Nontraumatic orbital hematoma secondary to labor and childbirth: a case report and review of the literature.

PURPOSE: Non-traumatic orbital hemorrhage without underlying vascular malformations or predisposing conditions is uncommon, and particularly rare in the context of maternal labor. This study combines a novel case report and retrospective review to analyze reported cases and propose insights. METHODS: This study is both a unique case report and literature review examining PubMed publications with articles traced back to original sources through citations for inclusion. Analysis included clinical presentation, visual examination, hematoma characteristics, neuroimaging, management strategies, and outcomes. RESULTS: We present a 37-year-old multigravida woman at 40 weeks gestation who developed acute right-sided proptosis, diplopia, retrobulbar pain, and periorbital edema during the second stage of labor. Computed tomography (CT) revealed a subperiosteal hemorrhage, with subsequent magnetic resonance imaging (MRI) excluding vascular anomalies. Symptoms resolved within two months. Only 14 cases of maternal orbital hematoma associated with labor have been reported. The average age was 28 with 42% (6/14) being primigravid. Including our case, forty percent (6/15) developed symptoms during the second stage of labor, 40% (6/15) immediately postpartum, and 20% (3/15) over 24 hours postpartum. Overall, 33% (5/15) had potentially contributing conditions including coagulopathies, delivery complications, or vascular malformations. Unilateral orbital hemorrhage occurred in 87% (13/15). Surgical intervention was necessary in 13% (2/15). Most (87%, 13/15) underwent observation or medical management with full recovery of symptoms. CONCLUSIONS: Non-traumatic orbital hematomas associated with maternal labor are rare and likely related to increased valsalva during delivery and heightened blood volume in pregnancy. Neuro-imaging and systemic workup are recommended to assess for vascular anomalies or underlying coagulopathies. The overall prognosis is favorable with most having full recovery.

Purification of swine carbonic anhydrase isoenzyme III and measurement of its levels in tissues and plasma.

The changes in the levels of carbonic anhydrase isozyme III (CA-III) in swine plasma and urine have not been previously determined or reported. CA-III is relatively specific to skeletal muscles, and should therefore be a useful diagnostic marker for muscle diseases. We isolated CA-III from swine muscle tissues and determined CA-III levels in the plasma and urine from both healthy and diseased pigs. The levels of CA-III in the tissues of female swine (age, 3 months) and plasma of young swine (age, 1-5 months) and adult female pigs (age, 2-3 years) were determined using the ELISA system for swine CA-III. The mean (± SD) levels of CA-III in the skeletal muscles were 3.8 ± 3.2 mg/g (wet tissue), and in the plasma, 230 ± 193 ng/ml at 1 month, 189 ± 208 ng/ml at 2 months, 141 ± 148 ng/ml at 3 months, 78 ± 142 ng/ml at 4 months and 53 ± 99 ng/ml at 5 months. The mean level of CA-III in the plasma samples from 2- to 3-year-old pigs was 18 ± 60 ng/ml. CA-III in the plasma samples was found to decrease from 1 month until 3 years of age (p < 0.01). We performed far-western blotting to clarify the cause of the observed decrease in CA-III in plasma. Our results demonstrated that CA-III is bound to the transferrin and albumin. In addition, we determined that the levels of CA-III in plasma and urine samples were higher in diseased swine compared with the healthy pigs.

A novel renal carbonic anhydrase type III plays a role in proximal tubule dysfunction.

Dysfunction of the proximal tubule (PT) is associated with variable degrees of solute wasting and low-molecular-weight proteinuria. We measured metabolic consequences and adaptation mechanisms in a model of inherited PT disorders using PT cells of ClC-5-deficient (Clcn5Y/-) mice, a well-established model of Dent's disease. Compared to cells taken from control mice, those from the mutant mice had increased expression of markers of proliferation (Ki67, proliferative cell nuclear antigen (PCNA), and cyclin E) and oxidative scavengers (superoxide dismutase I and thioredoxin). Transcriptome and protein analyses showed fourfold induction of type III carbonic anhydrase in a kidney-specific manner in the knockout mice located in scattered PT cells. Kidney-specific carbonic anhydrase type III (CAIII) upregulation was confirmed in other mice lacking the multiligand receptor megalin and in a patient with Dent's disease due to an inactivating CLCN5 mutation. The type III enzyme was specifically detected in the urine of mice lacking ClC-5 or megalin, patients with Dent's disease, and in PT cell lines exposed to oxidative stress. Our study shows that lack of PT ClC-5 in mice and men is associated with CAIII induction, increased cell proliferation, and oxidative stress.

