Relative potency of different generic brands of meropenem, colistin and fosfomycin: Implications for antimicrobial therapy and antimicrobial formulary

There is a need of a relatively simple and inexpensive method for the determination of relative potency of various generic brands of antibiotics in comparison to original products. The current study describes an agar diffusion method which can be performed in any microbiology laboratory, is cheap (costs $2 per test) and its results can be available after overnight incubation. The results show that neither all generics are reliable nor are all generic antibiotics of poor quality.

Cytomegalovirus reactivation and disease amongst patients with allogeneic haematopoietic stem cell transplantation in Eastern India: Epidemiology, outcome and healthcare cost

Purpose: Data from developing countries about incidence, prognosis and healthcare cost of cytomegalovirus (CMV) reactivation amongst patients with allogeneic hematopoietic stem cell transplantation (AHSCT) remain scarce. The purpose of the study was to describe the epidemiology, outcome and cost implications of CMV reactivation and CMV disease amongst patients with AHSCT in cancer hospital in Eastern India. Materials and Methods: The study design was a retrospective audit of clinical records. Results: Ninety-nine per cent of patients and 94% of the donors were found to be CMV seropositive. CMV reactivation rate was 43.8% amongst patients with AHSCT (n = 130 patients). CMV reactivation occurred 118 days after AHSCT (median; range: 28–943 days). Patients with any grade of graft-versus-host disease (GVHD) had higher CMV reactivation rate than patients without GVHD. Patients with CMV reactivation had more frequent GVHD than patients without CMV reactivation. Use of steroids was associated with CMV reactivation. We found no differences in overall survival of patients with or without CMV reactivation. The cost of in-house CMV-polymerase chain reaction at our centre was USD $57 (Rs. 3650), cost for intravenous ganciclovir was USD $26 (Rs. 1665) per infusion and oral valganciclovir USD $8 (Rs. 512)/900 mg tablet. The median duration of anti-CMV therapy was 14 days (interquartile range: 14–28 days) and the average cost per patient per month directed towards CMV management ranged between USD $800 and USD $1,300 (Rs. 51,238–Rs. 83,264). Three patients (2.3%) in this series had CMV disease, all of whom died. Conclusion: In an increasingly globalised world, where medical tourism is common, data from developing countries regarding cost and outcome of CMV infections in AHSCT patients are of relevance.

A high yield DNA extraction method for medically important Candida species: A comparison of manual versus QIAcube‑based automated system.

The prognosis of infected individuals with candidemia depends on rapid and precise diagnosis which enables optimising treatment. Three fungal DNA extraction protocols have been compared in this study for medically important Candida species. The quality and quantity of the DNA extracted by physical, chemical and automated protocols was compared using NanoDrop ND-2000 spectrophotometer. It was found that the yield and purity (260/230) ratio of extracted DNA was significantly high in the physical treatment‑based protocol as compared to chemical based or automated protocol. Extracted DNA‑based real time‑polymerase chain reaction showed an analytical sensitivity of 103 cfu/mL. The result of this study suggests physical treatment is the most successful extraction technique compared to other two protocols.

Molecular screening of the neutrophil elastase gene in congenital neutropenia.

Congenital neutropenia is a rare hematopoietic disease, which occurs sporadically or as an auto-somal dominant inherited disorder. Pathogenesis of congenital neutropenia can now be attributed to mutations of the ELA2 gene encoding neutrophil elastase. A child with severe congenital neutropenia with a heterozygous mutation G1887A in exon 2 of ELA2 gene is reported.

ATP7B mutations in families in a predominantly Southern Indian cohort of Wilson's disease patients.

