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Background Colorectal cancer (CRC) risk is increased in patients (pts) with long-standing colitis related to Inflammatory Bowel Disease (IBD). A higher frequency mucinous and signet-ring colonic adenocarcinoma has been suggested in IBD, but data regarding risk factors for these aggressive CRC are currently lacking. Primary aim was to assess the frequency of mucinous and signet-ring adenocarcinoma in IBD pts with CRC. Secondary aim was to assess risk factors for these histotypes of CRC. Methods From January 2002 to July 2022, all IBD pts with concomitant CRC were retrospectively enrolled. Inclusion criteria: 1) age ≥18; 2) well-defined diagnosis of IBD and CRC; 3) available histological and surgical report. Exclusion criteria: Missing data. Characteristics of IBD were reported according to standard criteria. Data were expressed as median [range]. Student-t Test and χ2 test were used for comparisons. Univariate logistic regression model was applied for assessing risk factors for mucinous and signet-ring adenocarcinoma (OR [95%CI]). Results The study population included 40 IBD pts with concomitant CRC: 24 (60%) with Ulcerative Colitis (UC) and 16 (40%) with Crohn’s Disease (CD). CRC included standard adenocarcinoma in 23 (57.5%) and mucinous or signet-ring in 17 (42.5%) pts. CD was more frequently stricturing in pts with standard adenocarcinoma (7 [77.8%] vs 1 [14.4%], p=0.04). CRC most frequently involved the rectum in pts with mucinous or signet-ring adenocarcinoma vs standard adenocarcinoma (4 [17.4%] vs 8 [47.1%]; p=0.04). Other IBD characteristics did not differ between standard and mucinous or signet-ring adenocarcinoma, including: age at CRC diagnosis (61 [30-80] vs 53 [29.80]; p=0.61), gender (F): 8 [34.8%] vs 5 [29.4%]; p=0.98), IBD duration at CRC diagnosis (14 [1-45] vs 17 [1-36]; p=0.74), smoking status (p=0.78), IBD type (UC: 14 [60.9%] vs 9 [56.3%]; p=0.84), UC extent and CD localization, frequency of perianal disease (p=0.37), thiopurine (p=0.55) or biologic (p=0.55) use. The proportion of pts surgically treated for CRC (20 [86.9%] vs 17 [100%], p=0.34) and the frequency of CRC-related death (3 [13.1%] vs 5 [29.4%]; p=0.37) were also comparable between groups. At diagnosis, CRC stage was comparable between pts with standard vs mucinous or signet-ring adenocarcinoma (stage I: 6 [26.1%] vs 1 [5.9%]; p=0.21; II: 9 [39.1%] vs 4 [23.5%]; p=0.48; III: 5 [21.7%] vs 7 [41.2%]; p=0.34); IV: 3 [13.1%] vs 2 [11.8%]; p=0.71). At univariate analysis, no specific risk factors for mucinous and signet-ring colonic adenocarcinoma were detected. Conclusion In the tested cohort of IBD patients with CRC, mucinous and signet-ring adenocarcinomas were observed in almost half of cases, although no specific risk factors were identified.

The properties of rotating turbulence driven by precession are studied using direct numerical simulations and analysis of the underlying dynamical processes in Fourier space. The study is carried out in the local rotating coordinate frame, where precession gives rise to a background shear flow, which becomes linearly unstable and breaks down into turbulence. We observe that this precession-driven turbulence is in general characterized by coexisting two dimensional (2D) columnar vortices and three dimensional (3D) inertial waves, whose relative energies depend on the precession parameter $Po$. The vortices resemble the typical condensates of geostrophic turbulence, are aligned along the rotation axis (with zero wavenumber in this direction, $k_z=0$) and are fed by the 3D waves through nonlinear transfer of energy, while the waves (with $k_z\neq0$) in turn are directly fed by the precessional instability of the background flow. The vortices themselves undergo inverse cascade of energy and exhibit anisotropy in Fourier space. For small $Po0.1$ turbulence is quasi-steady with only mild fluctuations, the coexisting columnar vortices and waves in this state give rise to a split (simultaneous inverse and forward) cascade. Increasing the precession magnitude causes a reinforcement of waves relative to vortices with the energy spectrum approaching Kolmogorov scaling and, therefore, the precession mechanism counteracts the effects of the rotation., 20 pages, 16 figures, 1 table, submitted to Physics of Fluids

The fluid flow in a precessing cylinder is investigated numerically with focus on the Ekman boundary layers in the strongly forced regime. Not surprisingly, in that regime, we find deviations from the linear theory due to significant modifications of the base flow in terms of an axisymmetric geostrophic mode whose rotation is opposite to that of the container. The transition of the bulk flow from a three-dimensional non-axisymmetric base flow to a geostrophic axisymmetric pattern is reflected in the scaling of both the sidewall boundary layers and the Ekman boundary layers on top and bottom of the cylinder. In our simulations, the Ekman layers surpass the threshold of the first instability (class A) and show an increase in the thickness together with a marked vertical flow advection inside the boundary layer in a limited range of the forcing magnitude. However, due to numerical restrictions in our simulations, which limit the range of achievable Ekman numbers, no developed boundary layer turbulence is found. An estimation by extrapolation shows that, for this purpose, Ekman numbers smaller by a factor of two have to be achieved.