Measurement of carbonic anhydrase isozyme VI (CA-VI) in bovine sera, saliva, milk and tissues.

Concentrations of bovine carbonic anhydrase isozyme VI (CA-IV) in bovine serum, saliva, normal milk, colostrum, submandibular gland, liver, and mammary gland were determined. CA-VI was purified from bovine saliva and an antibody to CA-VI was generated. The concentrations of CA-VI in the saliva (7.8 +/- 7.9 microg/ml), serum (2.1+/- 5.7 ng/ml), milk (7.9 +/- 12.1 ng/ml), submandibular gland (284.7 microg/g protein), liver (921.0 +/- 180.7 ng/g protein) and mammary gland (399.6 +/- 191.2 ng/g protein) were determined by ELISA. No seasonal change in CA-VI levels was observed in normal milk. The concentration of CA-VI in colostrum (day 1 post partum) was 119 ng/ml and decreased rapidly by 1 month following birth. Mammary gland contained much smaller amounts than the submandibular gland. CA-VI mRNA was detected in the liver and mammary gland of cow by RT-PCR. The ELISA used in this study proved to be a precise and sensitive method for determining CA-VI concentrations in saliva, serum, milk and tissue specimens from cows. The ELISA may enable the study of changes in CA-VI associated with hereditary or metabolic disorders of the salivary gland, mammary gland and liver using small samples of saliva, serum or milk.

Determination of carbonic anhydrase-III by enzyme-immunoassay in liver, muscle and serum of male rats with streptozotocin-induced diabetes mellitus.

Insulin has a plethora of metabolic effects but its action on carbonic anhydrase-III (CA-III), a key enzyme in acid-base regulation, has been little studied. The present studies examined the effects of streptozotocin induced diabetes on the concentrations of CA-III. The concentration of CA-III in the liver, muscles and serum of rats with experimental diabetes mellitus was measured by the method of enzyme-immunoassay. Streptozotocin-induced diabetes mellitus resulted in a reduction in concentration of CA-III in the liver and serum, but not in skeletal muscles, of adult male rats. A 98% reduction in hepatic CA-III content relative to control values was observed. The reduction in CA-III content in the liver was restored to control value by administration of insulin. The CA-III content in serum of diabetic rats declined to approx. 25% of control values, but the reduction was unaffected by administration of insulin. The concentration of CA-III in the liver and serum of diabetic rats was not influenced by administration of methyltestosterone. Although the content of CA-III in m. rectus femoris, m. tibialis craniaris and m. soleus differed, no significant difference of CA-III content was found between diabetes mellitus and control rats. The effect of chronic diabetes mellitus on CA-III content was obviously different between liver and muscle, suggesting that the regulation of CA-III biosynthesis differs between these two tissues. These results suggest that biosynthesis of CA-III in hepatocytes of rats is influenced by irregular patterns of GH secretion brought about by diabetes mellitus.

Immunohistochemical studies of the carbonic anhydrase isozymes in the bovine placenta.

Placentae from 11 cows, ranging from about 80 to 270 days gestation, were studied for immunohistochemical localization of carbonic anhydrase isozymes. CA isozymes were localized using the avidin-biotin-peroxidase complex (ABC) method in the bovine placenta. CA-II was found in the fetal trophoblastic cells with single nuclei as well as in erythrocytes of maternal and fetal blood. The percentage of positive cells for anti-CA-II in the trophoblastic cells did not change during development of gestation in the fetal bovine. CA-I and CA-III were not detected in the bovine placenta.

Distribution of carbonic anhydrase isozyme III (CA-III)-positive cells in duct segments of the bovine submandibular gland.

The distribution of carbonic anhydrase III isozyme (CA-III)-positive cells in bovine submandibular glands was studied by immunohistochemistry. CA-III showed strong immunoreactivity in basal cells and some lining cells of the intercalated ducts as well as in striated and interlobular ducts cells. No significant immunoreactivity could be found in other portions of the glands. Electron microscopically, the immunoreactive basal cells contained scattered tonofibrils in their cytoplasm. The distribution of CA-III suggests that the CA-III-positive basal cells may play a special physiological role, and that they do not only represent undifferentiated lining cells.

Carbonic anhydrase III in equine tissues and sera determined by a highly sensitive enzyme-immunoassay.

