Clinical Profile of Pediatric Cases of Dengue during the 2019 Epidemic in Bangladesh: A Multicenter Cross-Sectional Study.

Bangladesh experienced the largest dengue epidemic during 2019, with more than 100,000 confirmed cases and 164 deaths. Almost one-third of these cases were children. The present study aimed to investigate the clinical and hematological manifestations of pediatric dengue cases during the epidemic. This was a multicenter cross-sectional study conducted in Dhaka Medical College Hospital, Dhaka, Dr. Sirajul Islam Medical College Hospital and Tangail Sadar Hospital, Tangail, Bangladesh between the period of June 2019 and September 2019. The study included 208 pediatric patients (age <18 years) with confirmed dengue fever. Patient's demographics, clinical and laboratory features of dengue were collected through patients' interview, clinical examination and laboratory investigations. Descriptive statistics were used to represent the patients' socio-demographic information, clinical presentations and hematological parameters. The majority of the patients were aged between 6 and 17 years with male predominance. The most commonly presented clinical manifestations were fever (100.0%), headache (59.0%), myalgia (42.0%), rash (36.0%), retro-orbital pain (28.0%) and diarrhea (24.0%). Warning signs abdominal pain (40.0%) and persistent vomiting (29.0%), bleeding manifestations such as melena (17.0%), gum bleeding (7.0%) and epistaxis (6.0%) and evidence of plasma leakage such as oliguria (3.4%), ascites (2.4%), pleural effusion (1.4%), and shock (1.0%) were also present in the patients. Raised HCT levels, leucopenia and thrombocytopenia were present in almost 23.0%, 43.0% and 28.0% of children, respectively. Warning signs and plasma leakage were present in a substantial number of patients indicating potential risk of severe dengue. Prompt diagnosis and management based on best clinical judgment might prevent severe dengue at an early stage.

Nosocomial Infection Among Burn Patients Admitted to a Tertiary Care Hospital of Bangladesh: A Cross-Sectional Study.

Nosocomial infection is a major challenge for the appropriate management of burns. The present study aimed to investigate incidence, risk factors, and causative organisms of nosocomial infection in burn patients of Khulna, Bangladesh. This cross-sectional study was conducted among patients admitted to the Burn and Plastic Surgery Department of Khulna Medical College Hospital (KMCH) from January to December 2020. Relevant data were collected from the patients' hospital records. Samples of wound swabs and blood were collected and cultured in the microbiology laboratory of KMCH. Logistic regression models were used to determine risk factors for infective complications in burn patients. All statistical analyses were carried out using SPSS version 26.0. A total of 100 burn patients were included. Mean age was 29.2 years with a male-female ratio of 1.3:1. Flame burns were most prevalent among the patients (41%), followed by scald (23%) and electric burns (15%). Almost 40% patients had full thickness burn. The incidence of nosocomial infection was 42% (wound infection 33% and septicemia 9%). Total body surface area of burn >40% (OR 7.56, 95% CI 2.89-19.81), full thickness burn (OR 34.40, 95% CI 3.25-97.14) and prolonged hospital stay (aOR 1.31, 95% CI 1.15-1.51) were significant risk factors for nosocomial infection. Staphylococcus aureus was the most commonly isolated organism (45%), followed by Streptococcus (24%), Pseudomonas aeruginosa (19%) and Escherichia coli (12%). As the epidemiology of nosocomial infection is not the same in different health facilities, a facility-based comprehensive burn management protocol considering the local epidemiology and causative organisms of burn wound infection is crucial for the prevention and management of nosocomial infections in burn patients.

