Initial experience of laparoscopic retroperitoneal partial nephrectomy in an academic hospital in Malaysia.

Laparoscopic retroperitoneal partial nephrectomy (LRPN) is a technically demanding kidney surgery due to the limited space and unfamiliar approach in the retroperitoneal space. The aim of this study is to review the outcome of our initial experience in performing this procedure. All patients who underwent LRPN between 2019 to 2022 were included in this retrospective review. A total of 23 patients underwent LRPN. The mean operating time was 178±43 minutes and mean warm ischemia time was 20±5 minutes. The average estimated blood lost was 89±68ml and the mean postoperative hospital stay was 3.6±0.8 days. Two patients (11.1%) had positive margin and no local recurrence was seen after mean follow up of 15.8±12.0 months. Our initial experience on LRPN showed promising results to perform partial nephrectomy safely and effectively.

Villous adenoma of the prostatic urethra.

Primary villous adenoma originating from the urinary tract is an infrequent entity. We present a rare case of villous adenoma arising from a prostatic urethra with no sign of malignant transformation. Villous adenoma should be considered as one of the differential diagnoses of urethral lesions, especially if it has similar magnetic resonance imaging features as its colonic counterpart. Due to its potential for malignant transformation, its complete resection is mandatory.

A randomised controlled trial of a novel tramadol chewable tablet: pharmacokinetics and tolerability in children.

Tramadol is a bitter atypical opioid analgesic drug and is prescribed to treat postoperative pain in children. However, in many countries there is no licensed paediatric tramadol formulation available. We have formulated a novel chewable chocolate-based drug delivery system for the administration of tramadol to children. This pilot, single-centre, open-label, randomised clinical study assessed the taste tolerability and comparative population pharmacokinetics of the novel tramadol chewable tablet against a compounded tramadol hydrochloride oral liquid, at a dose of 1 mg.kg-1 . A 5-point facial hedonic scale was used by the children, parents and nurses to assess tolerability. One hundred and forty-one children aged 3-16 years were given tramadol 30 min before general anaesthesia. Blood samples were taken following the induction of anaesthesia and for up to 5 h following tramadol administration. Tramadol and its active metabolite O-desmethyltramadol were analysed using reversed-phase high-performance liquid chromatography. A population pharmacokinetic model was built using non-linear mixed effects modelling. The relative bioavailability for the tablet was 1.25 times higher (95%CI 1.16-1.35) than for tramadol hydrochloride oral liquid, while the absorption rate constant for the tablet was significantly lower (1.97 h-1 vs. 3.34 h-1 , p < 0.001). Larger inter-individual variability in absorption rates were observed with the liquid tramadol. The tramadol chewable tablet was more acceptable in taste to children when assessed by the children, parents and nurses (all p < 0.001). We conclude that the novel tramadol chewable tablet has favourable acceptability and more reliable relative bioavailability in children compared with tramadol hydrochloride oral liquid.

HPLC-UV assay of tramadol and O-desmethyltramadol in human plasma containing other drugs potentially co-administered to participants in a paediatric population pharmacokinetic study.

Multimodal analgesia is employed in paediatric pain management to maximise analgesia and minimise side effects. Tramadol is dosed at 1-1.5 mg/kg to treat severe pain in children but the assay for tramadol in plasma samples for pharmacokinetic and toxicology studies does not often consider concurrently administered medications. In this study we developed and validated an HPLC-UV method to quantify tramadol and its main metabolite (O-desmethyltramadol) in human plasma in the presence of seven potentially interfering drugs. Sample preparation method was developed by combining liquid-liquid extraction and protein precipitation. Chromatographic separation was achieved on a BDS-Hypersil-C18 column (5 µm, 250 × 4.6 mm) using a double gradient method. The limit of quantification was 6.7 ng/ml for both tramadol and ODT. The precision and accuracy were in compliance with ICH guidelines. This method was successfully employed to analyse the blood samples of 137 paediatric participants in a tramadol pharmacokinetic trial.

A novel, palatable paediatric oral formulation of midazolam: pharmacokinetics, tolerability, efficacy and safety.