OBJECTIVE: To analyze ATP7B mutations in Wilson's disease (WD) patients from the Indian subcontinent and to correlate these with WD phenotype. METHODS: We studied 27 WD patients from 25 unrelated families. Twenty-two families were from three southern Indian states - Tamil Nadu andhra Pradesh and Kerala. We applied conformation- sensitive gel electrophoresis (CSGE) to screen for the mutations in patients and their families. PCR products exhibiting aberrant patterns in CSGE were subjected to direct DNA sequencing. As siblings affected by WD within a family share identical ATP7B genotype, we compared WD phenotype among affected siblings within families. RESULTS: ATP7B mutations were detected in 22 of the 25 probands -13 were homozygotes and 9 were compound heterozygotes. Eleven novel mutations were detected. Only two common mutations were found: G3182A in 4 (16%) and C813A in 3 (12%) probands. 'Hot spots' for ATP7B mutations were exons 18 and 13. Lack of common dominant mutations prevented correlation of individual ATP7B mutations with WD phenotype. Symptomatic WD in a live sibling was not found in any family. In 8 families, a sibling died of presumed WD - in 6 of these, WD phenotype was identical to that in the proband. CONCLUSIONS: We describe the spectrum of ATP7B mutations including 11 novel mutations in Indian WD patients and document lack of a single dominant mutation. Identical WD phenotype among siblings in only 6 of 8 families with >1 child affected by WD suggests that factors other than ATP7B mutations influence WD phenotype.

Interhemispheric subdural hematoma: an uncommon sequel of trauma.

Interhemispheric subdural hematomas are relatively uncommon and usually seen in patients with bleeding disorders. They may present with signs of the falx syndrome or seizures. The management options range from craniotomy and evacuation to conservative management. We report such a case in a patient with normal bleeding parameters, which was managed with a twist drill craniostomy and drainage of the hematoma.

Hemorrhagic necrosis of pituitary adenomas.

A clinicopathological study of 41 cases of pituitary apoplexy in a series of 324 surgically treated pituitary adenomas is presented. In 23 patients, the predominant operative finding was hemorrhage with or without necrosis. However, there were 15 (37.7%) cases where pale, necrotic tissue with no evidence of hemorrhage was found at surgery. Pale, necrotic material was particularly found when there was a long interval between the acute clinical event and surgery. It is concluded that the pale, necrotic debris represents one stage in the resorption process of blood after hemorrhagic necrosis of pituitary adenomas. This entity needs to be kept in mind especially since the material closely resemble the pultaceous material seen in craniopharyngiomas and epidermoid cysts.

Management of symptomatic vertebral hemangiomas: review of 13 patients.

Thirteen consecutive cases with symptomatic vertebral hemangiomas, managed during a five year period from January 1995 at the Christian Medical College and Hospital, Vellore, were analysed. Twelve patients had lesions in the thoracic and 1 in the sacral region. Eight patients had multiple level involvement. Seven patients had laminectomy and soft tissue component excision, of which one had intraoperative injection of absolute alcohol and one had postoperative radiotherapy. One patient had vertebrectomy and stabilization with preoperative embolization. One patient underwent only endovascular embolization. However, the focus of this communication is on 4 patients who underwent a CT guided percutaneous transpedicular injection of absolute alcohol into the affected vertebral body. In the surgical group, 6 patients had cavernous type and 2 patients had mixed type of hemangiomas. Ten patients improved on the Ranawat grade by the time of discharge. On the MRC grade, 11 patients had improved, one was grade 5 pre and postoperative, while one did not improve. Several options are available for the management of symptomatic vertebral hemangiomas and multiple modalities may have to be used for a single patient. CT guided percutaneous transpedicular injection of absolute alcohol shows promising results. However, long term follow up is mandatory.

Midbrain venous angioma with obstructive hydrocephalus.

A rare case of a mid brain venous angioma with obstructive hydrocephalus is described. A dilated draining vein from the lesion in the aqueduct as the cause of the hydrocephalus is highlighted, and interesting features of the pathology of venous angiomas and associated cavernous hemangioma are described. The management of this interesting condition is discussed.

Transsphenoidal line of vision on MRI for pituitary tumor surgery.

