Short communication: quantitative trait loci affecting the somatic cell score on chromosomes 4 and 26 in Italian Holstein cattle.

This work aimed to confirm previously reported quantitative trait loci (QTL) affecting the somatic cell score (SCS) in dairy cattle on Bos taurus autosomes (BTA) 4 and 26. A granddaughter design with selective genotyping was implemented that included half-sib families from 12 male lines of Italian Holstein cattle. The animals were genotyped for 5 microsatellite markers each on regions of BTA 4 (average marker spacing 9.42 cM) and BTA 26 (average marker spacing 5.26 cM), previously reported by other authors as carrying QTL for somatic cell count. Quantitative trait loci analyses were performed using interval mapping by regressing sire breeding values for SCS onto genotype probabilities at 1-cM intervals along the 2 chromosome regions. Breeding values for SCS were estimated for the whole population using a test-day repeatability animal model. Results were not significant on a chromosome basis, but a possible QTL was found at BM4505 on BTA 26, confirming this region for further studies of QTL affecting SCS in the Italian Holstein population.

Liquid chromatography-tandem mass spectrometry for the identification of impurities in d-allethrin samples.

A previous GC/MS study highlighting the impurity profile of the synthetic pesticide d-allethrin is extended here to validate and confirm the impurities identity through the development of soft ionisation HPLC-MS methods. To accomplish this, we developed a reverse phase LC-MS analysis in gradient elution with two distinct soft ionisation techniques, the atmospheric pressure ionisation with electrospray source (API-ESI) and the chemical ionisation (APCI). A single quadrupole and an ion trap, which allowed the simultaneous determination of the molecular masses and structural information of the impurities by acquisition of collisionally induced (CID) product ions spectrum and in-source fragmentation, were employed as analysers. Single quadrupole and ion trap analysers resulted perfectly matching in the d-allethrin impurity fragmentation patterns. All the main impurities over 0.1% identified by GC/MS were confirmed. Results indicate that the proposed HPLC/MS method was found appropriate to confirm the presence of impurities such as chrysolactone, chloro allethrin derivatives, allethrolone and chrysanthemic acid, excluding their formation under GC/MS strong ionisation condition.

Stereoselective determination of allethrin by two-dimensional achiral/chiral liquid chromatography with ultraviolet/circular dichroism detection.

A two-dimensional achiral/chiral HPLC method with circular dichroism (CD) detection was optimized for the stereochemical resolution and determination of the elution order of the eight stereoisomers of synthetic allethrin. A monolithic silica HPLC column (Chromolith, Merck, 100 mm x 4.6 mm i.d.) was put orthogonally to an enantioselective OJ Daicel column (250 mm x 4.6 mm i.d.) by means of a switching valve. The resolution of cis and trans diastereoisomers on the silica column was obtained by using a mobile phase consisting of n-hexane:tert-butyl methyl ether (96:4) (v/v) at a flow rate of 1 ml min(-1). The cis and trans peaks were then switched to the enantioselective OJ column separately in two subsequent injections. The resolution of the four trans stereoisomers was accomplished by using n-hexane:tert-butyl methyl ether (90:10) (v/v), while the mobile phase composition for the four cis stereoisomers consisted of n-hexane:isopropanol (99.3:0.7) (v/v). The CD based detection system allowed the determination of the elution order on the basis of the CD signals of the single stereoisomers, together with the injection of pure stereoisomers. Under the final conditions, the validated method was applied to the determination of stereoisomeric composition and absolute configuration of the prevailing stereoisomers of real samples, i.e. commercial batches of different sources of d-allethrin.

Comparative FISH-mapping of the survival of motor neuron gene (SMN) in domestic bovids.

A comparative fluorescence in situ mapping of the SMN gene was performed on R-banded chromosome preparations of cattle (Bos taurus, BTA, 2n = 60), river buffalo (Bubalus bubalis, BBU, 2n = 50), sheep (Ovis aries, OAR, 2n = 54) and goat (Capra hircus, CHI, 2n = 60), as well as on those of a calf from Piedmont breed affected by arthrogryposis. SMN was located on BTA20q13.1, OAR16q13.1, CHI20q13.1 and BBU19q13. These chromosomes and chromosome bands are believed to be homeologous, confirming the high degree of chromosome homeologies among bovids. The position of SMN was refined in cattle, compared to the two previous localizations, while it is a new gene assignment in the other three bovids. A comparative fiber-FISH performed on extended chromatin of both normal cattle and calf affected by arthrogryposis revealed more extended FITC signals in the calf, compared to the normal cattle (control), suggesting a possible duplication of the SMN gene in the calf affected by arthrogryposis. .

