Spontaneous endometriosis in cynomolgus monkeys as a clinically relevant experimental model.

STUDY QUESTION: Does spontaneous endometriosis in cynomolgus monkeys have the characteristics required of a good experimental model? SUMMARY ANSWER: Spontaneous endometriosis in cynomolgus monkeys exhibited similar clinicopathological characteristics to the human disease and was useful as an experimental model. WHAT IS KNOWN ALREADY: The prevalence of endometriosis in autopsied cynomolgus monkeys (Macaca fascicularis) in a breeding colony was reported to be 28.7% in 1993. The histopathological findings we reported recently showed that components of spontaneous endometriosis were not only endometriotic epithelium and stromal cells (CD10-positive) with hemorrhage and inflammation, but also smooth muscle metaplasia and nerve fibers. STUDY DESIGN, SIZE, DURATION: During routine medical examinations at a research facility from 2008 to 2012, 614 female cynomolgus monkeys of reproductive age (6-25 years) were screened for endometriosis by the presence of regular menstrual bleeding, serum CA125 levels and palpation of the abdomen. In total, 29 monkeys were selected as subjects for the following study. PARTICIPANTS/MATERIALS, SETTING, METHODS: Of the 29 monkeys selected, 15 were diagnosed with endometriosis by laparoscopy and/or open surgery. The monkeys were monitored by observing their general condition, and eight of these were monitored using laparoscopy and MRI. In addition, to investigate appropriate screening parameters and endometriosis-associated biological parameters in monkeys, we retrospectively examined general laboratory parameters that correlate to the menstrual cycle and disease status. MAIN RESULTS AND THE ROLE OF CHANCE: The combination of CA125 serum levels (this was a useful marker for chocolate cysts), palpation of the abdomen, and fecal abnormalities was the most efficient screening method for diagnosing monkeys with endometriosis. Each animal could be diagnosed and assigned a disease stage by laparoscopy. While monitoring the disease stage by laparoscopy and/or MRI, disease status in individual monkeys was mainly stable or was progressive for 2-7 months. The detection rate by screening was low (15/614) but age-specific analysis suggests that screening would be more efficient if a colony for an endometriosis model is maintained with 11-20-year olds. As an endometriosis-associated biological parameter, the decrease in food consumption that coincided with menstruation was selected and correlated well (R2 value = 0.8239) with disease status (according to a modified adhesion revised American Fertility Society score). LARGE SCALE DATA: N/A. LIMITATIONS, REASONS FOR CAUTION: Peritoneal fluid was not analyzed because a smaller amount is produced in cynomolgus monkeys than in baboons. Although clinical endometriosis-associated pain is evaluated in women using a visual analog scale, pain could not be directly evaluated in this non-human primate model. WIDER IMPLICATIONS OF THE FINDINGS: Although cynomolgus monkeys are relatively small (2-5 kg) primates, laparoscopy and MRI make it possible to evaluate spontaneous endometriosis in these monkeys and to monitor its development over time. Spontaneous endometriosis in cynomolgus monkeys is a useful model for evaluating disease progress and drug efficacy because they have similar lesions to those in humans, and conventional laboratory methods and parameters for assessment are well established. STUDY FUNDING/COMPETING INTEREST(S): No external funds were used for this study. A.N.-K., K.T., H.T., A.K. and M.S. are full-time employees of Chugai Pharmaceutical Co., Ltd. R.K. received a consultancy fee from Chugai Pharmaceutical Co., Ltd. and lecture fees from Chugai Pharmaceuticals, Japan Vaccine Co. Ltd., Merck & Co., Mochida Co. Ltd., Roche Diagnostics, and BD, unrelated to the submitted work. S.N., S.O. and T.S. have nothing to declare.

Characteristics of histologically confirmed endometriosis in cynomolgus monkeys.