This study was done to determine the usefulness of the sagittal magnetic resonance image (MRI) in predicting the ease of resectability of pituitary adenomas operated through the transsphenoidal route. Tumors were graded according to Hardy's method and a new system proposed by us. In this system the amount of tumor above the line of vision (V-line) as seen on the sagittal MRI was estimated and correlated with the extent of excision assessed on the postoperative computed tomogram (CT). There were 7 Hardy's grade A (8.8%), 32 grade B (41.3%), 31 grade C (37.5%), 6 grade D (8.8%) and 2 grade E tumors (3.8%) among the 78 tumors studied. It was seen that most of the tumor volume was below the V-line in Hardy's grade A and B tumors. In grade C tumors 5 were < 25% above, 14 were 25-50% above and 12 were 50-75% above the V-line. A radical excision was possible in 15 of l9 grade C tumors in which < 50% of the tumor was above the V-line. However, only 5 of 12 with Hardy's grade C were radically excised when >50% of the tumor was above the V-line. In conclusion, Hardy's grade C tumors are not a homogenous group radiologically and using the V-line on MRI helps in predicting the case of respectability in a single stage.

Cytogenetic, fluorescent in situ hybridization & reverse transcriptase-polymerase chain reaction analysis in acute promyelocytic leukaemia patients.

BACKGROUND & OBJECTIVES: The presence of t(15;17) or PML-RAR alpha fusion transcript is the diagnostic hallmark of patients with acute promyelocytic leukaemia (APL). Cytogenetic (CG), fluorescent in situ hybridization (FISH) and reverse transcriptase-polymerase chain reaction (RT-PCR) are mainly the techniques used for detecting this abnormality. The objective of this study was to compare and assess the role of CG, FISH and RT-PCR in the diagnosis of APL. METHODS: CG, FISH and RT-PCR analysis were performed in 29 patients with APL (28 M3, 1 M3v; 27 studied at diagnosis and 2 at relapse). RESULTS: Karotypes obtained in 25 patients revealed t(15;17) in 21 normal karyotype in 3 and trisomy 8 in 1 patient. In 26 patients FISH was positive for PML-RAR alpha fusion in both interphase (IP) and metaphase, two were negative and one patient had no cells for FISH analysis. IP FISH confirmed the fusion of PML-RAR alpha in all patients with t(15;17) detected by CG. RT-PCR was positive in the 22 patients analyzed (7 patients did not have RT-PCR). PCR was positive in the 3 patients with cytogenetically normal karyotypes and in one patient when karyotyping was a failure. CG detected 21 (72.4%) patients with t(15;17) of which additional chromosomal abnormalities were detected in 20 per cent of patients with successful karyotype. INTERPRETATION & CONCLUSION: FISH and RT-PCR were useful in detecting PML-RAR alpha fusion in cytogenetically normal patients and those in when karyotyping was a failure and can be used in routine analysis for rapid confirmation of t(15;17) in patients with acute myeloid leukaemia.

Quantitative study of muscle spindles in suboccipital muscles of human foetuses.

The proprioceptive inputs from the cervical musculature play an important role in head-eye co-ordination and postural processes. Deep cervical muscles in humans are shown to have high spindle content. The density, distribution and morphology of muscle spindles were studied in superior oblique capitis, inferior oblique capitis and rectus capitis posterior major and minor three small suboccipital muscles. The muscles were obtained, post-mortem from stillborn human foetus. The spindle density was calculated as the ratio of mean spindle content to the mean wet weight of that muscle in grams. The distribution and arrangement of spindles within the muscle and their arrangement was studied. The spindle density of superior oblique muscle was found to be 190, that of inferior oblique was 242 and the rectus capitis posterior contained 98 spindles per gram of muscle. No tendon organs were seen. The serial transverse sections of inferior oblique muscle revealed muscle spindles of varying sizes, length varying between 100-650 microns and, diameter 50-250 microns. A complex parallel arrangements of group of large spindles were seen in the belly of the inferior oblique muscle, while the polar regions contain few small isolated spindles. The relevance of such high spindle receptor content in these tiny muscles is discussed.

