Ferning Patterns Research Articles

Ocular surface findings in patients with congential aniridia

Abstract Purpose to study the Schirmer´s test, tear Break‐up time, ocular tear ferning pattern test and impression cytology results in patients with congenital aniridia Methods 25 eyes of 25 patients with congenital aniridia underwent Schirmer´s test I, tear break‐up time, ocular ferning pattern test and impression cytology to study the ocular surface characteristics. Clinical findings were correlated with the results of the aforementioned tests. Results Aniridia related keratopathy was Grade 0 in 12%, Grade 1‐A in 52%, 1‐B in 20% and Grade 2 in 16% of eyes studied. Schirmers Test I was normal in 96% of eyes. Tear break‐up time was abnormal in 75% of patients. Ocular ferning pattern tests were as follows: Grade 1 (20%), grade 2 (50%), Grade 3 (20%) and Grade 4 (10%). Conjunctival squamous metaplasia revealed: Grade 0 (23%), Grade 3 (35%), Grade 5 (4%). Conjunctival keratinization was mild in 35% and moderate in 15%. Significant correlation was found between conjunctival keratinization and the degree of Anirida‐related keratopathy. Significant correlation was also found between ocular ferning pattern test and conjunctival keratinization. Conclusion Aniridia –related keratopathy is a muco‐deficient and lipo‐deficient, more than an aqueo‐deficient dry eye disease. Simple tests of the ocular surface must be done early on to direct the right kind of dry eye treatment in these difficult cases.

Salivary amino acids quantification using RP-HPLC during normal menstrual cycle

The present study was carried out to detect amino acids profile in women saliva in order to establish the qualitative and quantitative differences that might have potential value in detection of ovulation by noninvasive methods. For the collection of sample, the stages of menstrual cycle were decided by the physical and morphological examination of salivary fern pattern. The saliva from various reproductive phases (prepubertal, preovulatory, ovulatory, postovulatory phases and menopause) was collected and analyzed by reverse phase high performance liquid chromatography (HPLC) after precolumn derivitization of amino acids using O-Pthaldehyde (OPA) by means of RP-HPLC amino acid analyzer. Among the various amino acids identified the compounds such as tryptophan, arginine and phenylalanine were comparatively found to be higher during ovulatory phase when compared to that of other phases. The increase in amino acid concentration during ovulatory phase may be due to the circulation of steroid hormones. Thus, the presence of specific amino acids in ovulatory saliva makes the possibility to develop a biomarker for detection of ovulation by noninvasive methods. Key words: Ovulation steroid hormones, O-Pthaladehyde, chromatography, amino acids

Reprodutibilidade na classificação do teste de cristalização do filme lacrimal em pacientes com síndrome de Sjögren

To verify the reproducibility of Rolando's classification of the tear ferning test using five different examiners and to compare the patterns of crystallization found in Sjögren's syndrome patients and normal subjects. Tear ferning analysis of 29 patients with Sjögren's syndrome and of 45 patients without ocular disease were done using polarized light microscopy and the Rolando classification for tear ferning. Five examiners classified the ferning patterns of all the patients. ROC curve (Receiver Operating Characteristic) was used to find out the best score for the correct syndrome diagnosis. Kappa index (p<0.0001) was used to compare the results of the examiners among them and check the test's reproducibility. Charts were drawn to compare the two groups' results. Throught the ROC curve the score of 2.50 for diagnosis of Sjögren's syndrome was stablished. Considering the aggregated patterns I with II and III with IV, the examiners' level of pattern agreement was excellent (Kappa ranging from 0.82 to 0.97, p<0.0001). The group with Sjögren's syndrome was classified mostly as patterns III and IV and the patients without ocular disease mostly as I and II. The patterns associated with Sjögren's syndrome and normal patients matched the ones in the literature. The tear ferning test classification is reproductible when the Rolando classification was used for Sjögren's syndrome patients.

