MRI of oriental cholangiohepatitis.

Oriental cholangiohepatitis (OCH) also called recurrent pyogenic cholangitis is characterized by intrahepatic duct calculi, strictures, and recurrent infections. In turn cholangitis can result in multiple hepatic abscesses, further biliary strictures, and in severe cases, progressive hepatic parenchymal destruction, cirrhosis, and portal hypertension. Magnetic resonance cholangiopancreatography (MRCP) and conventional T1-weighted (T1W) and T2-weighted (T2W) magnetic resonance imaging (MRI) findings have been described in patients with OCH. MRCP findings include duct dilation, strictures, and calculi. MRCP can help to localize the diseased ducts and determine the severity of involvement. T1 and T2W sequences reveal the parenchymal changes of atrophy, abscess formation, and portal hypertension in addition to calculi. Post-treatment changes are also well depicted using MRI. Comprehensive, non-invasive assessment is achieved by using conventional MRI and MRCP in OCH providing a roadmap for endoscopic or surgical management.

Chemical activation of in vitro matured dromedary camel (Camelus dromedarius) oocytes: optimization of protocols.

Experiments were conducted to study the efficiency of sequential treatments of ionomycine and ethanol combined with phosphorylation inhibitor (6-dimethylaminopurine) or the specific maturation promoting factor inhibitor (roscovitine) in inducing artificial activation in dromedary M-II oocytes. Cumulus oocyte complexes (COCs), collected from slaughterhouse ovaries were cultured at 38.5 degrees C in an atmosphere of 5% CO2 in air for 24-48 h. In experiment 1, the COCs were either fertilized in vitro or activated with 5 microM ionomycine for 5 min or 7% ethanol for 7 min, both followed by exposure to 6-diethylaminopurine or roscovitine for 4h. After 14-15 h of in vitro culture, the oocytes were fixed and stained with 1% aceto-orcein to evaluate their nuclear status. In experiment 2, the oocytes were activated in the same manner as in experiment 1 but were cultured for 7 days to evaluate their post-parthenogenetic development. In experiment 3, oocytes were exposed to the ionomycine for 2, 3, 4 or 5 min to evaluate the better exposure time while as in experiment 4, the oocytes matured for 28-48 h were activated to see the effect of aging on post-parthenogenetic development. Higher proportion (P<0.01) of oocytes was activated in ionomycine/6-DMAP and ionomycine/roscovitine groups when compared with ethanol/6-DMAP, ethanol/roscovitine and in vitro fertilized groups. However, there was no difference (P>0.05) in the proportion of oocytes activated with ethanol when compared with in vitro fertilized group. No significant difference was seen on the proportion of morula on day 7 of culture, however the development to blastocyst stage was higher (P<0.01) in ionomycine/6-DMAP and ionomycine/roscovitine when compared with ethanol/6-DMAP and ethanol/roscovitine treated oocytes. A higher proportion of oocytes reached blastocyst stage when they were exposed to ionomycine for 3 min but they were not significantly different from the others (P>0.05). The proportion of blastocysts obtained was higher (P<0.05) in oocytes activated after 28 h of maturation when compared with oocytes activated after 32, 36, 40, 44 and 48 h of maturation. In conclusion, a protocol for chemical activation of dromedary camel oocytes with ionomycine/6-DMAP is demonstrated and optimized in the present study for further use in the development of assisted reproductive techniques in this species.

Allosteric Site Inhibitor Disrupting Auto-Processing of Malarial Cysteine Proteases.

Falcipains are major haemoglobinases of Plasmodium falciparum required for parasite growth and development. They consist of pro- and mature domains that interact via 'hot-spot' interactions and maintain the structural integrity of enzyme in zymogen state. Upon sensing the acidic environment, these interactions dissociate and active enzyme is released. For inhibiting falcipains, several active site inhibitors exist, however, compounds that target via allosteric mechanism remains uncharacterized. Therefore, we designed and synthesized six azapeptide compounds, among which, NA-01 & NA-03 arrested parasite growth by specifically blocking the auto-processing of falcipains. Inhibitors showed high affinity for enzymes in presence of the prodomain without affecting the secondary structure. Binding of NA-03 at the interface induced rigidity in the prodomain preventing structural reorganization. We further reported a histidine-dependent activation of falcipain. Collectively, for the first time we provide a framework for blocking the allosteric site of crucial haemoglobinases of the human malaria parasite. Targeting the allosteric site could provide high selectivity and less vulnerable to drug resistance.