Midazolam is one of many bitter drugs where provision of a suitable oral paediatric formulation, particularly in the pre-anaesthetic setting, remains a challenge. To overcome this problem, a novel chocolate-based tablet formulation has been developed with positive pre-clinical results. To further investigate the potential of this formulation, 150 children aged 3-16 years who were prescribed midazolam as a premedication were randomly assigned to receive 0.5 mg.kg-1 either as the novel formulation or an intravenous solution given orally, which is the current standard at our institution. Tolerability was assessed by each child, parent and nurse using a 5-point facial hedonic scale and efficacy was determined as the time to onset of sedation. Blood samples for midazolam and 1-hydroxymidazolam levels were analysed using high-performance liquid chromatography. Population pharmacokinetics were evaluated using non-linear mixed effects modelling. The novel formulation had significantly improved tolerability scores from children, parents and nurses (all p < 0.001). Time to effect was not different between the groups (p = 0.140). The pharmacokinetics of midazolam and 1-hydroxymidazolam were able to be modelled simultaneously. The novel formulation was subject to a higher estimated first-pass metabolism compared with the intravenous solution (8.6% vs. 5.0%) and a significantly lower relative bioavailability of 82.1% (p = 0.013), with no other significant differences. Exposure relative to dose was in the range previously reported for midazolam syrup. We conclude that the novel chocolate-based formulation of midazolam provides improved tolerability while remaining efficacious with suitable pharmacokinetics when used as a premedicant for children.

Tracheotomy: an alternative for tracheobronchial foreign body removal.

A 6 years old girl accidentally aspirated a plastic whistle while playing. Computed Tomography of thorax showed foreign body at carina level. Rigid bronchoscope under general anesthesia was attempted but unable to extract the whistle through vocal cord. Tracheostomy was later performed and foreign body was removed.

Mutant p53 mediates survival of breast cancer cells.

BACKGROUND: p53 is the most commonly mutated tumour-suppressor gene in human cancers. Unlike other tumour-suppressor genes, most p53 cancer mutations are missense mutations within the core domain, leading to the expression of a full-length mutant p53 protein. Accumulating evidence has indicated that p53 cancer mutants not only lose tumour suppression activity but also gain new oncogenic activities to promote tumourigenesis. METHODS: The endogenous mutant p53 function in human breast cancer cells was studied using RNA interference (RNAi). Gene knockdown was confirmed by quantitative PCR and western blotting. Apoptosis was evaluated by morphological changes of cells, their PARP cleavage and annexin V staining. RESULTS: We show that cancer-associated p53 missense mutants are required for the survival of breast cancer cells. Inhibition of endogenous mutant p53 by RNAi led to massive apoptosis in two mutant p53-expressing cell lines, T47D and MDA-MB-468, but not in the wild-type p53-expressing cells, MCF-7 and MCF-10A. Reconstitution of an RNAi-insensitive mutant p53 in MDA-MB-468 cells completely abolished the apoptotic effects after silencing of endogenous mutant p53, suggesting the specific survival effects of mutant p53. The apoptotic effect induced by mutant p53 ablation, however, is independent of p63 or p73 function. CONCLUSION: These findings provide clear evidence of a pro-survival 'gain-of-function' property of a subset of p53 cancer mutants in breast cancer cells.

Effect of chitosan salts and molecular weight on a nanoparticulate carrier for therapeutic protein.