Les infections nosocomiales sont une préoccupation majeure du traitement bien conduit des brûlés. Cette étude a eu pour but d'évaluer l'incidence, les facteurs de risque de survenue et les bactéries isolées d'infections nosocomiales survenues dans le CTB de Kulna (Bangladesh). Elle a étudié les dossiers l'ensemble des 100 patients admis dans le CTB du CHU de Kulna en 2020. Les analyses bactériologiques ont été réalisées dans le laboratoire du CHU. Une régression logistique a été utilisée pour déterminer les facteurs de risque d'infection. Toutes les analyses statistiques ont été réalisées avec SSPS 26.0. L'âge moyen était de 29,2 ans, le sex-ratio de 1,3H/1F. Les flammes représentaient 41% des causes, les liquides 23% et l'électricité 15%. Quasiment 40% des patients avaient des brûlures profondes. L'incidence des accidents infectieux était de 42% (cutanée 33%, bactériémies 9%). Les facteurs de risque indépendants de survenue d'une infection étaient une atteinte sur >40 % SCT (OR 7,56; IC95 2,89-19,81), une brûlure profonde (OR 34,40 ; IC95 3,25-97,14) et un séjour prolongé (OR 1,31; IC95 1,15-1,51). Les quatre bactéries les plus fréquentes étaient S. aureus (45%), Streptococcus spp (24%), P. æruginosa (19%), et E. coli (12%). Les épidémiologies bactériennes variant selon les services d'où elles sont issues, c'est sur l'épidémiologie locale que doivent se centre les mesures de contrôle des infections nosocomiales.

Protracted course of Krabbe disease in an adult patient bearing a novel mutation.

BACKGROUND: Krabbe disease, or globoid cell leukodystrophy, is an autosomal recessive disorder caused by the deficiency of galactocerebrosidase (GALC) activity. Although most cases are diagnosed in infancy and show a fatal outcome in childhood, adult patients have been identified, showing progressive spastic hemiparesis to tetraparesis, followed by optic atrophy, dementia, and neuropathy. The disease can be diagnosed by detecting the deficiency of GALC activity (less than 5% of normal) in any available tissue sample. The cloning of the human GALC gene allowed the molecular characterization of newly diagnosed patients. More than 75 disease-causing mutations and polymorphisms in this gene have been identified. OBJECTIVE: To describe a 28-year-old woman with Krabbe disease, correlating clinical and biochemical abnormalities to a novel mutation on the GALC gene. METHODS: Clinical investigation was enriched by neurophysiological and neuroimaging data. The activity of GALC was assayed in white blood cells using radiolabeled natural substrate. Genomic DNA was isolated from peripheral blood, and the GALC gene was sequenced. The mutated gene was expressed and GALC activity was measured in transfected COS-1 cells. RESULTS: The patient had progressive and bilateral amaurosis starting at 8 years of age. Although she was experiencing weakness in all her extremities, her intellect remained intact. She was found to be homozygous for a previously unreported missense mutation (T1886G), which leads to low, but not totally deficient, GALC activity. CONCLUSIONS: Expression of this mutation in COS-1 cells using the pcDNA3 expression vector (Invitrogen, Carlsbad, Calif) resulted in low, although not null, GALC activity, which can explain the protracted clinical course in this patient. Patients carrying the mutation described herein might be potential candidates for therapeutic trials, such as bone marrow transplantation or gene therapy.

Peripheral neuropathy with hypomyelinating features in adult-onset Krabbe's disease.

We describe three brothers suffering from Krabbe's disease with onset in the fifth decade. The proband showed a complete deficiency of leukocyte enzyme galactocerebrosidase and was found to be heterozygous for two previously described mutations: G > A809 and 502T/del consisting of a 30 kb deletion. In all three brothers the neurological examination showed features of asymmetrical peripheral neuropathy associated with pyramidal signs and the electrophysiological examination showed a generalized slowing of nerve conduction velocities. Two patients died at 59 and 61 years of age due to respiratory failure. Both the proband and his brother underwent a sural nerve biopsy. In the former the most striking finding was the presence of uniformly thin myelin sheaths without evidence of demyelination; a complete absence of fibers was found in the latter. Our findings confirm that peripheral neuropathy may be the presenting feature of late-onset Krabbe's disease. Hypomyelination rather than demyelination may represent the distinguishing pathological finding of this condition.

Leukocyte sonicates as a source for both enzyme assay and DNA amplification for mutational analysis of certain lysosomal disorders.

At present the identification of patients and carriers of most lysosomal disorders is accomplished by finding decreased activity of one enzyme in an easily obtained tissue sample such as leukocytes. As the genes for these enzymes are cloned and mutations identified, the use of molecular techniques to supplement enzyme testing will be warranted. To facilitate the implementation of such studies a simple method for isolating DNA from the remaining leukocyte sonicate, and using this DNA for polymerase chain reaction amplification of regions involved in three lysosomal disorders is described. The DNA from the sonicate was isolated without proteinase K digestion, was readily soluble in Tris-EDTA buffer and available for amplification almost immediately. The usefulness of the methods was confirmed by studies on patients and family members with three relatively common lysosomal disorders, metachromatic leukodystrophy. Gaucher disease and Tay-Sachs disease. This method allows immediate DNA analysis without the need for securing an additional blood sample.