Cranial Ultrasonography in Infantile Encephalitic Beriberi: A Useful First-Line Imaging Tool for Screening and Diagnosis in Suspected Cases.

BACKGROUND AND PURPOSE: Brain imaging is central to the diagnosis of infantile encephalitic beriberi. Because cranial sonography findings have not been described in infantile encephalitic beriberi, our aim was to investigate its role in the diagnosis of this condition. MATERIALS AND METHODS: We performed a retrospective review of head sonography of infants (admitted between November 1, 2014, and March 31, 2015) who presented with encephalopathy. Cranial ultrasonography scans were studied for the alteration of echogenicity of the basal ganglia. RESULTS: Of the 145 consecutive infants who presented with encephalopathy, 58 had thiamine-responsive encephalopathy (infantile encephalitic beriberi) and 87 had encephalopathy due to other causes. Forty-eight of 145 infants with encephalopathy showed hyperechoic basal ganglia. A hyperechoic appearance of the basal ganglia on cranial ultrasonography was found to have a sensitivity of 71% (41/58) and a specificity of 92% (80/87) in diagnosing infantile encephalitic beriberi. The sensitivity of cranial sonography increased with age. It was a maximum of 93% (14/15) in the 5 months and older age group. Specificity was a maximum of 100% (18/18) in infants older than 5 months of age. Sensitivity was maximum in Wernicke encephalopathy at 90% (18/20) and least in the acidotic form at 43% (10/23). Follow-up showed gradual normalization of the hyperechoic appearance of the basal ganglia during 8 weeks in 26/41 (63%), with mild atrophy of the basal ganglia in 6/41 (15%) CONCLUSIONS: Hyperechogenicity of the basal ganglia on cranial ultrasonography is a sensitive finding for the diagnosis of infantile encephalitic beriberi in infants who present with Wernicke encephalopathy.

Kinetics of nuclear maturation and effect of holding ovaries at room temperature on in vitro maturation of camel (Camelus dromedarius) oocytes.

Experiments were conducted to investigate kinetics of in vitro nuclear maturation and the effect of storing ovaries at room temperature on initial chromatin configuration and in vitro maturation of dromedary camel oocytes. Cumulus oocyte complexes (COCs) were collected from slaughterhouse ovaries and matured in vitro for 4-48h. At every 4h interval (starting from 0 to 48 h), groups of oocytes were fixed, stained and evaluated for the status of nuclear chromatin. Oocytes were categorized as germinal vesicle (GV), diakinesis (DK), metaphase-I (M-I), anaphase-I (A-I), metaphase-II (M-II) stage and those with degenerated, fragmented, activated or without a visible chromatin as others. At the start of culture, 74% (66/89) oocytes were at GV stage, 13% (12/89) at DK and 12% (11/89) were classified as others. Germinal vesicle breakdown started spontaneously in culture and at 20 h of culture 97% oocytes had already completed this process. After 8 and 16 h of maturation the highest proportion of oocytes (42%, 48/114 and 41%, 51/123) were at DK and M-I stage, respectively. The proportions of oocytes reaching M-II stage at 32 (42%, 50/118), 36 (45%, 47/104), 40 (49%, 57/117), 44 (52%, 103/198) and 48 h (46%, 55/120) of culture were not different from each other (P>0.05). The proportion of oocytes categorized as others, however, increased after 40 h of culture and was higher (P<0.05) at 48 h compared with other maturation periods. There was no difference (P>0.05) in the proportion of oocytes reaching M-II stage from the ovaries collected and stored in normal saline solution (NSS) at room temperature for 12h (43%, 64/148) and those collected in warm NSS (37 degrees C) and processed immediately after arrival in laboratory (49%, 57/117). However, low number of oocytes reached M-II stage from ovaries collected in warm NSS but stored at room temperature (29%, 37/128) compared with other two groups (P<0.05). It may be concluded that dromedary oocytes require 32-44h of in vitro culture to have an optimum number of oocytes in M-II stage. However, further studies are required to find out the most appropriate maturation period, which will result in the further development of these oocytes after IVF, ICSI, parthenogenetic activation or nuclear transfer. Ovaries can be collected and stored in normal saline solution at room temperature for 12h without any appreciable effect on the nuclear maturation of the oocytes.