Recombinant DRB sequences produced by mismatch repair of heteroduplexes during cloning in Escherichia coli.

Recombinant chimeric sequences originating from a mixture of the sequences of two different alleles are frequently found after amplification and cloning in Escherichia coli of exon 2 of the major histocompatibility complex (MHC) DRB genes. Several authors have suggested that the recombinant molecules result from in vitro recombination during PCR; nevertheless, a clear experimental demonstration of this hypothesis is lacking. In order to understand the mechanism producing the chimeric sequences, we set up a simple experiment based on the different restriction patterns of parental and recombinant sequences. Our data demonstrate that in the analysed case most of the recombinant variants were not produced by in vitro recombination during PCR, but were the result of the mismatch repair of heteroduplex molecules during cloning in E. coli. The high mutation rate in the alpha-helix region of DRB expressed genes, both after cloning in E. coli and after the germ-line differentiation process in vertebrates, suggests that the observed mutations are the result of similar gene conversion processes, probably favoured by chi-dependent microrecombination events.

Typhlitis (neutropenic enterocolitis) after a single dose of vinorelbine.

We report a case of a 50-year-old man with pretreated adenocarcinoma of the lung, who developed fatal neutropenic enterocolitis (typhlitis) after a standard dose of the cytotoxic drug vinorelbine. Blood cultures were negative for all microorganisms tested for. Stool cultures were negative for enteric rods but direct examination of fresh stool revealed the presence of Giardia lamblia. Abdominal pain and diarrhoea developed very rapidly while the patient was only moderately neutropenic. Metronidazole was prescribed without clinical benefit: the abdominal pain remained stable. The duration of neutropenia was very short (4 days). The abdominal catastrophe ending in shock occurred after complete recovery of the neutrophil count. Neutropenic colitis has been reported with increasing frequency in solid tumours after the introduction of taxanes. This complication has been observed mainly in phase I studies, near the maximally tolerated doses (MTD). The combined use of vinorelbine has recently been reported to exacerbate the toxic effects of taxane on the colon. The case presented here demonstrates that typhlitis can occur even with vinorelbine alone, used at a standard recommended dose (30 mg/m2).

Chromosome analysis and molecular characterization of highly repeated DNAs in the aphid Acyrthosiphon pisum (Aphididae, Hemiptera).

Despite the interest in aphid biology, information on chromatin organization of their holocentric chromosomes is still limited to few species. In order to fill this gap, we have performed an extensive survey on pea aphid mitotic chromosomes using both classical and molecular cytogenetic techniques. Our results after silver, CMA3 and DAPI-staining, C-banding and fluorescent in situ hybridization (FISH) using 28S rDNA and 5S rDNA as probes evidenced a tendency of repetitive DNAs to be concentrated on the X chromosomes. FISH experiments with the telomeric probe (TTAGG)n revealed bright hybridization signals on each telomere of all Acyrthosiphon pisum chromosomes. No interstitial signals were seen.

[Giant benign gastric ulcer penetrating into the liver, pancreas and mesocolon]. / Ulcera gastrica gigante benigna penetrante nel fegato, nel pancreas e nel mesocolon.

Penetration of the liver, pancreas and transverse mesocolon by a giant benign gastric ulcer is relatively uncommon, and literature contains a few reports of this complication. The preoperative histological diagnosis may be difficult or impossible. A 63-year-old female patient with a history of seven months of lack of appetite, asthenia, epigastric pain, a remarkable weight decrease, presenting at physical examination a large, smooth margins, not pulsating, quite fixed abdominal mass, is reported. Echography confirmed the presence of a mass of approximately 14 x 19 cm, with solid and liquid content. Biopsy showed inflammatory elements and cellular detriti. Barium enema showed that the mass compressed the descendent colon, which appeared dislocated. Tumor markers (CEA, CA 19-9, alpha-fetoprotein) where in the normal range. Endoscopy showed a giant angular ulcer whose bottom was represented by necrotic material (after the definitive histological examination it proved to be hepatic tissue). At TC scan of the abdomen, a remarkable thickening of the gastric wall was present. At surgery the stomach appeared increased in volume, with remarkably thickened walls, tenaciously sticking to II and III hepatic segments, to the pancreas and transverse mesocolon. A total gastrectomy was performed because of the depth of the ulcer penetration and the extension of the alteration of the gastric wall, even if the giant gastric ulcer, in the literature, is more frequently benign than malignant.

