Biomedical subjects

Federica Morandi

Publications and source records attributed to Federica Morandi.

14 recordsLinked to original sources

PubMed2006

Otoscopic, cytological, and microbiological examination of the equine external ear canal.

Otoscopic examination and cytology of the equine ear would be beneficial in diseases such as head trauma, headshaking, otitis externa secondary to otitis media, vestibular disease, aural neoplasia and aural pruritus secondary to parasites. In practice, otic examinations of horses are rarely done due to the perceived difficulty in visualizing the equine external ear canal and tympanic membrane, as well as the need for chemical restraint. In this study, the proximal external ear canal was examined in live horses using a handheld otoscope and in cadaver heads using video otoscopy. Visualization of the proximal ear canal of the sedated horse could be done with a handheld otoscope, but more sedation or general anaesthesia and a video otoscope would be required to adequately visualize the tympanic membrane in the live horse. The proximal ear canals of 18 horses were examined cytologically and cultured aerobically. In three horses, both ears were sampled. No cells or organisms were seen on cytological examination of 11/21 ears. Nine of the 21 ears were sterile when cultured. Ten of the 21 ears had mixed growth with low numbers of organisms (Corynebacterium sp. being most common). Two of the 21 ears had heavy growth of a single organism (Corynebacterium sp. and Staphylococcus intermedius, respectively). Equine cadaver heads were examined in cross-section by computed tomography (CT) imaging and histopathology in order to further understand the anatomy of the equine external ear canal. Equine practitioners should be aware that otic examination is possible and may provide important diagnostic information.

Animals↗

PubMed2006

Segmental tracheal dysplasia in a mixed breed dog.

A 6-month-old, spayed female, mixed breed dog was referred for respiratory difficulty. Radiographic and bronchoscopic evaluations showed tracheal narrowing from the thoracic inlet to its bifurcation. Under anesthesia, the trachea ruptured and the dog died. Microscopic evaluation revealed underlying segmental tracheal and bronchial cartilaginous malformation (dysplasia).

Animals↗

PubMed2005

Structure-based optimization of a non-beta-lactam lead results in inhibitors that do not up-regulate beta-lactamase expression in cell culture.

Bacterial expression of beta-lactamases is the most widespread resistance mechanism to beta-lactam antibiotics, such as penicillins and cephalosporins. There is a pressing need for novel, non-beta-lactam inhibitors of these enzymes. One previously discovered novel inhibitor of the beta-lactamase AmpC, compound 1, has several favorable properties: it is chemically dissimilar to beta-lactams and is a noncovalent, competitive inhibitor of the enzyme. However, at 26 microM its activity is modest. Using the X-ray structure of the AmpC/1 complex as a template, 14 analogues were designed and synthesized. The most active of these, compound 10, had a K(i) of 1 microM, 26-fold better than the lead. To understand the origins of this improved activity, the structures of AmpC in complex with compound 10 and an analogue, compound 11, were determined by X-ray crystallography to 1.97 and 1.96 A, respectively. Compound 10 was active in cell culture, reversing resistance to the third generation cephalosporin ceftazidime in bacterial pathogens expressing AmpC. In contrast to beta-lactam-based inhibitors clavulanate and cefoxitin, compound 10 did not up-regulate beta-lactamase expression in cell culture but simply inhibited the enzyme expressed by the resistant bacteria. Its escape from this resistance mechanism derives from its dissimilarity to beta-lactam antibiotics.

Bacterial Proteins↗

PubMed2004

Correlation of helical and incremental high-resolution thin-section computed tomographic and histomorphometric quantitative evaluation of an acute inflammatory response of lungs in dogs.