Conversion of regional to general anaesthesia at caesarean section: increasing the use of regional anaesthesia through continuous prospective audit.

BACKGROUND: Anaesthetic-related maternal deaths have largely been attributed to complications of general anaesthesia. In our unit a retrospective audit conducted between 1997 and 2002 showed a 9.4% conversion rate to general anaesthesia for caesarean sections amongst women with epidural catheters in-situ. The Royal College of Anaesthetists has stated that <3% of cases should need conversion to general anaesthesia. To improve our figures, from 2004 to 2007 we prospectively audited all caesarean sections requiring general anaesthesia. METHODS: Data were collected on the number of caesarean sections, initial anaesthetic technique used, need for conversion either pre- or intra-operatively and the use of labour epidural analgesia, where an epidural had been in-situ. RESULTS: There were 2273 caesarean sections during the audit period. Neuraxial anaesthesia rates were for elective cases 95.3% (2004), 96.3% (2005), 98.3% (2006) and 98.2% (2007) and for emergency cases 82.3% (2004), 88.6% (2005), 87.0% (2006) and 85.7% (2007). Common reasons given for not using a regional technique were urgency of delivery (category 1) or anticipated large blood loss. Conversion rates from regional to general anaesthesia for elective cases were 0.8% (2004), 2.5% (2005), 0.5% (2006) and 0% (2007), and for emergencies 7.8% (2004), 2.7% (2005), 3.7% (2006) and 5.4% (2007). Improvements were seen in all but category-1 caesarean sections. CONCLUSIONS: Prospective audit has been associated with improved rates for neuraxial anaesthesia and reduced need for conversion to general anaesthesia in all but category-1 caesarean sections. The Royal College of Anaesthetists standards may need to be reviewed to become category-specific.

Characterization of the large deletion in the GALC gene found in patients with Krabbe disease.

Globoid cell leukodystrophy (GLD) of Krabbe disease results from mutations in the galactocerebrosidase (GALC) gene. Previously, we had identified a large deletion in the GALC gene together with a C to T polymorphism at cDNA position 502 in a significant number of cases of infantile Krabbe disease; however, the deletion breakpoint had not been found. In this paper we show that the deletion is approximately 30 kb starting near the middle of the 12 kb intron 10, and includes all of the coding region through exon 17 plus an additional 9 kb. The deletion junction contains a 4 bp direct repeat and is preceded by sequence identified as belonging to the Alu family of interspersed repetitive elements. Using genomic DNA and a PCR-based test to detect normal and deleted sequences at that location, a large number of patients with all clinical types of GLD were analyzed. Of 21 infantile patients found to be heterozygous for the 502T polymorphism reported previously, 15 had the deletion, one could not tested and five, including a Hmong child, did not have the deletion. Sixteen other infantile patients previously tested were found to be either homozygous (10) or heterozygous (6) for the deletion. In addition, five patients with juvenile and adult GLD were found to be heterozygous for the deletion. In every case tested, the deletion was always found on the same allele as the 502T polymorphism. However, other disease-causing mutations have been found on the 502T allele. With careful genotype analysis these families can receive improved genetic information including patient and carrier identification and preimplantation diagnosis.

Structure and organization of the human galactocerebrosidase (GALC) gene.

The deficiency of galactocerebrosidase (GALC; EC 3.2.1.46) is responsible for globoid cell leukodystrophy (GLD, Krabbe disease) in humans and certain animals. This enzyme catalyzes the lysosomal hydrolysis of specific galactolipids including galactosylceramide (galactocerebroside) and galactosylsphingosine (psychosine), among others. Recently we cloned the full-length human GALC cDNA using amino acid sequence information obtained from GALC purified from human urine and brain. In this communication we describe the organization of the human GALC gene. The gene, of nearly 60 kb, consists of 17 exons, which, aside from the first and last, are relatively small, ranging from 39 to 181 nucleotides. The 16 introns range from 247 nucleotides to nearly 12 kb. The 5' untranslated region is GC-rich, containing no perfect CAAT or TATA sequences, similar to genes for other lysosomal proteins. This information will be useful for studies to identify mutations causing low GALC activity in all patients with GLD and to identify the homologous gene in the important animal models.