Extracellular vesicles isolated from follicular fluid of antral and preovulatory follicles present a different miRNA profile related to follicular development and breeding season in buffalo (bubalus bubalis)

Biological cells are utilized for diverse biotechnological and bioengineering purposes ranging from the production of biopharmaceuticals, to cell therapy, "human-on-a-chip" drug and toxicology assays, and drug-resistance tests. In these and other applications, it is critical to quantify the levels of not only viable but also non-viable cells. While traditional off-line cell-staining methods are available for counting of non-viable cells, many applications cannot periodically remove cells for their off-line analysis because of the risk of contamination or workflow logistics. Here we show in-situ label-free quantitation of viable and non-viable cells with multivariable multi-resonant sensors. We used Chinese hamster ovary (CHO) cells in suspension culture in single-use bioreactors as a representative example. The resonant sensor design strategy permitted enhanced sensor sensitivity versus conventional non-resonant measurements and probed the spectral dispersion of viable and non-viable cells with multiple resonances. These capabilities of label-free in-situ analysis of cell viability can be attractive in diverse cell applications such as cell suspensions, adhered cells, and their 3D assemblages.

Bitto Storico cheese is a raw milk seasonal cheese produced in the Orobic Alps, whose production is closely associated to local traditional activities and alpine pastures. The aim of this work was to characterize the microbiota diversity and the volatile fractions of cheese produced in different alpine pasture areas. Fifty-four Bitto Storico cheeses samples, produced in six different Alpine pastures, were collected and processed. Bacterial DNA was extracted using an optimized protocol and 16S rRNA gene amplicons on V3-V4 region analyzed by Miseq (Illumina). The volatile fraction was characterized by means of Solid Phase Extraction-Gas Chromatography-Mass Spectrometry (SPME-GC-MS). The microbial community of ripened cheese was mainly dominated by Firmicutes phylum with a high abundance of Streptococcaceae and Lactobacillaceae. Each Alpine area was characterized by peculiar bacterial communities, due, probably, to the pasture composition and the cheesemaking process. A total of 24 volatile compounds resulting from microbial activity and diet were found, the formers being mostly alcohols and esters, the latter being terpenes derived directly from the pasture.

Agilent Tape station profile of small RNA library (152 bp) obtained from pool of 24 samples concentrated with magnetic beads and size selected with Pippin Prep. 128 bp size peak represents primer dimmers co-purified with the library. (DOCX 40Â kb)

Individual bulls differ in their ability to fertilize oocytes due to physiological and molecular characteristics of spermatozoa. The combined use of advanced techniques, as standard semen quality assessment together with sperm molecular investigation, is a promising approach to achieve a better understanding of sperm functions and to predict bull fertility. The aim of this study was to evaluate the relationships among frozen bull sperm parameters, assessed by computer-assisted semen analysis (CASA) and flow cytometry (FCM), and miRNAs expression in high and low fertility bulls. Ten frozen semen doses from 10 Italian Holstein-Friesian bulls, 5 of high and 5 of low fertility according to the estimated relative conception rate (ERCR), were analyzed in two different breeding seasons (Sep-Oct and Feb-Mar), by using CASA system for motility and sperm kinetic parameters assessment and FCM for sperm viability, acrosomal status and DNA integrity evaluation. The semen doses were thawed and pooled; high (HM) and low (LM) motile sperm fractions were isolated by Percoll gradient. For each bull total RNA was extracted from HM fraction and small RNA libraries were generated using Illumina Truseq Small RNA Preparation kit. Libraries were sequenced on a single lane of Illumina Hiseq 2000. Sperm cells were successfully fractionated in HM and LM populations achieving a better quality (P

Many biopharmaceutical companies have replaced or are planning to replace traditional multi-use facilities (fixed-in-place stainless-steel fermenters, tanks, downstream equipment, and associated piping) with single-use systems to improve flexibility and cost. This article will describe a recently-completed comparative study of the environmental impacts of producing monoclonal antibodies using either single use or traditional process technology. The study was performed using Life cycle assessment methodology in which environmental impacts across the entire life cycle of each process component, from materials extraction and refining through component manufacturing, packaging, distribution, use, and disposal at end-of life are all considered. The assessment looked at the production of monoclonal antibodies (MAb) over a 10-batch campaign at three production scales chosen to reflect the clinical phase, scale-up phase, and production phase. The entire process trains were modeled including upstream and downstream processes from N-2 seed fermentation through product purification. Inventory data were derived mainly from Biopharm Services Ltd., developer of BioSolve, an industry standard bioprocess model that includes standard benchmark operations and costs that can be used to build any process including those for manufacture of mAbs, vaccines and bacterial based products. The results of this study indicate that the single-use process train exhibited lower environmental impacts compared to the traditional fixed-in-place process train in each environmental impact category studied. This is primarily due to a reduced need for the energy intensive water-for-injection, process water and clean steam that are required to perform cleaning and sterilization between batches for traditional fixed-in-place equipment.

Summary: Frozen bull semen was analyzed after fixation with glutaraldehyde (0.2% sol. in PBS) by clear field microscopy (after staining with Rose Bengal and Victoria blue B), phase contrast microscopy and differential-interference-contrast microscopy performed by two observers who analysed sets of 100 and 200 spermatozoa. The results obtained with phase-contrast microscopy dit not differ significantly from those obtained using interference contrast microscopy, performed by two observers on sets on 100 and 200 spermatozoa, concerning the following sperm abnormalities: abnormal detached heads, acrosome ruptures, tail abnormalities and total abnormalities. In view of the importance of extending the evaluation of sperm cytomorphology among artificial insemination centres, and the fact the phase-contrast microscopy system is less expensive than a differential-interference-contrast system and easier to operate, the authors recommend the use of phase-contrast microscopy for the routine study of sperm cytomorphology. Zusammenfassung: Nach Fixierung mit Goutaraldehyd (0,2% in PBS) wurden lichtmikroskopische Untersuchungen (Farbung mit Bengalrot und Victoriablau B) sowie phasenkontrastmikroskopische Untersuchungen (DH) und Differentialinterferenz-Kontrast-Mikroskopie-Untersuchungen (DIC) an Bullenspermatozoen vorgenommen, die tiefgerfroren waren. Die Studie wurde von zwei Untersuchern vorgenommen, die jeweils 100 und 200 Spermatozoen analysierten. Die Ergebnisse zeigten keine Differenzen bei DH und DIC im Hinblick auf abnormal losgeloste Kopfe, Akrosomrupturen, Schwanzabnormalitaten und Anomalien insgesamt. Unter Berucksichtigung der Bedeutung einer zunehmenden Notwendigkeit der Auswertung einer Spermatozoencyomorphologie gerade bei artifizieller Insemination verweisen die Autoren darauf, das wegen der hohen Kosten bei der DIC und der gleichwertigen PH eine Phasenkontrastmikroskopie absolut ausreichend sei fur die Routineuntersuchung in den genannten Fallen.

