Analysis of human rabies deaths reported at two hospitals in Karachi, Pakistan: a call to save lives by reforming rabies prevention facilities.

BACKGROUND: Rabies is endemic in low- and middle-income countries. It is caused mainly by the bite of a rabid dog and is fatal if not treated effectively and in a timely manner with quality post-exposure prophylaxis. Despite a profusion of private and public healthcare centres in Sindh province, most are ill-equipped to treat dog bites. METHODS: We analysed 129 human deaths from rabies who presented at the emergency departments of two tertiary care hospitals in Karachi over 10 y. Demographic data, time, location of the bite and distance travelled to report symptoms of rabies were recorded. RESULTS: Most victims were male, and children were more often affected; almost none had received post-exposure prophylaxis. A total of 12% of bites were on the face, head or neck. The mean incubation period was 56 d. Most (60%) of the rabies victims travelled long distances, hoping to receive treatment. CONCLUSIONS: Rabies deaths were either due to a lack of awareness or the non-availability of rabies immunobiologicals within easy reach. Public health services must raise awareness, conduct surveillance and provide appropriately spaced centres for free treatment of dog bites. This lethal disease must be prevented at all costs.

A quartet pollen phenotype identified in a population of Brassica interspecific hybrids shows incomplete penetrance and variable response to temperature.

Quartet pollen, where pollen grains remain attached to each other post-meiosis, is useful for tetrad analysis, crossover assessment and centromere mapping. We observed the quartet pollen phenotype for the first time in the agriculturally significant Brassica genus, in an experimental population of allohexaploid Brassica hybrids derived from the cross (Brassica napus × B. carinata) × B. juncea followed by two self-pollination generations. Quartet pollen production was assessed in 144 genotypes under glasshouse conditions, following which a set of 16 genotypes were selected to further investigate the effect of environment (warm: 25 °C and cold: 10 °C temperatures) on quartet pollen production in growth cabinets. Under glasshouse phenotyping conditions, only 92 out of 144 genotypes produced enough pollen to score: of these, 30 did not produce any observable quartet pollen, while 62 genotypes produced quartet pollen at varying frequencies. Quartet pollen production appeared quantitative and did not clearly fall into phenotypic or qualitative categories indicative of major gene expression. No consistent effect of temperature on quartet pollen production was identified, with some genotypes producing more and some producing less quartet pollen under different temperature treatments. The genetic heterogeneity and frequent pollen infertility of this population prevents strong conclusions being made. However, it is clear that the quartet phenotype in this Brassica population does not show complete penetrance and shows variable (likely genotype-specific) response to temperature stress. In future, identification of quartet phenotypes in Brassica would perhaps best be carried out via screening of diploid (e.g. B. rapa) TILLING populations.

Reducing Cost of Rabies Post Exposure Prophylaxis: Experience of a Tertiary Care Hospital in Pakistan.

BACKGROUND: Rabies is a uniformly fatal disease, but preventable by timely and correct use of post exposure prophylaxis (PEP). Unfortunately, many health care facilities in Pakistan do not carry modern life-saving vaccines and rabies immunoglobulin (RIG), assuming them to be prohibitively expensive and unsafe. Consequently, Emergency Department (ED) health care professionals remain untrained in its application and refer patients out to other hospitals. The conventional Essen regimen requires five vials of cell culture vaccine (CCV) per patient, whereas Thai Red Cross intradermal (TRC-id) regimen requires only one vial per patient, and gives equal seroconversion as compared with Essen regimen. METHODOLOGY/PRINCIPAL FINDINGS: This study documents the cost savings in using the Thai Red Cross intradermal regimen with cell culture vaccine instead of the customary 5-dose Essen intramuscular regimen for eligible bite victims. All patients presenting to the Indus Hospital ED between July 2013 to June 2014 with animal bites received WHO recommended PEP. WHO Category 2 bites received intradermal vaccine alone, while Category 3 victims received vaccine plus wound infiltration with Equine RIG. Patients were counseled, and subsequent doses of the vaccine administered on days 3, 7 and 28. Throughput of cases, consumption utilization of vaccine and ERIG and the cost per patient were recorded. CONCLUSIONS/SIGNIFICANCE: Government hospitals in Pakistan are generally underfinanced and cannot afford treatment of the enormous burden of dog bite victims. Hence, patients are either not treated at all, or asked to purchase their own vaccine, which most cannot afford, resulting in neglect and high incidence of rabies deaths. TRC-id regimen reduced the cost of vaccine to 1/5th of Essen regimen and is strongly recommended for institutions with large throughput. Training ED staff would save lives through a safe, effective and affordable technique.