A sensitive sandwich enzyme immunoassay (EIA) for measuring equine carbonic anhydrase III (CA-III) was established using a microplate as a solid-phase and peroxidase as a labelling enzyme. The assay can detect concentrations as low as 5 ng/ml using 20 microliters of sample sera. Within-run coefficients of variation obtained using standard equine CA-III were less than 5 per cent. CA-III levels in equine serum ranged from 5 to 50 ng/ml (n = 370), and apparently abnormal levels of CA-III from 100 to 1900 ng/ml (n = 27) were observed. The concentrations of immunoreactive CA-III in the extracts of various equine tissues were also determined; it was present at high concentrations in skeletal muscle and liver and to a much lesser extent in the thymus. Other tissues contained much smaller amounts.

Immunocytochemical localisation of carbonic anhydrase isozyme III in equine skeletal muscle.

The location of carbonic anhydrase III (CA-III) in frozen sections of biopsies of Thoroughbred horse skeletal muscle was studied. Fibre types were determined by ATP-ase and succinate dehydrogenase staining. CA-III isozyme was detected using a peroxidase conjugated anti-CA-III antibody. CA-III was found to be localised in slow twitch oxidative fibres (ST), but was also present in fast twitch oxidative (FTH) fibres in small amounts. Fast twitch glycolytic (FT) fibres were stained lightly compared with control sections. The concentrations of CA-III in muscle and liver were 70 micrograms/mg protein and 4 micrograms/mg protein, respectively. CA-I and CA-II were not found in muscle extracts by the double immunodiffusion method.

Radiotherapy for squamous cell carcinoma of the nasal cavity.

The management of squamous cell carcinoma of the nasal cavity, a relatively rare tumor, is described, with results of long-term follow-up. We treated five such patients by means of radiotherapy combined with surgery in all but one case. Three patients received a dose of 40 Gy preoperatively, one received 60 Gy of radical radiotherapy, and one received 38 Gy of postoperative radiotherapy. During the follow-up period, one patient developed a local recurrence and two patients developed distant metastases. Nasal cavity tumors in our patients were diagnosed early, and local control using a combination of radiotherapy and surgery was relatively effective. Distant metastases occurred in two of five patients during the follow-up period, suggesting that the possibility of distant metastases during follow-up was considered.

Pharmacokinetics and radiosensitizing effect of PR-350.

PURPOSE: The in vivo pharmacokinetics and sensitizing effects of PR-350, a newly developed, highly water-soluble radiosensitizer were examined. MATERIALS AND METHODS: C3H/HeJ mice bearing SCC-VII tumor cells were used in the experiments. Results were compared with those of PR-28 and SR-2508. RESULTS: The intratumoral concentration of PR-350 reached the maximum value of 77.7 mg/kg 20 min after intravenous injection. The concentration in the brain was below the detection limit. The enhancement ratios of PR-350 calculated using the growth delay method were 0.9 for PR-350 at 50 mg/kg and 1.4 for PR-350 at 100 mg/kg, compared with 1.4 for SR-2508 at 100 mg/kg and 1.0 for PR-28 at 100 mg/kg. CONCLUSION: PR-350 and SR-2508 revealed similar sensitizing effects. The side effects of PR-350 are expected to be equal to or less than those of SR-2508. PR-350 seems to be a promising radiosensitizer.

Intraluminal irradiation for T2M0 esophageal cancer: effect of patient selection on prognosis.

The contribution of intraluminal irradiation to the prognosis of T2M0 esophageal cancer was examined with reference to patient selection. To determine the effect of patient selection, we used esophagrams to assess the potential difficulty of inserting a tube for intraluminal irradiation in 39 patients who were treated with external irradiation prior to 1982. Twenty-two patients were assessed as likely to have been able to undergo intraluminal irradiation (group 1) while 17 were assessed as unlikely to have been able to undergo the procedure (group 2). Of 36 patients treated after 1983, 19 patients were treated with a combination of intraluminal irradiation and external beam therapy (group 3) and 17 by external irradiation alone (group 4). The median survival times of groups 1, 2, 3, and 4 were 10.8, 6.4, 12.0, and 8.5 months, respectively. The prognosis of patients treated with combination therapy (group 3) was superior to that of those treated before 1982 (groups 1 + 2). However, there was no improvement in the prognosis of T2M0 esophageal cancer treated by radiotherapy after the introduction of intraluminal irradiation, and the difference between the survival curves of patients treated by combination therapy (group 3) and with a wide esophageal lumen (group 1) was not significant. Our study thus showed that the selection of patients according to the insertability of the intraluminal therapy tube affected the prognosis.

Study on the pathogenesis of porcine serum-induced liver fibrosis in rats with special reference to the effects of hypertension.

