Long-term Results of Splenectomy in Transfusion-dependent Thalassemia.

Splenectomy is indicated in transfusion-dependent thalassemia (TDT) only in certain situations. This study aimed to present the effectiveness, complications, and long-term follow-up results of splenectomy in children with TDT. We performed a 30-year single-institution analysis of cases of splenectomy for TDT between 1987 and 2017 and their follow-up until 2021. A total of 39 children (female/male: 24/15) were included. The mean age at splenectomy was 11.2±3.2 years, and their mean follow-up duration after splenectomy was 21.5±6.4 years. Response was defined according to the patient's annual transfusion requirement in the first year postsplenectomy and on the last follow-up year. Complete response was not seen in any of the cases; partial response was observed in 32.3% and no response in 67.6%. Thrombocytosis was seen in 87% of the patients. The platelet counts of 7 (17.9%) patients were >1000 (10 9 /L), and aspirin prophylaxis was given to 22 (56.4%) patients. Complications were thrombosis in 2 (5.1%) patients, infections in 11 (28.2%) patients, and pulmonary hypertension in 4 (10.2%) patients. Our study showed that after splenectomy, the need for transfusion only partially decreased in a small number of TDT patients. We think splenectomy can be delayed with appropriate chelation therapy up to higher annual transfusion requirement values.

Assessments of some trace metals in water samples of nursery pond of Grass Carp (Ctenopharyngodon idella, Valenciennes, 1844) in Bannu Fish Hatchery of Khyber Pakhtunkhwa, Pakistan / Avaliações de alguns vestígios de metais em amostras de água de viveiro de carpa capim (Ctenopharyngodon idella, Valenciennes, 1844) em Bannu Fish Hatchery de Khyber Pakhtunkhwa, Paquistão

The present investigation was aimed to examine the concentrations of trace metals including e.g copper (Cu), manganese (Mn), nickel (Ni), and zinc (Zn) in water samples collected from nursery pond of grass carp (Ctenopharyngodon idella) in Bannu Hatchery of Khyber Pakhtunkhwa during the period from April 2018 to January 2019. The temperature and pH of each water sample were measured for the whole study duration. The concentration of Copper (Cu), Manganese (Mn), Nickel (Ni), and Zinc (Zn) in collected water samples were measured in mg/liter by using Atomic Absorption Spectrophotometer. The blank and standard solutions for device calibration Standard solutions i.e., 2.0 mg, 4.0 mg, and 6.0 were used to measure the concentration of these metals in water samples to verify the measurements. The data was statistically analyzed on descriptive statistics (estimation of proportions and standard deviation) used to summarize mean concentration. The results obtained of both temperature and pH of water samples were found in ranged 10 to 36 0C and 7.0 to 8.44; whereas the size of fry stages was ranged from 4.0 to 56.0 mm in total length. The results of investigated metals found in pond water samples are in order of Zn>Mn>Ni>Cu, respectively. As optimum temperature and pH for grass carp were mostly between 15 0C and 30 0C and pH 6.5 to 8.0. It was concluded from obtained results that temperature, pH, and trace metals were found appropriate for the growth of Ctenopharyngodon idella from fry to fingerling stages, but the highest amount of zinc can cause its mortality. It is a preliminary study on grass carp culturing in Bannu fish hatchery so, it would provide useful information for model fish seed production unit in a hatchery.

