Zeolite-based, dry formulations for conservation and practical application of Paraburkholderia phytofirmans PsJN.

AIMS: Producing dry, zeolite-based granular and powder inoculants of the Gram-negative, plant growth-promoting bacterium Paraburkholderia phytofirmans PsJN. Key aspects were maintenance of cell viability during desiccation and throughout storage at ambient conditions. METHODS AND RESULTS: Twenty additives and exopolysaccharide (EPS) produced by PsJN were screened for conserving cell viability of PsJN in air-drying and lyophilization. Suitable combinations (e.g. skimmed milk + air-drying) increased survival of PsJN up to 100 000-fold and maintained it for >7 months. EPS performed as good as skimmed milk during air-drying, but was second-rank regarding shelf life. Combinations of zeolite, skimmed milk and gelatin as a film-forming agent were extruded and processed into granules and powders, both displaying relatively stable viability for over 4 weeks at ambient conditions. Gelatin promoted brittleness of zeolite-based inoculants. CONCLUSIONS: Viability of highly sensitive PsJN was successfully conserved in dry formulations, taking into account the interplay between carrier, protectants, drying method and coating agent. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first study to provide ways of maintaining viability of PsJN during desiccation stress and to investigate the applicability of its EPS as a protectant, thus ultimately facilitating successful plant inoculation especially under field conditions.

[Scanning laser ophthalmoscope multifocal electroretinography and microperimetry in patients with Stargardt's disease]. / SLO-mfERG-Kampimetrie und SLO-Mikroperimetrie bei Morbus Stargardt.

PURPOSE: We used a scanning laser ophthalmoscope (SLO) evoked multifocal electroretinography (mf-ERG) to evaluate retinal function in patients with Stargardt's disease. SLO microperimetry could demonstrate the size of central retinal scotoma very well in these patients. The aim of the examination was to correlate the results of SLO mf-ERG and SLO microperimetry. METHODS: In four patients with Stargardt's disease SLO mf-ERG and SLO microperimetry were performed. The area of measurement in the SLO mfERG had a 24 degrees diameter (12 degrees visual angle) at the posterior pole of the eye. Stimulation was done using a helium-neon laser (632.8 nm). Simultaneous control of fixation was made using a infrared laser (730 nm). SLO microperimetry was performed with stimuli having the size of Goldmann III stimuli and the intensities 0 dB, 12 dB and 20 dB. In this study the reduction of SLO mfERG amplitudes was correlated to graded stimulus intensities in the SLO microperimetry. RESULTS: The area of reduced retinal function in the SLO mf-ERG measurement could be well correlated to the size of the scotoma in the SLO microperimetry, using the stimulus Goldmann III with the intensity 20 dB. CONCLUSION: SLO mfERG and SLO microperimetry are sensitive methods for quantifying functional deficits and are therefore useful for performing a detailed examination of the retina.

Detecting chloroquine retinopathy: electro-oculogram versus colour vision.

AIM: To investigate the relative sensitivity and specificity of two tests of retinal function (the electro-oculogram (EOG) and a computerised colour vision test) in screening for ocular toxicity caused by chloroquine and hydroxychloroquine. METHODS: 93 patients with rheumatic diseases receiving long term chloroquine and hydroxychloroquine therapy were followed for an average of 2.6 years. Clinical examination, an EOG, and a quantitative test of colour vision were carried out every 6 months. RESULTS: Mild fundus changes were observed in 38 patients. Four patients developed typical bull's eye maculopathy, three of whom had received 250, 365, and 550 g total dose of chloroquine, and one 1500 g of hydroxychloroquine. Statistical analysis of all patients showed that for those with no fundus changes or stippled pigmentation a number showed elevation of tritan threshold, so that if macular stippling is a sign of mild retinopathy the test on tritan changes has a 64% sensitivity and 63% specificity for an upper threshold value of 7%. All four patients with bull's eye lesions showed a marked disturbance of tritan colour vision, with a threshold of 14.8%, a sensitivity of 75%, and a specificity of 94%. For protan colour vision a threshold of 10% gives 75% sensitivity and 91% specificity. By contrast, neither an absolute nor a relative EOG reduction was a valid criterion for early or late chloroquine retinopathy. In advanced retinopathy an Arden coefficient (AQ) <180% yields 50% sensitivity and 54% specificity. When AQ <160% is the threshold, sensitivity does not increase but specificity rises to 82%. Occurrence of marked corneal deposits on clinical examination yields 50% sensitivity and 90% specificity in this situation. CONCLUSION: Screening for chloroquine retinopathy can be improved by using a sensitive colour test. Disturbance of the tritan axis appears to occur first. A normal test result on computerised colour testing virtually excludes any retinopathy by antimalarials. The EOG is of little diagnostic value.

