hydrophilic proteins of lung surfactant as a prognostic marker in experimental pneumonia

Background: SP-A and SP-D are hydrophilic proteins which regulate the inflammatory response of the lung. Pasteurella multocida is one of the most common bacteria isolated from calves suffering from shipping fever pneumonia, one of the majorproblems in dairy herds

Objectives: evaluation of surfactant content may provide a valuable diagnostic tool for detection of calf pneumonia due to Pasteurella multocida and also state of treatment

Methods: ten Holstein-Frisian bull calves aged 4 months with body weight of 120 +/- 5 kg were selected for study in two groups. The Pasteurella multocida [PMC66 Razi] was used in the present study for inducing pneumonia. The Bronchoalveolar lavage [BAL] process was done in selected calves. BAL fluid was collected and centrifuged and finally the sediment [crude surfactant] was reserved at -20degreeC. The cytological evaluation and surfactant content was assayed by ELISA, TPL kit assay and HPLC

Results: the serum levels of SP-A and SP-D in pneumonic group were significantly elevated. Although the increased Bronchoalveolar lavage fluid [BALF] level of SP-A in pneumonic cases was found as compared with the control animals, the statistical analysis did not show any significant differences between two groups. The level of SP-D in BALF of pneumonic group significantly elevated. The amount of Dipalmitoylphosphatidylcholine [DPPC] in pneumonic group decreased significantly in comparison with control group

Conclusions: pasteurella inducing pulmonary can change the major component of lung surfactant, evaluation of these markers can be helpful as an appropriate tool in diagnostic state of pneumonia and healing

Changes in clinical signs after treatment in calves with experimental colisepticemia with Escherichia coli

Background: Colisepticemia is an acute fatal disease in farm animal neonates. Clinical finding of septicemia is non-specific and cannot be differentiated from signs of non-infectious disease or disease with local infection such as diarrhea

Objectives: Evaluation of clinical signs variations in calves with experimental septicemia with Escherichia coli 01 11 :H8

Methods: Colisepticemia was experimentally induced in ten Holstein bull calves after an adaptation period. Vital signs and 7 clinical criteria were recorded from 24 h before septicemia until 48 h after that. Blood culture was performed and treatment was done based on antibiogram from 24 h after challenge

Results: Changes of suckling reflex and shock were not significant. Changes of appetite, dehydration, behavior, standing ability, total score from 24 h before the challenge to 24 h after treatment were significant [p=0.00l]. Fecal consistency altered with treatment [p<0.04]. Heart rate [p=0.04 and p=0.033, respectively], respiratory rate [p=0.009 and p=0.001, respectively] and body temperature [p0.00l and p=0.004, respectively] have significant changes till 24 h after challenge and till 24 h after starting treatment. Blood cultures were positive except for 0 h and 48 h after challenge

Conclusions: The present study indicated clinical signs changed unfavorably following septicemia that were dissolved approximately during 24 h, depending on treatment in appropriate time and drug choice. Thus, a targeted scoring system will be useful in clinical evaluation of septicemia, quantifying the changes procedure and treatment efficacy

Effects of different culture media on optimization of primary neuronal cell culture for in vitro models assay

BACKGROUND: In vitro model studies are becoming increasingly popular for experimental research designs. They include isolation and expansion of cells of a particular tissue, such as the nervous tissue which contributes to understanding the underlying mechanisms in many pathologies. It enables the scrutinization of intracellular signaling pathways responsible for cell death.

OBJECTIVES: In the literature, there are different methods for the isolation and culture of rat embryonic cortical neurons. However, this study developed a feasible, rapid and easily performable method.

METHODS: Isolation of neurons was performed without using enzymatic digestion. Primary cortical cultures neurite outgrowth and neuron numbers per field of common mediums were compared for neuronal cells isolation and expansion. In this study, three different culture mediums were considered: Medium I: Neurobasal medium, B-27 and L-glutamine; Medium II: DMEM, FBS and L-glutamine; and Medium III: DMEM/F-12, FBS and L-glutamine.

RESULTS: High survival rate and number of neurons was obtained with the current method. The best neuronal growth was achieved by Medium I, while Medium II and III had moderate effect on the neurite outgrowth.