Molecular genetics of Krabbe disease (globoid cell leukodystrophy): diagnostic and clinical implications.

Galactocerebrosidase (GALC) is a lysosomal beta-galactosidase responsible for the hydrolysis of the galactosyl moiety from several galactolipids, including galactosylceramide and psychosine. The deficiency of this enzyme results in the autosomal recessive disorder called Krabbe disease. It is also called globoid cell leukodystrophy (GLD), because of the characteristic storage cells found around cerebral blood vessels in the white matter of affected human patients and animal models. Although most patients present with clinical symptoms before 6 months of age, older patients, including adults, have been diagnosed by their severe deficiency of GALC activity. More than 40 mutations have been identified in patients with all clinical types of GLD. While some mutations clearly result in the infantile type if found homozygous or with another severe mutation, it is difficult to predict the phenotype of novel mutations or when mutations are found in the heterozygous state. A high incidence of polymorphic changes on apparent disease-causing alleles also complicates the interpretation of the effects of mutations. The detection of mutations has greatly improved carrier identification among family members and will permit preimplantation diagnosis for some families. The molecular characterization of the naturally occurring mouse, dog, and monkey models will permit their use in trials to evaluate different modes of therapy.

Multiple mutations in the GALC gene in a patient with adult-onset Krabbe disease.

A 53-year-old man was diagnosed 8 years earlier with globoid cell leukodystrophy (GLD, Krabbe disease) by his severe deficiency of galactocerebrosidase (GALC) activity. He was found to have eight nucleotide changes on the two copies of his GALC gene, including two in the leader sequence, four considered polymorphisms, and two unique mutations.

Cloning of the canine GALC cDNA and identification of the mutation causing globoid cell leukodystrophy in West Highland White and Cairn terriers.

Globoid cell leukodystrophy, or Krabbe disease, is a severe, autosomal recessive disorder resulting from a deficiency of galactocerebrosidase (GALC) activity. GALC is responsible for the lysosomal catabolism of certain galactolipids, including galactosylceramide and psychosine. In addition to the human patients, there are several naturally occurring animal models for this disease, including the twitcher mouse, West Highland White terriers (WHWT), and Cairn terriers. All species have deficient GALC activity and have the characteristic pathological findings in the nervous system. We now describe the cloning of the canine GALC cDNA and the identification of the disease-causing mutation in both terrier breeds. The 2007-bp open reading frame is 88% identical to that in human, and the deduced amino acid sequence is about 90% identical. However, the 3'-untranslated region is about 1 kb shorter than that in the human. Two nucleotide changes were found in affected dogs, an A to C transversion at cDNA position 473 (Y158S) and a C to T transition at position 1915 (P639S). Expression studies in COS-1 cells demonstrated that the A to C change at 473 is the disease-causing mutation. A rapid test for the identification of the genotype at that position has been developed, and over 100 WHWT and Cairn terriers have been screened. This will allow breeders to mate their dogs selectively and will permit the establishment of a colony of dogs for use in therapy trials.

Mutations in the lysosomal beta-galactosidase gene that cause the adult form of GM1 gangliosidosis.

Three adult patients with acid beta-galactosidase deficiency/GM1 gangliosidosis who were from two unrelated families of Scandinavian descent were found to share a common point mutation in the coding region of the corresponding gene. The patients share common clinical features, including early dysarthria, mild ataxia, and bone abnormalities. When cDNA from the two patients in family 1 was PCR amplified and sequenced, most (39/41) of the clones showed a C-to-T transition (C-->T) at nucleotide 245 (counting from the initiation codon). This mutation changes the codon for Thr(ACG) to Met(ATG). Mutant and normal sequences were also found in that position in genomic DNA, indicating the presence of another mutant allele. Genomic DNA from the patient in family 2 revealed the same point mutation in one allele. It was determined that in each family only the father carried the C-->T mutation. Expression studies showed that this mutation produced 3%-4% of beta-galactosidase activity, confirming its deleterious effects. The cDNA clones from the patients in family 1 that did not contain the C-->T revealed a 20-bp insertion of intronic sequence between nucleotides 75 and 76, the location of the first intron. Further analysis showed the insertion of a T near the 5' splice donor site which led to the use of a cryptic splice site. It appears that the C-->T mutation results in enough functional enzyme to produce a mild adult form of the disease, even in the presence of a second mutation that likely produces nonfunctional enzyme.