Il Bitto Storico è un formaggio d'alpeggio a latte crudo, prodotto nei mesi estivi sulle Prealpi Orobie, in alpeggi che praticano il pascolo turnato. La produzione di questo formaggio è strettamente legata ad attività tradizionali che valorizzano la biodiversità alpina delle zone di produzione. Il disciplinare di produzione prevede, infatti, l'utilizzo di Iatte crudo che deve essere lavorato a caldo entro 1 ora dal termine della mungitura e vieta di integrare l'alimentazione del bestiame con mangimi e insilati, e di utilizzare fermenti lattici durante il processo di caseificazione. Tali pratiche svolgono un ruolo basilare nella conservazione dell'ambiente e della biodiversità alpina garantendo una certa variabilità microbica e di conseguenza organolettica del formaggio, grazie alla flora batterica locale caratteristica di ogni produttore-alpeggio. Obiettivi del presente lavoro sono: 1) caratterizzare la variabilità microbica del formaggio Bitto Storico mediante tecnologia Next Generation Sequencing (NGS); 2) caratterizzare la frazione lipofila volatile (metaboloma volatile e composti terpenici) e non volatile (profilo acidico); 3) correlare quindi il microbiota alla frazione volatile del formaggio, e la componente terpenica e il profilo degli acidi grassi tenendo in considerazione il fattore produttore-alpeggio. Campioni di i formaggio sono stati prelevati da 54 forme di Bitto Storico prodotte in 6 diversi alpeggi in 3 zone geografiche lombarde (Val Gerola, Valli del Bitto, Valle Brembana), in tre periodi diversi (Giugno-Luglio-Agosto 2015). In ciascun periodo di lavorazione e per ciascun casaro sono state prelevate 3 repliche di formaggio nella stessa settimana di produzione. Per l'analisi metagenomica, il DNA batterico è stato estratto mediante un protocollo specifico ottimizzato in laboratorio ed utilizzato come templato per I'amplificazione del gene 16S rRNA (regione V3-V4 seguendo il protocollo standard 16S Metagenomic Sequencing Library Preparation e sequenziato su piattaforma Miseq (Illumina). La classificazione tassonomica dei microrganismi è stata effettuata utilizzando QllME (Caporaso et al., Nat Methods, 2010). La frazione lipofila volatile è stata caratterizzata mediante micro estrazione in fase solida-gascromatografia-spettrometria di massa (SPME-GC-MS) mentre il profilo in acidi grassi mediante gascromatografia ad alta risoluzione (HRGC). I risultati preliminari mostrano nei diversi campioni una comunità microbica dominata prevalentemente da Firmicutes con abbondanza di Streptococcaceae e Lactobacillaceae e una minor presenza di Enterococcaceae. Il metaboloma volatile mostra una variabilità coerente con una produzione casearia di montagna a latte crudo.

This paper presents a single case history of a drilled shaft constructed in the Atlantic Coastal Plain deposits for a bridge foundation that was subjected to axial loading. The predicted nominal axial capacity is estimated based on state of practice empirically derived methods specified in the current AASHTO LRFD Bridge Design Specifications. Predictions are compared to observed soil resistance derived from a static load test conducted on a full-size instrumented test shaft using the Osterberg Cell method. The results suggest that the AASHTO specified prediction methods should be applied cautiously for drilled shafts in the Atlantic Coastal Plain, incorporating an appropriate in situ testing program for evaluating soil design parameters, considering variations from the specific geologic environment and construction methodology used to develop the specified prediction methods, accounting for the load-deformation behavior of the shaft, and providing for instrumented static load testing to measure the actual behavior of the drilled shafts.

In the aim of studying possible relations between viruses detected in clinical specimens and the ones found in different environmental matrices, in the period May 2004 to April 2005, the collection of faecal samples from gastroenteritis cases and the monthly monitoring of raw and treated wastewater, river water, seawater and mussels were carried out. The viruses considered for environmental monitoring were adenovirus, rotavirus, enterovirus, norovirus, hepatitis A virus (HAV) and Torque teno virus (TTV): they were searched for with PCR and RT-PCR and confirmed by gene sequencing. Faecal coliforms and somatic coliphages' counts were also determined. The surveillance of case detected 45 positive faecal samples out of 255 (17.6%) while 35 of 56 environmental samples (62.5%) resulted positive for at least one of the considered viruses. The detection of the same viral strain in the faeces of gastroenteritis cases and in water was possible for adenovirus and rotavirus, which were also predominant in environmental matrices; thus they could be considered as a reference for risk assessment.

