Key Matrix Proteins Within the Pancreatic Islet Basement Membrane Are Differentially Digested During Human Islet Isolation.

Clinical islet transplantation achieves insulin independence in selected patients, yet current methods for extracting islets from their surrounding pancreatic matrix are suboptimal. The islet basement membrane (BM) influences islet function and survival and is a critical marker of islet integrity following rodent islet isolation. No studies have investigated the impact of islet isolation on BM integrity in human islets, which have a unique duplex structure. To address this, samples were taken from 27 clinical human islet isolations (donor age 41-59, BMI 26-38, cold ischemic time < 10 h). Collagen IV, pan-laminin, perlecan and laminin-α5 in the islet BM were significantly digested by enzyme treatment. In isolated islets, laminin-α5 (found in both layers of the duplex BM) and perlecan were lost entirely, with no restoration evident during culture. Collagen IV and pan-laminin were present in the disorganized BM of isolated islets, yet a significant reduction in pan-laminin was seen during the initial 24 h culture period. Islet cytotoxicity increased during culture. Therefore, the human islet BM is substantially disrupted during the islet isolation procedure. Islet function and survival may be compromised as a consequence of an incomplete islet BM, which has implications for islet survival and transplanted graft longevity.

Contrasting effects of A1 and A2b adenosine receptors on adipogenesis.

BACKGROUND: Adenosine mediates its actions through four G protein-coupled receptors, A1, A2a, A2b and A3. The A1 receptor (A1R) is dominant in adipocytes where it mediates many actions that include inhibition of lipolysis, stimulation of leptin secretion and protection against obesity-related insulin resistance. OBJECTIVE: The objective of this study is to investigate whether induced expression of A1Rs stimulates adipogenesis, or whether A1R expression is a consequence of cells having an adipocyte phenotype. METHODOLOGY: Human A1R and A2b receptors (A2bRs) were stably transfected into a murine osteoblast precursor cell line, 7F2. Adipogenesis was determined by lipid accumulation and expression of adipocyte and osteoblast marker molecules. Adenosine receptor expression and activation of associated signal molecules were also evaluated as 7F2 cells were induced to differentiate to adipocytes. RESULTS: 7F2 cells transfected with the A1R showed increased adipocyte marker mRNA expression; lipoprotein lipase and glycerol-3-phosphate dehydrogenase were both upregulated, whereas the osteoblast marker alkaline phosphatase (ALP) was downregulated. When cultured in adipocyte differentiating media, such cells also showed increased adipogenesis as judged by lipid accumulation. Conversely, A2bR transfection stimulated osteocalcin and ALP expression, and in addition, adipogenesis was inhibited in the presence of adipocyte differentiation media. Adipogenic differentiation of naive 7F2 cells also resulted in increased expression of the A1R and reduced or modified expression of the A2a and A2bR. The loss of A2 receptors after adipogenic differentiation was accompanied by a loss of cyclic adenosine monophosphate and ERK1/2 signalling. CONCLUSION: These data show that expression of A1Rs induced adipocyte differentiation, whereas A2bR expression inhibited adipogenesis and stimulated an osteoblastic phenotype. These data suggest that targeting A1 and A2bR could be considered in the management of obesity and diabetes. Targeting adenosine signal pathways may be useful in treatment strategies for diseases in which there is an imbalance between osteoblasts and adipocytes.

Staining characteristics of Lonchocarpus cyanescens leaf extract on the testis of Sprague-Dawley rats.

The use of nonallergic, nontoxic, and eco-friendly natural dyes has become a matter of significant importance due to increased environmental awareness on the need to avoid hazardous synthetic dyes. This study was to determine the staining properties of the dye extract of Lonchocarpus cyanescens on histomorphology of the testis. Freshly cut leaves of L. cyanescens obtained from Akpan Ifia Inan village in Ikono local government area of Akwa Ibom state (latitude 5° 10' 12″ N; longitude 7° 48' 0″ E) were put into a plastic jar and boiling water was poured to cover the leaves. It was covered and left for an hour. The liquid was strained and potassium hydroxide was added to the dye water mixture to reach a pH of 9. A whisk was used to mix air into the liquid, and the mixuture was then allowed to sit until the blue indigo had settled to the bottom of the container. The dye was diluted with 70% ethanol to a concentration of 0.1 g/mL and was used to stain sections of testes. Its potential for use as a counterstain was also investigated. The testes sections were stained in shades of blue. The dye overshadowed the colors of haematoxylin and eosin. Preliminary phytochemical screening of L. cyanescens revealed that it contains alkaloids, saponins, flavonoids, and tannins.