Motor and somatosensory evoked potentials in a primate model of experimental spinal cord injury.

Motor and somatosensory evoked potentials (MEP and SSEP) were compared after experimental spinal cord injury in Bonnet monkeys (macaca radiata). The MEP and SSEP changes following graded injuries were related to clinical outcome. Eight healthy mature monkeys with a mean weight of 4.2 + 0.9 Kg were chosen for the study. Graded spinal cord injury was caused using 50, 100, 200, 300 gm-cm force by modified Allens' weight drop device. MEP and SSEP recordings were done before injury and at 0, 2, 4 and 6 hours after injury and on the 7th postoperative day. Neurological assessment was done at 24 hours and on the 7th day following injury. 50, 100, 200 gm-cm force caused partial injuries and 300 gm-cm force caused severe spinal cord injury. The predictive value of MEP and SSEP following partial injuries was 80% and 66.67% respectively. Both MEP and SSEP were 100% predictive in severe injury. MEP and SSEP monitoring can therefore be complementary to each other in predicting the neurological outcome in partial injuries to the spinal cord.

A pilot study on the role of cytomegalovirus & human herpesvirus-6 infections in Indian bone marrow transplant recipients.

BACKGROUND & OBJECTIVES: Studies from Western transplant centers have shown the importance of cytomegalovirus (CMV) in infections among immunosuppressed post-transplant patients (both solid and bone marrow transplant recipients). Human herpesvirus-6 (HHV-6) infection is also important. Since such data are lacking from India, we carried out a pilot study to investigate the role of these two viruses in infections among Indian allogeneic bone marrow transplant (BMT) recipients. METHODS: A total of 21 BMT patients who developed acute graft versus host disease (GVHD), two patients who developed chronic GVHD, and eight recipients who did not develop GVHD but had skin rash/elevated liver enzymes, persistent cytopaenia or interstitial pneumonitis with a high clinical suspicion of possible CMV association were studied for markers of CMV and HHV-6 infections. RESULTS: CMV DNAemia was documented in 9 (42.8%) and CMV IgM in 4(19%) of the 21 patients with acute GVHD. HHV-6 DNAemia was not seen in any patient with acute GVHD but 2 (9.5%) had HHV-6 IgM. Of the 2 patients with chronic GVHD, 1 was positive for CMV DNA and IgM, and both were negative for HHV-6 markers. The lower incidence of CMV DNAemia in our recipients may be attributable to the presence of neutralizing antibody (anti gB/AD-1) among the 17 CMV and HHV-6 DNAemia negative recipients, 4(23.5%) had neutralizing antibodies (S/N ratio > or = 5). Of the 13 CMV DNAemia positive recipients, only one (7.7%) was positive for neutralizing antibodies. Among the 5 neutralizing antibody (S/N ratio > or = 5) positive recipients, 4 (80%) were negative for CMV DNAemia. The one nPCR positive was revealed only at high DNA (> 0.1 microgram) input indicating low CMV signal strength. INTERPRETATION & CONCLUSION: The present study shows the use of DNAemia in detecting CMV infections among BMT recipients. All recipients had high avidity CMV IgG (AI > 50%) confirming CMV reactivation or reinfection in these patients. There was evidence from this study suggesting that neutralizing antibodies may play a role in controlling CMV reactivation. We found no significant HHV-6 association with GVHD in Indian allogeneic BMT recipients.

Arsenic trioxide in the management of acute promyelocytic leukaemia.