Background: Inflammatory breast cancer (IBC) is a rare form of rapidly progressive breast cancer. We reviewed the diagnosis, treatment, and outcome of IBC in our inner city community-based hospital and compared results with previous published reports. Study Design: Twenty-five patients were diagnosed and treated for IBC at the Catholic Medical Center of Brooklyn and Queens during the 6-year period of January 1989 through December 1995. Criteria for inclusion in this study were clinical or histopathologic evidence, or both, of inflammatory carcinoma. Results: IBC comprised 2.0% (25 of 1,257) of all breast cancer patients initially diagnosed during this study. All presented with clinical signs of IBC. Invasion of dermal lymphatics by neoplastic cells was demonstrated in 68% (17 of 25) of biopsy specimens. Sixty-eight percent (17 of 25) of patients presented with metastatic (ie, stage IV) disease and 28% (7 of 25) with stage IIIb; one patient (4%) died before staging. Estrogen and progesterone receptor studies were done on 72% (18 of 25) of all specimens. Of those patients who died, 85% were estrogen and progesterone receptor negative; of those surviving, 60% were estrogen receptor positive. Twenty (80%) of the 25 patients died, after a mean survival of 11.8 months and 5 (20%) remain alive, with a mean survival of 44.8 months. Of those who died, 85% were stage IV at presentation. All five survivors were stage IIIb at presentation. Patients underwent a variety of multimodal therapies. Survival was significantly associated with earlier stage at diagnosis and estrogen receptor positivity. Conclusions: IBC is characterized by rapid progression and dismal outcome. Earlier stage at diagnosis and positive estrogen receptor status suggest a more favorable prognosis. Neoadjuvant chemotherapy, as part of a multimodal approach, has significantly improved the outcome for IBC, but this is limited to patients with stage IIIb disease. Most of our patients presented with stage IV disease. If improvement is to be realized at the community level, limited health care resources must be directed toward aggressive physician and public education.

Kisspeptin-1 receptor (Kiss-1R) expression has been reported in the placenta and a possible involvement of the Kiss-1R/Kisspeptins (Kiss-1R/Kps) system in regulating trophoblast invasion and proliferation has been hypothesized. The aim of the present study was to investigate whether Kiss-1R activation by Kisspeptin 10 (Kp-10) could modulate in vitro proliferation and progesterone (P4) secretion of bovine primary placental cell lines isolated from cotyledons of foetuses in the first pregnancy trimester. The involvement of Kiss-1R in observed cell responses was also analyzed. Uteri from cows at the first trimester of pregnancy were obtained from local abattoirs. Fetal cotyledon fragments were digested with collagenase in Low Glucose Dulbecco's Modified Eagle's Medium (LG-DMEM) and cells were collected and cultured in a LG-DMEM based expansion medium in presence of 0.01, 0.1, 1 and 10 µM Kp-10. Control cells were cultured in the absence of Kp-10. The total number of living cells was obtained every 3 days for each culture passage (P) from P1 to P10. Cells were tested for Kiss-1R mRNA expression analysis by Real Time RT-PCR and culture media were analyzed for P4 concentration by radioimmunoassay. Kp-10 modulated in vitro proliferation and P4 secretory activity of epithelial cell lines isolated from cotyledons recovered from bovine fetuses in the first trimester of pregnancy. Markedly different effects, inhibitory or stimulatory, of Kp-10 on cell proliferation were found in different cell lines and observed cell responses were found as being related to Kiss-1R mRNA levels and P4 secretion. Inhibition of cell proliferation matched with not significant variation of Kiss-1R expression but increased P4 secretion, whereas stimulation of cell proliferation was found as being related to Kiss-1R up-regulation and decreased P4 levels. These results allow to hypothesize that the Kiss-1R/Kps system could play a role in the fine developmental regulation of the bovine placental cotyledonal cell proliferation and P4 secretion in the first trimester of pregnancy.

The interest in developing assisted reproductive technologies (ARTs) and cryobanking for farm animal genetic resources conservation is recently increased. However, cryopreservation for ex-situ management of genetic diversity sometimes is not routinely feasible, due to the lack of facilities (AI centres, laboratories) and expertises near the local breeds farming area. In these cases epididymal sperm obtained from slaughtered or castrated animals, associated to the possibility of managing rather long periods between animal death, sperm recovery and freezing, would increase the opportunities to create semen storages. This investigation addresses the pre-freeze/post-thaw quality of goat epididymal sperm as function of testicles storage temperature (environment or +5°C) and time elapsed between animal's death and sperm recovery (0, 24, 48, 72 h) in order to establish the optimal protocols for the recovery and cryopreservation of epididymal sperm in this species. Testicles of 50 mature bucks collected at the abattoir were divided in two groups: half of testicles (n = 50) was transported to the laboratory at environment temperature (E), while the remaining half (n = 50) at a refrigeration temperature (R) of +5°C. In the two groups (E) and (R) one testicle from each pair was processed after slaughter forming the time 0 groups (0E and 0R). The contra-lateral testicle was processed after 24, 48 or 72 h of storage, at the corresponding temperature. Sperm motility and kinetics parameters, viability and morphology were assessed in pre-freeze and post-thaw samples. Until 48 h post mortem, both E and R temperatures are able to maintain good pre-freeze epididymal sperm quality. After 48 h post mortem, R temperature is fundamental to reduce epididymal sperm quality decay in pre-freeze samples. Moreover testicles refrigeration has a positive impact also on post-thaw samples, allowing a lower decline through time considering total motility, kinetics parameters, sperm viability and sperm abnormalities. Therefore when sperm cryopreservation is not immediately practicable goat testicles should be transported and stored at 5°C up to a maximum of 48 h post mortem to ensure an acceptable sperm quality.