A presente investigação teve como objetivo examinar as concentrações de metais traço, incluindo cobre (Cu), manganês (Mn), níquel (Ni) e zinco (Zn) em amostras de água coletadas de viveiro de carpa-capim (Ctenopharyngodon idella) em Incubatório de Bannu de Khyber Pakhtunkhwa durante o período de abril de 2018 a janeiro de 2019. A temperatura e o pH de cada amostra de água foram medidos durante todo o período do estudo. A concentração de cobre (Cu), manganês (Mn), níquel (Ni) e zinco (Zn) nas amostras de água coletadas foi medida em mg / litro usando espectrofotômetro de absorção atômica. As soluções em branco e padrão para as soluções padrão de calibração do dispositivo, ou seja, 2,0 mg, 4,0 mg e 6,0, foram usadas para medir a concentração desses metais em amostras de água para verificar as medições. Os dados foram analisados estatisticamente em estatística descritiva (estimativa de proporções e desvio padrão) usada para resumir a concentração média. Os resultados obtidos tanto de temperatura quanto de pH das amostras de água variaram de 10 a 36 0C e 7,0 a 8,44, enquanto o tamanho dos estágios de alevinos variou de 4,0 a 56,0 mm no comprimento total. Os resultados dos metais investigados encontrados em amostras de água de lagoas são da ordem de Zn > Mn > Ni > Cu, respectivamente. A temperatura e o pH ótimos para a carpa-capim foram principalmente entre 15 0C e 30 0C e pH 6,5 a 8,0. Concluiu-se, a partir dos resultados obtidos, que temperatura, pH e traços de metais foram considerados adequados para o crescimento de Ctenopharyngodon idella desde os estágios de alevinos até alevinos, mas a maior quantidade de zinco pode causar sua mortalidade. É um estudo preliminar sobre a cultura da carpa-capim em incubatório de peixes Bannu, portanto forneceria informações úteis para modelo de unidade de produção de sementes de peixes em um incubatório.

Assessments of some trace metals in water samples of nursery pond of Grass Carp (Ctenopharyngodon idella, Valenciennes, 1844) in Bannu Fish Hatchery of Khyber Pakhtunkhwa, Pakistan

Abstract The present investigation was aimed to examine the concentrations of trace metals including e.g copper (Cu), manganese (Mn), nickel (Ni), and zinc (Zn) in water samples collected from nursery pond of grass carp (Ctenopharyngodon idella) in Bannu Hatchery of Khyber Pakhtunkhwa during the period from April 2018 to January 2019. The temperature and pH of each water sample were measured for the whole study duration. The concentration of Copper (Cu), Manganese (Mn), Nickel (Ni), and Zinc (Zn) in collected water samples were measured in mg/liter by using Atomic Absorption Spectrophotometer. The blank and standard solutions for device calibration Standard solutions i.e., 2.0 mg, 4.0 mg, and 6.0 were used to measure the concentration of these metals in water samples to verify the measurements. The data was statistically analyzed on descriptive statistics (estimation of proportions and standard deviation) used to summarize mean concentration. The results obtained of both temperature and pH of water samples were found in ranged 10 to 36 0C and 7.0 to 8.44; whereas the size of fry stages was ranged from 4.0 to 56.0 mm in total length. The results of investigated metals found in pond water samples are in order of Zn>Mn>Ni>Cu, respectively. As optimum temperature and pH for grass carp were mostly between 15 0C and 30 0C and pH 6.5 to 8.0. It was concluded from obtained results that temperature, pH, and trace metals were found appropriate for the growth of Ctenopharyngodon idella from fry to fingerling stages, but the highest amount of zinc can cause its mortality. It is a preliminary study on grass carp culturing in Bannu fish hatchery so, it would provide useful information for model fish seed production unit in a hatchery.