Maturation rates of vitrified-thawed immature buffalo (Bubalus bubalis) oocytes: effect of different types of cryoprotectants.

Cryopreservation of oocytes collected from slaughtered animals of high genetic value, their subsequent utilisation for production of embryos for transfer may provide an opportunity to replenish the valuable germplasm lost. Experiments were conducted to study the effect of cryoprotectants, dimethyl sulfoxide (DMSO), ethylene glycol (EG), 1,2-propanediol (PROH) and glycerol at different concentrations (3.5, 4, 5, 6 and 7 M each with 0.5M sucrose and 0.4% BSA in DPBS) on morphological survival and in vitro maturation of vitrified-thawed immature buffalo oocytes. The cumulus oocyte complexes were harvested from the ovaries obtained from a local slaughterhouse by aspirating the visible follicles. Less number of oocytes reached metaphase-II stage from the oocytes cryopreserved in any of the concentrations of DMSO, EG, PROH and glycerol compared to fresh oocytes. Among the vitrified groups, highest maturation (40.3, 42.5, 40.4 and 23.5%) was obtained in 7 M DMSO, EG, PROH and glycerol, respectively. Oocytes reaching to M-II stage from the oocytes cryopreserved in 7 M glycerol were significantly lower than that of the oocytes vitrified in 7 M DMSO, EG and PROH. It can be concluded that 7 M solutions of DMSO, EG and PROH can be used for vitrification of immature buffalo oocytes for subsequent utilisation of these oocytes in IVM/IVF and embryo production for transfer.

Management of Mirizzi syndrome: a new surgical approach.

BACKGROUND: In a prospective study of a patient population of 1,340 with biliary calculus disease, that ran from January 1993 to December 1997, 34 patients (2.53%) were identified as having Mirizzi syndrome. Eight patients were found to have type I (A and B) and 26 patients were found to have type II Mirizzi syndrome. A history of recurrent biliary colic and jaundice was present in the majority of patients. METHODS: Ultrasonography was helpful in five patients and endoscopic retrograde cholangiopancreatography was helpful in 17 patients in the diagnosis of this condition. Because the amount of gall bladder tissue used in choledochoplasty is not yet standardized, a new policy regarding choledochoplasty was adopted. In type IA, retrograde cholecystectomy with simple closure of cystic duct was carried out. In type IB, retrograde cholecystectomy and choledochoplasty with 5 mm cuff of the gall bladder was carried out. In type II lesions the procedure depended on the size of fistula. Patients with fistula sizes of less than one-third of the common bile duct diameter underwent choledochoplasty with 5 mm cuff of the gall bladder, and patients with fistula sizes between one-third and two-thirds of the diameter of the common bile duct underwent choledochoplasty with 10 mm cuff of the gall bladder. Patients with fistula sizes of more than two-thirds of the common bile duct diameter underwent Roux-en-Y hepaticojejunostomy. RESULTS: There was no operative mortality and the complication rate was 17.64%. CONCLUSION: Although, out of 26 choledochoplasties, we encountered only one (3.84%) stump stone in a maximum follow-up period of 59 months, further long-term follow-up studies are required to prove the efficacy of the procedure.

Effect of cryoprotectants and their concentration on in vitro development of vitrified-warmed immature oocytes in buffalo (Bubalus bubalis).