A comparison of the anti-diabetic potential of d-ribose-l-cysteine with insulin, and oral hypoglycaemic agents on pregnant rats.

Over 18% of pregnant women are affected by diabetes mellitus (DM) and Insulin has been the commonest drug used in its treatment. There are reports of noncompliance to insulin due to trypanophobia, with suggestions for the use of oral hypoglycaemic agents (OHAs). However, the opposing views about the benefits and risk of oral hypoglycaemic agents (OHAs) warrant a continuous search for an alternative regimen. Therefore, this study is aimed at comparing the antidiabetic effects of d-ribose-l-cysteine (riboceine) with vildagliptin, glibenclamide, metformin, glipizide and insulin in diabetes in pregnancy. Forty (40) female Sprague-Dawley (SD) rats were mated with twenty (20) male SD rats. Diabetes was induced by streptozotocin and the female SD rats were divided into 8 groups of five (5) rats each. The animals were administered either of the OHAs vildagliptin, glibenclamide, metformin, glipizide and riboceine for a period of 19 gestational days. The results showed that streptozotocin (STZ) significantly (p < 0.05) decreased the weights of the animals, increased malondialdehyde, blood glucose levels and altered reproductive hormones. These effects of STZ were better ameliorated in animals that received insulin and riboceine compared to the other OHAs. While progesterone levels were significantly (p < 0.05) higher in animals that received riboceine compared to insulin. Glibenclamide increased (p < 0.05) foetal weights compared to non-diabetic animals. In conclusion, glibenclamide may be a threat to mother`s life in the management of diabetes in pregnancy however, riboceine as well as vildagliptin, metformin and glipizide are effective oral hypoglycaemic agents which could serve as a potent adjuvant comparable to insulin in the management of diabetes during gestation.

Disseminated Herpesvirus hominis 2 infection following drug overdose.

Fatal disseminated herpesvirus hominis type 2 infection developed in a 24-year-old woman with psychiatric disease. The primary site of virus entry appeared to be the upper respiratory tract, although the cervix and anus could have been primary sites. Dissemination of infection was facilitated by steroid therapy. An endotracheal tube may have increased spread to the lower respiratory tract and an intrauterine device may have aided spread to the uterus. Intranuclear inclusion body-bearing cells in the respiratory secretions should alert the clinician to possible herpesvirus disease, local and disseminated, even in the nonimmunecompromised patient.

Secondary solid cancer screening following hematopoietic cell transplantation.

Hematopoietic stem cell transplant (HCT) recipients have a substantial risk of developing secondary solid cancers, particularly beyond 5 years after HCT and without reaching a plateau overtime. A working group was established through the Center for International Blood and Marrow Transplant Research and the European Group for Blood and Marrow Transplantation with the goal to facilitate implementation of cancer screening appropriate to HCT recipients. The working group reviewed guidelines and methods for cancer screening applicable to the general population and reviewed the incidence and risk factors for secondary cancers after HCT. A consensus approach was used to establish recommendations for individual secondary cancers. The most common sites include oral cavity, skin, breast and thyroid. Risks of cancers are increased after HCT compared with the general population in skin, thyroid, oral cavity, esophagus, liver, nervous system, bone and connective tissues. Myeloablative TBI, young age at HCT, chronic GVHD and prolonged immunosuppressive treatment beyond 24 months were well-documented risk factors for many types of secondary cancers. All HCT recipients should be advised of the risks of secondary cancers annually and encouraged to undergo recommended screening based on their predisposition. Here we propose guidelines to help clinicians in providing screening and preventive care for secondary cancers among HCT recipients.

Perindopril ameliorates glomerular and renal tubulointerstitial injury in the SHR/N-corpulent rat.