Camels represent part of the Arab heritage. The interest in developing assisted reproductive technologies and cryobanking for the conservation of animal genetic resources has recently increased. However, semen collections in camelids present many problems as sitting position during copulation, slow ejaculation and difficult animal handling. In these cases epididymal sperm from slaughtered or recently died animals will increase the opportunities to create semen storages. The present work was designed for assess motility of camel epididymal sperm extended in Ovixcell® and in Tris-fructose-egg yolk semen extender. Spermatozoa were extracted from 16 epididymides using the retrograde flushing technique, washing sperm cells in a retrograde direction from the ductus deferens through the cauda epididymidis with a syringe loaded with warmed (37°C) extender. Total motility was evaluated after 15 minute of incubation in a water bath at 37°C under phase-contrast microscopy using a pre-warmed (37°C) Makler Chamber. Total motility was similar in Ovixcell® or Tris-fructose-egg yolk semen extender (52.8 ± 0.7% vs 41.22 ± 33.56%, respectively). Further studies aiming to the test the fertilizing capacity should be carried out in order to confirm the optimal testicles storage condition for the creation of semen cryo storages in camels.

Recognizing cultural diversity among local breed farmers is crucial for the successful development and implementation of farm animal genetic resources FAnGr conservation policies and programmes. In this study based on survey data collected in the EUropean REgional CAttle breeds project from six European countries, a typology of local breed farmers was designed and profiles for each of the farmer types were developed to assist these policy needs. Three main farmer types were constructed: production-oriented, product and service-oriented and hobby-oriented farmers. In addition, seven subtypes were characterized under the main types: sustainable producers, opportunists, multi-users, brand makers, traditionalists, pragmatists and newcomers. These types have many similarities to the 'productivist', 'multifunctional' and 'post-productivist' farmer types. The typology not only reveals the high level of diversity among local cattle breed farmers in Europe, which presents an opportunity for the in situ conservation of animal genetic resources, but also a challenge for policy to meet the differing requirements of the farmer types.

Flow cytometry is a useful tool that provides an accurate, objective and rapid evaluation of semen quality. The use of this technique could significantly improve the quality of buffalo semen samples used in artificial insemination. This study was carried out to evaluate, by flow cytometry, frozen-thawed buffalo spermatozoa quality parameters such as sperm viability by SYBR-14 / propidium iodide staining; mitochondrial function by JC-1 potentiometric probe; sperm chromatin stability (SCSA) by acridine orange; and acrosome reaction (AR) by FITC-PNA staining. Semen samples from five Italian Mediterranean buffalo bulls were used. Sperm viability was not different between bulls and ranged from 33.4% to 43.6%. A consistent rate (55.1 ± 10.8%) of sperm cells showed high mitochondrial membrane potential (Dwhigh), with no significant differences between subjects. Sperm chromatin structure assay differed significantly between the five buffalo bulls; moreover, data showed high stability within each buffalo. DNA fragmentation indexes (DFI), such as %-DFI, X-DFI, SD-DFI, were 11.2 ± 8.6, 153.3 ± 24.6 and 81.6 ± 21.2, respectively. Regarding AR, the percentage of acrosome-reacted live (ARL) and acrosome-reacted dead (ARD) spermatozoa was 0.3 ± 0.2 and 15.3 ± 5.5, respec- tively. This functional parameter differed significantly between buffalo bulls and showed high stability. Following to Ca2+ ionophore A23187 for 3 h, AR significantly differed between subjects and was characterized by an increase in both ARL (10.8%) and ARD population (22.0%). This study indicates that flow cytometry could be a useful tool for a quick multiparametric evaluation of sperm quality in buffalo. In particular, SCSA and AR resulted in sperm functional parameters sensitive enough for the diagnosis of frozen-thawed semen fertilizing potential.

Single-use biopharmaceutical manufacturing requires monitoring of critical manufacturing parameters. However, the lack of reliable single-use sensors prevents the biopharmaceutical industry from fully embracing single-use biomanufacturing processes. We report an approach for temperature-independent pressure sensing in single-use bioprocess components using passive radio-frequency identification (RFID) sensors. An RFID pressure sensor is fabricated by applying a pressure sensitive flexible membrane to an RFID-tag-based transducer and a layer that modulates the electromagnetic field (EMF) generated in the RFID sensor antenna. The sensor signal is modulated upon pressure-induced flexing of the membrane, providing a desired quantitative response of pressure of the fluid during the operation of the single-use component. We demonstrate a temperature-independent RFID pressure sensor that was tested to measure pressures from −5 to 33 psi with the ± 0.25 psi accuracy after gamma irradiation. Temperature-independent pressure response is provided from the multivariate analysis of the measured impedance of the sensor.

Single-use biopharmaceutical manufacturing requires monitoring of critical manufacturing parameters. We have developed an approach for passive radio-frequency identification (RFID)-based sensing that converts ubiquitous passive 13.56 MHz RFID tags into inductively coupled sensors. We combine several measured parameters from the resonant sensor antenna with multivariate data analysis and deliver unique capability of multiparameter sensing and rejection of environmental interferences with a single sensor. We demonstrate here the integration of these RFID sensors into single-use biopharmaceutical manufacturing components. We have tested these sensors for over 500 h for measurements of temperature and solution conductivity with the accuracy of 0.1°C (32-48°C range) and accuracy of 0.3-2.9 mS/cm (0.5-230 mS/cm range). We further demonstrate simultaneous temperature and conductivity measurements with an individual RFID sensor with the accuracy of 0.2°C (5-60°C range) and accuracy of 0.9 mS/cm (0.5-183 mS/cm range). Developed RFID sensors provide several important features previously unavailable from other single-use sensing technologies such as the same sensor platform for measurements of physical, chemical, and biological parameters; multi-parameter monitoring with individual sensors; and simultaneous digital identification.