Twenty-four-hour urine constituents in stone formers: a study from the northeast part of Peninsular Malaysia.

Urolithiasis is a common disease with increasing incidence and prevalence world-wide, probably more common in industrialized countries. The metabolic evaluation of 24-h urine collection has been considered as part of the management of urinary stone patients. The aim of this study was to evaluate the 24-h urine constituents in stone formers and its relation to demographic data in the northeast part of Peninsular Malaysia. One hundred and six patients were recruited in this study from two hospitals in the same geographical region; 96 patients fulfilled the inclusion criteria and an informed consent was obtained from all subjects. The 24-h urine was collected in sterile bottles with a preservative agent and calcium, oxalate, citrate, uric acid, magnesium and phosphate were tested using commercial kits on a Roche Hitachi 912 chemistry analyzer. The age (mean ± SD) of 96 patients was 56.45 ± 13.43 years and 82.3% of the patients were male while 17.7% were female. The 24-h urine abnormalities were hypercalciuria (14.5%), hyperoxaluria (61.4%), hypocitraturia (57.2%), hyperuricouria (19.7%), hypomagnesuria (59.3%) and hyperphosphaturia (12.5%). Hyperoxaluria (61.4%) was the most common abnormality detected during the analysis of 24-h urine constituents in contradiction to industrial countries, where hypercalciuria was the most common finding. The high frequencies of hypomagnesuria and hypocitraturia reflect the important role of magnesium and citrate in stone formation and their prophylactic role in the treatment of urinary stone disease in the given population.

Pneumatic ureterolithotripsy in paediatric and adolescent patients: a ten-year experience at the Hospital Universiti Sains Malaysia.

INTRODUCTION: Two to three percent of stone disease cases occur in the paediatric age group. It is common in some parts of the world, such as in Turkey, India and Thailand. More than 50 percent of stones in children are still managed through open surgery. Ureteroscopic intervention for children remains a challenging treatment option. However, in contemporary urology practice, this mode of intervention is becoming more common. In this retrospective study, we reviewed our experiences with ureteroscopy and pneumatic lithotripsy in the paediatric and adolescent age group. METHODS: A total of 13 patients at or below the age of 18 years underwent 17 retrograde semirigid ureteroscopy between 1998 and 2008. Their median age was 14 (range 3-18) years. The technique of ureterolithotripsy used for this age group was similar to that used among adults. The ureteric orifice and intramural part of the ureter were not dilated in all the patients. A double J stent was inserted into all the patients. RESULTS: The mean stone size was 7.9 (range 5-13) mm. The stone was in the distal ureter in eight patients, in the mid ureter in seven patients and at the ureterovesical junction in another two patients. The overall stone-free rate after one ureterolithotripsy procedure was 84.6%. We failed to clear the stones in two patients at the first sitting. CONCLUSION: Semi-rigid ureteroscopy and pneumatic ureterolithotripsy are safe and effective procedures that can be performed without ureteric dilation in experienced hands.

Increased levels of circulating Annexin A5 in Familial Mediterranean fever.