[Acute pancreatitis. Our experience during 20 years]. / Pancreatite acuta. La nostra esperienza nell'arco di venti anni.

BACKGROUND: On the basis of personal experience and studies, the importance to diagnose quickly the seriousness of an acute pancreatitis with the help of simple and credible criteria of evaluation is underlined. It's also underlined the help of endoscopic papillosphincterotomy in the initial phase of a biliary pancreatitis; in fact this exam permits to perform a laparoscopic cholecystectomy in a second time and reduce surgical trauma. METHODS: Personal experience with 288 cases of acute pancreatitis (AP), during a twenty-year period (1975-1996) is described. In 61% of cases the AP was associated with biliary illness, and in 13% with alcohol abuse. Real important for therapeutic implication and prognostic significance is the classification of severity of acute attack, by using clinical evaluation (pain, fever, jaundice, abdominal tenderness), multiparametric score (i.e. Imrie's score, Ranson's criteria, SAPS or APACHE II score system) and morphological evaluation of the pancreas (echo and CT scan). 151 patients have been admitted at different times to surgery, the cholecystectomy being the more common surgical procedure, with a surgical rate of 52%. RESULTS: The overall mortality has been 7.2%, ranging from 45.4% in 17 severe necrotic haemorrhagic AP to 2.1% for the mild one. CONCLUSIONS: The widespread use of endoscopic retrograde colangiopancreatography (associated to endoscopic sphincterotomy if necessary) and the recent but universally accepted diffusion of laparoscopic cholecystectomy have changed the modern approach to surgical treatment of biliary associated AP, ensuring mininvasive less traumatic surgical procedures.

Report of the first workshop on the genetic map of bovine chromosome 1.

A report of the first workshop on the genetic map of bovine chromosome 1 (BTA1) is presented. Five laboratories contributed 31,962 informative meioses from 70 loci. Thirty-two loci which had been typed by at least two laboratories were used to construct a framework genetic map with a likelihood ratio support of at least 1000:1 for locus order. The resulting sex-averaged framework map contained 26 loci and spanned 163.6 CM. The lengths of the female and male maps were 159.5 CM and 165.3 CM, respectively, and there was evidence for an expansion in the telomeric one-third of the male map. Of the four cases where order for closely linked loci differed among the maps produced for each of the contributing laboratories, a consensus order was obtained for three in the framework map. The average genetic distance between framework loci on the sex-averaged map was 6.3 CM.

Molecular definition of an allelic series of mutations disrupting the myostatin function and causing double-muscling in cattle.

We have determined the entire myostatin coding sequence for 32 double-muscled cattle sampled from ten European cattle breeds. Seven DNA sequence polymorphisms were identified, of which five would be predicted to disrupt the function of the protein, one is a conservative amino acid substitution, and one a silent DNA sequence variant. Four additional DNA sequence polymorphisms were identified in myostatin intronic sequences. In all but two breeds, all double-muscled animals were either homozygous or compound heterozygotes for one of the five loss-of-function mutations. The absence of obvious loss-of-function mutations in the coding sequence of the two remaining breeds points either towards additional mutations in unexplored segments of the gene, or towards locus heterogeneity of double-muscling.

Assessment of the RSCL quality of life instrument during chemotherapy in an Italian setting.