Single-use biopharmaceutical manufacturing requires monitoring of critical manufacturing parameters. However, the lack of reliable single-use sensors prevents the biopharmaceutical industry from fully embracing single-use biomanufacturing processes. We report here an approach for passive radio-frequency identification (RFID)-based sensing that does not rely on costly proprietary RFID memory chips with an analog input but rather implement ubiquitous passive 13.56 MHz RFID tags as inductively coupled sensors with 16-bit resolution provided by a sensor reader. Developed RFID sensors combine several measured parameters from the resonant sensor antenna with multivariate data analysis and deliver unique capability of multiparameter sensing and rejection of environmental interferences with a single sensor. In this study we are integrating these RFID sensors into single-use biopharmaceutical manufacturing components such as buffer bags. Performance of these sensors for simultaneous solution conductivity and temperature sensing is discussed.

The single-use bioprocessing is an attractive new approach of biopharmaceutical manufacturing. Digital identification of single-use bioprocess components is critical to facilitate their asset management, to document electronic pedigree, and to provide authentication. This identification can be achieved using radio frequency identification (RFID) tags. In this study, we critically analyze the challenges for the gamma-sterilizable RFID tag technology to retain the reliable read/write ability of the tags after their gamma irradiation. In RFID tags, the gamma radiation induced loss of device performance (i.e. the ability to reliably write and read data from an integrated circuit (IC) memory chip) originates from two independent sources such as (1) radiation effects on the non-charge-based storage memory material and (2) radiation effects on the performance of analog and digital circuit components of an IC memory chip device. Our interdisciplinary knowledge of product design, analytical instrumentation, RF engineering, and Six Sigma statistics has resulted in the design and implementation of the tag interrogation concept that insures the high reliability of tag read/write after the gamma irradiation.

Bullet embolism to the right heart from a distal vein is a rare event. Its diagnosis requires a high index of suspicion and use of various radiographic modalities. The many complications associated with a retained intracardiac foreign body argue strongly for its retrieval.

The routine use of exclusion angiography in trauma to the extremities has resulted in varying percentages of negative studies, mostly because of a great variability in location and type of injury. The authors reviewed 117 low-velocity gunshot wounds to the extremities. Twenty-six cases presented with hard signs of vascular injury and underwent immediate exploration. Ninety-one exclusion angiograms were performed (11 for soft signs and 80 for proximity only), 89 (98%) were considered negative. Two cases (2 % ) that presented without hard signs required vascular repair. Physical examination alone was 93 % sensitive and 99 % specific for recognition of vascular injuries. Routine angiography did not substantially improve the diagnostic accuracy and had a positive predictive value of only 2%. The authors conclude that angiography may be safely omitted in low-velocity gunshot wounds to the extremities if the absence of signs or symptoms of vascular injury can be firmly established.

Municipal and agricultural wastewater contain a variety of microorganisms and in particular enteric viruses. For the reuse of this treated wastewater it is important to ensure the efficiency of purification treatments and disinfection practices, that have often been insufficient to lower the viral load below the risk level. For this reason, for the routine analysis of recycled waters, the research into pathogenic viruses (e.g. HAV) and classical bacterial parameters (E. coli, enterococci and Salmonella) has to be associated with specific viral indicators such as somatic coliphages, adenovirus and TTV. The results of environmental monitoring, carried out in a wastewater treatment plant, showed the presence of adenovirus DNA in 100% of collected samples and TTV DNA in 95% (19/20) of raw sewage and in 85% (17/20) of the exit samples, while HAV was detected only in 2 samples over 40 (5%). The quantitative analysis has revealed an average reduction of 2 log for adenovirus and 1.58 log for TTV. The bacterial indicators were reduced by 1.74 log and 1.99 log respectively for E. coli and enterococci, while for somatic coliphages an average reduction of 2.2 log was observed. No significant correlation was shown between these parameters, confirming their inadequacy for the virological risk assessment. However the results of adenovirus confirm it as the best indicator to evaluate the efficacy of wastewater depuration plant in eliminating viruses.

Torque teno virus (TTV) is prevalent worldwide in general populations but at present is not related with any specific pathology. Its presence in faeces and its remarkable environmental stability suggest the possibility of using it as an indicator of faecal contamination in the environment. To evaluate the waterborne spread of TTV and its possible relationship with human pathogen enteric viruses, water samples were collected monthly for a year (May 2004–April 2005) from a river receiving the effluent of the treatment plant of the city of Pisa, concentrated and assayed with bimolecular tests (PCR, RT-PCR). TTV was detected in three samples (25%) while 16% of samples were positive for enteroviruses, 33% for rotaviruses, 8% for noroviruses genotype 1 and 25% for noroviruses genotype 2. Only two TTV samples (June and January) were also positive for rotavirus and norovirus, respectively. The detection of TTV in water confirmed its possible faecal-oral route of transmission but data are still insufficient to draw conclusions about the role of TTV as a viral indicator.

Background Colorectal carcinomas that present with perforation are stated in the literature to carry a poor prognosis. This study is to verify or refute the dismal connotation associated with perforated colorectal carcinomas (PCCs). Methods A retrospective analysis of 1551 patients with colorectal carcinoma revealed that 51 (3.3%) patients presented with perforation. Mary Immaculate and St. John's Queens Hospital Divisions of the Catholic Medical Center of Brooklyn and Queens chart and tumor board data were retrieved for the period 1983 through 1993. Results Localized perforation with abscess formation occurred in 31 (61%) patients, and free perforation with generalized peritonitis in occurred 20 (39%) patients. Sixteen (31%) patients had distant metastasis at diagnosis with a mean survival of only 6 months. Overall operative mortality rate was 12%, and overall 5-year survival rate was 32%. By excluding 16 patients with doc umented Stage IV disease at diagnosis and 6 operative mortalities (3 of whom also had Stage IV disease at diagnosis), the remaining 32 patients had a mean survival of 59 months and a 5-year survival of 58%. Conclusion In view of the 58% survival in our subset of patients, aggressive management is recommended. This includes management of sepsis and radical surgical resection of adjacent involved organs. A negative attitude associated with PCC is not substantiated in this retrospective 10-year study.