The objective of this study was to investigate the potential of chitosan salts as a carrier in the preparation of protein-loaded nanoparticles. Glutamic and aspartic acids were used to prepare chitosan salts of 35, 100, and 800 KDa. Nanoparticles of chitosan base, chitosan glutamate, and chitosan aspartate were produced by ionotropic gelation with sodium tripolyphosphate (TPP). Bovine serum albumin (BSA) was applied as a model protein at loading concentrations ranging from 0.2 to 2 mg/mL. The size of the nanoparticles, as measured by photon correlation spectroscopy, was in the range of 195 to 3450 nm, depending on type and molecular weight of chitosan. Nanoparticles prepared with higher molecular weight chitosan showed larger sizes. The encapsulation was controlled by the competition of BSA in forming ionic cross-linking with chitosan and by the entrapment of BSA during the gelation process. Higher BSA encapsulation efficiency (EE) was obtained for nanoparticles prepared with chitosan salts compared to those prepared with the base. The higher EE was a result of a higher degree of ionization, causing more active sites to interact with BSA. In addition, a higher and faster release of BSA from the nanoparticles into pH 7.4 buffer medium was observed for nanoparticles of the chitosan salts than was observed for nanoparticles of the chitosan base. The higher and faster release was attributed to higher EE and lower entrapment of BSA within the matrix of the nanoparticle during the gelation process. The influence of molecular weight on the property of nanoparticles exhibited different effects. The difference was a result of different organic acids used to prepare nanoparticles leading to the difference in polymer conformation and viscosity of organic acid solution. Therefore, this study showed that the characteristics of chitosan nanoparticles loaded with a protein drug could be readily modulated by changing the salt form or the molecular weight of the chitosan carrier.

Influence of dietary solvents on strength of nanofill and ormocer composites.

The objective of this study was to determine the influence of dietary solvents on the shear punch strength of nanofill (Filtek Supreme [FS], 3M-ESPE) and ormocer (Admira [AM], Voco) composites. The strength of these materials was also compared to a minifill composite (Z250 [ZT], 3M-ESPE), a compomer (F2000 [FT], 3M-ESPE) and a highly viscous glass ionomer cement (Ketac Molar Quick [KM], 3M-ESPE). Thirty-two specimens (8.7 mm diameter and 1-mm thick) of each material were made, randomly divided into four groups of eight and conditioned for one week as follows-Group 1 (control): distilled water at 37 degrees C; Group 2: 0.02M citric acid at 37 degrees C; Group 3: 50% ethanol-water solution at 37 degrees C and Group 4: heptane at 37 degrees C. After conditioning, the specimens were restrained with a torque of 2.5 Nm and subjected to shear punch strength testing using a 2-mm diameter punch at a crosshead speed of 0.5 mm/minute. The shear punch strength of the specimens was computed and data subjected to ANOVA/Scheffe's tests at significance level 0.05. With the exception of AM, the strength of all materials was not significantly influenced by dietary solvents. For AM, conditioning in heptane resulted in significantly higher shear strength values. The strength of the nanofill and ormocer composites was lower than the minifill composite but higher than the compomer and highly viscous glass ionomer cement investigated.

Characterization of chitosan acetate as a binder for sustained release tablets.

A chitosan derivative as an acetate salt was successfully prepared by using a spray drying technique. Physicochemical characteristics and micromeritic properties of spray-dried chitosan acetate (SD-CSA) were studied as well as drug-polymer and excipient-polymer interaction. SD-CSA was spherical agglomerates with rough surface and less than 75 microm in diameter. The salt was an amorphous solid with slight to moderate hygroscopicity. The results of Fourier transform infrared (FTIR) and solid-state (13)C NMR spectroscopy demonstrated the functional groups of an acetate salt in its molecular structure. DSC and TGA thermograms of SD-CSA as well as FTIR and NMR spectrum of the salt, heated at 120 degrees C for 12 h, revealed the evidence of the conversion of chitosan acetate molecular structure to N-acetylglucosamine at higher temperature. No interaction of SD-CSA with either drugs (salicylic acid and theophylline) or selected pharmaceutical excipients were observed in the study using DSC method. As a wet granulation binder, SD-CSA gave theophylline granules with good flowability (according to the value of angle of repose, Carr's index, and Hausner ratio) and an excellent compressibility profile comparable to a pharmaceutical binder, PVP K30. In vitro release study of theophylline from the tablets containing 3% w/w SD-CSA as a binder demonstrated sustained drug release in all media. Cumulative drug released in 0.1 N HCl, pH 6.8 phosphate buffer and distilled water was nearly 100% within 6, 16 and 24 h, respectively. It was suggested that the simple incorporation of spray-dried chitosan acetate as a tablet binder could give rise to controlled drug delivery systems exhibiting sustained drug release.