Evaluation of the quality of life of oncological patients is now perceived as a major element of cancer treatment, and various instruments for measuring it have been proposed. However, whatever instrument is used must be capable of reflecting the different ways in which different contexts and cultures respond to cancer. We administered the Italian version of the Rotterdam Symptom Checklist (RSCL) to 60 patients with advanced disease attending five oncological centres in Romagna (central Italy). The RSCL was found to be able to measure both psychological and physical well-being and to detect a difference between patients on low/medium and those on high-burden chemotherapy. The Italian version of the RSCL is an adequate scale that is capable of capturing patient perceptions and could be used for monitoring the quality of life of Italian patients participating in clinical trials.

Association of BoLA class II haplotypes with subclinical progression of bovine leukaemia virus infection in Holstein-Friesian cattle.

The influence of bovine lymphocyte antigen (BoLA) complex polymorphism on subclinical progression of bovine leukaemia virus (BLV) infection was investigated in 41 Holstein-Friesian cows from two herds in Italy. All cows were seropositive for BLV and 22 had persistent lymphocytosis (PL). BoLA-A specificities were defined by serology, and class II haplotypes were defined based on restriction fragment length polymorphism (RFLP) and polymerase chain reaction (PCR)-RFLP analysis of DQ and DR genes. Chi-square analysis revealed a significant and absolute association of haplotype DQA*3A-DQB*3A-DRB2*2A-DRB3.2*11 with resistance to PL (P chi 2 = 0.028, relative risk (RR) = 0.061). Consistent with this observation, multiple regression analysis revealed that animals carrying this haplotype had lower lymphocyte counts (P = 0.0057). By contrast, haplotype DQA*12-DQB*12-DRB2*3A-DRB3.2*8 was associated with susceptibility to PL (P chi 2 = 0.043, RR = 9.625) and increased lymphocyte counts (P = 0.0537). These results confirm the association of haplotype DQA*3A-DQB*3A-DRB2*2A-DRB3.2*11 with resistance to PL, and substantiate earlier findings of haplotype DQA*12-DQB*12-DRB2*3A-DRB3.2*8 as a risk factor for subclinical progression to PL in BLV-infected Holstein-Friesian cattle.

Improving the quality of health services organization structure by reengineering: circular design and clinical case impact in an academic medical center.

Innovation to improve the quality of structure and process in health care organization is reported in this case example of change in an academic medical center. Interactive planning and the circular organization design concept were the driving principles and methods. This report presents the needs for and initial obstructions to change, planning and project design work, a description of the change process, and illustrative accomplishments to date--two cases, one of conscious sedation policy and one of nuisance pages. Evaluative criteria for judging the progress and lessons of the project regarding key design characteristics also are included.

BoLA class I polymorphism and in vitro immune response to M. bovis antigens.