Experiments were conducted to study the effect of cryoprotectants, dimethyl sulfoxide (DMSO), ethylene glycol (EG), 1,2-propanediol (PROH), and glycerol at different concentrations (3.5, 4, 5, 6, and 7 M each with 0.5 M sucrose and 0.4% BSA in DPBS) on survival, in vitro maturation, in vitro fertilization, and post-fertilization development of vitrified-thawed immature buffalo oocytes. The COCs were harvested from the ovaries by aspirating the visible follicles. The recovery of post-thaw morphologically normal oocytes was lower in 3.5 and 4 M DMSO, EG, and PROH compared to 5, 6, and 7 M. In all the concentrations of glycerol, an overall lower numbers of oocytes recovered were normal compared to other cryoprotectants. Less number of oocytes reached metaphase-II (M-II) stage from the oocytes cryopreserved in any of the concentrations of DMSO, EG, PROH, and glycerol compared to fresh oocytes. Among the vitrified groups, highest maturation was obtained in 7 M solutions of all the cryoprotectants. The cleavage rates of oocytes vitrified in different concentrations of DMSO, EG, PROH, and glycerol were lower than that of the fresh oocytes. The cleavage rates were higher in oocytes cryopreserved in 6 and 7 M DMSO, EG, PROH, and glycerol compared with oocytes cryopreserved in other concentrations. However, the percentage of morula and blastocyst formation from the cleaved embryos did not vary in fresh oocytes and vitrified oocytes. In conclusion, this report describes the first successful production of buffalo blastocysts from immature oocytes cryopreserved by vitrification.

A study of colorectal adenocarcinoma.

One hundred and seven patients with resected colorectal adenocarcinoma were studied between 1983 and 1987. These formed 3.3% of all cancer cases and 19.4% of gastrointestinal malignancies seen during the period. A majority of patients (68.7%) were between 41 and 60 years of age; 31.8% were below the age of 40 years. There was a male preponderance (62.6%). Left sided cancers were more common (65.4%). The predominant histologic patterns were well differentiated adenocarcinoma (72%) and mucinous carcinoma (20.6%). Mucinous carcinoma was more common on the left side. A majority of cases (95.3%) were in Duke's stages B and C. Only 2.8% of cases were confined to the mucosa.

Leiomyoblastoma stomach.

Three patients with epithelioid leiomyomatous tumors (leiomyoblastomas) of the stomach are presented.

Evaluation of bacteria and biliary tract diseases.

Bile specimens were taken for culture from 100 patients who had disease originating in the biliary tract. Routine cases of chronic cholecystitis with cholelithiasis were found to have a low incidence of infected bile. An acute process appears to be needed in order to produce infected bile, and an acute obstruction of the bile duct due to gallstones produced infected bile. An effort should be made to select these patients and begin antibiotic therapy as soon as possible.

Pre-operative and post-operative evaluation of circulating immune complexes in patients with gastric adenocarcinoma.

Circulating immune complexes (CICs) in the sera of patients with histologically proven adenocarcinoma of stomach were sequentially studied. Serial CICs levels, quantitated using a sensitive method F(ab')2 anti-C3 ELISA, were measured before surgery and in a post-operative follow up. CICs could be detected in 85% of the patients pre-operatively, while ten days after surgery positivity decreased to 71%. Thirty days after surgery, the mean CIC levels decreased significantly and positivity fell to 46%. The results indicate that removal of primary tumor mass results in a sharp decline of CIC levels.

Fibrosarcoma breast. A case report.

The youngest case of Fibro-sarcoma of breast is reported. The review of literature and brief note of management is described.

Diffuse abdominal lipomatosis.

We report a 22-year male who developed progressive distension of abdomen, clinically diagnosed as ascites. A diagnosis of abdominal lipomatosis was made on the basis of CT evidence of excessive fatty tissue in abdominal cavity which was confirmed on laparotomy.

Primary carcinoma of gall bladder--a review of thirty six cases.