BACKGROUND: Familial Mediterranean fever is a genetic autoinflammatory disease most commonly affecting the ethnic groups originating from around the Mediterranean Sea. Apoptosis plays an important role in down-regulation of the inflammatory response by reducing the lifespan of activated immunocompetent cells. Thus, increased apoptosis may be associated with pathogenesis of familial Mediterranean fever. METHODS: In the present study we determined the serum levels of apoptotic marker, Annexin A5, in familial Mediterranean fever patients, within an attack and attack-free, in comparison to healthy subjects and assessed the influence of colchicine treatment on this parameter. In addition, in all study subjects serum levels of C-reactive protein and interleukine-1ß, and the total leukocyte count were also determined. RESULTS: Our results demonstrated that pathogenesis of familial Mediterranean fever is characterized by the increased levels of circulating Annexin A5, which is higher in patients within the attack and which associate with the increased levels of C-reactive protein and interleukine-1ß and total leukocyte count. CONCLUSIONS: The results obtained indicate elevated rates of apoptosis of subpopulations of leukocytes involved in autoinflammation and recurrent episodes of fever in familial Mediterranean fever. It was also revealed that regular colchicine treatment sufficiently decreases the rate of apoptosis in familial Mediterranean fever patients by affecting the intensity of autoinflammatory reactions.

Alterations in the complement cascade in post-traumatic stress disorder.

BACKGROUND: In the present study we assessed the functional state of the major mediator of the immune response, the complement system, in post-traumatic stress disorder (PTSD). METHODS: Thirty one PTSD patients within 13 years from traumatic event and the same number of sex- and age-matched healthy volunteers were involved in this study. In the blood serum of the study subjects hemolytic activities of the classical and alternative complement pathways, as well as the activities of the individual complement components have been measured. Correlation analysis between all measured parameters was also performed. RESULTS: According to the results obtained PTSD is characterized by hyperactivation of the complement classical pathway, hypoactivation of the complement alternative pathway and overactivation of the terminal pathway. CONCLUSIONS: The results obtained provide further evidence on the involvement of the inflammatory component in pathogenesis of PTSD.

Involvement of motility and flagella in Bacillus cereus biofilm formation.

Bacillus cereus is a food-borne pathogen and a frequent contaminant of food production plants. The persistence of this pathogen in various environments results from the formation of spores and of biofilms. To investigate the role of the B. cereus flagellar apparatus in biofilm formation, we constructed a non-flagellated mutant and a flagellated but non-motile mutant. Unexpectedly, we found that the presence of flagella decreased the adhesion of the bacterium to glass surfaces. We hypothesize that this decrease is a consequence of the flagella hindering a direct interaction between the bacterial cell wall and the surface. In contrast, in specific conditions, motility promotes biofilm formation. Our results suggest that motility could influence biofilm formation by three mechanisms. Motility is necessary for the bacteria to reach surfaces suitable for biofilm formation. In static conditions, reaching the air-liquid interface, where the biofilm forms, is a strong requirement, whereas in flow cells bacteria can have access to the bottom glass slide by sedimentation. Therefore, motility is important for biofilm formation in glass tubes and in microtitre plates, but not in flow cells. Motility also promotes recruitment of planktonic cells within the biofilm by allowing motile bacteria to invade the whole biofilm. Finally, motility is involved in the spreading of the biofilm on glass surfaces.

Molecular basis for group-specific activation of the virulence regulator PlcR by PapR heptapeptides.

The transcriptional regulator PlcR and its cognate cell-cell signalling peptide PapR form a quorum-sensing system that controls the expression of extra-cellular virulence factors in various species of the Bacillus cereus group. PlcR and PapR alleles are clustered into four groups defining four pherotypes. However, the molecular basis for group specificity remains elusive, largely because the biologically relevant PapR form is not known. Here, we show that the in vivo active form of PapR is the C-terminal heptapeptide of the precursor, and not the pentapeptide, as previously suggested. Combining genetic complementation, anisotropy assays and structural analysis we provide a detailed functional and structural explanation for the group specificity of the PlcR-PapR quorum-sensing system. We further show that the C-terminal helix of the PlcR regulatory domain, specifically the 278 residue, in conjunction with the N-terminal residues of the PapR heptapeptide determines this system specificity. Variability in the specificity-encoding regions of plcR and papR genes suggests that selection and evolution of quorum-sensing systems play a major role in adaptation and ecology of Bacilli.