Resumo A presente investigação teve como objetivo examinar as concentrações de metais traço, incluindo cobre (Cu), manganês (Mn), níquel (Ni) e zinco (Zn) em amostras de água coletadas de viveiro de carpa-capim (Ctenopharyngodon idella) em Incubatório de Bannu de Khyber Pakhtunkhwa durante o período de abril de 2018 a janeiro de 2019. A temperatura e o pH de cada amostra de água foram medidos durante todo o período do estudo. A concentração de cobre (Cu), manganês (Mn), níquel (Ni) e zinco (Zn) nas amostras de água coletadas foi medida em mg / litro usando espectrofotômetro de absorção atômica. As soluções em branco e padrão para as soluções padrão de calibração do dispositivo, ou seja, 2,0 mg, 4,0 mg e 6,0, foram usadas para medir a concentração desses metais em amostras de água para verificar as medições. Os dados foram analisados estatisticamente em estatística descritiva (estimativa de proporções e desvio padrão) usada para resumir a concentração média. Os resultados obtidos tanto de temperatura quanto de pH das amostras de água variaram de 10 a 36 0C e 7,0 a 8,44, enquanto o tamanho dos estágios de alevinos variou de 4,0 a 56,0 mm no comprimento total. Os resultados dos metais investigados encontrados em amostras de água de lagoas são da ordem de Zn > Mn > Ni > Cu, respectivamente. A temperatura e o pH ótimos para a carpa-capim foram principalmente entre 15 0C e 30 0C e pH 6,5 a 8,0. Concluiu-se, a partir dos resultados obtidos, que temperatura, pH e traços de metais foram considerados adequados para o crescimento de Ctenopharyngodon idella desde os estágios de alevinos até alevinos, mas a maior quantidade de zinco pode causar sua mortalidade. É um estudo preliminar sobre a cultura da carpa-capim em incubatório de peixes Bannu, portanto forneceria informações úteis para modelo de unidade de produção de sementes de peixes em um incubatório.

Assessments of some trace metals in water samples of nursery pond of Grass Carp (Ctenopharyngodon idella, Valenciennes, 1844) in Bannu Fish Hatchery of Khyber Pakhtunkhwa, Pakistan / Avaliações de alguns vestígios de metais em amostras de água de viveiro de carpa-capim (Ctenopharyngodon idella, Valenciennes, 1844) em Bannu Fish Hatchery de Khyber Pakhtunkhwa, Paquistão

Abstract The present investigation was aimed to examine the concentrations of trace metals including e.g copper (Cu), manganese (Mn), nickel (Ni), and zinc (Zn) in water samples collected from nursery pond of grass carp (Ctenopharyngodon idella) in Bannu Hatchery of Khyber Pakhtunkhwa during the period from April 2018 to January 2019. The temperature and pH of each water sample were measured for the whole study duration. The concentration of Copper (Cu), Manganese (Mn), Nickel (Ni), and Zinc (Zn) in collected water samples were measured in mg/liter by using Atomic Absorption Spectrophotometer. The blank and standard solutions for device calibration Standard solutions i.e., 2.0 mg, 4.0 mg, and 6.0 were used to measure the concentration of these metals in water samples to verify the measurements. The data was statistically analyzed on descriptive statistics (estimation of proportions and standard deviation) used to summarize mean concentration. The results obtained of both temperature and pH of water samples were found in ranged 10 to 36 0C and 7.0 to 8.44; whereas the size of fry stages was ranged from 4.0 to 56.0 mm in total length. The results of investigated metals found in pond water samples are in order of Zn>Mn>Ni>Cu, respectively. As optimum temperature and pH for grass carp were mostly between 15 0C and 30 0C and pH 6.5 to 8.0. It was concluded from obtained results that temperature, pH, and trace metals were found appropriate for the growth of Ctenopharyngodon idella from fry to fingerling stages, but the highest amount of zinc can cause its mortality. It is a preliminary study on grass carp culturing in Bannu fish hatchery so, it would provide useful information for model fish seed production unit in a hatchery.

