy is unknown. Viral load was measured in 22 chronic hepatitis C (genotype 1) patients before and 24 h after 9 MU IFN‐α‐2a (days 0 and 1), and before and during weekly 180 μg PEG‐IFN‐α‐2a (days 7, 8, 11, 14 and 21) administration. Thereafter, ribavirin (800 mg/d) was added for 6 months. Virological responders continued treatment for a further 6 months. Twenty‐eight patients treated with standard IFN/ribavirin therapy in a previous study using an analogous protocol served as historic controls. After 6 months 15 (68.2%) patients were (HCV‐RNA) negative, eight of whom (36.4%) had a sustained response. The decrease in viral load 24 h after 9 MU IFN‐α‐2a was greater in virological responders (1.05 log [0.25–1.67]) than in nonresponders (NR) (0.34 [0.14–0.65]; P=0.003). In contrast, viral decline was not different between responders and NRs during the first 2 weeks on PEG‐IFN‐α‐2a. All patients with an initial decline > 1.4 log became sustained responders. Five of 12 patients with a log change < 0.8 became end of treatment responders, two had a sustained response. Antiviral response to PEG‐IFN‐α‐2a is different to that on standard IFN. In spite of a lower initial response PEG‐IFN‐α‐2a/ribavirin combination therapy may overcome predicted IFN unresponsiveness.
Conservation status and causes of decline of the threatened New Zealand Long‐tailed Bat Chalinolobus tuberculatus (Chiroptera: Vespertilionidae)
Authors: O’Donnell Colin F. J.
Published: March 2003

Historical anecdotes and preliminary monitoring since 1990 indicate that New Zealand Long‐tailed Bats (Chalinolobus tuberculatus, Vespertilionidae) are now rare or absent at many sites where formerly they were common. Chalinolobus tuberculatus appeared to be common throughout New Zealand in the 1800s but by 1900–30 it was becoming scarce in many districts. Formal surveys in the South Island since 1990 either failed to find C. tuberculatus, or recorded bats in low numbers. Of eight sites where transect counts were undertaken, bats were recorded frequently at two sites (45–66% of counts; Eglinton and Dart Valleys), rarely at four sites (2.4–10.7% of counts), and were not recorded at the remaining two sites despite considerable survey effort. Of 10 sites where stationary counts using automatic detector units were used, no C. tuberculatus were recorded in three areas (153 nights combined), they were found rarely at six sites (2.1–21.0% of nights; 461 nights combined) and were recorded commonly only in the Eglinton Valley (85% of 120 nights). Assertions that C. tuberculatus are ‘common’ and that the conservation status is ‘secure’ are questionable and this review supports suggestions that the species should be classed as ‘Vulnerable’. Possible causes of decline have been suggested including clearance and logging of lowland forests, predation by introduced mammals and owls, competition for roost sites by introduced mammals, birds and wasps, and human interference and disturbance at roosting sites. However, authors’ claims have all been speculative and unsubstantiated. There has been no research undertaken to quantify these claims, and this is required urgently. The results of these preliminary surveys provide a new baseline against which future population trends might be compared. Increased effort using standardized monitoring techniques, applied at a national level, is required to confirm the possible trends and to help identify the best sites where conservation managers may attempt to restore the population level.
A review of the historical evidence of the habitat of the Pine Marten in Cumbria
Authors: Webster J. A.
Published: March 2003

Place names and evidence from the literature show the Pine Marten (Martes martes) to have been well distributed in Cumbria until late in the nineteenth century, with a core area in the central fells. The habitat available to these martens is assessed and the species' status and the reasons for its decline reviewed. Despite the modern emphasis on the absolute importance of woodland to the Pine Marten, it is argued that historical Cumbrian populations survived successfully for many generations in highly fragmented habitats.
Comparative kinetics of bloodmeal intake by Triatoma infestans and Rhodnius prolixus, the two principal vectors of Chagas disease
Authors: PEREIRA,  , PENIDO,  , MARTINS  , DIOTAIUTI
Published: March 2003