The study presents the clinicopathological findings in thirty six cases of primary carcinoma of gall bladder diagnosed at SKIMS Srinagar over a period of four years (1983-86). Particular attention was paid to age, sex, presenting symptoms, presence of gall stones and site of metastasis. Majority of patients were found to be middle aged females (24 out of 36 cases). Most common presenting symptom was abdominal pain and duration of symptoms was two months or less in majority of cases. Gall stones were found in 55.5% cases. Diagnosis of primary gall bladder carcinoma was an incidental histopathological finding in 22% patients. Well differentiated adenocarcinoma was found to be the commonest type of malignancy (77.7%). 33% cases had widespread metastatic disease at the time of operation and liver was the commonest site of invasion (47.2%). Follow up records of 21 out of 36 patients available show only two patients to be alive till date with more than two year follow up and in both these cases carcinoma was an incidental histopathological diagnosis at cholecystectomy. Rest of the patients had died within 5-6 months after surgery. This reinforces the poor outcome for primary gall bladder carcinoma patients irrespective of treatment unless diagnosed and treated at a very early stage. Long term survival can be expected in incidentally found carcinoma at cholecystectomy.

Bilateral quadriceps tendon rupture as the presenting manifestation of chronic kidney disease.

Bilateral painful knees with loss of extension in a patient with chronic kidney disease (CKD) may be due to spontaneous quadriceps tendon rupture. This rare complication is usually seen in patients on long term dialysis. We present a case of bilateral spontaneous quadriceps tendon rupture demonstrated by magnetic resonance imaging in a 20-year-old woman who on evaluation was found to have CKD.

Ultrasonographic-guided retrieval of cumulus oocyte complexes after super-stimulation in dromedary camel (Camelus dromedarius).

In Experiment 1, studies were conducted to apply the transvaginal ultrasound guided ovum pick-up (OPU) technique in dromedary camels after their ovarian super-stimulation and in vivo oocyte maturation. In Experiment 2, the developmental potential of two commonly used oocyte types, i.e., in vivo matured oocytes collected by OPU and abattoir derived in vitro-matured oocytes was compared after their chemical activation. In Experiment 3, developmental competence of oocytes collected from super-stimulated camels by OPU, matured either in vivo or in vitro, was compared after their chemical activation. Mature female dromedary camels super-stimulated with a combination of eCG and pFSH were given an injection of 20 microg of the GnRH analogue, buserelin 24, 26, or 28 h before the scheduled OPU. For collection of cumulus oocyte complexes (COCs) the transducer was guided through the vulva into the cranial most portion of the vagina and 17-gauge, 55 cm single-lumen needle was placed in the needle guide of the ultrasound probe and advanced through the vaginal fornix and into the follicle. Follicular fluid was aspirated using a regulated vacuum pump into tubes containing embryo-flushing media. Aspirates were searched for COCs using a stereomicroscope, and they were then denuded of cumulus cells by hyaluronidase and repeated pipetting. The oocytes were classified as mature (with a visible polar body), immature (with no visible polar body), activated (with divided or fragmented ooplasm) and others (degenerated and abnormal). Overall an average of 12.12 +/- 7.9 COCs were aspirated per animal with an oocyte recovery rate from the aspirated follicles of about 77%. The majority (> 90%) of the collected COCs by OPU were with loose and expanded cumulus cells. The proportion of matured oocytes obtained at 28-29 h (91.2 +/- 4.1) and 26-27 h (82.1 +/- 3.4) were higher (P < 0.005) when compared with those obtained at 24-25 h (40.4 +/- 16.3) after GnRH administration. In Experiment 2, a higher proportion (P < 0.05) of in vivo matured oocytes cleaved (84.6 +/- 2.1 vs. 60.9 +/- 6.6) and developed to blastocyst stages (52.4 +/- 4.1 vs. 30.5 +/- 3.3) when compared with in vitro matured oocytes collected from slaughterhouse ovaries. In Experiment 3, no difference was observed between the developmental competences of oocytes, collected from super stimulated camels, matured in vitro with those collected after their in vivo maturation. In conclusion, about 80-90% mature oocytes can be collected by ultrasound guided transvaginal ovum pick-up from super-stimulated dromedary camels 26-28 h after GnRH administration. The developmental response, to chemical activation, of in vivo matured oocytes collected by ultrasound guided transvaginal OPU is better than in vitro matured oocytes obtained from slaughterhouse ovaries. However, no difference was observed in the developmental competence of oocytes collected by OPU whether they were matured in vivo or in vitro.