SUMMARY: From a sample of 119 Friesian calves, serologically typed for BoLA class I, 47 subjects were chosen expressing 9 different MHC types (A6, A6.9, A10, A11, A14, A15, A30, W16, M103) with the same age and reared in the same farm conditions. The animals were s.c. injected with a water in oil suspension of killed M. bovis and the treatment was repeated two days later. Before the treatment and 21 days later, calves were bled and on PBM (peripheral blood mononuclear leucocytes) were performed the following tests: 1. Lymphocyte Stimulation with bovine and avian PPDs (Purified protein derivative of Mycobacterium bovis and Mycobacterium avium, respectively). 2. Phagocytic activity towards M. bovis. 3. Class II molecules expression on cell surface. 4. Percentage of leucocyte populations and subpopulations. In the in vitro Lymphocyte Stimulation test, all the animals and classes were responders. Animals bearing A10 BoLA class I presented c.p.m. (counts per minute) and index values higher than the other cattle; these values were significantly positively related both to bovine and avian PPDs (P < .01). By variance analysis A14 BoLA type showed a slight positive significant correlation with more efficient phagocytic activity. BoLA class I type did not seem to significantly affect percentage of class II positive cells and leucocyte percentages on PBM. ZUSAMMENFASSUNG: Der BoLA Klasse I Polymorphismus und in vitro immunologische Antwort gegen die Antigene von M. bovis Aus einer Stichprobe von 119 für BoLA Klasse I serologisch typisierten Friesian Kälber, wurden 47 Subjekte ausgewählt, die 9 verschiedene MHC Typen ausdrückten (A6, A6.9, A10, A11, A14, A15, A30, W16, M103). Alle waren gleich alt und in gleichen Haltungsbedingungen. Die Tiere wurden mit einer Wasser-in-Ol Suspension abgetöteter M. bovis subkutan injiziert und die Behandlung nach zwei Tagen wiederholt. Vor und 21 Tage nach Behandlung wurden die folgenden Tests ausgeführt: 1. Lymphozyten-Stimulationstest mit bovinen und Geflügel PPDs. 2. Phagozyten Aktivität gegen M. bovis. 3. Zeil-Oberflächen, Expression der Klasse II Moleküle. 4. Anteile der Lymphozyten Populationen und Subpopulationen. Im in vitro Lymphozyten-Stimulationstest waren alle Tiere und Klassen responder. Tiere mit A10 BoLA I zeigten höhere c.p.m. und Indexwerte als die anderen; diese Werte waren in signifikant positiver Beziehung mit der PPD von M. bovis und auch mit M. avium (P < .01). BoLA Typ A14 zeigte leicht signifikant positive Korrelation mit wirksamerer Phagozyten Aktivität. BoLA Klasse I Typ scheint nicht den Prozentsatz der positiven Zellen der Klasse II und der Leukozyten der PBM signifikant zu beeinflussen. RESUMEN: Polimorfismo de BoLA clase I y immunidad a los antigenos del M. bovis Se escojeron 47 novillos dentro de un grupo de 119 animales que segun analisis previamente hecha tenian BoLA de clase I. Estos 47 novillos fueron escojidos de manera que tuvieran 9 distintos tipos de MHC (A6, A6.9, A10, A11, A14, A15, A30, W16, M103), la misma edad, las mismas condiciones de cria. Estos animales fueron inoculados subcutaneo con M. bovis matados en una suspension oleosa y la misma inoculacion fue repetida una secunda vez despues de dos dias. Por cada animal se tomaron muestras de sangre antes y 21 dias despues de la inoculacion de arriba. Las muestras de sangre fueron pruebaoas con: 1. Stimulacion Lymhocitaria con PPD bovina y avicola. 2. Actividad phagocitaria a M. bovis. 3. Expresion sobre la superficie celular de moleculas de clase II. 4. Porcentaje de poblaciones y de subpob-laciones de leucocitos. Todos los animales y todos los tipos de MHC dieron respuestas positivas en las pruebas de Stimulacion Lymphocitaria. Los animales que tenian la BoLA A10 presentaron valores de c.p.m. y indices mas altos de los demas animales. Estos valores se encontraron significativamente y positivamente relacionados sea a la PPD bovina que a la PPD avicola. Por medio de la analisis de varianzas se encontro que el tipo BoLA A14 muestraba una correlacion significativa y algo positiva con una mejor actividad fagocitaria. Los tipos de clase BoLA I no parecieron que influenzaran de manera appreciable el porcentaje de positividad por la clase II y el porcentaje de leucocitos en la sangre PBM.

Exposure to low-frequency pulsed electromagnetic fields increases mitogen-induced lymphocyte proliferation in Down's syndrome.

We previously reported that exposure of human mitogen-stimulated peripheral blood lymphocytes (PBL) to extremely low frequency pulsed electromagnetic fields (PEMFs) could restore the defective proliferative capability of PBL from aged subjects. The effects of exposure to PEMFs were studied in PBL from 25 patients with Down's syndrome (DS), a syndrome of premature aging characterized by precocious immune system derangement, including age-related defective PBL proliferative capability. PBL were stimulated with different doses of phytohemagglutinin, and cell proliferation was assessed by measuring the incorporation of tritiated thymidine. After PEMF-exposure, a significant increase in cell proliferation was observed in cells from DS children and young adults, but it was much more evident in PBL from relatively aged DS patients. The age-related effect of PEMFs on DS lymphocytes demonstrates that age must be considered a major variable when studies on DS are performed, and confirms that DS must be regarded as a syndrome of accelerated aging.

Dressing for success: measuring productivity can ensure continuing success.

Both managerial and financial benefits can be gained by implementing and monitoring productivity standards in the health-care organization. However, these standards must be reasonable and acceptable to both management and staff if they are to be effective. By implementing a logging system to determine actual daily activity, Elizabethtown Hospital was able to develop reasonable productivity standards that allowed department managers to manage their operations more efficiently.