Evaluation of oxytetracycline leaching from pregelatinized starch-coated medicated fish feed.

The rapid growth and intensification of aquaculture industries have led to an increased use of antibiotics. Consequently, growing concerns have mounted over the environmental contamination of these drugs from medicated feeds and the risk that this poses for antimicrobial resistance. To circumvent environmental leaching, farmers topcoat medicated feeds with oil; however, this only partially addresses the issue. This study investigated the potential of food-grade pregelatinized corn starch (PGS) as a second top-coating agent to reduce oxytetracycline (OTC) leaching from the hand-mixed medicated feed. We immersed top-coated medicated feeds for different periods of time and measured the concentration of OTC in the water to determine leaching. We found a significantly lower level of OTC in water samples collected from the PGS-coated medicated feed compared to the non-PGS-coated medicated feed, with concentrations of OTC approximately 4 and 2.6 times the latter after 5 min and 2 h of water immersion, respectively. We also fed PGS-coated antibiotic feed to jade perch to determine if fish accepted the top-coating and whether they absorbed the OTC. Results from a feeding trial suggested no difference in palatability between PGS and non-PGS-coated medicated feed. We also found that muscle tissue from fish fed with the aforementioned diets had similar levels of OTC concentrations, suggesting that PGS coating does not alter the gastrointestinal absorption of this medication. From our experiment, we conclude that PGS is potentially a new top-coating agent to reduce leaching in hand-mixed OTC medicated feed.

Does REM sleep behavior disorder change in the progression of Parkinson's disease?

OBJECTIVES/BACKGROUND: Rapid eye movement (REM) Sleep Behavior Disorder (RBD) in Parkinson's disease (PD) may be associated with a malignant phenotype. Despite its prognostic value, little is known about the time course of RBD in PD. In this study, we aimed to ascertain whether or not RBD is a stable feature in PD. In this study, we prospectively evaluated clinical and neurophysiological features of RBD, including REM Sleep Without Atonia (RSWA), in PD patients with RBD at baseline and after three years then assessed whether the changes in measures of RSWA parallel the progression of PD. PATIENTS/METHODS: In sum, 22 (17M, mean age 64.0 ± 6.9 years) moderate-to-advanced PD patients (mean PD duration at baseline:7.6±4.8 years) with RBD, underwent a video-polysomnography (vPSG) recording and clinical and neuropsychological assessment at baseline and after three years. RESULTS: At follow-up, the self-assessed frequency of RBD symptoms increased in six patients, decreased in six and remained stable in 10, while RSWA measures significantly increased in all subjects. At follow-up, patients showed worse H&Y stage (p = 0.02), higher dopaminergic doses (p = 0.05) and they performed significantly worse in phonetic and semantic fluency tests (p = 0.02; p = 0.04). Changes in RSWA correlated significantly with the severity in levodopa-induced dyskinesia (r = 0.61,p = 0.05) and motor fluctuation (r = 0.54,p = 0.03) scores, and with the worsening of executive functions (r = 0.78,p = 0.001) and visuo-spatial perception (r = -0.57,p = 0.04). CONCLUSION: Despite the subjective improvement of RBD symptoms in one-fourth of PD patients, all RSWA measures increased significantly at follow-up, and their changes correlated with the clinical evolution of motor and non-motor symptoms. RBD is a long-lasting feature in PD and RSWA is a marker of the disease's progression.

Bactérias zoonóticas isoladas de Passeriformes silvestres recuperados do tráfico de animais no estado do Ceará/Brasil / Zoonotic bacteria isolated from wild Passeriformes recovered from animal trafficking in the State of Ceará/Brazil