Scanning laser ophthalmoscope-evoked multifocal-ERG (SLO-m-ERG) by using short m-sequences.

PURPOSE: To demonstrate the possibility of topographic mapping of retinal function under simultaneous control of fixation in humans, by scanning laser ophthalmoscope evoked multifocal electroretinography (SLO-m-ERG). METHODS: A confocal scanning laser ophthalmoscope was used as a stimulator and trigger unit to take m-ERGs. Short m-sequences based on a modified algorithm were used, with the advantage that each measurement cycle can be evaluated separately. We examined 78 normal subjects; in 62 a distortion factor of 1:1 was applied, and a factor of 1:4 in 16. RESULTS: The recorded amplitudes decreased with eccentricity, approximately following the decrease of retinal cone density. Amplitudes were higher in the central hexagonal element in the group with 1:4 distortion than in the group with the 1:1 distortion setting. CONCLUSIONS: SLO-m-ERG is a reliable technique for topographic mapping of retinal function under simultaneous control of fixation.

[Topographic mapping of retinal function with a scanning laser ophthalmoscope and multifocal electroretinography using short M-sequences]. / Topograficheskoe kartirovanie funktsii setchatki pri ispol'zovanii skaniruiushchego lazernogo oftal'moskopa i M-posledovatel'nostei dlia mul'tifokal'noi élektroretinografii.

A new method of multifocal electroretinography making use of scanning laser ophthalmoscope with a wavelength of 630 nm (SLO-m-ERG), evoking short spatial visual stimuli on the retina, is proposed. Algorithm of presenting the visual stimuli and analysis of distribution of local electroretinograms on the surface of the retina is based on short m-sequences. Mathematical cross correlation analysis shows a three-dimensional distribution of bioelectrical activity of the retina in the central visual field. In normal subjects the cone bioelectrical activity is the maximum in the macular area (corresponding to the density of cone distribution) and absent in the blind spot. The method detects the slightest pathological changes in the retina under control of the site of stimulation and ophthalmoscopic picture of the fundus oculi. The site of the pathological process correlates with the topography of changes in bioelectrical activity of the examined retinal area in diseases of the macular area and pigmented retinitis detectable by ophthalmoscopy.

X-linked ocular albinism (Nettleship-Falls): a novel 29-bp deletion in exon 1. Carrier detection by ophthalmic examination and DNA analysis.

BACKGROUND: Mutations in the OA1 gene on the short arm of the X chromosome are known to cause X-linked ocular albinism (x1OA) in males. A four-generation family with this disorder, including asymptomatic carrier females, was investigated by molecular analysis of the OA1 gene. METHODS: DNA samples were available from 22 individuals of this family, including 6 affected males and 6 obligate carriers. The nine exons of the OA1 gene were amplified and further analyzed by SSCP and sequencing. RESULTS: A detailed clinical examination of the index patient and two female carriers showed the typical signs of ocular albinism. Visual evoked potential responses showed markedly asymmetrical responses from the two hemispheres in the affected person as well as in the carriers, as a result of misrouting and decussation of optic nerve fibers. Molecular genetic analysis demonstrated a previously undescribed 29-bp deletion at position 225-253 in exon 1 of the OA1 gene, which segregated in the family. CONCLUSION: Clinical examination combined with molecular genetic analysis enhances the potential for a precise diagnosis for persons at risk of x1OA and provide an accurate basis for genetic counseling.