Ultrasonication of chitosan and chitosan nanoparticles.

The objective of this study was to evaluate the effects of ultrasonication on chitosan molecules and nanoparticles. Molecular weight (M(v)) of chitosan HCl (M(v) 146 kDa and degree of deacetylation (DD) 96%) decreased linearly with increasing duration and amplitude of ultrasonication. DD and FTIR absorption were unaffected. X-ray diffraction (XRD) analysis suggested greater chain alignment in the ultrasonicated chitosan samples. Chitosan nanoparticles had mean diameter of 382 nm, polydispersity of 0.53 and zeta potential of 47 mV. Ultrasonication administered at increasing duration or amplitude decreased the mean diameter and polydispersity of the nanoparticles. Zeta potential and FTIR absorbance were unaffected, while XRD suggested a greater disarray of chain alignment in the nanoparticle matrix. Under the transmission electron microscope (TEM), freshly prepared nanoparticles were dense spherical structures which became fragmented after ultrasonication for 10 min at amplitude of 80. Untreated nanoparticle formulation turned turbid upon storage for 3 weeks at ambient conditions due to substantial swelling of the nanoparticles. Ultrasonicated nanoparticle formulation remained clear on storage. Although the particles had also swelled, they were no longer spherical, assuming instead an irregular shape with branching arms. In conclusion, high-intensity ultrasonication induced considerable damage on the chitosan nanoparticles which could affect their function as drug carriers.

Chitosan-alginate-CaCl(2) system for membrane coat application.

Water-based formulations are preferred for membrane coat application because they do not require the use of noxious solvents. A novel aqueous chitosan-alginate-CaCl(2) system was evaluated as a potential formulation to produce water-insoluble membranes of biodegradable polymers. Chitosan-alginate coacervates were prepared by controlled reaction of chitosan (0.25% w/v) and sodium alginate (0.25% w/v) solutions. Coherent membranes were obtained by casting and drying the coacervates suspended in aqueous CaCl(2) solutions (0.05-0.07% w/v). Increasing the calcium content did not modify membrane thickness (25-26 microm), but reduced the water vapor transmission rate from 658 to 566 g/m(2)/day, and improved the tensile strength of the membranes from 9.33 to 17.13 MPa. Differential scanning calorimetry, Fourier transform infrared spectroscopy, and elemental analyses of the chitosan-alginate coacervates indicated they were stable for up to 4 weeks of storage in distilled water at ambient temperature. Membranes of the stored coacervates required less calcium to attain maximum mechanical strength. They also had higher water vapor transmission rates than corresponding films prepared from fresh coacervates. On the basis of the properties of the cast film and its storage stability, the chitosan-alginate-CaCl(2) system can be considered for potential membrane coat application.

Chitosan-alginate films prepared with chitosans of different molecular weights.

Chitosan-alginate polyelectrolyte complex (CS-AL PEC) is water insoluble and more effective in limiting the release of encapsulated materials compared to chitosan or alginate. Coherent CS-AL PEC films have been prepared in our laboratory by casting and drying suspensions of chitosan-alginate coacervates. The objective of this study was to evaluate the properties of the CS-AL PEC films prepared with chitosans of different molecular weights. Films prepared with low-molecular-weight chitosan (Mv 1.30 x 10(5)) were twice as thin and transparent, as well as 55% less permeable to water vapor, compared to films prepared with high-molecular-weight chitosan (Mv 10.0 x 10(5)). It may be inferred that the low-molecular-weight chitosan reacted more completely with the sodium alginate (M(v) 1.04 x 10(5)) than chitosan of higher molecular weight. A threshold molecular weight may be required, because chitosans of Mv 10.0 x 10(5) and 5.33 x 10(5) yielded films with similar physical properties. The PEC films exhibited different surface properties from the parent films, and contained a higher degree of chain alignment with the possible formation of new crystal types. The PEC films exhibited good in vitro biocompatibility with mouse and human fibroblasts, suggesting that they can be further explored for biomedical applications.

Concurrent production of chitin from shrimp shells and fungi.