A ordem dos Passeriformes é uma das mais pressionadas pelas ações antrópicas, especialmente as relativas ao tráfico de animais, que, devido às más condições de manejo e higiênico-sanitárias, favorecem a infecção dos espécimes por patógenos virulentos e zoonóticos, como cepas de Escherichia coli e Salmonella spp., cujo isolamento em suabes cloacais, bem como a análise dos genes de virulência das cepas de E. coli foram objetivos do estudo. Para isso, 120 Passeriformes silvestres nativos, recebidos pelo Cetas/CE, foram avaliados individualmente. As cepas isoladas foram submetidas a teste de disco difusão para determinação da sensibilidade aos antimicrobianos. Em etapa posterior, foi realizada PCR para a detecção de oito genes de virulência dos principais patotipos diarreiogênicos de E. coli. Quanto aos resultados, nenhuma cepa de Salmonella spp. foi isolada, no entanto a ocorrência de E. coli foi de 40,8%. Foi observada elevada resistência, principalmente aos antimicrobianos tetraciclina, ampicilina e sulfazotrim, ocorrendo multirresistência em 42,8% das cepas. Pela análise molecular, foram diagnosticados quatro entre os nove genes pesquisados, com a identificação de EPEC típicas, EPEC atípicas, ETEC, EHEC e EAEC. Os resultados apontam para a importância de Passeriformes como possíveis disseminadores de zoonoses.(AU)

The order Passeriformes is one of the most pressured by anthropic actions, especially those related to animal trafficking. Due to poor sanitary and hygienic conditions, the infection of the specimens is favored by virulent and zoonotic pathogens such as strains of Escherichia coli and Salmonella spp., whose isolation in cloacal swabs as well as the analysis of the virulence genes of E. coli strains were the objectives of the study. For this, 120 native wild Passeriformes, received by CETAS/CE were individually evaluated. The isolated strains were submitted to diffusion disc test to determine sensitivity to antimicrobials. In a later stage, PCR was performed for the detection of eight virulence genes from the main E. coli diarrhoeagenic pathogens. Regarding the results, no strain of Salmonella spp. was isolated; however, the occurrence of E. coli was 40.8%. High resistance was observed, mainly to the antimicrobials Tetracycline, Ampicillin and Sulfazotrim, with multi-resistance in 42.8% of the strains. By molecular analysis, four of the nine genes were diagnosed, identifying typical EPEC, atypical EPEC, ETEC, EHEC and EAEC. The results point to the importance of Passeriformes as possible disseminators of zoonoses.(AU)

Relationship between clinical symptomatology on the isolation of Salmonella Gallinarum of japanese quail (Coturnix coturnix) / Relação entre a sintomatologia clínica no isolamento de Salmonella Gallinarum de codornas japonesas (Coturnix coturnix)

Salmonella Gallinarum is capable of causing high mortality in birds of the order Galliformes. This study aimed to relate the presence of clinical signs with the recovery of Salmonella Gallinarum from organs and c loacal swabs of Japanese quails (Coturnix coturnix) experimentally infected. A total of 70 female quails were housed in a pair per cage and divided in two groups (IG: quails inoculated with 1.5x106 CFU of Salmonella Gallinarum Nalr/mL and CG: control group). After the inoculation, birds were evaluated three times a day to verify the presence of clinical signs. Birds that presented ruffled feathers, eyes closed and remained quiet in the cage were removed for euthanasia, as well as the same number of birds from the inoculated groups that presented no clinical signs and from the control group. Cloacal swabbing was performed following euthanasia for the sampling of liver, spleen, caeca, ovarian follicles and lung for microbiological procedure. Quails with clinical signs and quails found dead presented positivity of 100%. While inoculated quails with no clinical signs presented a lower positivity (38.5%). Therefore, quails with septicemia caused by SG present clinical signs of the disease and the pathogen can be isolated and quantified in the organs.(AU)