OBJECTIVE: To compare quantitative densitometric computed tomography (CT), morphometric, and histologic data of normal lungs in dogs with similar parameters obtained after induction of an acute inflammatory response and determine whether CT densitometry correlated with histopathologic changes. ANIMALS: 6 healthy adult dogs. PROCEDURE: After initial CT, 1 mL of 0.1 M hydrochloric acid (HCI) and 3 mL of autologous blood were instilled into the right middle (RM) and caudal segment of the left cranial (LCCd) lung lobes, respectively. Immediately and 24 hours after instillation, CT was repeated. At 24 hours, dogs were euthanatized and lungs were fixed and sampled for morphometric and histologic evaluation. The CT data were compared with lung morphology and morphometry by use of unpaired t tests. Comparison with lungs from control dogs was performed using Spearman rank correlation coefficients. RESULT: Mean Hounsfield units (HU) from control and baseline HU from experimental dogs were identical. Immediately after instillation of HCI or blood, there was increased attenuation in both lobes. Autologous blood initially induced severe changes that almost completely resolved at 24 hours; HCI induced severe changes at 24 hours. Significant increases in percentage of parenchymal airspace and alveolar diameter resulted in decreased surface area-to-volume ratio in lobes receiving HCl. Histologic scores were significantly higher in the RM lobe, compared with controls. CONCLUSIONS AND CLINICAL RELEVANCE: Computed tomography attenuation correlated well with histomorphometry and histologic findings in this model. Lung lesions after autologous blood were transient and of limited severity. Lesions induced by HCl were severe; alterations in morphometric and histologic parameters were reflected in CT attenuation measurements.

Acute-Phase Reaction↗

PubMed2003

Nanomolar inhibitors of AmpC beta-lactamase.

beta-lactamases are the most widespread resistance mechanism to beta-lactam antibiotics, such as the penicillins and the cephalosporins. In an effort to combat these enzymes, a combination of stereoselective organic synthesis, enzymology, microbiology, and X-ray crystallography was used to design and evaluate new carboxyphenyl-glycylboronic acid transition-state analogue inhibitors of the class C beta-lactamase AmpC. The new compounds improve inhibition by over 2 orders of magnitude compared to analogous glycylboronic acids, with K(i) values as low as 1 nM. On the basis of the differential binding of different analogues, the introduced carboxylate alone contributes about 2.1 kcal/mol in affinity. This carboxylate corresponds to the ubiquitous C3(4)' carboxylate of beta-lactams, and this energy represents the first thermodynamic measurement of the importance of this group in molecular recognition by class C beta-lactamases. The structures of AmpC in complex with two of these inhibitors were determined by X-ray crystallography at 1.72 and 1.83 A resolution. These structures suggest a structural basis for the high affinity of the new compounds and provide templates for further design. The highest affinity inhibitor was 5 orders of magnitude more selective for AmpC than for characteristic serine proteases, such as chymotrypsin. This inhibitor reversed the resistance of clinical pathogens to the third generation cephalosporin ceftazidime; it may serve as a lead compound for drug discovery to combat bacterial resistance to beta-lactam antibiotics.

Anti-Bacterial Agents↗

PubMed2003

Correlation of helical and incremental high-resolution thin-section computed tomographic imaging with histomorphometric quantitative evaluation of lungs in dogs.

OBJECTIVE: To develop protocols for helical computed tomography (CT) and axial high-resolution CT (HRCT) of lungs and correlate densitometric CT values with morphometric and histologic data for normal pulmonary tissue in dogs. ANIMALS: 8 healthy adult dogs. PROCEDURE: 2 dogs were used to establish a protocol for helical CT and HRCT of lungs. Six dogs were used to acquire densitometric CT data regarding normal lungs. After the dogs were euthanatized, their lungs were fixed and sampled for morphometric and histologic evaluation. Four CT acquisitions were compared by means of paired t tests. RESULTS: For normal lung tissue of dogs, mean densitometric CT value obtained during helical CT scans reconstructed in a sharp algorithm was -846 Hounsfield units. Values obtained via helical CT or HRCT acquisitions and reconstructed with sharp or standard algorithms did not differ significantly. Morphometric analysis was used to determine the proportion of lung parenchymal (82%) and non-parenchymal tissue (18%). Alveolar size, estimated by mean linear intercept, was approximately 172 microm, and alveolar surface area-to-volume ratio was 0.024 to 0.026 microm(-1). Histologic evaluation confirmed the presence of normal lung tissue. CONCLUSIONS AND CLINICAL RELEVANCE: Correlation of densitometric CT data with morphometric and histologic findings and the establishment of helical CT and HRCT protocols were attained; clinical use of this information may facilitate investigation of pulmonary disease in dogs. Sharp helical CT acquisitions were preferred because of better lung parenchyma detail and rapid image acquisitions, compared with HRCT.