STUDY QUESTION: What are the characteristics of spontaneous endometriosis in cynomolgus monkeys? SUMMARY ANSWER: Spontaneous endometriosis in cynomolgus monkeys exhibited similar characteristics to the human disease. WHAT IS KNOWN ALREADY: One previous report described the prevalence and the basic histopathology of spontaneous endometriosis in cynomolgus monkeys. STUDY DESIGN, SIZE, DURATION: Endometriotic lesions that had been histologically confirmed in 8 female cynomolgus monkeys between 5 and 21 years old were subjected to study. PARTICIPANTS/MATERIALS, SETTING, METHODS: The monkeys died of, or were sacrificed because of, sickness consequent on endometriosis. Specimens were evaluated histopathologically with haematoxylin and eosin staining, iron staining and immunohistochemistry (CD10, CD31, α-SMA and PGP9.5), and by observing them under a microscope. MAIN RESULTS AND THE ROLE OF CHANCE: Endometriotic and stromal cells (CD10-positive) with haemorrhage and inflammation were observed. Smooth muscle metaplasia and nerve fibres were also noted in the endometriotic lesions. Endometriotic lesions in lymph nodes were incidentally found. LIMITATIONS AND REASONS FOR CAUTION: Since laparoscopic analysis for monitoring the disease state was not set as a parameter of the current study, time course changes (progression) of the disease were not assessed. WIDER IMPLICATIONS OF THE FINDINGS: Further investigation of spontaneous endometriosis in cynomolgus monkeys may contribute to better understanding of the disease pathobiology. STUDY FUNDING/COMPETING INTERESTS: No external funds were used for this study. A.N.K., S.M., S.H., T.I., O.K., A.K. and M.S. are full-time employees of Chugai Pharmaceutical Co., Ltd. R.K. received lecture fees from Chugai Pharmaceutical Co., Ltd., unrelated to the submitted work. S.N., S. O., L.Y., K.Y. and T.S. have nothing to declare. TRIAL REGISTRATION NUMBER: N/A.

Neuroimaging and neuropathologic findings in AIDS patient with cytomegalovirus infection.

This 21-year-old male with hemophilia A developed cytomegalovirus (CMV) retinitis associated with acquired immunodeficiency syndrome (AIDS). He had a history of numerous blood transfusions. Serum antibody titers became positive for human immunodeficiency virus (HIV), when the patient was 18 years of age. Three years later, he developed CMV retinitis due to his immunosuppression. Ganciclovir (DENOSINE, TANABE SEIYAKU CO., LTD., Osaka, Japan) therapy given for 4 weeks produced a marked improvement in the ocular fundal findings, but the neurologic signs and symptoms, including headache, hypoesthesia, disorientation, and dementia became worse. T2-weighted magnetic resonance imaging (MRI) demonstrated a diffuse high intensity area in the periventricular white matter and small focal or patchy lesions in the hippocampus, basal ganglia, midbrain, medulla oblongata and the nucleus dentatus. The patient died of HIV encephalopathy and CMV infection. Characteristic CMV intranuclear inclusion bodies were observed histologically in most sites of the brain including the hippocampus, white matter, basal ganglia, midbrain, medulla oblongata, nucleus dentatus and the retina. Infiltration by monocyte-macrophage and multinucleated giant cells, which are characteristic of HIV encephalopathy, were observed in the periventricular white matter and the hippocampus. In this patient, the neuroimaging findings were compatible with the neuropathologic observations. MR imaging proved useful in detecting the central nervous system (CNS) lesions of AIDS and CMV infection.

Living related partial liver transplantation for primary biliary cirrhosis--a case report.

An adult living related partial liver transplantation was performed on a 49 year old female with terminal hepatic failure due to primary biliary cirrhosis (PBC). The donor was her 53 year-old sister. A sufficient volume of graft tissue was obtained, which comprised 1.5% of the body weight of the recipient. The recipient had an excellent recovery without any major complications, and was discharged 35 days after the operation. At 15 months after the operation, the patient has shown no signs of rejection while using FK506 and prednisolone as immunosuppressants. The progression of symptomatic PBC can be predicted, and the timing of the transplantation can be easily determined. In addition, the results of liver transplantation for PBC are good. Therefore, adult living related partial liver transplantation is an excellent treatment for primary biliary cirrhosis.

[Pharmacokinetic and clinical studies on cefprozil granules in the pediatric field].