Arsenical compounds were used as early as 2000 BC, both as medicines as well as poisons. Arsenicals gained importance in the beginning of the last century as the primary mode of treating syphilis. In 1931, Folkner and Scott used an arsenical preparation called Fowler's solution in the treatment of chronic myeloid leukaemia. This continued to be used until the introduction of busulphan in 1953. In the 1970s, arsenic trioxide was introduced for the treatment of acute promyelocytic leukaemia in China and was found to be extremely effective in treating this condition. Since then, numerous in vitro and in vivo studies have confirmed this observation. This article reviews the pathogenesis of acute promyelocytic leukaemia, the possible mechanism of action of arsenic trioxide in this condition and the literature on its use in the treatment, with special reference to the clinical and molecular response rates, toxicity and pharmacology of this compound. It also attempts to address the role of arsenic trioxide in the present algorithm for the treatment of acute promyelocytic leukaemia.

Determination of Ki-67 labeling index in pituitary adenomas using MIB-1 monoclonal antibody.

Proliferative activity of 94 pituitary adenomas was assessed by the determination of the growth fraction, using MIB-1 monoclonal antibody in formalin fixed, paraffin embedded sections. This index was correlated with clinical and radiological evidence of invasiveness. The mean Ki-67 labeling index for all pituitary adenomas was 0.84% (range 0-17.45%). Hardy stage E tumours (1.44%) had a higher Ki-67 labeling index (LI) as compared with Hardy stage 0 tumours (0.36%). The difference in the Ki-67 labeling indices between invasive and non-invasive adenomas was not statistically significant. Hence, the Ki-67 labeling index is not a reliable indicator of invasiveness in pituitary adenomas.

Effect of dichloracetate on infarct size in a primate model of focal cerebral ischaemia.

Acidosis is a major contributing factor towards spread of the ischaemic focus in the brain. Drugs that increase pyruvate dehydrogenase activity could decrease the formation of lactic acidosis. The sodium salt of dichloracetic acid (DCA) has been found to be effective in reducing lactate. This study was undertaken to study the efficacy of DCA in reducing infarct size in experimental focal ischaemia in monkeys. Macaca radiata monkeys in the treatment group were given 35 mg per kilogram of dichloracetate intravenously immediately before occluding and interrupting the middle cerebral artery, and the control group was given saline as placebo under similar conditions. Mean infarct size expressed as a percentage of the size of the hemisphere in all the three brain slices was 35.38 in the control group as against l2.06 in the treated group (p=0. 0008).

Haemostasis with cryoprecipitate in patients undergoing surgery for severe von Willebrand disease.

BACKGROUND: There are limited data on the management of haemostasis in patients with severe von Willebrand disease undergoing major surgery. Data on the use of cryoprecipitate in this setting are even more limited. In many developing countries cryoprecipitate is often the only available source of factor replacement. The minimum factor levels required for maintaining haemostasis after surgery have never been carefully evaluated. METHODS: Data from 3 patients with severe von Willebrand disease who underwent 4 major surgical procedures at our institution, using lower than standard recommended doses of cryoprecipitate were analysed for adequacy of factor replacement and complications. RESULTS: The average preoperative cryoprecipitate infusion was 22.5 i.u. of factor VIII/kg (range: 15-25). The bleeding time done by the modified Ivy method, 30 minutes after infusion, was normal in all these patients. The average cryoprecipitate support for days 1-3 was 16.5 i.u. of factor VIII/kg/day (range: 12.5-25) and for days 4-10 was 12.4 i.u. of factor VIII/kg/day (range: 8.3-16). The mean duration of factor replacement was 12 days (range: 7-17). Two patients had delayed bleeding, one on day 3 attributed to the inadvertent use of a non-steroidal anti-inflammatory drug and the second on day 10 which was probably secondary to septicaemia. Bleeding resolved in both these patients as soon as the precipitating factors were relieved. CONCLUSION: The total amount of factor replaced in our patients is approximately half of what would have been used if the usual recommendations were followed. The data suggests that lower doses of cryoprecipitate could be adequate for major surgery and wound healing in severe von Willebrand disease. This will lead to lowering of costs and reducing the risk of transfusion-associated virus infection.