Crustacean shells constitute the traditional and current commercial source of chitin. Conversely, the control of fungal fermentation processes to produce quality chitin makes fungal mycelia an attractive alternative source. Therefore, the exploitation of both of these sources to produce chitin in a concurrent process should be advantageous and is reported here. Three proteolytic Aspergillus niger (strains 0576, 0307 and 0474) were selected from a screening for protease activity from among 34 zygomycete and deuteromycete strains. When fungi and shrimp shell powder were combined in a single reactor, the release of protease by the fungi facilitated the deproteinization of shrimp-shell powder and the release of hydrolyzed proteins. The hydrolyzed proteins in turn were utilized as a nitrogen source for fungal growth, leading to a lowering of the pH of the fermentation medium, thereby further enhancing the demineralization of the shrimp-shell powder. The shrimp-shell powders and fungal mycelia were separated after fermentation and extracted for chitin with 5% LiCl/DMAc solvent. Chitin isolates from the shells were found to have a protein content of less than 5%, while chitin isolates from the three fungal mycelia strains had protein content in the range of 10-15%. The relative molecular weights as estimated by GPC for all chitin samples were in the 10(5) dalton range. All samples displayed characteristic profiles for chitin in their FTIR and solid-state NMR spectra. All chitin samples evaluated with MTT and Neutral Red assays with three commercial cell lines did not display cytotoxic effects.

Preparation and characterization of chitin beads as a wound dressing precursor.

Chitin was dissolved in N, N-dimethylacetamide/5% lithium chloride (DMAc/5%LiCl) to form a 0.5% chitin solution. Chitin beads were formed by dropping the 0.5% chitin solution into a nonsolvent coagulant, ethanol. The beads were left in ethanol for 24 h to permit hardening, consolidation, and removal of residual DMAc/5%LiCl solvent in order to give spherical chitin beads uniform size distribution. The ethanol-gelled chitin beads had an average diameter of 535 microm. The chitin beads were subsequently activated in 50% (w/v) NaOH solution and reacted with 1.9 M monochloroacetic acid/2-propanol solution to introduce a carboxymethylated surface layer to the chitin beads. The bilayer character of the surface-carboxymethylated chitin (SCM-chitin) beads was verified by Fourier transform infrared (FTIR) spectroscopy, thermogravimetric analysis (TGA), and confocal microscopy. The bilayered SCM-chitin beads were found to absorb up to 95 times their dry weight of water. These SCM-chitin beads have potential as a component of wound dressings.

PEC films prepared from Chitosan-Alginate coacervates.

Chitosan-alginate polyelectrolyte complex (PEC) have been prepared in situ in beads and microspheres. This study examines the preparation of suitable chitosan-alginate coacervates for casting into homogeneous PEC films for potential applications in packaging, controlled release systems and wound dressings. Coacervation between chitosan and alginate was rapid, but the rate may be controlled with the addition of water miscible organic solvents. Compared with ethanol and PEG200, acetone was the more promising solvent moderator. Suspensions of fine, uniformly dispersed coacervates were produced by a dropwise addition of 0.25% w/v chitosan solution (solvent: 1: 1 v/v of 2% acetic acid and acetone) into 0.25% w/v sodium alginate solution in water under rapid agitation. The PEC films were transparent and flexible. They exhibited high permeability to water vapor, but resisted complete dissolution in 0.1 M HCI, distilled water and pH 7.4 phosphate buffer solution. Microscopic heterogeneity in the films could be reduced by immersion in aqueous media, but this was accompanied by modifications in the thickness, permeability and mechanical property of the films.

Structure-activity relationships for G2 checkpoint inhibition by caffeine analogs.

Caffeine inhibits the G2 checkpoint activated by DNA damage and enhances the toxicity of DNA-damaging agents towards p53-defective cancer cells. The relationship between structure and G2 checkpoint inhibition was determined for 56 caffeine analogs. Replacement of the methyl group at position 3 or 7 resulted in loss of activity, while replacement at position 1 by ethyl or propyl increased activity slightly. 8-Substituted caffeines retained activity, but were relatively insoluble. The structure-activity profile did not resemble those for other known pharmacological activities of caffeine. The active analogs also potentiated the killing of p53-defective cells by ionizing radiation, but none was as effective as caffeine.