Resumo A presente investigação teve como objetivo examinar as concentrações de metais traço, incluindo cobre (Cu), manganês (Mn), níquel (Ni) e zinco (Zn) em amostras de água coletadas de viveiro de carpa-capim (Ctenopharyngodon idella) em Incubatório de Bannu de Khyber Pakhtunkhwa durante o período de abril de 2018 a janeiro de 2019. A temperatura e o pH de cada amostra de água foram medidos durante todo o período do estudo. A concentração de cobre (Cu), manganês (Mn), níquel (Ni) e zinco (Zn) nas amostras de água coletadas foi medida em mg / litro usando espectrofotômetro de absorção atômica. As soluções em branco e padrão para as soluções padrão de calibração do dispositivo, ou seja, 2,0 mg, 4,0 mg e 6,0, foram usadas para medir a concentração desses metais em amostras de água para verificar as medições. Os dados foram analisados ​​estatisticamente em estatística descritiva (estimativa de proporções e desvio padrão) usada para resumir a concentração média. Os resultados obtidos tanto de temperatura quanto de pH das amostras de água variaram de 10 a 36 0C e 7,0 a 8,44, enquanto o tamanho dos estágios de alevinos variou de 4,0 a 56,0 mm no comprimento total. Os resultados dos metais investigados encontrados em amostras de água de lagoas são da ordem de Zn > Mn > Ni > Cu, respectivamente. A temperatura e o pH ótimos para a carpa-capim foram principalmente entre 15 0C e 30 0C e pH 6,5 a 8,0. Concluiu-se, a partir dos resultados obtidos, que temperatura, pH e traços de metais foram considerados adequados para o crescimento de Ctenopharyngodon idella desde os estágios de alevinos até alevinos, mas a maior quantidade de zinco pode causar sua mortalidade. É um estudo preliminar sobre a cultura da carpa-capim em incubatório de peixes Bannu, portanto forneceria informações úteis para modelo de unidade de produção de sementes de peixes em um incubatório.

Hepatitis-Associated Aplastic Anemia: Etiology, Clinical Characteristics and Outcome.

Hepatitis-associated aplastic anemia (HAA) is a form of acquired aplastic anemia (AA) in which bone marrow failure develops after an acute attack of hepatitis. Bone marrow failure leading to AA is generally severe in cases of HAA and fatal if left untreated. This retrospective multicenter study investigated clinical and laboratory characteristics, possible causes, treatment, and outcome of HAA in children. Twenty patients from 8 centers were included in the study. Aspartate aminotransferase and alanine aminotransferase were <3 to 5×upper limit of normal (ULN) in 2 patients, <5 to 10×ULN in 2 patients, and >10×ULN in 16 patients. Acute liver failure developed in 5 (29%) patients. Pancytopenia was simultaneously present in 6 of 20 (30%) patients. Eleven of the 20 patients (55%) were alive, in remission and transfusion free. Those who were alive either had undergone hematopoietic stem cell transplantation and/or immunosuppressive treatment, except 1 patient who had received no treatment. Patients with the diagnosis of acute hepatitis should be evaluated and followed up carefully for presence of cytopenia, so that definitive treatment of AA can be initiated in a timely and appropriate manner when needed.

Clinical Features and Outcome of Children With Hereditary Spherocytosis.

OBJECTIVE: The aim of this study was to evaluate the demographics, clinical, and laboratory findings and treatment responses of patients with hereditary spherocytosis (HS). MATERIALS AND METHODS: Data of children with HS were examined. Diagnosis was based on clinical history, physical examination, family history, presence of spherocytes on peripheral blood smear, and osmotic fragility test. RESULTS: A total of 101 patients were included. The median (range) age at diagnosis was 38.0 (1 to 188) months. Mild, moderate, and severe forms of HS were present in 29 (28.7%), 15 (14.9%), and 57 (56.4%) patients, respectively. Family history was available in 73 patients and 56 of these (76.7%) had a positive family history for HS. Forty-five (44.5%) patients needed regular transfusions and all of these had severe disease. Although most patients did not require transfusion postsplenectomy, 2 of 45 (4.4%) patients continued to require transfusion. Transfusion dependence was significantly (P<0.001) higher in patients with severe spherocytosis. CONCLUSIONS: In HS, splenomegaly, pallor, and jaundice are the most common clinical features. Splenectomy dramatically reduces hemolysis in most cases and virtually abolishes further requirement for transfusion.

The Efficacy of Oral Vitamin B12 Replacement for Nutritional Vitamin B12 Deficiency.