Reproductive performance of crossbred dairy cows reared under traditional low input production system in the eastern Himalayas

Reproductive performance of crossbred cows reared under traditional low input production system was assessed. A total number of 160 farmers were surveyed and traditional fodders were evaluated for proximate composition. Total protein, albumin and cholesterol concentrations in blood serum were analyzed. The age at first service, age at first calving, calving interval, calving to first service and calving to conception interval were calculated from records of 261 crossbred dairy cows. The crude protein, crude fiber, nitrogen free extract, ether extract and total ash content varied from 7.2 to 13.9, 18.2 to 34.4, 39.1 to 59.2, 2.1 to 4.1 and 7.2 to 17.9%, respectively. The total protein and albumin concentrations in blood serum were 7.6 +/- 0.3 and 4.3 +/- 0.3 g/dl, respectively. The cholesterol concentration was 221.1 +/- 8.2mg/dl. The mean age at first service and age at first calving was 28.6 +/- 1.0 and 40.7 +/- 1.1months, respectively. The mean values for calving to first service and calving to conception intervals were 182 +/- 14.5 and 224 +/- 9.0days, respectively. The conception rate was significantly high among the cows that showed typical fern pattern of cervical mucus (56.6%). The conception rate based on 1(st) insemination was 44.4%. The incidence of repeat breeders and anestrus among the crossbred cows were 12.8 and 14.1%, respectively.

Tear Osmolality and Ferning Patterns in Postmenopausal Women

To compare tear osmolality and ferning patterns in postmenopausal women (PMW) with and without dry eye symptoms. Thirty-seven healthy PMW (>50 years of age), not on hormone replacement therapy, were categorized as being symptomatic or asymptomatic of dry eye based on their responses to an Allergan "Single-Item Score Dry Eye Questionnaire" (SIDEQ). They subsequently completed the Allergan "Ocular Surface Disease Index" (OSDI) questionnaire. Tear samples were collected from participants to evaluate osmolality and ferning patterns. A novel freezing point depression osmometer (Advanced Instruments Inc., Model 3100 Tear Osmometer), was used to measure the osmolality of the tear film. The tear ferning test was performed and evaluated for the quality of ferning based on the Rolando grading system. SIDEQ responses revealed 21 symptomatic and 16 asymptomatic participants. The OSDI total score was 6.5 +/- 5.9 for the non-dry-eyed (NDE) group and 25.7 +/- 12.4 for the dry-eyed (DE) group. The subscores for the DE group were significantly greater than the NDE group (p < 0.001). Osmolality values in DE individuals were significantly different from NDE (328.1 +/- 20.8 vs. 315.1 +/- 11.3 mOsm/kg; p = 0.02). Fifty percent of the DE participants showed type II ferning patterns and 29% of the DE participants showed type III ferning patterns, whereas the NDE participants showed either type I (44%) or II (66%) ferning patterns. There was a significant difference between the DE and NDE participants for the ferning patterns (p = 0.019). There was no significant correlation between tear osmolality and tear ferning (DE: r = 0.12; p > 0.05, NDE: r = -0.17; p > 0.05). Osmolality in mild and moderately DE PMW is higher than in NDE PMW and tear ferning is a rapid, simple, noninvasive laboratory procedure that indicates altered tear quality in PMW with symptoms of dry eye.

Lightning and Lichtenberg figures

The Lichtenberg figure (LF), also known as a ferning pattern, is a pathognomonic sign, found only in victims of lightning strikes. Pathognomonic signs are valuable because they confirm the diagnosis. LF is a recognizable skin pattern that was found on two lightning-strike patients reported here.

Immunodacryochemistry (immunoferning): a new screening method for proteins in tears.