This article presents the production traits and performance (meat and carcass quality) of Cinta Senese * Large White progeny and purebred Nera Siciliana. The reproductive traits of five Italian local pig breeds (Calabrese, Casertana, Cinta Senese, Nera Sicilian and Mora Romagnola) were also presented.

BACKGROUND: The epidemiology of penetrating abdominal trauma is changing to reflect an increasing incidence of multiple injuries. Not only do multiple injuries increase the risk of infection, a very high risk of serious infection is conferred by immunosuppression from hemorrhage and transfusion and the high likelihood of intestinal injury, especially to the colon. Optimal timing and choice of presumptive antibiotic therapy has been established for penetrating trauma, but duration has not been studied extensively in such seriously injured patients. The purpose of this study was to test the hypothesis that 24 hours of antibiotic therapy remains sufficient to reduce the incidence of infection in penetrating abdominal trauma. METHODS: Three hundred fourteen consecutive patients with penetrating abdominal trauma were prospectively randomized into two groups: Group I received 24 hours of intravenous cefoxitin (1 g q6h) and group II received 5 days of intravenous cefoxitin. The development of a deep surgical site (intra-abdominal) infection as well as any type of nosocomial infection, as defined by the Centers for Disease Control and Prevention, (ie, surgical site infections, catheter-related infections, urinary tract, pneumonia), was recorded. Hospital length of stay was a secondary endpoint. Statistical analysis included chi-square tests for coordinate variables and two-tailed unpaired t tests for continuous variables. The independence of risk factors for the development of infection was assessed by multivariate analysis of variance. Significance was determined when P RESULTS: Three hundred patients were evaluable. There was no postoperative mortality, and no differences in overall length of hospitalization between groups. The duration of antibiotic treatment had no influence on the development of any infection ( P = 0.136) or an intraabdominal infection ( P = 0.336). Only colon injury was an independent predictor of the development of an intraabdominal infection ( P = 0.0031). However, the overall infection incidence was affected by preoperative shock ( P = 0.003), colon ( P = 0.0004), central nervous system (CNS) injuries ( P = 0.031), and the number of injured organs ( P = 0.026). Several factors, including intraoperative shock ( P = 0.021) and injuries to the colon ( P = 0.0008), CNS ( P = 0.0001), and chest ( P = 0.0006), were independent contributors to prolongation of the hospital stay. CONCLUSIONS: Twenty-four hours of presumptive intravenous cefoxitin versus 5 days of therapy made no difference in the prevention of postoperative infection or length of hospitalization. Infection was associated with shock on admission to the emergency department, the number of intra-abdominal organs injured, colon injury specifically, and injury to the central nervous system. Intra-abdominal infection was predicted only by colon injury. Prolonged hospitalization was associated with intraoperative shock and injuries to the chest, colon, or central nervous system.

Crack, the free-base form of cocaine, was introduced as an illicit street drug in 1986. Since then, we have noted a significant increase in acute gastroduodenal perforations. Between 1982 and 1986, we treated 11 patients with such perforations. This represents a constant occurrence rate of 6% of hospital admissions for peptic ulcer disease. Since 1986 we have treated 16 patients with gastroduodenal perforation, which yields an occurrence rate of 16%. Nine of the 16 patients had a close temporal relationship between the use of crack and the onset of their perforation. This group was younger and disproportionately comprised of male patients. These findings led us to believe that there may be a pathogenic relationship between the use of crack and acute gastroduodenal perforation, and the clinician should be aware of the various potential complications of this new drug. This relationship also raises questions about the exact pathophysiology of peptic ulcer disease.

Juxtahepatic inferior vena cava injuries are often lethal. Various operative strategies have been used to improve outcome, but the mortality rate reported in the literature is 80 percent or more. The atriocaval shunt has been advocated for isolation of bleeding retrohepatic vena cava, but recent reports suggest that mortality might be even higher in patients selected for shunting, perhaps owing to ongoing hemorrhage because of indecision and delay prior to insertion, or to technical difficulty with insertion. A series of patients with juxtahepatic inferior vena cava injuries treated successfully with total vascular isolation and occlusion were studied.Consecutive series of 10 patients with penetrating injuries to the juxtahepatic inferior vena cava were treated at an urban, university-affiliated Level I trauma center. A rapid and direct approach was used along with isolation techniques similar to those used in liver transplantation and elective resection for neoplasm. As resuscitation continued, repair of the inferior vena cava was accomplished in a bloodless field, created by manual compression of the liver, wide exposure, portal inflow occlusion, and proximal and distal control of the inferior vena cava. Aggressive fluid resuscitation and transient aortic cross-clamping controlled resulting systemic hypotension.Mean injury severity score was 26 and mean penetrating abdominal trauma index score was 28. After exposure, three patients had tangential injuries controlled by undersewing a partially occluding clamp. Subdiaphragmatic aortic cross-clamping was performed if total occlusion of the inferior vena cava reduced systolic blood pressure to 60 mm Hg, which was necessary in the remaining seven patients. Nine patients survived surgery, and seven of nine survived to hospital discharge. One postoperative death was a result of multiple organ dysfunction syndrome, and the other of necrotizing bacterial pneumonia.Total vascular occlusion with selective use of aortic cross-clamping yielded 70 percent survival in an injury that historically has been associated with survival of 20 percent or less. Minimization of visceral ischemia is accomplished by occluding the aorta only after complete isolation of the inferior vena cava.