We compared the effects of long-term treatment with the angiotensin-converting enzyme inhibitor perindopril and triple therapy (hydrochlorothiazide, reserpine, and hydralazine) on the metabolic and renal features in the SHR/N-corpulent (cp) rat, a genetic model of non-insulin-dependent diabetes mellitus and hypertension. Obese male SHR/N-cp rats (4 to 6 weeks old) were fed a 54% carbohydrate diet containing 18% sucrose and 36% starch. After 2 months on the diet, rats were assigned to one of three groups: one group (n=8) received perindopril (PE); the second group (n=8) received triple therapy (TT); and the third group (n=8) did not receive therapy. Treatment was maintained for 3 to 4 months. Body weight, food intake, and fasting levels of serum glucose and insulin did not differ among the three groups. Control rats exhibited progressive proteinuria in parallel with the rise in systolic blood pressure (SBP). Both PE and TT equally lowered SBP to normal levels and reduced proteinuria in treated rats. However, the reduction of proteinuria was greater and more sustained with PE than with TT (P<.05), whereas the effect of TT on proteinuria was delayed. Plasma renin activity was increased in PE and TT rats compared with control rats (P<.02). Semiquantitative analysis of renal lesions showed that the percentage of glomeruli with mesangial expansion and sclerosis and the tubulointerstitial score (an index of severity of tubulointerstitial lesions, namely tubular atrophy, inflammatory cellular infiltrates, and interstitial fibrosis) was reduced in both PE and TT rats. However, the reduction of glomerulosclerosis and tubulointerstitial lesions was greater in PE than in TT rats (P<.01). The percentage of glomerular sclerosis was positively correlated with the severity score of tubulointerstitial lesions (r=.60, P<.01). We conclude that PE is more effective than TT in halting the progression of proteinuria in the SHR/N-cp rat with non-insulin-dependent diabetes mellitus and hypertension. The antiproteinuric effect of PE is associated with significant reduction in glomerulosclerosis and tubulointerstitial lesions, independent of the effect of treating hypertension.

Pancreatic isleitis with coxsackie virus B5 infection.

Coxsackie group B virus infection may be responsible for some cases of insulin-dependent diabetes mellitus. However, Coxsackie B5 virus is rarely implicated in this respect. The authors observed striking pancreatic islet cell damage in an infant who died of a Coxsackie B5 virus infection. The inflammatory response consisted of mononuclear cells, and the acinar tissue was completely uninvolved.

Development and characteristics of a new strain of obese hyperinsulinemic and hyperlipidemic Dahl salt-sensitive rat. The Dahl salt-sensitive/NIH-corpulent rat.

A new congenic rat strain, the Dahl salt-sensitive/NIH-corpulent (DSS/N-cp) rat, has been developed to study the role of obesity and type of dietary carbohydrate in the development of hypertension and its complications. Three groups (n = 6) of young male obese and lean DSS/N-cp rats were fed diets containing either 54% sucrose, 18% sucrose plus 36% starch, or 54% starch, with 0.1% dietary sodium for 12 weeks. Regardless of the diet, obese and lean rats showed mildly elevated systolic blood pressure (SBP), being significantly higher in obese than in lean rats (SBP 156 +/- 5 mm Hg v 141 +/- 3 mm Hg, P < .05). However, SBP was not different between the three diet groups. Levels of serum insulin, triglyceride, and cholesterol as well as urinary protein excretion were significantly higher in obese than in lean rats. Obese rats fed the sucrose diets as compared to the starch diet, had higher serum insulin and lipid levels, but had lower body weights and higher serum creatinine levels. Histopathologic examination of tissues from different organs revealed a vasculopathy seen almost exclusively in obese rats fed the sucrose diets. Vascular lesions were characterized by subintimal fibrin deposition, fibrinoid necrosis, and cell proliferation with "onion skinning" in small arteries and arterioles of kidneys, intestine, pancreas, and testes.(ABSTRACT TRUNCATED AT 250 WORDS)

Bullous dermatosis and systemic lupus erythematosus in a 15-year-old boy.