Cefprozil (CFPZ), a newly developed oral cephalosporin in a fine granular form for pediatric use, was administered to children with bacterial infections. MICs were determined for 6 drugs including CFPZ, cephalexin (CEX), cefaclor (CCL), ampicillin (ABPC), methicillin (DMPPC) and cloxacillin (MCIPC) against the following 84 strains isolated from cases to which CFPZ was administered; 55 strains of Gram-positive cocci (GPC) including 2 strains of Staphylococcus aureus, 49 strains of Streptococcus pyogenes, 4 strains of Streptococcus pneumoniae, and 29 strains of Gram-negative bacilli (GNB) including 10 strains of Haemophilus influenzae, 18 strains of Escherichia coli, and 1 strain of Proteus mirabilis. MIC determination of these strains was done with an inoculum size of 10(6) CFU/ml. In pharmacokinetic studies, serum concentrations, urinary concentrations and urinary recovery rates were investigated using bioassay and high-performance liquid chromatography (HPLC). CFPZ was orally administered 30 minutes before meals to 9 children with ages ranging from 7 years and 1 month to 12 years and 3 months. Three groups of 3 children were tested with doses of 4.0, 7.5 and 15.0 mg/kg, respectively. In addition to the above, clinical and bacteriological studies were performed in a total of 160 cases consisting of children with ages ranging 5 months to 12 years and 5 months. A mean dose of 8.6 mg/kg in 3-4 divided doses (130 cases of t.i.d. and 30 cases of q.i.d.) was administered for an average of 7 days. The 160 cases included 34 cases of pharyngitis, 5 cases of tonsillitis, 8 cases of acute bronchitis, 8 cases of pneumonia, 52 cases of scarlet fever, 4 cases of acute purulent otitis media, 47 cases of urinary tract infection, 1 case of purulent lymphadenitis and 1 case of posthitis. Adverse reactions and abnormal clinical laboratory test results were also examined in 166 cases, including 6 cases excluded from the evaluation of clinical efficacy. The results obtained are summarized as follows: 1. With regard to GPC, MICs of CFPZ against 2 strains of S. aureus were 0.78 or 1.56 micrograms/ml and CFPZ showed the second highest activity to MCIPC. MICs of CFPZ against 49 strains of S. pyogenes were all less than 0.025 micrograms/ml.(ABSTRACT TRUNCATED AT 400 WORDS)

[Experimental and clinical studies on cefodizime in pediatrics].

Cefodizime (CDZM), a newly developed injectable cephem antibiotic, was given via bolus intravenous injection at each of 3 dose levels of 10, 20 and 40 mg/kg to each 3 children, and serum and urinary levels and urinary recovery rates were followed. A total of 57 patients received CDZM in the following regimen via bolus intravenous injection, and clinical efficacies, and microbial responses were evaluated. Mean dosage per application: 20.9 mg/kg, number of application per day: between 2 and 4 (2 times for 3 patients, 3 for 26 patients and 4 for 28 patients), mean duration of the therapy: 1 week. Patients consisted of 1 case of peritonsillar abscess, 2 acute bronchitis, 38 pneumonia, 8 urinary tract infection, 1 staphylococcal scalded skin syndrome, 2 cellulitis, 4 purulent lymphadenitis and 1 typhoid fever. In addition to the patients mentioned above 6 patients who dropped out were involved in the evaluation of adverse reactions and influence of the drug on laboratory test data, and the following results were obtained. 1. Five minutes after bolus intravenous injection in doses of 10, 20 and 40 mg/kg, serum levels determined by the bioassay method were at their maxima, i.e. 114.0, 264.6 and 461.6 micrograms/ml, respectively. Serum levels of drugs were dose-dependent throughout all the dosage levels tested. Mean serum half-lives of the drug were 1.757, 1.552 and 1.668 hours, respectively, for the 3 dose levels. Serum levels of the drug determined by the HPLC method were similar to those by the bioassay method: The maximum serum levels occurred at 5 minutes after administration, mean maximum concentrations were 105.5, 264.0 and 461.7 micrograms/ml for the 3 dose levels, and a dose response was noted for the 3 dose levels. The half-lives were 1.755, 1.598 and 1.668 hours, respectively. 2. Mean maximum concentrations in urine determined by bioassay for 2 of 3 cases received 10 mg/kg and 3 cases each given 20 and 40 mg/kg of CDZM were 884.3, 3,061 and 7,352 micrograms/ml, respectively, in the first 2 hours after administration. These levels were also dose-dependent. Mean recovery rates were 74.4, 78.4 and 71.5%, respectively, in the first 8 hours after administration. Mean maximum concentrations in urine measured by HPLC were similar to those determined by bioassay, i.e. 962.3, 3,404 and 7,899 micrograms/ml in the first 2 hours. They were, also, dose-dependent. Mean recovery rates were 82.1, 86.0 and 76.5%, respectively, in the first 8 hours after administration. The HPLC determinations gave slightly higher levels than the bioassay.(ABSTRACT TRUNCATED AT 400 WORDS)

Periventricular leucomalacia (PVL)-like lesions in two neonatal cynomolgus monkeys (Macaca fascicularis).