CONCLUSIONS: Enzyme-free treatment was introduced and Medium I was used as an alternative method for optimal neuron isolation and expansion. The neuronal cultures are similar to nervous tissue in physiological aspects. Hence, Medium I is more similar to the in vivo condition compared to Mediums II and III.

effect of butyric acid with autogenous omental graft on healing of experimental achilles tendon injury in rabbits

In this study, the role of local injection of butyric acid [BA] with autogenous omental graft was evaluated in healing of experimental Achilles tendon injury in rabbits. Nine adult male New Zealand rabbits were anesthetized and a partial thickness tenotomy was created on both hindlimbs. In treated group, omental graft was secured in place using BA soaked polygalactin 910 suture. In control group, the graft was sutured without BA. Butyric acid and normal saline were injected daily to treatment and control groups for three days, respectively. Based on the findings, on day 15 after injury, the tendon sections showed that healing rate in BA treated group was higher than that in control group. Furthermore, at days 28 and 45, comparison between BA treated and control groups demonstrated that BA increased the healing rate but with no significance. In summary, results of this study show that application of BA with autogenous omental graft can improve healing process of damaged Achilles tendon

[Comparison of diagnostic accuracy of ultrasonography and radiology in radial critical-sized defects healing process in rabbit]

Healing of Critical-Sized Defects [CSDs] is one of the major challenges facing orthopedic surgeons. To assess the bone healing process usually plain radiography is used. Serial radiography results in certain side effects and recent findings are indicating the early detection of bone healing via ultrasonography. The purpose of current study is to compare the diagnostic accuracy of radiography and ultrasonography in healing process of radial CSDs in rabbit. Sixteen New Zealand White Rabbits were used in this study. The radial CSDs of 15 mm size were created in a routine surgical procedure described previously and the two diagnostic tools were compared 12 weeks post-surgery. In case of obtaining different results from radiology and ultrasonography, to determine which diagnostic imaging method is of more accuracy, the histopathologic results of samples from the same day were used. In 15 cases of 16, ultrasonography findings were confirmed by radiography and only in one case they were in contradiction, in which radiographs showed an Atrophic Nonunion while Ultrasound detected an outstanding filled defect with a reverberation artifact underneath. In histopathology, the defect was filled with an osteoid matrix of high cellularity and calcification was obvious in some regions, confirming the ultrasound results. This study indicates that in diagnostic imaging of CSDs, the ultrasonography and radiography are usually consonant and even ultrasound is more accurate than radiology. In addition, in case of detection of a reverberation artifact and lack of any healing-related finding in radiography, this artifact may be an indication of osteoid matrix formation

Effects of mesenchymal stem cells with injectable scaffold on cardiac function in myocardial infarction in Rabbit

Bone marrow-derived mesenchymal cells can transdifferentiate into Cardiomyocyte cells and improve heart function after transplantation. Since biomaterials can improve the cell retention in the site, cell survival and differentiation, heart tissue engineering is now being explored as an applied solution to support cell-based therapies and increase their efficacy for myocardial diseases. Chitosan in combination with Glycerol Phosphate [GP] can produce a thermo sensitive material that in body temperature can form a jellylike material. The aim of this study was to evaluate the effects of a combination of autologous undifferentiated bone marrow mesenchymal stem cells [MSCs] and injectable scaffold on cardiac function improvement in rabbits after inducing myocardial infarction. The Left Anterior Descending [LAD] coronary artery was ligated by No. 6-0 poly amide suture material, and autologous MSCs with injectable scaffold were injected into the margins of the infarcted zone at the time of surgery. At 4 weeks after transplantation, the cardiac function and structure was detected using echocardiography. There was no significant difference among the three groups [MI only, MI Scaffold, and MI+Scaffold+MSCs] in the Echocardio-graphic parameters including, heart rate [HR], Ejection Fraction [EF], Fractional Shortening [FS], Left Ventricular Diameter [LVD] and Left Ventricular Parietal Wall Diameter [LVPW]. A combination of autologous undifferentiated bone marrow MSCs and injectable scaffold made of Chitosan+ Glycerol Phosphate in echocardiographic evaluation did not have a positive influence on achieving functional improvement

Assessment of avian osteoporosis by a quantitative radiographic method

The aim of this study was to develop a quantitative radiographic method for assessment of experimental osteoporosis in Leghorn breed laying hens. Three groups of 24 Leghorn pullets were reared in cage and fed a ration containing different calcium levels, including 3.55, 2.075 and 0.6 percent [for groups 1-3, respectively]. The hens were fed this diet from 17 weeks of age to the end of the experiment. At 20, 28 and 36 weeks of age, 8 hens from each group were selected randomly. Radiographs were obtained from the tibiotarsus and the humerus of each hen. Radiographs were digitized using a camera and assessed by "Image J" software. Bone radiopacities and bone cortex/diameter [C/D] ratios were measured. The hens were sacrificed and the bone ash and calcium contents were measured. Bone densities of the birds in different groups were significantly different at just 36 weeks of age; they were greater in the tibiotarsus bone of the control group than in the tibiotarsus bone of median [P=0.02] and with the low calcium [P=0.007] groups. Humerus densities were also greater in control group compared with that of median [P=0.04] and with low calcium [P=0.0004] diet group. Cortex/diameter index of the tibiotarsus bone was different in all three stages between control and the two other groups, while there were no significant differences between the humeri C/D indices and the three groups in the first stage. Humeri C/D indices of the second and third stages had significant differences between control and the two other groups [P?0.05]. This study showed that radiographic evaluation of bone density is valuable just in progressed osteoporosis, while C/D index can be used for diagnosis of osteoprotic bones in earlier stages