Animals↗

PubMed2002

Structure-based discovery of a novel, noncovalent inhibitor of AmpC beta-lactamase.

beta-lactamases are the most widespread resistance mechanisms to beta-lactam antibiotics, and there is a pressing need for novel, non-beta-lactam drugs. A database of over 200,000 compounds was docked to the active site of AmpC beta-lactamase to identify potential inhibitors. Fifty-six compounds were tested, and three had K(i) values of 650 microM or better. The best of these, 3-[(4-chloroanilino)sulfonyl]thiophene-2-carboxylic acid, was a competitive noncovalent inhibitor (K(i) = 26 microM), which also reversed resistance to beta-lactams in bacteria expressing AmpC. The structure of AmpC in complex with this compound was determined by X-ray crystallography to 1.94 A and reveals that the inhibitor interacts with key active-site residues in sites targeted in the docking calculation. Indeed, the experimentally determined conformation of the inhibitor closely resembles the prediction. The structure of the enzyme-inhibitor complex presents an opportunity to improve binding affinity in a novel series of inhibitors discovered by structure-based methods.

Ampicillin↗

PubMed2002

Using steric hindrance to design new inhibitors of class C beta-lactamases.

beta-lactamases confer resistance to beta-lactam antibiotics such as penicillins and cephalosporins. However, beta-lactams that form an acyl-intermediate with the enzyme but subsequently are hindered from forming a catalytically competent conformation seem to be inhibitors of beta-lactamases. This inhibition may be imparted by specific groups on the ubiquitous R(1) side chain of beta-lactams, such as the 2-amino-4-thiazolyl methoxyimino (ATMO) group common among third-generation cephalosporins. Using steric hindrance of deacylation as a design guide, penicillin and carbacephem substrates were converted into effective beta-lactamase inhibitors and antiresistance antibiotics. To investigate the structural bases of inhibition, the crystal structures of the acyl-adducts of the penicillin substrate amoxicillin and the new analogous inhibitor ATMO-penicillin were determined. ATMO-penicillin binds in a catalytically incompetent conformation resembling that adopted by third-generation cephalosporins, demonstrating the transferability of such sterically hindered groups in inhibitor design.

Acylation↗

PubMedDate not supplied

Evaluation of the feline pancreas using computed tomography and radiolabeled leukocytes.

This study was designed to test the feasibility and utility of computed tomography and radiolabeled granulocytes in evaluating the feline pancreas in six normal cats. Autologous granulocytes were labeled with 99mTc-hexamethylpropyleneamine oxime (HMPAO) and injected into each cat. Whole body scintigraphic images were acquired at 1, 5, 15, and 30 minutes, and 1, 2, and 4 hours following injection. The following day, each cat was anesthetized and computed tomographic images of the abdomen were acquired both pre- and post-contrast. Following CT, a surgical pancreatic biopsy was collected. Feline granulocytes were successfully labeled with 99mTc-HMPAO with a labeling efficiency of 15-42% (average of 27%). An average of 5.42 x 10(7) cells in a 2 mL volume were injected into each cat. Less than 1 minute was required to acquire 500,000 count images. Granulocytes distributed predominantly to the lung, spleen and liver in order of decreasing activity. Only background activity was identified in the region of the pancreas. The pancreas was easily identified on CT images of the abdomen. The pancreas was hypoattenuating relative to both the spleen and liver. The pancreas enhanced with the administration of contrast medium, peaking immediately, then gradually clearing over the 30-minute test period. Following contrast medium administration the pancreas remained hypoattenuating relative to the spleen. All biopsies confirmed the absence of pancreatic inflammation in the study cats and no adverse effects were recognized as a result of pancreatic biopsy. Both computed tomography and radiolabeled granulocytes appear to hold promise as imaging procedures for the detection of feline pancreatitis. We predict that these described normal parameters may be altered in the face of inflammation, facilitating detection of feline pancreatitis. Data from cases of suspect feline pancreatitis are needed to evaluate these methods for clinical utility.

Animals↗

PubMedDate not supplied

Combined use of ultrasonography and contrast enhanced computed tomography to evaluate acute necrotizing pancreatitis in two dogs.