The ferning test of tears has been an intensively discussed method for rough but quick visualization of general tear fluid composition. Regarded as crystallization process resulting from simple water subtraction (Battaglia and Giusto 1993), it has useful features, amongst them the minimal equipment required, but standardization is a known problem. The classification according to Rolando (1984) seems to significantly increase the clinical applicability of this technique (Pensyl and Dillehay 1998). Even though the ferning test has been proposed for predicting contact lens tolerance (Ravazzoni et al. 1998), the underlying mechanisms of the ferning pattern is still not clearly understood (Kogbe et al. 1991). Recently, it has been evaluated as a method to develop a rough picture of the crystallization process, reflecting the qualitative composition of tears (Sommer et al. 1994). The accuracy of the method, however, in clinical conditions such as dry eyes seems to be inferior to impression cytology (Jackson and Perrigin 1999), but it may show clear pathological changes (Norn 1994, Vaikoussis et al. 1994, Maragou et al. 1996). Today, with the current means of interpretation, the ferning test is seen as a practical, quick, easy and reliable complement to other tests (Albach et al. 1994). In the present study we develop a further use of the ferning procedure.

Use of vaginal electrical resistance (VER) to predict estrus and ovarian activity, its relationship with plasma progesterone and its use for insemination in buffaloes

In three experiments we studied the baseline and changes in VER during different natural estrous cycle stages (n=146) in ovarian structures and in plasma progesterone during estrus induced by prostaglandin injection (n=16) and the VER at insemination (n=90) in an attempt to predict estrus, ovulation and the best VER range for inseminating buffaloes for optimum conception. The baseline VER was classified on the basis of ovarian findings and estrous cycle stages. The mean VER during estrus, metestrus, diestrus, proestrus and anestrus was 32.68 ± 0.46, 41.26 ± 1.17, 50.23 ± 0.55, 43.20 ± 0.64 and 55.86 ± 0.57 ohms, respectively. There was a significant difference (P<0.01) between the VER except those between metestrus and proestrus. The ANOVA for VER over estrous cycle stages showed a highly significant (P<0.01) effect of stage of estrous cycle on VER in buffaloes. The percent decrease in VER was more pronounced from diestrus to estrus. In the second part of the study plasma progesterone profiles and the appearance of estrus in buffaloes induced to estrus using two dose schedules and routes of PGF 2α administration showed that luteolysis and estrus induction was slower in the 10 mg IVSM route (Intra Vulvo Submucosal) (only 60% animals evinced estrus in 48 to 72 hours) as compared to the 25 mg IM route (83.33% evidenced estrus in 48 to 72 hrs). Fall in plasma progesterone was synchronous to a fall in VER, the correlation (0.65) between them being positive and significant (P<0.01). After ovulation the VER started rising, showing a distinct relationship between VER and ovulation. By using VER, an additional 36.6% of the buffaloes could be detected in estrus. In the third part of the study, insemination of buffaloes induced to estrus (n=11) and normal-estrus buffaloes (n=79) showed that the overall conception rates to single insemination when the buffaloes were inseminated at the VER range of 26 to 30, 31 to 35 and 36 to 40 ohms were 81.48, 58.97 and 16.66%, respectively. Buffaloes showing VER from 31 to 35 ohms and 36 to 40 ohms also evidenced atypical and Null fern pattern in the cervicovaginal mucus. The study proved that VER can be used successfully to predict the stage of estrous cycle, ovarian status and ovulation; and insemination at a low VER distinctly improves the conception rates in buffaloes.

Ocular Ferning Test – Effect of Temperature and Humidity on Tear Ferning Patterns