Sample preparation for beta-exotoxin determination in Bacillus thuringiensis cultures by reversed-phase high-performance liquid chromatography.

Beta-exotoxin is a nucleotide analogue produced by the entomopathogenic bacterium Bacillus thuringiensis. We have defined two new HPLC procedures for quantification of this exotoxin in culture supernatants of B. thuringiensis grown in poor or rich medium. The sample is prepared either by precipitation in solvent or by solid-phase extraction. Solvent precipitation is achieved treating the sample with acetone and acetonitrile. Solid-phase extraction is performed with a C18 and an anion-exchange cartridge. Reversed-phase HPLC with gradient elution of the prepared samples gives a limit of quantitation of 2 microg/ml for samples prepared by solvent precipitation and of 0.3 microg/ml for samples prepared by solid-phase extraction.

[The effects of estrogen in the cardiovascular system]. / Die Wirkungen von Ostrogen im kardiovaskulären System.

Estrogen replacement therapy (ERT) has been established for the treatment of perimenopausal symptoms and postmenopausal prevention of osteoporosis. Clinical (not randomized) cohort studies have shown an association of a significantly reduced cardiovascular mortality with ERT. However, a first randomized, double-blind and placebo-controlled study (HERS) could not support these findings. However, this study is limited by the use of (heterogeneous) conjugated estrogens and combined progestin treatment because animal experiments demonstrated an inhibition of protective estrogen effects by progesterone. On the other hand, experimental and clinical findings demonstrated beneficial estrogen effects on lipid metabolism, lipid-peroxidation, smooth-muscle-cell proliferation, hemostasis, and vasomotion. Actually, several authors are discussing the mediation of estrogen's effects by vascular estrogen receptors. Recent findings on different subtypes of estrogen receptors (ER-alpha and ER-beta) may explain some antagonistic effects as proliferation in the one (endometrium) and anti-proliferation in the other (vascular wall) tissue. In this context, the exact detection of the mechanism(s) of estrogen action may probably lead to new approaches in the treatment of coronary artery disease.

Development and field performance of a broad-spectrum nonviable asporogenic recombinant strain of Bacillus thuringiensis with greater potency and UV resistance.

The main problems with Bacillus thuringiensis products for pest control are their often narrow activity spectrum, high sensitivity to UV degradation, and low cost effectiveness (high potency required). We constructed a sporulation-deficient SigK(-) B. thuringiensis strain that expressed a chimeric cry1C/Ab gene, the product of which had high activity against various lepidopteran pests, including Spodoptera littoralis (Egyptian cotton leaf worm) and Spodoptera exigua (lesser [beet] armyworm), which are not readily controlled by other Cry delta-endotoxins. The SigK(-) host strain carried the cry1Ac gene, the product of which is highly active against the larvae of the major pests Ostrinia nubilalis (European corn borer) and Heliothis virescens (tobacco budworm). This new strain had greater potency and a broader activity spectrum than the parent strain. The crystals produced by the asporogenic strain remained encapsulated within the cells, which protected them from UV degradation. The cry1C/Ab gene was introduced into the B. thuringiensis host via a site-specific recombination vector so that unwanted DNA was eliminated. Therefore, the final construct contained no sequences of non-B. thuringiensis origin. As the recombinant strain is a mutant blocked at late sporulation, it does not produce viable spores and therefore cannot compete with wild-type B. thuringiensis strains in the environment. It is thus a very safe biopesticide. In field trials, this new recombinant strain protected cabbage and broccoli against a pest complex under natural infestation conditions.

Histopathological effects of silicone rubber 'Ovabloc' on the human fallopian tube.

OBJECTIVE: To study the histopathological effects of silicone rubber on the human fallopian tube. METHODS: A prospective longitudinal study at Benha University Hospital and Boulak Eldakrour General Hospital, Egypt. Nine patients who were on the waiting list for hysterectomy and requested postponing of their operation for personal reasons. RESULTS: Our results suggest that silicone rubber induces histopathological changes in the form of ciliary loss and intracellular changes. CONCLUSIONS: These effects tend to increase with the increase of the duration of tubal plugging and are best seen by electron microscopy.