Standard treatment of vitamin B12 deficiency has not been well established in childhood, the ideal amount of supplemental vitamin B12 is not clear. Vitamin B12 deficiency is classically treated with intramuscular injections. In this study, we aimed to investigate the efficacy of oral therapy in children with vitamin B12 deficiency. Patients with serum cobalamin concentrations <300 pg/mL aged between 6 months to 18 years were included in this prospective study. Children were treated orally either with a combination of multivitamin tablet daily or vitamin B12 ampules. Serum specimens were obtained at the end of first and third months of treatment for vitamin B12 levels. A total of 79 patients were included in the study. The mean pretreatment vitamin B12 level increased from 182±47.6 pg/mL to 482±318 pg/mL after 1 month of treatment in the whole cohort. Comparison of the pretreatment vitamin B12 levels with first and third month posttreatment values showed significant difference (P-value, 0.001 and 0.028, respectively). In this study, oral cyanocobalamin was found effective for the treatment of vitamin B12 deficiency in children.

Utility of baseline serum phosphorus levels for predicting remission in acromegaly patients.

PURPOSE: High GH and IGF I levels increase tubular phosphate reabsorption in patients with acromegaly. We aimed to investigate the utility of serum phosphorus levels as an indicator for predicting chance of remission in acromegaly patients. DESIGN: Fifty-one patients (n: 51; F: 24, M: 27) with diagnosis of acromegaly were included in the study. Plasma IGF-1, Phosphorus (P) and nadir GH levels on oral glucose tolerance test (OGTT) at the time of diagnosis were analysed retrospectively. Patients were classified into two groups according to their plasma P levels; P ≤ 4.5 mg/dl (Group-1, n: 23, 45.1%), P > 4.5 mg/dl (Group-2, n: 28, 54.9%). Two groups were compared according to remission status; remission (n: 27) and non-remission (n: 24). Remission was defined with absence of clinical symptoms, normal plasma IGF-1 (adjusted for age and gender) and GH levels (<1 mcg/dl) at least 3 months after initial treatment. RESULTS: Serum P levels decreased significantly after treatment in both groups (p < 0.001). There was a significant correlation between baseline phosphorus levels and remission rates, nadir GH in OGTT, pituitary adenoma size and Ki-67 scores (p = 0.001, r: -0.51; p = 0.01, r: 0.44; p = 0.001, r: 0.52; p = 0.02, r: 0.71, respectively). Mean baseline P levels were significantly higher in patients with non-remission (4.8 vs 4.2, P < 0.001). Logistic regression analysis did not reveal an independent effect on remission with any of these risk factors. CONCLUSION: High serum P levels may be an indicator for a low likelihood of onset of remission in acromegaly patients. Further studies with wider spectrum are needed to make specific suggestions.

INCIDENTAL PAGET'S DISEASE DISGUISED AS BONE METASTASIS IN A PATIENT WITH ENDOMETRIUM CARCINOMA.

Paget's disease is a disorder of aging bone which occurs in the setting of accelarated bone remodelling. In the presented case we discuss the difficulties in the diagnosis of Paget's disease in a 77 year old patient with coexisting endometrium carcinoma. The patient was initially diagnosed with metastatic bone disease due to endometrium adenocarcinoma when she was admitted to oncology clinic with pelvic pain. Bone scintigraphy with Tc99 and (18)F fluorodeoxyglucose positron emission tomography/CT revealed an increased uptake on the bone lesions which were reported as metastatic bone involvement. Although the (18)F-FDG uptake was much higher than the levels that would generally be anticipated in a case with Paget's disease, high levels of bone turnover markers indicated further evaluation in the differential diagnosis and the definitive diagnosis of Paget's disease was established with the pathological evaluation of bone biopsy.

Identification of ATP8B1 as a blood-brain barrier-enriched protein.