Salmonella Gallinarum pode causar alta mortalidade em aves da ordem Galliformes. Objetivou-se neste estudo relacionar a presença de sinais clínicos com a recuperação de Salmonella Gallinarum de órgãos e swabs cloacais de codornas japonesas (Coturnix coturnix) experimentalmente infectadas. Um total de 70 codornas fêmeas foram alojadas em par por gaiola e divididas em dois grupos (IG: codornas inoculadas com 1,5x106 UFC de Salmonella Gallinarum Nalr / mL e CG: grupo controle). Após a inoculação, as aves foram avaliadas três vezes ao dia para se verificar a presença de sinais clínicos. As aves que se apresentaram com penas eriçadas, olhos fechados e permaneciam imóveis na gaiola foram removidas para a eutanásia, assim como o mesmo número de aves dos grupos inoculados que não apresentaram sinais clínicos e do grupo controle. O swab cloacal foi realizado após a eutanásia para a amostragem de fígado, baço, ceco, folículos ovarianos e pulmão para procedimento microbiológico. As codornas com sinais clínicos e as encontradas mortas apresentaram positividade de 100%, enquanto as codornas inoculadas sem sinais clínicos apresentaram menor positividade (38,5%). Portanto, codornas com septicemia causada por SG apresentam sinais clínicos da doença e o patógeno pode ser isolado e quantificado em diversos órgãos.(AU)

Simultaneous quantitation of sphingoid bases by UPLC-ESI-MS/MS with identical 13C-encoded internal standards.

Free sphingoid bases (lysosphingolipids) of primary storage sphingolipids are increased in tissues and plasma of several sphingolipidoses. As shown earlier by us, sphingoid bases can be accurately quantified using UPLC-ESI-MS/MS, particularly in combination with identical 13C-encoded internal standards. The feasibility of simultaneous quantitation of sphingoid bases in plasma specimens spiked with a mixture of such standards is here described. The sensitivity and linearity of detection is excellent for all examined sphingoid bases (sphingosine, sphinganine, hexosyl-sphingosine (glucosylsphingosine), hexosyl2-sphingosine (lactosylsphingosine), hexosyl3-sphingosine (globotriaosylsphingosine), phosphorylcholine-sphingosine) in the relevant concentration range and the measurements show very acceptable intra- and inter-assay variation (<10% average). Plasma samples of a series of male and female Gaucher Disease and Fabry Disease patients were analyzed with the multiplex assay. The obtained data compare well to those earlier determined for plasma globotriaosylsphingosine and glucosylsphingosine in GD and FD patients. The same approach can be also applied to measure sphingolipids in the same sample. Following extraction of sphingolipids from the same sample these can be converted to sphingoid bases by microwave exposure and subsequently quantified using 13C-encoded internal standards.

Reciprocal interference of experimental dyslipidemia and food allergy in the evolution of both diseases.

Background. Food allergies have been shown to reduce serum triacylglycerol, glucose, cholesterol, and free fatty acid levels in mice. In turn, dyslipidemias, especially dyslipidemias presenting with low levels of HDL cholesterol, are important risk factors for the development of atherosclerosis. However, the consequences of food allergies on dyslipidemia and atherosclerosis have not been fully investigated. Methods. Food allergy was induced using an egg white solution (EWS) in ovalbumin- (OVA-) sensitized C57BL/6 and low-density lipoprotein receptor knockout mice (LDLr(-/-)) for 5 weeks and was confirmed by the high production of anti-OVA IgE and IgG1 antibodies in both mouse strains. Results. The allergic C57BL/6 mice exhibited EWS aversion that was associated with less visceral fat and high levels of anti-Ova IgE antibodies after 5 weeks of EWS intake compared to controls. However, LDLr(-/-) allergic mice showed reduced anti-Ova IgE levels that were similar to the nonsensitized group. The LDLr(-/-) allergic mice also demonstrated a reversal of food aversion and sustained visceral fat after 5 weeks of allergy. Although HDL cholesterol levels were reduced in both sensitized mouse strains, lipid deposition in thoracic and abdominal aorta as well as area and composition of atherosclerotic plaques as unaffected by chronic ingestion of EWS. Conclusion. LDLr(-/-) mice develop an attenuated food allergy, as they showed a reversal of food aversion and lower IgE production after 5 weeks of induced allergy. The development of atherosclerosis, in turn, was not accelerated in the allergic LDLr(-/-) group despite the more atherogenic lipid profile.

Association of apoliprotein E polymorphisms and metabolic syndrome in subjects with extreme obesity.