Mutational analysis in a patient with a variant form of Gaucher disease caused by SAP-2 deficiency.

It is now clear that the lysosomal hydrolysis of sphingolipids requires both lysosomal enzymes and so-called sphingolipid activator proteins (SAPs). One gene, called prosaposin, codes for a precursor protein that is proteolytically cut into four putative SAPs. These four SAPs, of about 80 amino acids, share some structural features but differ somewhat in their specificity. Domain 3 of prosaposin mRNA contains the coding region for SAP-2, an activator of glucocerebrosidase. While most patients with Gaucher disease store glucosylceramide due to defects in glucocerebrosidase, a few patients store this lipid in the presence of normal enzyme levels. In this paper we describe the identification of a point mutation in domain 3 of a patient who died with this variant form of Gaucher disease. Polymerase chain reaction amplification was performed in the small amount of genomic DNA available using primers generated from the intronic sequence surrounding domain 3. The patient was found to have a T-to-G substitution at position 1144 (counting from the A of ATG initiation codon) in half of the M13 recombinant clones. This changes the codon for cysteine382 to glycine. His father and unaffected brother also had this mutation, but his mother did not. She was found to have half of the normal amount of mRNA for prosaposin in her cultured skin fibroblasts. Therefore, this child inherited a point mutation in domain 3 from his father and a deficiency of all four SAPs coded for by prosaposin from his mother.

Cloning and expression of cDNA encoding human galactocerebrosidase, the enzyme deficient in globoid cell leukodystrophy.

Globoid cell leukodystrophy (Krabbe disease) is an autosomal recessive disorder resulting from the deficiency of galactocerebrosidase (GALC) activity. GALC is responsible for the lysosomal catabolism of galactosylceramide, a major lipid in myelin, kidney and epithelial cells of small intestine and colon. We describe the molecular cloning of human GALC cDNA and its expression in COS-1 cells. Degenerate PCR primers, derived from N-terminal amino acid sequence from the 51 kDa band from human brain, were used to amplify cat testes RNA, and the resulting product was used to screen human testes and brain libraries. Two overlapping clones contained the total protein coding region, while additional clones and PCR amplification were needed to obtain the complete 3' end of the cDNA. The 3795 bp obtained include 47 bp 5' to the initiation start site, 2007 bp of open reading frame (coding for 669 amino acids), and 1741 bp of 3' untranslated sequence. Modification of the sequence surrounding the initiation codon to one more favorable for expression, resulted in a 6-fold increase in GALC activity in transfected COS-1 cells. The isolation of this clone will permit investigations into the causes for GALC deficiency in humans and available animal models, development of more accurate tests for patient and carrier identification, and evaluation of methods for effectively treating GALC deficiency, initially using the animal models.

Regional mapping of the human galactocerebrosidase gene (GALC) to 14q31 by in situ hybridization.

The cDNA for human galactocerebrosidase (GALC) has recently been cloned and expressed. A portion of this cDNA was used for in situ hybridization, and the region of strongest signal corresponded to human chromosome region 14q31. This agrees with recent linkage studies that localized Krabbe disease (globoid cell leukodystrophy) to the same region. This information will be useful in future studies for mapping this gene in animal models of GALC deficiency.

The mechanism for a 33-nucleotide insertion in mRNA causing sphingolipid activator protein (SAP-1)-deficient metachromatic leukodystrophy.

Metachromatic leukodystrophy is a severe autosomal recessive disorder caused by accumulation of sulfatide resulting from deficient lysosomal degradation. While most patients have mutations in the lysosomal enzyme arylsulfatase A, some patients have mutations in a required heat stable sphingolipid activator protein, we call SAP-1. One patient with SAP-1 deficiency was previously demonstrated to have a 33-nucleotide insertion in her mRNA. This resulted in the production of mature SAP-1 with 11 extra amino acids, which was unstable during intracellular processing. In this manuscript we demonstrate that the 33 nucleotides are present near the middle of a 4-kb intron, and that a single base change, c to a, in the second position preceding the 33-nucleotide insertion, coupled with the presence of a string of pyrimidines immediately upstream from this change, creates a new 3' splice junction. The presence of a string of pyrimidines within the 33-nucleotide insertion, which has three cag trinucleotides near the 3' end, leads to alternative splicing in normal people as found in this laboratory and by others. The insertion region is followed by a gt dinucleotide that is spliced to a typical 3' consensus sequence. The single nucleotide change, c to a, was confirmed by identifying normal and mutant sequence in the consanguineous parents and a sister, previously identified as a carrier of this disorder.