To investigate the pathogenesis of porcine serum (PS)-induced liver fibrosis in rats, two experiments were carried out, taking into consideration of hypertension and vascular changes. In Experiment I, spontaneous hypertensive rats (SHRs), two-kidney, one clip hypertensive F344 rats (2K1C rats), and normotensive F344 rats were given an intraperitoneal injection of PS of 0.5 ml twice a week for 8 weeks. Histopathological, immunohistochemical, and electron microscopical examinations were performed on the liver from each rat. Histological features of liver fibrosis in hypertensive and normotensive rats were essentially identical. However, in the PS-treated SHRs, 2 of 5 animals showed the most severe fibrosis in all PS-treated groups. Electron microscopically, degranulated mast cells, eosinophils, and macrophages engulfing apoptotic cells were rarely observed in the late stage of fibrous septa (FS) in the PS-treated SHR liver. In Experiment II with normotensive F344 rats, histopathological features of early FS in the liver were compared with those of late FS observed in Experiment I using serial sections, and we found that FS developed along the wall of newly formed vessels to connect between neighboring central veins. However, the effect of hypertension on this fibrosis could not be clearly demonstrated in the present study using SHRs and 2K1C rats.

Purification of carbonic anhydrase isozyme VI (CA-VI) from swine saliva.

Salivary or secreted carbonic anhydrase (CA), which constitutes a new class of CA, designated CA-VI, was isolated. Swine CA-VI purified from swine saliva by inhibitor-affinity chromatography and ion exchange chromatography had a specific activity of 5,468 units/mg. The molecular weight was 250,000, as determined by gel filtration under non-denaturing conditions, and the subunit molecular weight was found to be 37,000 by sodium dodecyl sulfate polyacrylamide gel electrophoresis, indicating that swine CA-VI consists of 7 subunits. The treatment of the enzyme with endo-N-acetylglucosaminidase F reduced its subunit molecular weight from 37,000 to 35,000 and 32,000. We raised a rabbit antibody against purfied swine CA-VI. Double immunodiffusion showed that anti-swine CA-VI serum reacted with swine CA-VI and swine saliva, but not with hemolysate (containing CA-I and CA-Il) or muscle extracts (containing CA-III). The concentration of CA-VI in swine saliva, measured using single radial immunodiffusion, was 0.027 +/- 0.017 mg/mg total protein.

Comparison of the distribution of carbonic anhydrase isozymes (CA-I, CA-II, CA-III) in the rat gastrointestinal tract.

The present paper described the immunohistochemical distributions of carbonic anhydrase (CA) isozymes. CA-I, CA-II and CA-III, in the epithelium lining the rat gastrointestinal tract, with rabbit antibodies to equine CA-I, CA-II and CA-III. Prior to the immunohistochemical examinations, the crossreactivities of these antibodies to the rat-antigens were confirmed in this study. In the stomach, surface epithelial cells and parietal cells of the glandular region showed an immunoreactivity only to CA-II. In the large intestine, each immunoreactivity to CA isozyme (CA-I, CA-II and CA-III) was localized in the upper portion of intestinal glands, and decreased toward the distal digestive tract, but absent in the small intestine. The present histological findings suggested that the CA isozymes might play a role in the ion-transportation during the water absorption in the rat large intestine.

Immunohistochemical localization of carbonic anhydrase isoenzymes in salivary gland and intestine in adult and suckling pigs.

Localizations of carbonic anhydrase isoenzymes (CA I, CA II and CA III) were investigated immunohistochemically in the salivary glands and intestine of mature and suckling pigs. Carbonic anhydrase isoenzymes were not detected in the salivary glands of sucklings, but were present in the adult. Bicarbonate ion in saliva might be important for the digestion of solid foods in mature pigs, but unnecessary for the digestion of milk in sucklings. Expressions of CA I and CA II were detected strongly in the large intestine of the adult and sucklings, and faintly only at duodenum in the small intestine. CA I and CA II isoenzymes in the large intestine may be involved, at least in part, in ion absorption and water metabolism during digestion and absorption of milk in suckling pigs. In addition, CA I and CA II expression in the duodenal villus enterocyte may support the process of bicarbonate absorption in the duodenum.

Determination of carbonic anhydrase III isoenzyme concentration in sera of racehorses with exertional rhabdomyolysis.