BACKGROUND: Obesity has been linked to metabolic syndrome (MS), which increases the risk of cardiovascular disease (CVD). Polymorphisms of Apolipoprotein E have also been associated with increased CVD risk. Therefore, this study investigated the association between MS and Apo E polymorphisms. METHODS: We measured anthropometric and biochemical variables and determined the Apo E genotype of 147 grade III obese patients. RESULTS: The percentage of female subjects was 86.4%. The mean age and BMI of the subjects were 41 years and 53.5 kg/m(2), respectively. MS had been diagnosed in 79% of the subjects. The proportions of those exhibiting MS risk factors were as follows: 100% had a high BMI, 80% had hypertension, 65% had low levels of high-density lipoprotein (HDL), 38% had diabetes, and 39% had hypertriglyceridemia. We found five genotypes for which the allelic distribution was different in the MS group compared to the general population. The ε4 allele was more frequent in the group with neither MS nor hypertension. CONCLUSIONS: The morbidly obese patients exhibited a higher incidence of MS and a different allelic distribution when compared with other populations. The ε4 allele was associated with the absence of MS and hypertension.

Are there differences between cephalic and extracephalic cutaneous allodynia in migraine patients?

Cutaneous allodynia (CA), pain in response to innocuous cutaneous stimuli, is recognized as a sign of central sensitization during migraine episodes. It is either restricted within the pain area on the ipsilateral head, or extends within and outside the head. Moreover, CA can be elicited in response to thermal (heat or cold) and/or mechanical stimuli. This raises the question as to whether cephalic and extracephalic CAs share the same properties. We assessed cephalic and extracephalic CAs in migraine episodic patients using a questionnaire completed at home during migraine attacks. A total of 67 episodic migraine patients (58 women, nine men; 4013 years old) addressed all questions in the questionnaire. Forty-nine patients (73%) cited one or more allodynic symptoms during or immediately after the migraine attack. Almost all 49 patients reported cephalic CA, whereas 24 (49%) also reported extracephalic CA. Occurrence and extension of CA correlated (P = 0.005) with headache intensity. Modalities of cephalic and extracephalic CA were different (chi2 = 12.03; P = 0.002), extracephalic CA being mostly thermal (75%) whereas cephalic CA was mostly mechanical (92%). This suggests that cephalic and extracephalic CAs involve different mechanisms.

Cloning, expression, purification, crystallization and preliminary structure determination of glucose-1-phosphate uridylyltransferase (UgpG) from Sphingomonas elodea ATCC 31461 bound to glucose-1-phosphate.

The cloning, expression, purification, crystallization and preliminary crystallographic analysis of glucose-1-phosphate uridylyltransferase (UgpG) from Sphingomonas elodea ATCC 31461 bound to glucose-1-phosphate are reported. Diffraction data sets were obtained from seven crystal forms in five different space groups, with highest resolutions ranging from 4.20 to 2.65 A. The phase problem was solved for a P2(1) crystal form using multiple isomorphous replacement with anomalous scattering from an osmium derivative and a SeMet derivative. The best native crystal in space group P2(1) has unit-cell parameters a = 105.5, b = 85.7, c = 151.8 A, beta = 105.2 degrees . Model building and refinement are currently under way.

A vaccine to prevent herpes zoster and postherpetic neuralgia in older adults.