Ultradian variations in sensitivity of rat aorta rings to noradrenaline.

The in vitro sensitivity of the rat aorta to the vasoconstrictor effect of noradrenaline was found to vary according to the time the animals were killed, with a minimum at 10:00 h and a maximum at 16:00 h. This ultradian rhythm was not influenced by the presence of the endothelium but was modified by some, as yet unidentified, circulating factor(s). The results also strongly suggested an ultradian rhythm of in vitro sensitivity to the beta-adrenoceptor agonists, salbutamol and isoprenaline. This rhythm appears to be opposite to that for the sensitivity of aortic alpha-adrenoceptors to stimulation. In conclusion, our results demonstrate that the regulation of aortic tone follows an ultradian rhythm.

Removal of intrauterine devices with missing tails during early pregnancy.

During a 70-month period, CO2 hysteroscopy was carried out on 52 pregnant women wearing IUDs with retracted tails. Ultrasonography was done prior to hysteroscopy to determine the size of the gestational sac and the site of the device. Successful removal of the device occurred in 46 cases. The technique, the findings of the study, and the limitations of the procedure are described.

PIP: IUDs were successfully removed from 46 of 52 pregnant women bearing IUDs with retracted tails, from December 1985 to November 1991, at the Department of Gynecology and Obstetrics, Benha University Hospital, Egypt. The presence and location of the IUD was confirmed with ultrasonography, excluding 3 patients who did not have retracted IUD tails, and 8 who were 12 weeks' gestation. After paracervical block or short-acting IV anesthesia, the 7 mm Storz hysteroscope was introduced, using cervical dilatation only if needed. CO2 gas, up to 200 ml, was injected into the internal os with a Hysteroflator (Wiest, Berlin). The IUD was carefully removed with a grasping forceps. The women received the antiprostaglandin endomethasin (sic) by suppository the night before, and after the procedure. Patients were observed for 5 days, and ultrasound was repeated 2 weeks later to confirm fetal viability. There were 6 failures to remove an IUD, 2 involving disturbance of the gestational sac, and 2 where the IUD lay beneath the sac. 2 spontaneous abortions resulted in the series. 24 Lippes Loops, 12 Copper Ts, 8 Copper 7s, and 6 Nova T devices were removed. It is recommended that the operator gain experience on women having their pregnancy terminated before attempting to remove an IUD from a pregnant woman wishing to continue her pregnancy.

The role of copper(I)-nicotinic acid complex on kojic acid biosynthesis by Aspergillus flavus.

Addition of cooper-monovalent-nicotinic acid complex to a synthetic medium specific for kojic acid production by Aspergillus flavus enhanced the production by about 47%. The substance is proposed to act via a biochemical utilization of the copper(I)-B3 complex in a manner similar to that of the naturally utilized nicotinic acid. NAD and NADP like carriers with higher reactivity have been predicted. According to this prediction the biosynthetic route of kojic acid has been interpreted on the basis of a model proposed by Bajpai et al. (1981). In this model the enzymes participating are dependent on NAD and NADP (glucose dehydrogenase and gluconate dehydrogenase) as well as on other reduction processes.

Diurnal rhythms in the synthesis and release of haemolymph proteins in the crustacean Isopoda, Porcellio dilatatus (Brandt), with special reference to vitellogenin.

Synthesis and release of vitellogenin in the crustacean Isopoda, Porcellio dilatatus, were maximal during premolt. At that time, diurnal variations occurred in the haemolymph protein level and in the synthesis and release of vitellogenin and other proteins in the haemolymph. The pattern of variations was always identical and bimodal: the minima were at dusk and at dawn when the maxima occurred in the middle of the day and of the night. These variations were highly significant, and the diurnal means were significantly higher than the nocturnal means.