Effect of alcohol and light on the retinal pigment epithelium of normal subjects and patients with retinal dystrophies.

BACKGROUND: Light absorbed by photoreceptors causes oscillations in the voltage across the retinal pigment epithelium (RPE). This is the basis of the clinical test, electro-oculography (EOG). We have previously shown that alcohol causes a sequence of voltage changes which are so precisely the same as those caused by light that they must be produced by the same RPE machinery. There is good evidence that alcohol produces its effect by a direct action on the RPE. Consequently, in diseases associated with loss of photoreceptors, alcohol should continue to produce the voltage changes of the EOG unless secondary changes have occurred in the RPE. METHODS: The alcohol response in patients with retinitis pigmentosa (RP) was investigated using EOG. RESULTS: In no patient with RP was there any alcohol rise. CONCLUSION: In patients with RP secondary abnormalities of function of the RPE must occur.

[Color vision in relation to age: a study of normal values]. / Farbensehen in Abhängigkeit vom Alter: eine Normwertstudie.

BACKGROUND: It is difficult to quantify thresholds in most colour vision tests, and this is especially the case for tritan hues, where a strong age-related increase of threshold has been reported. With the development of computer-graphic methods it is possible to remove brightness clues caused by lens absorption. This study attempts to give normative values for colour contrast thresholds and assess the age related changes therein. PATIENTS AND METHODS: 115 patients aged between 6 & 71 years were tested for central and peripheral colour contrast sensitivity. No patient had any systemic or eye disease. As a preliminary, heterochromatic flicker balance between the luminosities of the R and G and B and G phosphors was established, so that all colours subsequently generated were isoluminant for the person tested. Then, using a modified binary search technique, colour contrast thresholds were established using both 2 degree optotypes, for central vision, and a ring, 12.5 degrees in radius for peripheral vision. In the latter case, the observer had to name the position of the missing quadrant in the ring. Stimuli were presented for 200 msec at 1 Hz. Colours were modulated on protan, deutan or tritan colour axis. RESULTS: No correlation between age and central colour vision thresholds was observed. By contrast a significant but only minor increase of peripheral colour vision threshold was observed for the peripheral protan and tritan axis. DISCUSSION: The present system removes luminance clues from colour vision tests and permits both central and peripheral retina to be tested. The results are simple in that the influence of age can be neglected. The variability of threshold results is small, and it is easy to detect the relatively large changes associated with disease. Since high-quality monitors are standardised and calibrated, providing the stimulus parameters described are adhered to, the results given here for upper limits of normal may be used for other similar systems.

S-cone ERGs elicited by a simple technique in normals and in tritanopes.

PURPOSE: To measure changes in the relative spectral sensitivities of the dark adapted and light adapted ERG and thus to establish the possible contribution of rods to the 'blue cone' ERG elicited by flashes of blue light. BACKGROUND: Short wavelength stimuli in the light-adapted eye evoke small rounded b-waves which have been considered to be S-cone responses. We have recorded such responses from tritanopes, which called the assumptions into question. METHODS: Small ERGs were recorded to blue and green flashes. The stimulus was a Ganzfeld which employed light emitting diodes. ERGs were obtained in both the dark-adapted eye and after light adaptation to intense orange light (peak wavelength 610 nm). The change in sensitivity with light adaptation and the relative spectral sensitivity was determined from the voltage/log light intensity functions, using a 10 microV criterion. RESULTS: (1) peak times and changes in sensitivity did not help distinguish light-adapted rod from possible S-cone responses; (2) analysis of the change in the ratio of blue:green sensitivity from darkness to 4.4 log Td. 610 nm background suggests that in seven normal subjects, 90% or more of the ERG evoked by 440 nm flashes is generated by S-cones; (3) three tritanopes have insignificantly reduced S-cone responses. CONCLUSIONS: (1) clinical techniques used to isolate S-cone ERGs are appropriate; (2) there are at least two types of tritanope and in those we investigated, functional S-cones are probably displaced into the retinal periphery.