The concentration of carbonic anhydrase III isoenzyme (CA-III) in serum samples from 216 clinically normal Thoroughbreds was determined by use of an enzyme immunoassay. The concentration range of CA-III was from 16.0 to 254.5 ng/ml (mean, 56.5 +/- 11.9 ng/ml). Significant differences were not detected according to age or sex. To confirm whether serum CA-III concentration was high in horses with muscle disease, serum samples of 11 horses with exertional rhabdomyolysis were analyzed by enzyme immunoassay. Their serum CA-III concentration was about 56 times (3,136 +/- 2,610 ng/ml) that of healthy Thoroughbreds. Concentration of CA-III was higher in horses with rhabdomyolysis that had been transiently recumbent than in horses with mild disease that were reluctant to move. Blood samples obtained serially from 6 horses with exertional rhabdomyolysis were studied. Serum activities of aldolase, creatine kinase, aspartate transaminase, and lactate dehydrogenase were high. Increases and decreases in concentration of CA-III were more rapid than that for aldolase, creatine kinase, aspartate transaminase, and lactate dehydrogenase activities; thus, CA-III may be clinically applicable as a diagnostic marker for muscle disease in horses.

Isolation and measurement of carbonic anhydrase isoenzymes in erythrocytes of dogs.

OBJECTIVE: To purify canine carbonic anhydrase (CA) isoenzymes CA-I and CA-II and to determine concentrations of CA-I and CA-II in erythrocytes of Beagles and dogs native to Japan. SAMPLE POPULATION: Blood samples from 116 Beagles, including 24 pregnant Beagles, and blood samples from 29 dogs native to Japan. PROCEDURE: Canine CA-I and CA-II were purified by use of column chromatography. Concentrations of CA-I and CA-II in erythrocytes of dogs were determined, using an ELISA. RESULTS: Mean (+/- SD) concentrations of CA-I and CA-II in erythrocytes of Beagles were 3.21+/-0.86 and 1.63+/-0.39 mg/g of Hb, respectively. Mean concentration of CA-I was greater in male Beagles than female Beagles. In contrast, mean concentration of CA-II was greater in female Beagles than male Beagles. Furthermore, concentration of CA-II was greater in pregnant female Beagles than male or nonpregnant female Beagles. Mean concentrations of CA-I and CA-II in erythrocytes of dogs native to Japan were 11.03+/-4.39 and 3.29+/-0.91 mg/g of Hb, respectively. Mean concentration of CA-I was greater in male dogs from Japan than female dogs from Japan. CONCLUSIONS AND CLINICAL RELEVANCE: The ELISA used in this study proved to be precise and sensitive for determining CA-I and CA-II concentrations in dogs. The ELISA may enable study of changes in isoenzymes associated with hereditary or metabolic disorders of blood or other body fluids, using only a small sample. Measurement of the concentrations of CA isoenzymes in dogs may be of diagnostic value.

Immunohistolocalization of the carbonic anhydrase isoenzymes (CA-I, CA-II, and CA-III) in the reproductive tract of male horses.

OBJECTIVE: To elucidate locations of cytosolic carbonic anhydrase isoenzyme (CA-I, CA-II, and CA-III)-positive epithelial cells in equine male reproductive organs. DESIGN: Descriptive and immunohistochemical study. ANIMALS: 4 clinically normal male horses. PROCEDURE: The testis (seminiferous tubules, rete tubules), epididymis (initial, middle, and terminal segments), proximal and distal portions of the ductus deferens, ampulla ductus deferentis, seminal vesicle, prostate, and bulbourethral gland were excised from euthanatized horses after administration of an overdose of pentobarbital. The tissue specimens were quickly placed in fixative solution, dehydrated in ethanol, and embedded; then thin sections were cut. For immunohistochemical staining, antibodies against purified equine CA-I, CA-II, and CA-III were raised in rabbits. After examination of the specificity of each antiserum, the monospecific antisera against carbonic anhydrase isoenzymes were used to localize the isoenzymes. RESULTS: Specific staining for CA-III was found in the Sertoli and basal cells of the ductus deferens. Most of the testicular and epididymal tissue, as well as ductus deferens, were virtually negative for the enzymes when stained with the antibody to CA-I and CA-II. In the initial segment of the epididymis, a few principal cells had intense cytoplasmic staining with anti-CA-II. In the male accessory glands, CA-I, CA-II, and CA-III were detected in the epithelial cells of the seminal vesicle, prostate, and bulbourethral gland. CONCLUSIONS: In the equine male reproductive tract, the bicarbonate in semen originates mainly from accessory reproductive glands. All 3 isoenzymes may have central roles in the regulation of bicarbonate concentration in seminal plasm and, accordingly, regulate seminal plasma pH. Distribution of CA-III in Sertoli and basal cells of the ductus deferens suggests other specialized physiologic roles.