Background: The ocular ferning test is used as a diagnostic aid to evaluate patients with dry-eye disease. The ferning phenomenon is a dendritic growth form of dried tear fluid. The influence of temperature and relative humidity (rH) on the morphology of tear ferning patterns was investigated. Methods: Two microliters of tear fluid of 5 volunteers with normal tear function were dropped onto a microscope slide with a pipette and were dried in a climatic chamber at four different temperatures (20, 22, 24 and 26°C) and at four different grades of rH (30, 40, 50 and 60%) at each temperature. The dried droplet was divided according to distinct morphological features into a center, transition zone and margin. The center of the dried droplet was classified into four types according to the system of Rolando. Results: In the temperature range between 20 and 26°C, rH has a strong influence on the ferning patterns. At these temperatures, increasing rH resulted in a deterioration of type I ferning obtained at 30% rH to type II or type III at 60% rH in all subjects. Otherwise, at rH of 30, 40 and 50%, no definite influence of temperature between 20 and 26°C was observed. At an rH of 60%, the ferning patterns changed with increasing temperature from 20 to 26°C from type II to type III. Conclusion: High humidity can modify and deteriorate the ferning patterns of tear fluid from subjects with normal tear function. To obtain reproducible results using the ocular ferning test, stable conditions are necessary. An rH not higher than 50% at a temperature range between 20 and 26°C produced ferning patterns without interstitial spaces which could be related to the best quality according to the system of Rolando.

Ocular ferning during the menstrual cycle in healthy women.

The aim of the study is to investigate whether tear ferning patterns change during different phases of the menstrual cycle. The tear ferning test was performed on twelve normal women of childbearing age at three day intervals throughout one complete menstrual cycle. Serum hormone levels (progesterone, estrogen, testosterone) were measured. Eight women showed type I ferning, and the other four had type II ferning initially. These patterns did not change during the menstrual cycle. Serum hormone levels were all in the normal range. Since no change in ferning pattern was detected during the menstrual cycle, the ferning test can be done at any time in women. This study showed no effect of different menstrual cycle phases on tear ferning patterns.

Quality of salivary tears following autologous submandibular gland transplantation for severe dry eye.

This study aimed to characterise the composition of the pre-ocular fluid after transplantation of the autologous submandibular gland (SMG) for patients with severe dry eye. Stimulated and unstimulated pre-ocular fluid from 15 patients (17 eyes) with a viable SMG graft ("SMG-salivary tears"), as well as normal tears and SMG saliva (20 normal subjects/20 eyes), was sampled. As global tear parameters, fern pattern analysis and SDS gel electrophoresis were performed. As specific quality parameters, total protein content, secretory immunoglobulin A (SIgA), lysozyme, amylase, sodium, potassium and osmolality were measured using routine laboratory methods. The flow rate of SMG-salivary tears was determined in 5 patients by means of sequential scintillography. The fern pattern of SMG-salivary tears was coarse and thus more similar to normal SMG saliva than tears. SDS gel electrophoresis of the SMG-salivary tears showed albumin and two unidentified proteins in addition to the normal tear pattern. Osmolality and total protein content of SMG-salivary tears were higher than in normal SMG saliva, but still lower than in normal tears. High activities of normal tear antibacterial proteins (SIgA, lysozyme and amylase) were detected in the salivary tears. Stimulation of the secretion did not alter the composition of SMG-salivary tears. The flow rate of SMG-salivary tears was closer to that of normal tears than normal SMG saliva. Salivary tears resulting from SMG-transplantation represent condensed SMG saliva. Thus their quality is intermediate between normal tears and normal SMG saliva. High levels of secretory proteins demonstrate that the gland maintains an active function. Surgical denervation and residual tear components from the ocular surface are the most likely factors to cause the complex differences between normal SMG saliva and SMG-salivary tears. The effects of this secretion on the ocular surface are currently being evaluated in a clinical and laboratory study.

The repeatability of tear mucus ferning grading.