Caffeine and nicotinamide enhances the aqueous solubility of the antimalarial agent halofantrine.

The aqueous solubility of the antimalarial agent halofantrine in phosphate buffers pH 5.9 and 7.0 (ionic strength 0.08) is increased by the addition of caffeine and nicotinamide. Solubility is increased to a greater extent in the presence of caffeine (12.5-125 mM) than nicotinamide (125 mM -2.0 M). The greatest increase in solubility was observed at pH 5.9 where the basal solubility of halofantrine rose from 0.91 to 435 microM when 125 mM caffeine was added. Phase solubility studies support the formation of a 1:1 complex between caffeine and halofantrine which is characterised by a K(1:1) constant of 2.75x10(3) M(-1) (pH 5.9). A less stable 1:1 complex is formed at pH 7.0 (K(1:1)=6.37x10(3) M(-1)). Differential scanning calorimetry of solid mixtures of caffeine and halofantrine showed the absence of the endotherms of the two drugs and the appearance of a distinct endotherm (with a smaller enthalpy) characteristic of the complex. An analysis of the 1H-NMR spectra of mixtures of caffeine and halofantrine revealed perturbations in the chemical shifts of the methyl group and proton at positions 4 and 8 of caffeine, and a change in splitting pattern of the H(9) proton of the phenanthrene ring in halofantrine.

Storage of partially deacetylated chitosan films.

Chitosan has wide-ranging applications as a biomaterial, but its stability in storage is not widely known. The objective of this study was to evaluate the storage stability of films prepared from chitosan of 77% deacetylation. Both the neutralized and acetate films were evaluated, as chitosan salts offer the advantage of being soluble in water at the neutral-to-basic pH range. Aqueous solutions containing 0.5-5% acetic acid were used as solvents. The X-ray diffraction pattern, the IR spectrum, water uptake, and solubility of the films were influenced by the presence of the N-acetyl functionality, the acetate ions, and storage of the films. The anhydrous chitosan crystal in the neutralized films was unstable to storage at 4 degrees C and 28 degrees C. Its formation, as well as that of the hydrated crystal, were further hindered by the presence of even small quantities of the acetate ions. The resultant amorphous nature of the acetate films, coupled with the acidifying action of the acetic acid, led to greater water uptake and solubility compared to the neutralized films. Storage reduced the differences between the neutralized and acetate films. It also minimized the influence of the initial acetic acid content on the IR absorption and water uptake of the acetate films, exerting its leveling effects mainly within the first week of storage. Using a lower storage temperature of 4 degrees C or heating the films for 2 h at 120 degrees C prior to storage did not significantly modify the results. A pertinent factor appears to be the degree of deacetylation of the chitosan that was used to prepare the films.

The antimalarial agent halofantrine perturbs phosphatidylcholine and phosphatidylethanolamine bilayers: a differential scanning calorimetric study.

The interaction of halofantrine with phosphatidylcholine and phosphatidylethanolamine bilayers has been investigated by differential scanning calorimetry. Halofantrine caused a broadening of the gel to liquid crystalline phase transition endotherm of the phosphatidylcholines. A decrease in the transition temperature Tm and enthalpy (delta H) of transition was also observed. This varied with the chain length of the phospholipid and was more pronounced with short chain members. Halofantrine-induced changes to the thermotropic characteristics of dipalmitoylphosphatidylcholine (DPPC)/cholesterol bilayers suggested that the penetration of halofantrine into the bilayer was diminished in the presence of cholesterol. A more complex calorimetric profile was observed in the interaction of halofantrine with phosphatidylethanolamines and the results suggested that halofantrine did not disrupt the cooperativity of the phosphatidylethanolamine bilayers to the same extent as that observed with the phosphatidylcholines. Halofantrine caused significant perturbation of phospholipids and this property might have an important bearing on its pharmacodynamic effects.