BACKGROUND: The incidence and severity of herpes zoster and postherpetic neuralgia increase with age in association with a progressive decline in cell-mediated immunity to varicella-zoster virus (VZV). We tested the hypothesis that vaccination against VZV would decrease the incidence, severity, or both of herpes zoster and postherpetic neuralgia among older adults. METHODS: We enrolled 38,546 adults 60 years of age or older in a randomized, double-blind, placebo-controlled trial of an investigational live attenuated Oka/Merck VZV vaccine ("zoster vaccine"). Herpes zoster was diagnosed according to clinical and laboratory criteria. The pain and discomfort associated with herpes zoster were measured repeatedly for six months. The primary end point was the burden of illness due to herpes zoster, a measure affected by the incidence, severity, and duration of the associated pain and discomfort. The secondary end point was the incidence of postherpetic neuralgia. RESULTS: More than 95 percent of the subjects continued in the study to its completion, with a median of 3.12 years of surveillance for herpes zoster. A total of 957 confirmed cases of herpes zoster (315 among vaccine recipients and 642 among placebo recipients) and 107 cases of postherpetic neuralgia (27 among vaccine recipients and 80 among placebo recipients) were included in the efficacy analysis. The use of the zoster vaccine reduced the burden of illness due to herpes zoster by 61.1 percent (P<0.001), reduced the incidence of postherpetic neuralgia by 66.5 percent (P<0.001), and reduced the incidence of herpes zoster by 51.3 percent (P<0.001). Reactions at the injection site were more frequent among vaccine recipients but were generally mild. CONCLUSIONS: The zoster vaccine markedly reduced morbidity from herpes zoster and postherpetic neuralgia among older adults.

Pre-treatment and post-treatment assessment of the C(6) test in patients with persistent symptoms and a history of Lyme borreliosis.

It was recently reported that antibody to C(6), a peptide that reproduces an invariable region of the VlsE lipoprotein of Borrelia burgdorferi, declined in titer by a factor of four or more in a significant proportion of patients after successful antibiotic treatment of acute localized or disseminated Lyme borreliosis. The present study evaluated the C(6) test as a predictor of therapy outcome in a population of patients with post-treatment Lyme disease syndrome. The serum specimens tested were from patients with well-documented, previously treated Lyme borreliosis who had persistent musculoskeletal or neurocognitive symptoms. All of the patients had participated in a recent double-blind, placebo-controlled antibiotic trial in which serum samples were collected at baseline and 6 months thereafter, i.show $132#e. 3 months following treatment termination. In this patient population no correlation was found between a decline of C(6) antibody titer of any magnitude and treatment or clinical outcome. Antibodies to C(6) persisted in these patients with post-treatment Lyme disease syndrome following treatment, albeit at a markedly lower prevalence and titer than in untreated patients with acute disseminated Lyme disease. The results indicate that C(6) antibody cannot be used to assess treatment outcome or the presence of active infection in this population.

Underexpression of peroxisome proliferator-activated receptor (PPAR)gamma in PAX8/PPARgamma-negative thyroid tumours.

The expression of peroxisome proliferator-activated receptor (PPAR)gamma in thyroid neoplasias and in normal thyroid (NT) tissues has not been fully investigated. The objectives of the present work were: to study and compare the relative expression of PPARgamma in normal, benign and malignant thyroid tissues and to correlate PPARgamma immunostaining with clinical/pathological features of patients with thyroid cancer. We analysed the expression of PPARgamma in several types of thyroid tissues by reverse transcription-polymerase chain reaction (RT-PCR), interphase fluorescent in situ hybridisation, real-time RT-PCR and immunohistochemistry. We have demonstrated that NT tissues express PPARgamma both at mRNA and at protein level. PAX8-PPARgamma fusion gene expression was found in 25% (six of 24) of follicular thyroid carcinomas (FTCs) and in 17% (six of 36) of follicular thyroid adenomas, but in none of the 10 normal tissues, 28 nodular hyperplasias, 38 papillary thyroid carcinomas (PTCs) and 11 poorly differentiated thyroid carcinomas (PDTCs). By real-time RT-PCR, we observed that tumours negative for the PAX8-PPARgamma rearrangement expressed lower levels of PPARgamma mRNA than the NT. Overexpression of PPARgamma transcripts was detected in 80% (four of five) of translocation-positive tumours. Diffuse nuclear staining was significantly (P<0.05) less prevalent in FTCs (53%; 18 of 34), PTCs (49%; 19 of 39) and PDTCs (0%; zero of 13) than in normal tissue (77%; 36 of 47). Peroxisome proliferator-activated receptorgamma-negative FTCs were more likely to be locally invasive, to persist after surgery, to metastasise and to have poorly differentiated areas. Papillary thyroid carcinomas with a predominantly follicular pattern were more often PPARgamma negative than classic PTCs (80% vs 28%; P=0.01). Our results demonstrated that PPARgamma is underexpressed in translocation-negative thyroid tumours of follicular origin and that a further reduction of PPARgamma expression is associated with dedifferentiation at later stages of tumour development and progression.