Chagas disease vector insects Triatoma infestans and Rhodnius prolixus (fifteen stage III nymphs per 4 litre cage) were allowed to feed on anaesthetized mice for 1 h (control group), or on active non‐anaesthetized mice (NAM) for 2, 4 or 8 h exposure. The bloodmeal size (weight increase) for both species was proportional to the duration of contact with NAM, due to ingestion of multiple small bloodmeals, up to 142% of control weight for T. infestans with 8 h exposure to NAM. The mean weight increase of T. infestans nymphs after 4 h contact with NAM was similar to that of the control group, whereas for R. prolixus, 8 h contact with NAM gave only 64% of the control value. For both species of insect, within 4 h of feeding, >20% of the bloodmeal weight was lost by defaecation and diuresis. The proportions of unfed nymphs and mortality during 2 h contact with NAM were significantly higher for R. prolixus, demonstrating better exploitation of the host blood source by T. infestans, apparently because during blood‐feeding the latter insect species caused less irritation to the host.
Sandfly midgut lectin: effect of galactosamine on Leishmania major infections
Authors: VOLF PETR , KIEWEGOVÁ  , SVOBODOVÁ
Published: March 2003

Galactosamine, which has been shown in vitro to specifically inhibit sandfly midgut lectin activity, was fed to Phlebotomus duboscqi females with blood containing promastigotes of Leishmania major. Non‐inhibitory sugar, galactose, was added in controls. For two strains of L. major (LV 561 and Neal‐P), galactosamine substantially enhanced the establishment of infection in the sandfly posterior midgut and significantly increased parasite loads after defaecation, but did not affect anterior migration of Leishmania. On day 3 post‐infection, most infections in galactosamine‐fed sandfly groups (92% of LV 561 and 100% of Neal‐P) were found in the ectoperitrophic space of the posterior midgut, whereas most infections in the galactose‐fed groups of sandflies (85% in LV 561 and 96% in Neal‐P) were restricted to the peritrophic sac. On day 9, however, the proportion of infections colonizing the stomodeal valve was similar in both dietary groups of sandflies for both strains of L. major. The addition of galactosamine prevented the decrease of parasite loads which occurred in controls between days 3 and 6 post‐infection. On days 6 and 9, heavy infections were observed almost exclusively in galactosamine‐fed females. Differences between groups were more pronounced for the Neal‐P strain, which normally developed poorly in sandflies. Morphology of L. major LV 561 was not affected by galactosamine supplement: the lengths of parasite body and flagellum were similar in both sandfly groups. Two hypotheses are considered for the role of sandfly midgut lectin in Leishmania development in the vector midgut. One proposes that sandfly lectin kills Leishmania promastigotes, the other assumes that lectin blocks LPG‐mediated binding of promastigotes to sandfly midgut microvilli.
Comparative scanning electron microscopy of third‐instar Hypoderma spp. (Diptera: Oestridae)
Authors: COLWELL DOUGLAS D. , MARTÍNEZ‐MORENO  , MARTÍNEZ‐MORENO  , HERNÁNDEZ‐RODRÍGUEZ  , DE LA FUENTE‐LÓPEZ  , ALUNDA  , HALL
Published: March 2003

Scanning electron microscope study of third‐instar larvae of four species of Hypoderma revealed differences among species in the pattern of spination, spine morphology and morphology of the spiracular plates. These observations identify characters that enable the differentiation of Hypoderma actaeon and H. diana, parasitizing red deer (Cervus elaphus) in Europe, and provide additional characters for differentiating H. bovis and H. lineatum parasitizing cattle.
Serum and skin surface antibodies and their associations with sheep biting lice, Bovicola ovis, on experimentally infested sheep
Authors: JAMES  , MOON  , RAGSDALE
Published: March 2003

The sheep biting louse (Bovicola ovis) feeds superficially on the skin of sheep but appears to stimulate an immune response. In this study we examined the association between louse infestation and serum and skin surface antibodies. Louse numbers were monitored on experimentally infested Polypay and Columbia ewes for two years and on their lambs in the second year. Serum and skin wash samples were tested for antibodies to soluble extracts of B. ovis, Stomoxys calcitrans and Musca autumnalis by enzyme linked immunosorbent assay (ELISA). In addition, the effects of skin wash extracts on B. ovis were examined in vitro.

The titre of anti‐B. ovis antibodies in the serum did not differ significantly between infested and naive ewes. However, there was an increase in serum antibody titre which coincided with periods of high louse density in ewes with high louse counts. Infested lambs had higher serum antibody levels than naive lambs. Substantial cross reactivity was evident among extracts of the different insects.