Correction of sulfatide metabolism after transfer of prosaposin cDNA to cultured cells from a patient with SAP-1 deficiency.

The lysosomal removal of the sulfate moiety from sulfatide requires the action of two proteins, arylsulfatase A and sphingolipid activator protein-1 (SAP-1). Recently, patients have been identified who have a variant form of metachromatic leukodystrophy which is characterized by mutations in the gene coding for SAP-1, which is also called "prosaposin." All of the mutations characterized in these patients result in (a) deficient mature SAP-1, as determined by immunoblotting after SDS-PAGE of tissue and cell extracts, and (b) decreased ability of cultured skin fibroblasts to metabolize endocytosed [14C]-sulfatide. We now report the insertion of the full-length prosaposin cDNA into the Moloney murine leukemia virus-derived retroviral vector, pLJ, and the infection of cultured skin fibroblasts from a newly diagnosed and molecularly characterized patient with SAP-1 deficiency. The cultured cells infected with the prosaposin cDNA construct now show both production of normal levels of mature SAP-1 and completely normal metabolism of endocytosed [14C]-sulfatide. These studies demonstrate that the virally transferred prosaposin cDNA is processed normally and is localized within lysosomes, where it is needed for interaction between sulfatide and arylsulfatase A. In addition, normal as well as mutant sequences can now be found by allele-specific oligonucleotide hybridization of PCR-amplified genomic DNA by using exonic sequences as primers.

A large deletion together with a point mutation in the GALC gene is a common mutant allele in patients with infantile Krabbe disease.

Galactocerebrosidase (GALC) activity is deficient in all patients with globoid cell leukodystrophy (GLD). While most patients have the severe infantile form of this autosomal recessive disorder (Krabbe disease), patients up to 50 years of age have been diagnosed in this laboratory. With the cloning of the GALC cDNA and availability of information regarding the gene organization, patients can be evaluated for their disease-causing mutations. We now report that a large deletion, together with a polymorphic C to T transition at position 502 of cDNA (counting from the A of the initiation codon), is responsible for a large number of disease-causing alleles in patients with Krabbe disease. Of 48 patients evaluated, 10 were found to be homozygous for the 502/del allele, five patients were heterozygous for this allele, 21 patients were heterozygous for the 502 mutation (presence of the deletion could not be confirmed), and one infantile patient was homozygous for the 502 mutation but at least one allele was not deleted. No patient was found to have the deletion without the 502 polymorphism. The delineation of mutations causing infantile Krabbe disease will provide new information regarding structure-function relationships in this multi-subunit enzyme and will improve the identification of patients and carriers in some families.

Characterization of the rhesus monkey galactocerebrosidase (GALC) cDNA and gene and identification of the mutation causing globoid cell leukodystrophy (Krabbe disease) in this primate.

Krabbe disease or globoid cell leukodystrophy (GLD) is a severe lysosomal disorder resulting from the deficiency of galactocerebrosidase (GALC) activity. This deficiency results in the insufficient catabolism of several galactolipids that are important in the production of normal myelin. Since the cloning of the human GALC cDNA and gene many disease-causing and polymorphic changes have been identified. This autosomal recessive disease has been reported to occur in several animal species, and recently the murine and canine GALC genes have been cloned. We now describe the cloning of the GALC cDNA and gene from the rhesus monkey and the identification of the mutation causing GLD in this species. The nucleotide sequence of the coding region and the gene organization were nearly identical to human. The deduced amino acid sequence of the monkey GALC was compared to the human, dog, and mouse, and it was found to be 97, 87, and 83% identical, respectively. The mutation causing GLD in the rhesus monkey is a deletion of AC corresponding to cDNA positions 387 and 388 in exon 4. This results in a frame shift and a stop codon after 46 nucleotides. A rapid method to detect this mutation was developed, and when 45 monkeys from this colony were tested, 22 were found to be carriers. The availability of this nonhuman primate model of GLD will provide unique opportunities to evaluate treatment for this severe disease.