A 15-year-old boy had a bullous eruption suggestive of bullous pemphigoid and established systemic lupus erythematosus (SLE). Direct immunofluorescence studies of the bullae and adjacent skin revealed the linear deposition of IgG and complement localized to the basement membrane zone. Indirect immunofluorescence examination of the serum failed to reveal circulating basement membrane zone antibodies. The differential diagnosis of the bullous eruption is reviewed, and the problem of diagnosis in cases of coexistent bullous disease and SLE is discussed.

Retinitis secondary to acquired systemic toxoplasmosis with isolation of the parasite.

A 43-year-old woman developed a heterophile-negative infectious mononucleosis syndrome for which no cause was apparent. During her illness she developed subjective changes in the central vision of her right eye and had focal retinal inflammation, which suggested the diagnosis of toxoplasmosis. The clinical course was accompanied by an increased titer to Toxoplasma gondii. Organisms were isolated from lymph node tissue and injected into the peritoneum of mice from which organisms were harvested.

A comparison of the usefulness of electron microscopy and immunohistochemistry. One laboratory's experience.

As a quality assurance measure, the usefulness of transmission electron microscopy (EM) and immunohistochemistry (IHC) in our surgical pathology laboratory was compared. The surgical pathology reports from 150 consecutive neoplasms that were examined by both EM and IHC were reviewed. Based on the reported clinical histories, final diagnoses, light microscopy results, and the findings by EM or IHC, the contributions of EM and IHC were classified as "helpful" or "not helpful" for each specimen. Electron microscopy was helpful (92%) more often than was IHC (73%). Electron microscopy was most useful in further classifying poorly differentiated carcinomas, while IHC was particularly useful in classifying poorly differentiated neoplasms. Electron microscopy and IHC were of limited value in identifying the origin of metastatic carcinomas of an uncertain primary. All cases determined to be "not helpful" by either modality were further analyzed to establish a reason for the lack of information provided in each case. This analysis demonstrated a need for improved technical quality and ordering patterns of immunohistochemical stains in our laboratory.

Design of a self-cleaning thermoresponsive nanocomposite hydrogel membrane for implantable biosensors.

Following implantation of a biosensor, adhesion of proteins and cells and eventual fibrous encapsulation will limit analyte diffusion and impair sensor performance. A thermoresponsive nanocomposite hydrogel was developed as a self-cleaning biosensor membrane to minimize the effect of the host response and its utility for an optical glucose sensor, demonstrated here. It was previously reported that thermoresponsive nanocomposite hydrogels prepared from photopolymerization of an aqueous solution of N-isopropylacrylamide (NIPAAm) and polysiloxane colloidal nanoparticles released adhered cells with thermal cycling. However, poly(N-isopropylacrylamide) hydrogels exhibit a volume phase transition temperature (VPTT) of approximately 33-34 degrees C, which is below body temperature. Thus, the hydrogel would be in a collapsed state in vivo, which would ultimately limit diffusion of the target analyte (e.g., glucose) to the encapsulated sensor. In this study, the VPTT of the nanocomposite hydrogel was increased by introducing N-vinylpyrrolidone (NVP) as a comonomer, so that the hydrogel was in the swollen state in vivo. This thermoresponsive nanocomposite hydrogel was prepared by the photopolymerization of an aqueous solution of NIPAAm, NVP, and polysiloxane colloidal nanoparticles. In addition to a VPTT a few degrees above body temperature, the hydrogel also exhibited good mechanical strength, glucose diffusion, and in vitro cell release upon thermal cycling. Thus, this nanocomposite hydrogel may be useful as a biosensor membrane to minimize biofouling and extend the lifetime and efficiency of implantable glucose sensors and other biosensors.

Effect of the juice of lime (Citrus aurantifolia) on estrous cycle and ovulation of Sprague-Dawley rats.