Changes in the proportions of the various lipid components in spermatozoa were investigated throughout the reproductive period (24-72 wk of age) of male chickens. Sperm motility and in vivo fertility were also measured, and correlation coefficients with the lipid values were determined. The proportion of total phospholipid (PL) increased to reach a maximum value at 39 wk and decreased significantly thereafter. The relative content of free cholesterol and triacylglycerols showed no change in spermatozoa during aging or in relation to fertility values; free fatty acids and cholesterol esters increased continuously with age. Of the various PL classes, phosphatidylserine and phosphatidylcholine displayed a pattern of changes with age positively and negatively, respectively, in relation to the changes of fertility. The proportion of phosphatidylethanolamine had significantly decreased by the end of the reproductive period. The proportions of C16:0, C18:0, and C18:1n-9 within the PL of the spermatozoa increased with age, and those of C20:4n-6, C22:4n-6, and C22:6n-3 decreased. Positive correlations were found between fertility and total PLs, phosphatidylserine, and PL-bound C20:4n-6 and C22:4n-6; a negative correlation was found between fertility and phosphatidylcholine. Motility was positively correlated with the level of PL and negatively with that of free cholesterol; it was also positively correlated with the levels of C22:4n-6 and C22:6n-3 and negatively with those of C16:0, C18:0, and C18:1n-9. The results suggest that the lipid and fatty acid compositions of spermatozoa may be important predictors of fertility.

The lack of reliable single-use sensors prevents the biopharmaceutical industry from fully embracing single-use biomanufacturing processes. Sensors based on the same detection platform for all critical parameters in single-use bioprocess components would be highly desirable to significantly simplify their installation, calibration and operation. We review here our approach for passive radio-frequency identification (RFID)-based sensing that does not rely on costly proprietary RFID memory chips with an analog input but rather implements ubiquitous passive 13.56 MHz RFID tags as inductively coupled sensors with at least 16 bit resolution provided by a sensor reader. The developed RFID sensors combine several measured parameters from the resonant sensor antenna with multivariate data analysis and deliver unique capability of multiparameter sensing and rejection of environmental interferences with a single sensor. This general sensing approach provides an elegant solution for both analytical measurement and identification and documentation of the measured location.

A rare anatomical variation was encountered during a laparoscopic cholecystectomy. The right hepatic duct emptied into the infundibulum of the gallbladder. This confluence then joined the left hepatic duct to form the common bile duct. The right hepatic duct was transected between the gallbladder and the common bile duct as a normal cystic duct would have been isolated and divided laparoscopicaly. This anatomic variant was recognized after further mobilization of the gallbladder from the liver bed. A laparotomy was performed to create a right hepatic duct enteric anastomosis. The case illustrates one possible pitfall that may be encountered during laparoscopic cholecystectomy.

Gallbladder agenesis is an extremely rare disease. Necropsy incidence has been reported to be 0.016%. Failure to locate the gallbladder at the time of a planned cholecystectomy can be very challenging. We report such a case during a laparoscopic cholecystectomy. The indication for surgery in these patients are complaints of gallbladder symptoms along with a false-positive ultrasound study. During exploration, an abnormal location of the gallbladder has to be excluded. Ectopic gallbladder locations include intrahepatic, lesser omentum, retroperitoneal, retrohepatic, within the falciform ligament, retroduodenal, and retrohepatic areas. Thorough exploration and cholangiography are essential. Embryologically, the gallbladder and cystic duct arise from the caudal portion of the hepatic bud. All of the previously reported cases of gallbladder agenesis have shown an absence of both the gallbladder and cystic duct. We report an embryological oddity wherein a patent cystic duct was found along with an agenetic gallbladder. This is the first case report of this finding, along with this being the first absent gallbladder discovered laparoscopically.

The smuggling of illicit drugs by concealing them within the human body has become a widespread practice. Those individuals who transport packaged drugs are commonly known as "body packers" or "mules". "Body stuffers," on the other hand, are individuals who emergently place the contraband in a body orifice when they sense apprehension is imminent. In the latter instance, the drugs are not well packaged for transportation by human "consumption," hence the high risk for leakage. These individuals require prompt surgical attention under two circumstances: when they are found to suffer from drug overdosage caused by inadvertent leakage or when obstruction in the body is caused by the drug-laden bags. Two such cases are reported. The first patient presented with acute drug overdose and required an emergency laparotomy. The second patient presented with pyloric obstruction and was treated by endoscopic removal of the bag. One must be aware that these patients are walking time bombs, carrying drugs that may be well packed but have the potential to deliver a lethal dose without warning. Knowledge of the type of drug and type of packaging are essential in managing these patients. The overall plan should be close observation, careful monitoring, conservative therapy, and expectant rapid surgical intervention as needed.

A technique for electrocardiographic (ECG) guided percutaneous placement of central venous catheters (CVC) was studied in a prospective, randomized manner. In 34 patients, 51 ECG guided percutaneous CVC were compared with 29 blind percutaneous CVC in 23 patients. Thirty-nine percent of CVC placements were changes over a guide wire. Ideal catheter tip location at the atriocaval junction was achieved in 96 percent of the patients in the study versus 59 percent of those in the control group (p less than 0.001). In addition, we report 25 patients with open placement of CVC using intraoperative ECG guidance and fluoroscopic confirmation. Ideal location of the catheter tip was achieved in 100 percent of these patients. ECG guided CVC placement using the technique described herein obviates the need for catheter repositioning, repeat roentgenographic studies and intraoperative fluoroscopic imaging, along with the attendant costs and radiation exposure to staff and patient. Aberrant catheter tip placement and the associated morbidity are also eliminated.

We report a gastric duplication in a 23-year-old man. The case is unique because the stomach is the rarest location for enteric duplication, the duplication almost never occurs in adults, and the duplication consisted exclusively of pancreatic tissue. This extreme example illustrates the close embryologic and anatomic association between stomach and pancreas. A case is made for the use of the general term of foregut duplication to avoid confusing nomenclature.