Rolando's classification system for tear mucus ferning patterns subjectively assigns grades based on the size and spacing between ferns. Grade 1 and 2 patterns are considered "normal," whereas grade 3 and 4 patterns are often associated with keratoconjunctivitis sicca. This study was designed to examine the intraobserver and interobserver repeatability of Rolando's system. Photographic slides (N = 418) of portions of tear ferning patterns were randomly assembled, numbered, and graded by two investigators (I1 and I2). I1 graded the slides twice; the slides were remixed under masked conditions between the first and second runs. I2 graded the slides once, independent of I1. For the four-grade system, intraobserver agreement was 85.41% (simple kappa = 0.75; 95% confidence interval ]CI] = 0.69-0.82). Interobserver agreement was 80.62% (kappa = 0.67; CI = 0.60-0.74) and 86.12% (kappa = 0.75; CI = 0.68-0.83) for the first and second runs of I1, respectively. Analysis of the ability to simply classify normal (grade 1 and 2) from abnormal (grade 3 and 4) patterns revealed intraobserver repeatability of 94.50% (kappa = 0.76; CI = 0.67-0.86). Interobserver agreement was 92.10% (kappa = 0.65; CI = 0.56-0.75) and 94.26% (kappa = 0.71; CI = 0.62-0.81) for the first and second runs, respectively. Based on the high rate of agreement between intraobserver and interobserver trials, Rolando's grading system appears to be an easy and consistent method for the classification of tear ferning patterns.

Evaluation of the Lady Free Biotester® in determining the fertile period

Small hand-held microscopes have been developed for self-observation of salivary ferning patterns to detect the fertile time of a woman’s menstrual cycle. The purpose of this study was to evaluate one such microscope, the Lady Free Biotester®, by comparing it with the self-observation of cervical-vaginal mucus and with the self-detection of luteinizing hormone (LH) in the urine. Twelve natural family planning teachers (average age 34.6 years) observed their cervical-vaginal mucus, tested their urine for LH, and observed salivary and cervical-vaginal mucus ferning patterns (with the Lady Free Biotester) for two menstrual cycles. Data indicated that there was a strong correlation between the LH in the urine and the peak in self-observed cervical-vaginal mucus ferning (r = 0.99, p ≤0.001) and salivary ferning (r = 0.98, p ≤0.001). However, it was difficult to assess the beginning and end of the fertile time based on the salivary ferning patterns. Further testing of salivary ferning patterns is recommended before widespread use of these devices for family planning.

Tear functions in patients with pterygium.

In the etiology of pterygium abnormalities in tear functions have also been emphasized. In this study, tear function tests are evaluated in patients with pterygium. Schirmer's test 1, tear film break-up time and mucus fern patterns were evaluated in 70 eyes with pterygium and in 70 eyes of the age matched control group. Marginal tear films were also assessed. Tear film break-up time was significantly reduced in the pterygium group. Mucus fern patterns and marginal tear films were found to be markedly abnormal in the eyes with pterygium, however, there was no significant difference in Schirmer's test 1. Tear function tests disclosed disrupted tear film stability which is more likely to be due to the altered mucin. This change may either be the primary factor inducing pterygium formation or reflect an existing pathology in the cells lining the ocular surface.

Clinical status, ocular surface changes and tear ferning in patients with cystic fibrosis

Twenty-three cystic fibrosis patients and 20 controls were examined for ocular surface changes and tear fluid ferning characteristics. The patients were also evaluated systemically and given numerical scores according to Schwachman's scoring system. Frequency of blepharitis, fluorescein staining, and the Schirmer's test values did not differ between patients and controls. Patients with cystic fibrosis had a higher frequency of altered ferning pattern that corresponded to the disease severity as measured by the Schwachman score (r = -0.48, p < 0.05). Although altered ferning pattern was frequent, the rate of type I ferning was also high (30%) in patients with cystic fibrosis. We conclude that the ferning test should not be used as an aid in diagnosis of cystic fibrosis but it may be used as an indicator of clinical status during follow-up.

Ferning of aqueous humor in the pseudoexfoliation syndrome

The authors report on the results obtained in the crystallization of the aqueous humor in 6 patients suffering from cataract with pseudoexfoliation syndrome. The principle characteristics of the aqueous in these cases are described and are compared with the features of aqueous humor in 20 cases of uncomplicated cataract. Tetragonal shaped masses of material with a distinct tendency to localization in the intersections of fern patterns were demonstrated in the crystallized aqueous from patients with pseudoexfoliation syndrome. These aggregates most probably correspond to exfoliation material.