In order to define the molecular anatomy of the blood-brain barrier (BBB) that may be relevant to either barrier or transport function, proteins that are overexpressed in the cerebral microvessels should be identified. We used differential display to identify novel proteins that are overexpressed or unique to the BBB. DNA sequence analysis is one of the differentially expressed transcripts showed that it is highly homologous with the ATPase class I, type 8B, and member 1 (ATP8B1) protein and contains an ATPase domain and a phospholipid-binding domain. ATP8B1 is expressed in the BBB microvessels but not brain tissue lacking microvessels. Likewise, ATP8B1 was enriched in BBB microvessels similar to glucose transporter 1. Immunohistochemistry using an ATP8B1-specific antibody demonstrated preferential staining of the microvessels within the cerebral tissue. These results suggest that ATP8B1, a P-type aminophospholipid translocase, is enriched in cerebral microvessels and may have a role in plasma membrane lipid transport.

A dose-finding study of temsirolimus and liposomal doxorubicin for patients with recurrent and refractory bone and soft tissue sarcoma.

There are few effective therapies for high-risk sarcomas. Initial chemosensitivity is often followed by relapse. In vitro, mammalian target of rapamycin (mTOR) inhibition potentiates the efficacy of chemotherapy on resistant sarcoma cells. Although sarcoma trials using mTOR inhibitors have been disappointing, these drugs were used as maintenance. We conducted a Phase I/II clinical trial to test the ability of temsirolimus to potentiate the cytotoxic effect of liposomal doxorubicin and present here the dose-finding portion of this study. Adult and pediatric patients with recurrent or refractory sarcomas were treated with increasing doses of liposomal doxorubicin and temsirolimus using a continual reassessment method for escalation, targeting a dose-limiting toxicity rate of 20%. Blood samples were drawn before and after the first dose of temsirolimus in Cycles 1 and 2 for pharmacokinetic analysis. The maximally tolerated dose combination was liposomal doxorubicin 30 mg/m(2) monthly with temsirolimus 20 mg/m(2) weekly. Hematologic toxicity was common but manageable. Dose-limiting toxicities were primarily renal. Concurrent administration of liposomal doxorubicin resulted in increased exposure to sirolimus, the active metabolite of temsirolimus. Thus, the combination of liposomal doxorubicin and temsirolimus is safe for heavily pretreated sarcoma patients. Co-administration with liposomal doxorubicin did not alter temsirolimus pharmacokinetics, but increased exposure to its active metabolite.

The effects of human amniotic membrane and periosteal autograft on tendon healing: experimental study in rabbits.

In this study, the effects of periosteal autograft and human amniotic membrane on tendon healing were compared. Forty-two New Zealand rabbits were divided into three groups. Flexor digitorum fibularis tendons were cut and repaired with a modified Kessler technique and circumferential sutures in all groups. Tendon repair alone was carried out in group 1, tendon repair and application of human amniotic membrane was done in group 2 and tendon repair and application of periosteal autograft was done in group 3. Biomechanical and histopathological examinations were done 2 and 6 weeks postoperatively. Biomechanical examination showed that group 3 was the strongest at weeks 2 and 6. Adhesion, inflammation and new bone formation showed no difference between groups at week 2. However, adhesion formation was found to be less in groups 2 and 3 than group 1 at week 6. The application of periosteal autograft might be useful in repair of tendon injuries.

Lipopolysaccharide alters the blood-brain barrier transport of amyloid beta protein: a mechanism for inflammation in the progression of Alzheimer's disease.