The imaging findings in two miniature schnauzers with acute necrotizing pancreatitis are described. Both dogs were treated previously for diabetes mellitus and hyperlipidemia. Vomiting, anorexia, and lethargy were observed in both dogs at presentation. Laboratory evaluations supportive of pancreatitis included left shift, abnormally high serum amylase and lipase activities, hypocalcemia, and abnormally high serum activities of liver enzymes. Sonographically, both dogs had diffusely enlarged hypoechoic pancreatic tissue with anechoic foci compatible with necrosis, abscessation, phlegmon, and pseudocysts formation. Contrast-enhanced computed tomography (CT) findings in both dogs were compatible with pancreatic necrosis. Dog 1 was managed medically for 11 days. Follow-up CT scan in this dog disclosed decreased pancreatic size and increased contrast enhancement compatible with partial resolution of pancreatitis.

Animals↗

PubMedDate not supplied

Diagnosis of congenital cardiac right-to-left shunts with 99mTc-macroaggregated albumin.

Diagnosing right-to-left congenital cardiac shunts can be difficult. Cardiac catheterization and angiocardiography represent the traditional gold standard for diagnosis, but they are invasive. Nuclear scintigraphy using 99mTc-macroaggregated albumin (MAA) has been employed in humans as an alternate method of diagnosis. This study reviews eight dogs presented for evaluation of a suspect right-to-left cardiac shunt that were examined using 99mTc-MAA. In all, 2-4 mCi (74-148 MBq) of reduced particle 99mTc-MAA were injected IV in a cephalic vein and static images of the whole body, including right and left lateral, dorsal, and ventral views, were acquired for 60 s and stored into a 256 x 256 x 16 matrix. Shunt fractions were calculated. One dog with radiopharmaceutical distribution limited to the lungs did not have a shunt. Seven dogs had distribution of the radiopharmaceutical outside the pulmonary capillary bed, indicating bypassing of the pulmonary capillary circulation due to a right-to-left shunt. Four dogs had 99mTc-MAA within the brain. Three dogs that did not have brain uptake, but instead had a sharp cutoff of radioactivity at the level of the front limbs and neck, were diagnosed with reverse patent ductus arteriosus (PDA). The asymmetric distribution of the radiopharmaceutical is due to the location of the shunt, distal to the brachiocephalic trunk and left subclavian artery. Shunt fractions of dogs with extrapulmonary radioactivity ranged from 40% to 59%. Nuclear scintigraphy with 99mTc-MAA is a quick alternative method of diagnosing right-to-left cardiac shunts that permits quantification of shunt fraction. Distinguishing between reverse PDA and other right-to-left shunts may be possible based on the radiopharmaceutical distribution.

Animals↗

PubMedDate not supplied

Trans-splenic portal scintigraphy in normal dogs.

The purpose of this study was to (1) establish a technique for ultrasound-guided trans-splenic portal scintigraphy (TSPS) using 99mTcO4(-), (2) evaluate portal vein morphology, (3) compare the radiation exposures for TSPS vs. per-rectal portal scintigraphy (PRPS), and (4) compare the quality of numerical data from the TSPS vs. PRPS. Eight juvenile dogs underwent PRPS and TSPS (minimum of 48h between studies) after initial screening tests. PRPS was done according to established protocol using 425 +/- 36MBq (mean +/- SD) of 99mTcO4(-). TSPS was done with the dogs in right lateral recumbency over the gamma camera. 99mTcO4(-) (57 +/- 13.9 MBq) was injected into the spleen 1-2s following initiation of the dynamic acquisition. The frame rate was 4 frames/s for 5 min. There was significantly lower radioactivity of 99mTcO4(-) given and significantly higher total counts recorded in the liver and heart during the TSPS compared with PRPS. The total counts for the TSPS and PRPS were 7120 +/- 4386 and 830 +/- 523, respectively. Percent absorption from the spleen was 52.5 +/- 19.1% compared with 9.2 +/- 5.7% for the colon. Calculated transit time for the TSPS studies was 7 +/- 2.3s. In TSPS studies, the splenic and portal veins were clearly identified. Radiation exposure levels of the dogs were significantly lower following TSPS than after PRPS. TSPS appears superior to PRPS as a method to image the portal venous system representing a valid alternative diagnostic test for animals with suspected portosystemic shunts.