Equine marrow-derived mesenchymal stem cells: isolation, differentiation and culture optimization

Most studies regarding the marrow-derived equine mesenchymal stem cells [MSCs] have mainly focused on the cell transplantation without considering the capacity of differentiation and in vitro requirements of the cells. These concerns were investigated in the present study. Equine MSCs were isolated from the sternal marrow aspirates and expanded through two successive subcultures. Passage-2 equine MSC cultures were then treated with appropriate supplements in order to examine the cell osteogenic, chondrogenic and adipogenic differentiation potential. Furthermore, the culture of the cells was investigated in terms of the optimal concentration of fetal bovine serum [FBS] and the initial cell-seeding density. Additionally, a growth curve was plotted for the cells to study their growth characteristics. According to our findings, equine MSCs were easily generated specialized bone, cartilage and adipose cell lineages as confirmed by specific staining and RT-PCR analysis. Moreover, the cells exhibited rapid expansion when being cultivated in the medium with 15% FBS at 100 cells/cm[2]. Growth curves indicated that these cells rapidly entered the log phase after a brief lag [adaptation] period. In summary, marrow-derived equine MSCs possess tripotent differentiation capacity and rapid growth rate in the appropriate culture conditions

Adhesion and proliferation of horse fibroblast and keratinocyte cells in fibr in glue

Evaluation of autogenous fibrinogen extracted from horse blood in fibrin glue and it's effects on horse's fibroblast and keratinocyte cells were preformed. Fibrinogen has been obtained by glycine precipitation technique from horse's blood. Fibroblast cells were separated by explant culture method from neck skin and keratinocytes were separated by enzyme digestion from lib samples. Autologous fibrinogen, horse`s fibroblast and keratinocyte cells separately mixed with bovine thrombin, calcium gluconate and tranexamic acid. This mixture was uniformly spread on cell culture plate and evaluated for six days. After one day of culture the thickness of fibrin glue were observed by invert microscope, fibroblast cells in elongated shape were observed in the fibrin glue in all thickness and keratinocyte cells were flattened and adherent on surface of plate. Both of them proliferated after 3 days. Fibrinogen that was obtained from horse blood by glycin precipitation technique can support horse fibroblasts as a scaffold and support horse keratinocyte proliferation in vitro

Comparision of the validity of different diagnostic methods in determining ghlo infections in cats

The aim of the present study was to compare different diagnostic methods in the determination of gastric helicobacter infection. 30 healthy stray and 27 domestic cats. were studied The procedure involved taking biopsy samples from antrum and body by pediatric gastroscope in off feed animals. Diagnosis of gastric helicobacter infection by cytology, rapid urease test, histopathology and PCR was also conducted. In all diagnostic methods, helicobacter infection was found to be more prevalent in domestic cats than the stray ones. None of the cats were infected with H. pylori and most of them had mixed infections with H. felis and H. heilmanii. Sensitivity and specificity of different diagnostic methods in comparison with PCR were 93 and 44%, respectively in rapid urease test, 97 and 92% respectively in histopathology and 98 and 94%, respectively in cytology. Based on result of the study most of the cats may be infected with helicobacter spp without any significant clinical signs. Cytology was the most simple and fastest screening methods in the diagnosis of helicobacter infection. With respect to morphologic similarities of H. felis and H. heilmanii and inability to culture H. heilmanii, PCR can be considered as one of the best methods in the identification of infecting helicobacter strains in cats

Growth characteristics of isolated fibroblasts from the distal limb of thoroughbred horses and caspian miniature horses