Crystallization patterns in anterior vaginal fluid from bitches in oestrus.

Bitches exhibited a characteristic arborization pattern of the fluid from the anterior vagina during pro-oestrus and oestrus. These changes were monitored together with conventional vaginal cytology and plasma oestrogen and progestagen concentrations. A classical ferning pattern, similar to that seen in bovine cervical mucus at oestrus, occurred after the peak in plasma oestrogen concentrations. Ferning was most intense after the second peak of cornification of vaginal epithelial cells. It is suggested that a 'Ferning Index', when combined with conventional vaginal cytology, can be of use in determining the optimum mating time in the bitch.

Oestrus during pregnancy in the cow

Forty-three oestruses were observed in 35 pregnant cows in one beef suckler herd and 17 dairy herds; at each oestrus the pregnant cow stood willingly to be mounted by another cow or bull. Such oestruses were observed at all stages of pregnancy, although more often between 121 and 240 days, occurred more than once per pregnancy and were also seen in successive pregnancies. On one farm where the dairy cows were observed for four 30 minute periods daily, oestrus was recorded in 5.7 per cent of pregnant cows. The behavioural signs associated with oestrus during pregnancy were indistinguishable from those of true oestrus in non-pregnant animals and although its duration was shorter (mean 5.6 hours), its intensity was comparable to that of the true oestrus. Pregnant cows showing oestrus were usually seen interacting with other oestrous cows in the sexually active group. Pregnant cows showing oestrus had a higher mean condition score (3.9 +/- 0.64) than control pregnant cows (3.0 +/- 0.36). Physiological changes in the genital tract normally associated with true oestrus were not observed in pregnant cows showing oestrus. There was no ovulation or metoestrous bleeding. The characteristics of cervical mucus, including ferning patterns, were similar to those of pregnant cows at the same stage of pregnancy. Hormonal changes associated with oestrus in non-pregnant cows were not observed in the pregnant cows exhibiting oestrus. Seven of nine pregnant cows at oestrus stood willingly to be mounted by a bull. On seven occasions, bulls exposed only to cervical mucus from pregnant cows showing oestrus did not display flehmen.(ABSTRACT TRUNCATED AT 250 WORDS)

An interesting use of the study of tear ferning patterns in contactology.

A study of tear ferning patterns in normal and keratoconjunctivitis sicca eyes led to the classification into 4 types by Rolando [1984]. We practised this in our laboratory and observed specific and distinct tear ferning patterns for electrophoretic types 1 and 2 which Liotet et al. [1983] described as abnormal tear proteins modification found in patients who repeatedly and rapidly clog their hydrophilic lenses. In a general way, a study of tear ferning patterns can result in an appreciation of tear quality in contactology.

A note on the concentrations of plasma oestradiol-17β and progesterone around the time of puberty in heifers

ABSTRACTIn heifers, plasma progesterone levels were low (&lt;0·3μg/l) from the 59th to about the 15th day before puberty (defined as the day when oestrus was first detectable by the fern pattern of dried cervical mucus). Plasma oestradiol-17β levels varied randomly within the range 1 to 4 ng/l over the same period, but then showed three peaks of about 6, 9 and 4 ng/l, respectively 8 days before puberty, on the day of puberty and 4 days later. The pre-pubertal oestradiol-17p peak was followed by a short period of elevated progesterone levels. Whether this oestradiol-17β peak induced ovulation is uncertain; it is however clear that a corpus luteum of normal life-span was not formed. The oestradiol-17β peaks on the day of puberty and 4 days later, when progesterone levels were rising rapidly, were typical both in magnitude and duration of those occurring before and after ovulation in post-pubertal oestrous cycles.