Periventricular leucomalacia (PVL) is a lesion of immature cerebral white matter that occurs in the perinatal period. In man, PVL is the predominant form of brain injury and a cause of cerebral palsy and cognitive deficits in premature infants. PVL affects fetuses and newborns, particularly those who have undergone oxygen deprivation as may occur in premature birth. Many clinical and pathological studies of PVL have been performed in man, but there is no clear definition of PVL in animals. A few spontaneous PVL-like cases in puppies or experimental cases in other animal species have been reported. The present study reports the histopathological and immunohistochemical features of PVL-like lesions in two neonatal cynomolgus monkeys. In both cases, there was cerebral white matter necrosis with marked infiltration of lipid-laden phagocytes and a reduction of neurons in the cerebral cortex. In case 1 there was extensive cavitation of the cerebral white matter. In case 2 there was reactive astrocytosis associated with a decrease in oligodendroglial cells and a decrease in cerebral white matter myelin. To our knowledge, this is the first report of PVL-like leucoencephalomalacia in non-human primates.

Congenital cystic adenomatoid-like malformation in a cynomolgus monkey (Macaca fascicularis).

Congenital cystic adenomatoid malformation (CCAM) is a developmental lung abnormality characterized by abnormal proliferation of mesenchymal elements and failure of bronchiolar structures to mature, ultimately resulting in the compression of normal pulmonary tissue and mediastinal shift with rapid expansion of cysts. Although various clinical and pathologic studies of CCAM in humans exist, CCAM has yet to be reported in animals, even in nonhuman primates. In the present study, histopathologic analyses of a neonatal cynomolgus monkey that died 17 days after birth revealed that normal lung architecture was replaced by disorganized overgrowths of cysts lined with simple cuboidal epithelium. The epithelium projected a few ciliates into the air spaces and produced mucus. To our knowledge, this is the first case study describing CCAM or a CCAM-like lesion in nonhuman primates.

Transthyretin amyloidosis and two other aging-related amyloidoses in an aged vervet monkey.

An aged male vervet monkey showed severe cardiac arrhythmia for more than 3 years. A multifocal amyloid consisting of transthyretin was deposited in all areas of the heart wall, especially in the extracellular stroma among muscle fibers and external tunica of arterioles. Moreover, the amyloid was deposited in the stroma and arterioles of other systemic organs except the liver and spleen. These characteristics are consistent with senile systemic amyloidosis in humans. A second amyloid consisting of amyloid beta protein was in senile plaques and cerebral amyloid angiopathy in the cerebral cortex. A third amyloid consisting of islet amyloid polypeptide was deposited in islets of the pancreas. Apolipoprotein E and amyloid P component colocalized with the 3 amyloids. Thus, 3 different aging-related amyloids were found in an aged vervet monkey. In particular, to our knowledge, this is the first report on spontaneous transthyretin amyloidosis in animals.

Cerebral fat embolism studied by magnetic resonance imaging, transcranial Doppler sonography, and single photon emission computed tomography: case report.

Cerebral fat embolism syndrome is an uncommon complication of trauma. We present a patient who developed cerebral fat embolism syndrome secondary to long-bone fractures. Although computed tomography of the brain failed to show any intracranial lesion, magnetic resonance imaging (MRI) detected scattered, high-signal-intensity lesions on T2-weighted images. 99mTc-d, 1-hexamethyl-propylene amine oxine single photon emission computed tomography (99mTc-HMPAO SPECT) and transcranial Doppler sonography (TCD) demonstrated low cerebral blood flow in the acute stage. MRI, 99mTc-HMPAO SPECT, and TCD correlated well with the clinical course of cerebral fat embolism syndrome.