In this study, in vitro growth characteristics of fibroblasts of metacarpal region of horses and Caspian miniature horses were evaluated. Under general anesthesia and aseptic condition, A full thickness skin incision was created on the lateral aspect of mid-third of left metacarpal region of 4 mixed thoroughbred horses and 4 caspian miniature horses and 3 grams of subcutaneous tissue was harvested and placed in culture medium [RPMI- 1640] in an incubator at 37[degree sign] C [5%CO2]. After growth of fibroblasts, count of cells was performed for 8 days and growth rates and percentage viability of fibroblasts were recorded. There were no significant differences in the growth rates and viability rate of fibroblasts between horses and Caspian horses. It was therefore concluded that due to Genetic similarity between horses and Caspian horses, growth characteristics of fibroblasts in these two groups are the same, but Further In vivo research is needed to identify the wound healing pattern in Caspian horses in compare with horses

[Effect of aloe-vera extract on experimentally induced open wound healing in sheep]

To determine the effects of Aloe-vera extract on skin wounds healing. Experimental study. Four fat tailed sheep with 1-1.5 years old and 25-30 kg weighing. Bilateral skin wounds [two 2x2 cm full thickness as rectangular shape] were made in the cranial and caudal parts of chest. These four areas in each animal were treated individually with Aloe-vera extract [9% in Euserin], Phenytoine [1% in Euserin], pure Euserin and normal saline two times per day. In order to eliminate the effects of anatomical factors on wounds healing drugs were different from one sheep to another. Gross photographs were taken from wounds, scanned and computerized. Then total wound area, granulation of tissue and new epithelialization were measured carefully by Scion Image software. The percentages of wound healing, wound contraction and epithelialization were calculated. Paired t-test. While Aloe-Vera did not show any signifcant effect on the wound geometrical parameter. On the other hand in microscopical evaluations, reepithelialization, tissue granulation and proliferation of connective tissue in wounds were seen in response to Aloe-vera. Furthermore, collagenous fibers were thicker and well arranged in these groups too. While quantitative measurements did not show supportive effects of Aloe- vera on wound geometrical parameters, qualitative observations showed it can improve healing process and wound appearance

Cervical sialoceles [salivary mucoceles] in German shepherd dogs: Review of 3 cases

To identify clinical, ultrasonographic andhistopathological features of cervical sialoceles [salivarymucoceles] in dogs. Retrospective study. Three German shepherd dog. Case records of three dogs that underwentsurgery to remove a cervical sialocele at the University ofTehran Small Animal Hospital were reviewed forsignalment, clinical signs, Ultrasonographic findings andhistopathology. The dogs were male German shepherd withaverage age of 4 years. In all dogs, the masses werelocated in the right cranioventral cervical region.Ultrasonography showed echo texture masses withaccumulation of fluid. The masses removed surgically.Macroscopically, the masses were round, relativelylabulated, and fluctuant with a thick, fibrous capsule.Microscopic examination revealed parotid sialocele inone case and mandibular sialocele in the other two cases.Coagulative necrosis in one case and secondary infectionin the other were observed. There were no recurrences ofthe masses after at least 6 month to 2 years from theoperation. Comparison of these results withstudies by other authors showed that surgical excision ofaffected salivary glands is the treatment of choice forsialocele. Histopathologically, the wall of sialocelesconsists of an outer, highly vascularized layer ofimmature connective tissue and an inner zone of looselyarranged fibroblasts. A pleocellular inflammatoryreactionis evident in the central area, which also containsmuch amorphous acidophilic or amphophilic debris

[Retrospective study of radiography in diagnosis of diseases and disorders in equidae]

To determine the frequency of diagnosed disorders or anomalies in different organs and systems and also effects of breed, age and sex on their incidence rate.Retrospective study.32 horses, 7 donkeys and 5 mules referred to the radiology division.Data were collected from 107 equidae radiographs which had been taken in the Department of Radiology University of Tehran during 4- years [1998-2001]. Radiographs were evaluated for finding injuries and showing relation of disease with breed, sex, age and site of injuries. Information was gathered in the specific charts for determining of frequency of each condition. Diseases and disorders were fractures, exostosis, osteomylitis, osteoarthrosis, arthritis, navicular syndrome, pneumonia, pulmonary hypervascularization, soft tissue swelling, laminitis, guttural pouches disease, abscess and calcification. Fractures [18.18%] and exostosis [18.18%] had the highest frequency in horses. The most frequent disorders were diagnosed in phalangeal area of horse [31.25%]. Furthermore, the first and second most frequent bone disorders in horses were exostosis [25%] and comminuted fracture [18.75%]. Fractures and guttural pouches disease were seen with equal frequency [25%]. In mule, fractures [60%], exostosis [20%] and soft tissue swelling [20%] were diagnosed.This study showed that radiography is a valuable method for diagnosing of equidae disordered organs and systems [in relation with factors such as sex, breed, ageandsite] andalsotheirfrequency