Effects of refractive blur on the multifocal electroretinogram.

A significant difference in the response density of the MF-ERG response has been suggested for every 2 diopter change of refraction. The influence of refractive blur on the MF-ERG was studied in 8 healthy volunteers using either the VERISTM system (Group A: n=5) or Retiscan(TM) (Group B: n=3). For each eye recordings were obtained with a corrective lens of -3 dpt, 0 dpt, +3 dpt and +6 dpt placed in front of the contact lens electrode. The viewing distance was adjusted to compensate for the induced changes in the retinal image size. When the changes in retinal image size due to the refractive lens were compensated for, no influence due to refraction was observed in either latencies or amplitudes of (KI (P > 0.05). This held true for the central response average (four degrees) as well as for the outer 6-25 degrees. In KII.1 only the peripheral amplitudes of Group B showed an influence due to refraction (P < or = 0.05). This may be due to adaptation as the recordings of group B were obtained in succession. As expected, significant differences were observed when the recordings obtained with the different systems were compared (P < or = 0.05).

Solar retinopathy. A study from Nepal and from Germany.

319 patients with a solar retinopathy were seen in an eye clinic in Nepal within 20 months. All patients had either a positive history of sun-gazing or typical circumscribed scars in the foveal area. In more than 80% of the patients the visual acuity was 6/12 or better and did not deteriorate over time. 126 (40%) patients had a history of gazing at the sun during an eclipse, 33 (10%) were sun worshipers and 4 (1%) were in both categories. Three years later 29 patients were re-examined in a follow-up study. Only 16 had had visual disturbances directly after they had gazed into the sun. No colour vision defects were seen in any of the 44 affected eyes, when tested with Panel D 15, while four patients (6 eyes) had some uncertainty with the tritan plates of the Ishihara test charts. Metamorphopsia were recorded in 11 eyes. Five German patients with solar retinopathy were examined in more detail. Colour contrast sensitivity (CCS) was tested for the central and the peripheral visual field. CCS for tritan axis was raised in all patients for the central visual field, while it was normal for the peripheral visual field.

HIV-related ocular microangiopathic syndrome and color contrast sensitivity.

PURPOSE: Color vision deficits in patients with acquired immunodeficiency syndrome (AIDS) or human immunodeficiency virus (HIV) disease were reported, and a retinal pathogenic mechanism was proposed. The purpose of this study was to evaluate the association of color vision deficits with HIV-related retinal microangiopathy. METHODS: A computer graphics system was used to measure protan, deutan, and tritan color contrast sensitivity (CCS) thresholds in 60 HIV-infected patients. Retinal microangiopathy was measured by counting the number of cotton-wool spots, and conjunctival blood-flow sludging was determined. Additional predictors were CD4+ count, age, time on aerosolized pentamidine, time on zidovudine, and Walter Reed staging. The relative influence of each predictor was calculated by stepwise multiple regression analysis (inclusion criterion; incremental P value = < 0.05) using data for the right eyes (RE). The results were validated by using data for the left eyes (LE) and both eyes (BE). RESULTS: The only included predictors in multiple regression analyses for the RE were number of cotton-wool spots (tritan: R = .70; deutan: R = .46; and protan: R = .58; P < .0001 for all axes) and age (tritan: increment of R [Ri] = .05, P = .002; deutan: Ri = .10, P = .004; and protan: Ri = .05, P = .002). The predictors time on zidovudine (Ri = .05, P = .002) and Walter Reed staging (Ri = .03, P = .01) were additionally included in multiple regression analysis for tritan LE. The results for deutan LE were comparable to those for the RE. In the analysis for protan LE, the only included predictor was number of cotton-wool spots. In the analyses for BE, no further predictors were included. The predictors Walter Reed staging and CD4+ count showed a significant association with all three criteria in univariate analysis. Additionally, tritan CCS was significantly associated with conjunctival blood-flow sludging. CONCLUSION: CCS deficits in patients with HIV disease are primarily associated with the number of cotton-wool spots. Results of this study are in accordance with the hypothesis that CCS deficits are in a relevant part caused by neuroretinal damage secondary to HIV-related microangiopathy.