Alzheimer's disease (AD) brains are characterized by accumulation of amyloid beta protein (Abeta) and neuroinflammation. Increased blood-to-brain influx and decreased brain-to-blood efflux across the blood-brain barrier (BBB) have been proposed as mechanisms for Abeta accumulation. Epidemiological studies suggest that the nonsteroidal anti-inflammatory drug (NSAID) indomethacin slows the progression of AD. We hypothesized that inflammation alters BBB handling of Abeta. Mice treated with lipopolysaccharide (LPS) had increased brain influx and decreased brain efflux of Abeta, recapitulating the findings in AD. Neither influx nor efflux was mediated by LPS acting directly on BBB cells. Increased influx was mediated by a blood-borne factor, indomethacin-independent, blocked by the triglyceride triolein, and not related to expression of the blood-to-brain transporter of Abeta, RAGE. Serum levels of IL-6, IL-10, IL-13, and MCP-1 mirrored changes in Abeta influx. Decreased efflux was blocked by indomethacin and accompanied by decreased protein expression of the brain-to-blood transporter of Abeta, LRP-1. LPS paradoxically increased expression of neuronal LRP-1, a major source of Abeta. Thus, inflammation potentially increases brain levels of Abeta by three mechanisms: increased influx, decreased efflux, and increased neuronal production.

Signal sequence mutation in autosomal dominant form of hypoparathyroidism induces apoptosis that is corrected by a chemical chaperone.

Autosomal dominant familial isolated hypoparathyroidism (AD-FIH) is caused by a Cys --> Arg mutation (C18R) in the hydrophobic core of the signal peptide of human preproparathyroid hormone (PPTH). Although this mutation impairs secretion of the hormone, the mechanism by which one mutant allele produces the autosomal-dominant disease is unexplained. Using transfected HEK293 cells, we demonstrate that the expressed mutant hormone is trapped intracellularly, predominantly in the endoplasmic reticulum (ER). This ER retention was found to be toxic for the cells, which underwent apoptosis, as evident from the marked increase in the number of cells staining positive for Annexin V binding and for the TUNEL reaction. The cells producing mutant hormone also had marked up-regulation of the ER stress-responsive proteins, BiP and PERK, as well as the proapoptotic transcription factor, CHOP. Up-regulation of these markers of the unfolded protein response supported a causal link between the ER stress and the cell death cascade. When the C18R PPTH was expressed in the presence of 4-phenylbutyric acid, which is a pharmacological chaperone, intracellular accumulation was reduced and normal secretion was restored. This treatment also produced remarkable reduction of ER stress signals and protection against cell death. These data implicate ER stress-induced cell death as the underlying mechanism for AD-FIH and suggest that the pharmacological manipulation of this pathway by using chemical chaperones offers a therapeutic option for treating this disease.

Carbonic anhydrase XIV in skeletal muscle: subcellular localization and function from wild-type and knockout mice.

The expression of carbonic anhydrase (CA) XIV was investigated in mouse skeletal muscles. Sarcoplasmic reticulum (SR) and sarcolemmal (SL) membrane fractions were isolated from wild-type (WT) and CA XIV knockout (KO) mice. The CA XIV protein of 54 kDa was present in SR and SL membrane fractions as shown by Western blot analysis. CA activity measurements of WT and KO membrane fractions showed that CA XIV accounts for approximately 50% and 66% of the total CA activities determined in the SR and SL fractions, respectively. This indicates the presence of at least one other membrane-associated CA isoform in these membranes, e.g., CA IV, CA IX, or CA XII. Muscle fibers of the extensor digitorum longus (EDL) muscle were immunostained with anti-CA XIV/FITC and anti-sarco(endo)plasmic reticulum Ca(2+)-ATPase 1/TRITC, with anti-CA XIV/FITC and anti-ryanodine receptor/TRITC, or with anti-CA XIV/FITC and anti-monocarboxylate transporter-4/TRITC. CA XIV was expressed in the plasma membrane and in the longitudinal SR but not in the terminal SR. Isometric contraction measurements of single twitches and tetani and a fatigue protocol applied to fiber bundles of the fast-twitch EDL and of the slow-twitch soleus muscle from WT and KO mice showed that the lack of SR membrane-associated CA XIV did not affect maximum force, rise and relaxation times, and fatigue behavior. Thus, it is concluded that a reduction of the total SR CA activity by approximately 50% in CA XIV KO mice does not lead to an impairment of SR function.