OBJECTIVE: To determine the effect of lime juice on the estrous cycle and ovulation of cyclic female rats. METHODS: Twenty-five adult female Sprague-Dawley rats were used. The study was divided into 2 experiments (I and II). In experiment I, 15 rats were randomly subclassified into 3 groups (Ia, Ib, and Ic) of 5 rats each. The estrous cycles of the rats were studied for the first 16 days to establish cyclicity, after which lime juice was administered by gastric gavage for the next 24 days. Rats in group Ia received 1 mL of undiluted lime juice, rats in group Ib received 1 mL of 50% diluted lime juice, and rats in group Ic (control animals) received only distilled water. In experiment II, 10 female rats were used and were categorized into 2 groups (IIa and IIb), with 5 rats in each group. Rats in group IIa received 1 mL of undiluted lime juice during the morning of proestrus, and those in group IIb received only distilled water on the day of proestrus. The rats were killed the next day with use of chloroform anesthesia. The upper parts of the oviducts were excised and examined under the light microscope for assessment of the number of ova shed. RESULTS: There was an irregular pattern in all phases of the estrous cycle of 100% of the rats given undiluted lime juice and in 80% of those given 50% diluted lime juice. There was a significant (P = .001) reduction in the number of ova shed in rats administered undiluted lime juice in comparison with the control animals. Ovulation was partially blocked, as shown by the reduced number of ova observed in the oviducts from the rats given undiluted lime juice (5.10 +/- 2.37) in comparison with the control rats (12.70 +/- 1.14). CONCLUSION: In rats, lime juice causes irregularity of the estrous cycle, partially blocks ovulation, and may possibly compromise fertility.

Antiovulatory and anti-implantation potential of the methanolic extract of seeds of Abrus precatorius in the rat.

OBJECTIVE: To investigate the effect of the methanolic extract of seeds of Abrus precatorius on the estrous cycle, ovulation, and implantation of fetuses in Sprague-Dawley rats. METHODS: Cyclic female rats were randomly classified into 4 groups (A through D). Treated rats in group A had daily vaginal smears for a total of 64 consecutive days while being fed A precatorius extract for the first 32 of those days. Treated rats in group B received a single oral dose of the extract on the day of proestrus and were killed the following morning so that shed ova could be counted. Treated rats in group C received A precatorius extract from postcoital day 1 to 10 and were killed on day 12 to assess for anti-implantation effect, whereas the treated dams in group D received the extract from the 6th to the 19th day of gestation. The control animals in all 4 groups received an equal volume of distilled water. RESULTS: The methanolic extract of A precatorius caused a reversible disruption in the estrous cycle of the regularly cyclic rats and completely blocked ovulation in all the treated rats. Despite successful mating of the female rats with male rats of proven fertility, uterine dissection on postcoital day 12 revealed neither implantation nor resorption sites in all the animals treated with A precatorius. The extract of A precatorius caused a decrease in mean body weight, mean crown-rump length, and mean tail length of fetuses of the treated rats. CONCLUSION: There is a need to continue the search for new antifertility agents that have minimal side effects and widespread acceptability in addition to being reversible, affordable, and accessible. In this study, methanolic extract of A precatorius seeds caused reversible alterations in the estrous cycle pattern and completely blocked ovulation in Sprague-Dawley rats. In addition, the extract demonstrated anti-implantation activity and the potential to affect gross fetal morphometry in rats.

Enterovirus infection in a semi-closed community.

Infection with enteroviruses was studied over a 61-week period (during 1960-61) in a semi-closed child community in the Detention Home of the Allegheny County Juvenile Court in Pittsburgh, Pennsylvania. While most of the viruses isolated were known adeno- and enterovirus types, three apparently represent new enterovirus types or 'prime strains'. Viruses were isolated in all but 6 weeks of the 61-week study period from the gastro-intestinal tracts of 110 children out of a total population of 514 (21%); of these 110, 24 children (22%) excreted virus at the time of admission. The population averaged 37 children (more than half of whom were under 5 years of age), with a turnover of about nine per week. Spread of infection in this community on introduction of a new virus was demonstrated, with virus shedding of variable duration after infection. Thirteen of the 110 positive children (12%) showed mixed virus infections. During the entire study period, no clinical diarrheal illness was found associated with the viral infections detected.

Pancreatic necrosis in progressive systemic sclerosis.

Fatal pancreatic necrosis, secondary to extensive acute arteritic changes, is reported in a case of progressive systemic sclerosis. The patient presented first with hypertension and renal involvement, with active vascular lesions demonstrated by biopsy. The renal lesion at necropsy was inactive, showing the characteristic concentric fibrosis only, while the pancreatic vascular lesions were both chronic proliferative and acute in type.