Impairment of colour contrast sensitivity and neuroretinal dysfunction in patients with symptomatic HIV infection or AIDS.

Ophthalmic and neurological complications are frequent findings in patients with AIDS. Little is known about neuroretinal dysfunction in patients with HIV infection. The purpose of this study was to measure and evaluate colour vision in patients with HIV infection or AIDS. Colour contrast sensitivity tests were performed on 75 patients (150 eyes) in different stages of HIV infection. A highly sensitive computer graphics system was used to measure tritan, deutan, and protan colour contrast thresholds. Patients were classified into three clinical groups: (a) asymptomatic HIV infection, (b) lymphadenopathy syndrome or AIDS-related complex, and (c) AIDS. Overall, tritan (p < 0.0001), deutan (p = 0.003), and protan (p = 0.009) colour contrast sensitivities were significantly impaired in patients with HIV infection compared with normal controls. Colour thresholds in patients with asymptomatic HIV infection (mean tritan threshold: 4.33; deutan: 4.41; protan: 3.97) were not impaired compared with normal controls. Colour vision was slightly impaired in patients with lymphadenopathy syndrome or AIDS-related complex (tritan: 6.25 (p < 0.0001); deutan: 4.99 (p = 0.02); protan: 4.45 (p = 0.05)). In patients with AIDS the impairment was even more marked (tritan: 7.66 (p < 0.0001); deutan: 5.15 (p < 0.0009); protan: 4.63 (p = 0.004)). Analysis of covariance controlling for age demonstrated a close association between impairment of tritan colour contrast sensitivity and progression of HIV disease (p < 0.0001). Following Köllner's rule, our study suggests that neuroretinal dysfunction occurs in patients with symptomatic HIV infection or AIDS. This is emphasised by the finding that the relative impairment in tritan vision compared with deutan/protan vision might reflect the difference in the number of cones or receptive fields. Measurement of tritan colour contrast sensitivity appears to be an appropriate and easily applicable method to detect early neuroretinal dysfunction in patients with HIV disease.

[Color vision defect. A early sign of Sorsby retinal dystrophy?]. / Farbsinnstörung. Ein Frühzeichen bei Sorsby-Netzhautdystrophie?

In 34 asymptomatic patients from three families at 50% risk of developing Sorsby's fundus dystrophy, colour contrast sensitivity was measured. In 16 the thresholds--mainly of the tritan axis--were raised above the normal values. It is concluded that testing of colour vision is useful in detecting the abnormal genotype. Raised colour contrast sensitivity was not observed before the first fundus or fluorescein angiographic changes in two of the families, while the colour defect occurred in the absence of ophthalmoscopic abnormality in the third family. Therefore, our results support the clinical suggestion that Sorsby's fundus dystrophy is more than one disease.

Leber's hereditary optic neuroretinopathy and the X-chromosomal susceptibility factor: no linkage to DXs7.

Leber's hereditary optic neuroretinopathy (LHON) was the first human disease for which mitochondrial inheritance was demonstrated. Analysis of genealogies, however, suggests the existence of an interacting X-linked factor, and linkage to DXS7 was recently described. We tested this location in four LHON families, with DXS7 and two flanking markers, OTC and DXS426. We found recombinations with DXS7 in two families and with DXS426 in one. The two point lod scores to DXS7 were negative with all the allele frequencies for the X-linked factor tested (q = 0.5; 0.35; 0.05).