Densities of lice on the ewes during population decline were negatively related to the titre of skin surface antibodies. Skin washings collected from sheep during B. ovis population decline reduced the number of louse progeny when incorporated into louse diet. These results indicate that B. ovis stimulates an immune response in sheep and suggest that compounds on the skin surface may play a role in the regulation of louse populations.
Colonization and bionomics of the sandfly Phlebotomus kazeruni from Sinai, Egypt
Authors: Hanafi H. A. , Jr W. W. Kanour , Beavers G. M. , Tetreault G. E.
Published: March 2003

Phlebotomus kazeruni (Diptera: Psychodidae) females were collected by light‐trap in southern Sinai, Egypt, and this sandfly species was colonized for the first time as a laboratory strain, maintained by the procedures of
Modi & Tesh (1983). Laboratory‐reared females did not lay eggs autogenously; they blood‐fed more readily (P = 0.02) on a hamster (37%) than a human (22%) during 1 h exposure. Fecundity of hamster‐fed females was significantly greater than for those fed on human blood: 69.4 ± 5.8 vs. 45.2 ± 8.1 eggs/female from the first gonotrophic cycle. Pre‐oviposition and egg incubation periods were significantly less for females fed on hamster compared with human blood, but the larval development and pupation periods were not affected by this difference in bloodmeal source. Egg to adult survival was equivalent (38%) for progeny of females blood‐fed on hamster or human. The mean generation time of progeny from females fed on hamster (51.9 ± 1.0 days) or human (53.3 ± 1.7 days) was not significantly different. The sex ratio of adult male:female progeny was similar (P = 0.2) for both hosts: 42 : 58% from hamster, 46 : 54% from human blood‐fed female parents. Evidently P. kazeruni from Sinai is sufficiently anthropophagic to be a potential vector of Leishmania from rodents to humans.
Genetic differentiation of someGlossina morsitans morsitanspopulations
Authors: Wohlford D. L. , Krafsur E. S. , Griffiths N. T. , Marquez J. G. , Baker M. D.
Published: March 2003

To study the population structure ofGlossina morsitans morsitansWestwood (Diptera: Glossinidae), polymerase chain reaction (PCR) and single‐strand conformational polymorphism (SSCP) methods were used to estimate mitochondrial DNA diversity at four loci in six natural populations from Zambia, Zimbabwe and Mozambique, and in two laboratory cultures. The Zambian and Zimbabwean samples were from a single fly belt. Four alleles were recorded at12S and 16S1, and five alleles at16S2andCOI. Nucleotide sequencing confirmed their singularities. Chi‐square contingency tests showed that allele frequencies differed significantly among populations. Mean allele diversities in populations averaged over loci varied from 0.14 to 0.61. Little loss in haplotype diversity was detected in the laboratory cultures thereby indicating little inbreeding. Wright’s fixation indexFSTin the natural populations was 0.088 ± 0.016, the correlation of haplotypes within populations relative to correlations in the total. A function of its inverse allows an estimate of the mean equivalent number of females exchanged per population per generation, 5.2. No correlation was detected between pairwise genetic distance measures and geographical distances. Drift explains the high degree of differentiation.
Extensive multiple test centre evaluation of the VecTestTM malaria antigen panel assay
Authors: RYAN J. R. , DAVÉ K. , COLLINS K. M. , HOCHBERG L. , SATTABONGKOT JETSUMON , COLEMAN R. E. , DUNTON R. F. , BANGS M. J. , MBOGO C. M. , COOPER R. D. , SCHOELER G. B. , RUBIO‐PALIS Y. , MAGRIS M. , ROMERO L. I. , PADILLA N. , QUAKYI I. A. , BIGOGA J. , LEKE R. G. , AKINPELU O. , EVANS B. , WALSEY M. , PATTERSON P. , WIRTZ R. A. , CHAN A. S. T.
Published: March 2003

Abstract To determine which species and populations of Anopheles transmit malaria in any given situation, immunological assays for malaria sporozoite antigen can replace traditional microscopical examination of freshly dissected Anopheles. We developed a wicking assay for use with mosquitoes that identifies the presence or absence of specific peptide epitopes of circumsporozoite (CS) protein of Plasmodium falciparum and two strains of Plasmodium vivax (variants 210 and 247). The resulting assay (VecTestTM Malaria) is a rapid, one‐step procedure using a ‘dipstick’ test strip capable of detecting and distinguishing between P. falciparum and P. vivax infections in mosquitoes. The objective of the present study was to test the efficacy, sensitivity, stability and field‐user acceptability of this wicking dipstick assay. In collaboration with 16 test centres world‐wide, we evaluated more than 40 000 units of this assay, comparing it to the standard CS ELISA. The ‘VecTestTM Malaria’ was found to show 92% sensitivity and 98.1% specificity, with 97.8% accuracy overall. In accelerated storage tests, the dipsticks remained stable for >15 weeks in dry conditions up to 45°C and in humid conditions up to 37°C. Evidently, this quick and easy dipstick test performs at an acceptable level of reliability and offers practical advantages for field workers needing to make rapid surveys of malaria vectors.