Cloning and sequence analysis of the ces10 gene encoding a Sphingomonas paucimobilis esterase.

The ces10 gene of the gellan gum-producing strain Sphingomonas paucimobilis ATCC 31461 was cloned and sequenced. Multi-sequence alignment of the deduced protein indicated that Ces10 belongs to the serine hydrolase family with a potential catalytic triad comprising Ser(153) (within the G-X-S-X-G consensus sequence), His(75) and Asp(125). The mixed block results obtained following pattern search and the low identities detected in a BLAST analysis indicate that Ces10 is significantly different from other characterised bacterial esterases/lipases. Nevertheless, the Ces10 amino acid sequence showed 45% similarity with Rhodococcus sp. heroin esterase and 48% with Bacillus subtilis p-nitrobenzyl esterase. Ces10, with a predicted molecular mass of 30,641 Da, was overproduced in Escherichia coli and purified to homogeneity in a histidine-tagged form. Enzyme assays using p-nitrophenyl-esters (p-NP-esters) with different acyl chain-lengths as the substrate confirmed the anticipated esterase activity. Ces10 exhibited a marked preference for short-chain fatty acids, yielding the highest activity with p-NP-propionate (optimal pH 7.4, optimal temperature 37 degrees C).

Characterization of the ugpG gene encoding a UDP-glucose pyrophosphorylase from the gellan gum producer Sphingomonas paucimobilis ATCC 31461.

The ugpGgene, which codes for a UDP-glucose pyrophosphorylase (UGP) (or glucose-1-phosphate uridylyltransferase; EC 2.7.7.9) in Sphingomonas paucimobilis ATCC 31461, was cloned and sequenced. This industrial strain produces the exopolysaccharide gellan, a new commercial gelling agent, and the ugpG gene may convert glucose-1-phosphate into UDP-glucose in the gellan biosynthetic pathway. The ugpG gene is capable of restoring the capacity of an Escherichia coli galU mutant to grow on galactose by functional complementation of its deficiency for UDP-glucose pyrophosphorylase activity. As expected, the predicted gene product shows strong homology to UDP-glucose pyrophosphorylases from several bacterial species. The N-terminal region of UgpG exhibits the motif GXGTRXLPXTK, which is highly conserved among bacterial XDP-sugar pyrophosphorylases, and a lysine residue (K(192)) is located within a VEKP motif predicted to be essential for substrate binding or catalysis. UgpG was purified to homogeneity as a heterologous fusion protein from crude cell extracts prepared from IPTG-induced cells of E. coli, using affinity chromatography. Under denaturing conditions, the fusion protein S-UgpG-His(6) migrated with an estimated molecular mass of 36 kDa [corresponding to the predicted molecular mass of native UgpG (31.2 kDa) plus 5 kDa for the S and histidine tags). Kinetic analysis of UgpG in the reverse reaction (pyrophosphorolysis) showed a typical Michaelis-Menten substrate saturation pattern. The apparent K(m) and V(max) values estimated for UDP-glucose were 7.5 microM and 1275 micromol/min/g.

Gellan gum biosynthesis in Sphingomonas paucimobilis ATCC 31461: genes, enzymes and exopolysaccharide production engineering.