Hypoglycemia due to ectopic release of insulin from a paraganglioma.

Insulin-secreting pancreatic tumors and insulin-like growth hormone-secreting non-islet cell tumors can cause hypoglycemia. However, insulin-releasing paraganglioma or pheochromocytoma has almost never been reported. A 67-year-old female patient was admitted to our hospital because of headache, palpitation, perspiration, faintness, frequent sense of hunger and absent-mindedness. These intermittent symptoms had begun approximately a year before admission. On physical examination, she had high blood pressure of 150/90 mm Hg. Hormonal studies demonstrated increased urinary norepinephrine levels, and hyperinsulinemic hypoglycemia was confirmed while the patient was symptomatic. Abdominal MRI revealed a retroperitoneal mass measuring 4.5 cm in the pancreatic region. She was treated with an alpha-blocking agent to control blood pressure preceding the removal of the mass. Histopathological diagnosis was paraganglioma, and immunohistochemically insulin staining in the neoplastic cells was demonstrated. Her blood pressure normalized and hypoglycemia relieved after the operation. The patient did not have recurrence of hypoglycemia after a year of follow-up. Paraganglioma is a rare tumor of the neural crest, and co-secretion of insulin and catecholamines has been reported only by a single case report in the literature. The present patient is another case with this co-secretion.

Management of tibial fractures with Orthofix axial external skeletal fixator in 15 dogs.

In the present study, the applicability of fixator, surgical time, time to weightbearing, the healing time and complications of tibial fractures in dogs after fixation with Orthofix axial external skeletal fixator were evaluated. The study was performed on 15 cases which were presented to the Clinics of Small Animal Surgery. After the determination of the fracture type, fracture location, appropriate screw diameter and the location of the first screw on the fragment according to preoperative radiographs, the dogs were placed under general anaesthesia and the fixator was applied with cranio-medial approach to the tibia. Clinical and radiological examinations were continued until the removal of the external skeletal fixators at three different times (at approx. days 45, 60 and 75) according to the clinical and radiological fracture healing. The treatment results were determined by the Modified Hannover Veterinary Medicine Faculty Small Animal Fracture Evaluation Scale. The results were excellent in eight cases (53.3%), good in four cases (26.7%), fair in one case (6.7%) and poor in two cases (13.3%). Based on this study, the use of external skeletal fixation with Orthofix axial external skeletal fixator can be recommended for dogs with tibial fractures as long as careful pin tract management is performed. Further investigation is needed to determine the use of Orthofix axial external skeletal fixator for the management of other long bone fractures.

Carbonic anhydrase II deficiency syndrome (osteopetrosis with renal tubular acidosis and brain calcification): novel mutations in CA2 identified by direct sequencing expand the opportunity for genotype-phenotype correlation.

The carbonic anhydrase II (CA II) deficiency syndrome is an autosomal recessive disorder that produces osteopetrosis, renal tubular acidosis, and cerebral calcification. Other features include developmental delay, short stature, cognitive defects, and a history of multiple fractures by adolescence. With one exception, all patients with osteopetrosis and renal tubular acidosis examined have proven to have CA II deficiency. All CA II-deficient patients analyzed have been found to have mutations in the CA2 gene. Previously, we used single strand conformational (SSCP) analysis to identify exons to be sequenced from CA II-deficient patients. In this report, we amplified all seven exons by PCR from genomic DNA and directly sequenced the amplified products. Application of this method allowed identification of eleven new mutations in 21 patients referred for confirmation of the diagnosis of CA II deficiency. These mutations were scattered over the genome from exon 2 to 7. In two opportunities for prenatal diagnosis, one from cultured amniocytes and one from chorionic villus biopsy, we demonstrated the general utility of the direct sequencing method for prenatal DNA diagnosis. These studies expand our knowledge of the heterogeneity in mutations underlying the CA II deficiency syndrome.