Detection of the G to A mitochondrial DNA mutation at position 11778 in German families with Leber's hereditary optic neuropathy.

Leber's hereditary optic neuropathy (LHON) is characterized by acute or subacute bilateral (usually permanent) loss of central vision, caused by neuroretinal degeneration. The maternal inheritance is explained by the mitochondrial origin of the disease. Recently, a single mitochondrial DNA (mtDNA) mutation, a G to A substitution at position 11778 that converts a highly conserved arginine to histidine, has been associated with LHON. The mutation eliminates an SfaNI restriction enzyme recognition site and thus provides a method for detection of the mutation by amplification, enzyme digestion and agarose gel electropheresis of polymerase chain reaction (PCR) products. Leukocyte mtDNA from 7 German families with LHON, diagnosed by clinical criteria, was tested for the presence of the G to A mutation at bp 11778. The mtDNa mutation, detected as a loss of the SfaNI site, was seen in one family. The G to A mtDNA mutation is the only known gene alteration associated with LHON so far. It has been identified in patients of different ethnic origin and recent reports strongly support the hypothesis that it represents the most frequent cause of LHON. Identification of the mtDNA replacement mutation using PCR and restriction enzyme digestion requires only a small amount of blood and can be performed rapidly. This method is thus a useful tool in the diagnosis of LHON.

A survey of color discrimination in German ophthalmologists. Changes associated with the use of lasers and operating microscopes.

Color vision tests were performed on 211 German ophthalmologists during their annual meeting at Essen. The subjects also answered detailed questionnaires about their use of lasers and operating microscopes, and their ocular and general health. It was found that 33% of doctors who use lasers or operating microscopes have decreased color discrimination for colors in a tritan color-confusion axis (greater than 2 standard deviations above normal). There is a relationship between number of patients treated and the degree of threshold elevation. Thirty hours of using the operating microscope produces an increase in tritan threshold equivalent to one panretinal photocoagulation.

Colour contrast sensitivity changes caused by peripheral retinal laser photocoagulation.

Macular phototoxicity is known to occur with laser use, and there is evidence that the wavelength of the light used influences this effect. In this study, a computer based colour contrast sensitivity test was used to assess the immediate macular effects of photocoagulation of peripheral flat retinal holes in otherwise normal retinas, using blue-green (488 and 514 nm), yellow (577 nm), orange (595 nm) or red (647 nm) laser light. The laser aiming beam was not allowed to traverse the macula at any stage during treatment. No protan or deutan axis threshold changes were noted in the 17 patients tested irrespective of the laser wavelength used. Tritan axis sensitivity was significantly reduced one hour after treatment with the blue-green laser, but no tritan axis change was found after treatment with longer wavelength lasers. The effect was no longer present the day after treatment in the subjects tested. The results show that even peripheral retinal treatment with blue-green laser can cause acute macular phototoxicity.

Electrophysiology and colour perimetry in dominant infantile optic atrophy.

A typical finding in dominant infantile optic atrophy (DIOA) is the variation of the phenotypic expression of the DIOA gene even within one family. It is of special interest for genetic consultation to evaluate an examination method for detecting subclinically involved patients. Seven patients of two families were examined. Three of them had the typical symptoms of DIOA: reduced visual acuity, tritan defect, temporal pallor of both optic discs, and a relative central scotoma for white test spots. In visual evoked cortical potentials (VECP) the amplitudes were reduced, and in one patient the latencies were slightly delayed and two patients considerably so. The amplitude of the negative component of the PERG was markedly reduced, while the positive component was normal. In the remaining four family members normal retinal and cortical responses were recorded under standard conditions and visual fields and colour vision (FM 100 hue) were also normal. However, static perimetry with blue test spots showed in two family members enlarged central scotomas, thus proving that they had subclinical DIOA.