The commercial gelling agent, gellan, is an extracellular polysaccharide (EPS) produced by Sphingomonas paucimobilis ATCC 31461. In recent years, significant progress in understanding the relationship between gellan structure and properties and elucidation of the biosynthesis and engineering of this recent product of biotechnology has been made. This review focuses on recent advances in this field. Emphasis is given to identification and characterization of genes and enzymes involved, or predicted to be involved, in the gellan biosynthetic pathway, at the level of synthesis of sugar-activated precursors, of the repeat unit assembly and of gellan polymerization and export. Identification of several genes, biochemical characterization of the encoded enzymes and elucidation of crucial steps of the gellan pathway indicate that possibilities now exist for exerting control over gellan production at any of the three levels of its biosynthesis. However, a better knowledge of the poorly understood steps and of the bottlenecks and regulation of the pathway, the characterization of the composition, structure and functional properties of gellan-like polymers produced either by the industrial strain under different culture conditions or by mutants are still required for eventual success of the metabolic engineering of gellan production.

Antibody response to IR6, a conserved immunodominant region of the VlsE lipoprotein, wanes rapidly after antibiotic treatment of Borrelia burgdorferi infection in experimental animals and in humans.

Invariable region (IR)(6), an immunodominant conserved region of VlsE, the antigenic variation protein of Borrelia burgdorferi, is currently used for the serologic diagnosis of Lyme disease in humans and canines. A longitudinal assessment of anti-IR(6) antibody levels in B. burgdorferi-infected rhesus monkeys revealed that this level diminished sharply after antibiotic treatment (within 25 weeks). In contrast, antibody levels to P39 and to whole-cell antigen extracts of B. burgdorferi either remained unchanged or diminished less. A longitudinal analysis in dogs yielded similar results. In humans, the anti-IR(6) antibody titer diminished by a factor of > or =4 in successfully treated patients and by a factor of <4 in treatment-resistant patients. This result suggests that the quantification of anti-IR(6) antibody titer as a function of time should be investigated further as a test to assess response to Lyme disease therapy or to determine whether a B. burgdorferi infection has been eliminated.

Lyme disease: an update.

Lyme disease is a multisystem illness caused by the spirochete Borrelia burgdorferi, and it is the most common vector-borne illness in the United States. Lyme disease is also endemic in Europe and Asia. There have been major advances in the field since the disease was first described, including the sequencing of the B. burgdorferi genome; an increase in understanding of the interactions among the spirochete, the tick, and the mammalian host; new and improved laboratory tests; and a vaccine for prevention of the disease. Still, the diagnosis of Lyme disease remains based on history and clinical findings, supplemented by careful use of laboratory tests, and requires that the physician be familiar with the disease's clinical manifestations and the shortcomings of the available diagnostic tests.

Medullary carcinomas of the thyroid: a monoclonal origin.

We studied the clonality of medullary thyroid carcinomas (MTC) from 16 female patients by determining X chromosome inactivation by polymerase chain reaction (PCR) amplification of a CAG repeat in exon 1 of the human androgen-receptor gene. One patient with sporadic medullary thyroid carcinoma (MTC) was homozygous for this microsatellite and was not considered for the assessment of clonality. Sixteen tumor samples from the informative 15 patients were studied: 11 were from sporadic cases and 5 were from familial cases (3 cases of multiple endocrine neoplasia type 2A [MEN 2A]; 1 case of familial medullary thyroid carcinoma [FMTC]). Fourteen tumor samples (10/11 sporadic, 3/4 MEN 2A and 1/1 FMTC) were clearly monoclonal with allelic cleavage ratios between 2.5 and 49.1. Sixty-four percent of these cases (9/14) had the preferential amplification of the shorter allele while 36 percent (5/14) had the preferential amplification of the longer allele. Two frozen tumor samples (1 sporadic and 1 MEN 2A) were polyclonal. However, the corresponding tumor embedded in paraffin from the sporadic case was monoclonal. The other polyclonal tumor was found in the right thyroid lobe of a patient with MEN 2A who had a monoclonal tumor in the left lobe. Our results clearly demonstrate that MTC have a monoclonal origin in the majority of the cases.

Evaluation of a new culture medium for Borrelia burgdorferi.

We evaluated the new MPM medium for the growth of Borrelia burgdorferi. All 18 blood samples from 17 patients with Lyme disease were negative. Growth studies showed that by day 4, most organisms in MPM were not viable. Our results reinforce the use of BSK medium as the primary choice for growing B. burgdorferi.