Animals↗

PubMedDate not supplied

Use of 99mTCO4(-) trans-splenic portal scintigraphy for diagnosis of portosystemic shunts in 28 dogs.

Ultrasound-guided percutaneous trans-splenic portal scintigraphy (TSPS) using 99mTcO4(-) has been used to image the portal venous system in normal dogs. Compared with per-rectal portal scintigraphy, it provides higher count density, consistent nuclear venograms of the splenic and portal vein, and significantly decreased radiation exposures. This paper describes the use of TSPS for the diagnosis of portosystemic shunts in 28 dogs. TSPS was performed injecting 70 +/- 28 MBq of 99mTcO4(-) (mean +/- SD) into the splenic parenchyma with ultrasound guidance. A dynamic acquisition at a frame rate of four frames/s for 5 min was initiated after placement of the needle and approximately 2s prior to injection. All dogs had diagnoses confirmed via exploratory laparotomy or ultrasonographic identification of the shunting vessel(s). Three studies (10.7%) were nondiagnostic because of intraperitoneal rather than intrasplenic injection of the radionuclide. Three pathways were recognized on the scintigraphic images: (1) portoazygos shunts--the 99mTcO4(-) bolus traveled dorsally, running parallel to the spine and entering the heart craniodorsally; (2) single portocaval or splenocaval shunts--the 99mTcO4(-) bolus ran from the area of the portal vein/splenic vein junction in a linear fashion toward the caudal vena cava entering the heart caudally; (3) internal thoracic shunt-the 99mTcO4 bolus traveled ventrally along the thorax and abdomen entering the cranial aspect of the heart. Single and multiple shunts were easily distinguished. There were no distinguishing features between single intra and extrahepatic portocaval shunts.

Animals↗

PubMedDate not supplied

Quantitative assessment of hepatic function by means of 99mTc-mebrofenin in healthy horses.

99mTc-mebrofenin is used in humans and small animals to assess hepatic function. This study was undertaken to measure hepatic clearance of 99mTc-mebrofenin in healthy horses and to determine whether feed deprivation and increased serum total bilirubin (TBIL) concentration alter 99mTc-mebrofenin clearance. Plasma clearance of 99mTc-mebrofenin was determirned in 7 healthy horses at 0, 48, and 96 hours of feed withholding. Serum TBIL and nonesterified fatty acid (NEFA) concentrations were measured every 24 hours. 99mTc-mebrofenin (4.16 +/- 0.62 mCi, mean +/- SD) was injected into a jugular vein, and blood samples were retrieved from the contralateral jugular vein. A plasma time-activity curve of 99mTc-mebrofenin was generated, from which the area under the curve (AUC) and the T1/2 of the fast-phase (T1/2) and slow-phase (T1/2f) were calculated. Mean +/- SD AUC was 17,700 +/- 4,257, 18,616 +/- 8,078, and 16,168 +/- 6,031 counts per minute (cpm) at 0, 48, and 96 hours, respectively; mean +/- SD T1/2f was 2.80 +/- 0.38 minutes, 3.52 +/- 1.46 minutes, and 3.82 +/- 1.29 minutes at 0, 48, and 96 hours, respectively; median T1/2s was 63.9, 49.2, and 45.8 minutes at 0, 48, and 96 hours, respectively. No difference was detected between the values of AUC, T1/2f, and T1/2s at 0, 48, and 96 hours. There was a significant increase in TBIL with fasting, with a mean +/- SD of 6.3 +/- 1.3 mg/dL at 26 hours. NEFAs increased, reaching a plateau at 48 hours (650 +/- 152 micromol/L). Plasma TBIL concentrations did not correlate with AUC or T1/2s but correlated weakly with T1/2f (r = 0.50). Plasma NEFA concentrations did not correlate with AUC, T1/2s, or T1/2f values. This study suggests that 99mTc-mebrofenin plasma clearance is not affected by feed withholding and that hyperbilirubinemia associated with feed withholding does not affect the hepatic extraction efficiency of this radiopharmaceutical.

Aniline Compounds↗